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Mining Candidate Genes for Litter Size Traits in English Springer Spaniel Bitches Based on Whole Genome Resequencing GAO Yilong, HE Xingliang, ZHOU Xiaopeng, LI Dawei, BAO Xijun, LI Laiyou Laboratory Animal and Comparative Medicine    DOI: 10.12300/j.issn.1674-5817.2025.149 Online available: 23 December 2025

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Study on the 90-day Feeding Experimental Background Data of SD Rats for Drug Safety Evaluation QIN Chao, LI Shuangxing, ZHAO Tingting, JIANG Chenchen, ZHAO Jing, YANG Yanwei, LIN Zhi, WANG Sanlong, WEN Hairuo Laboratory Animal and Comparative Medicine    2025, 45 (4): 439-448.   DOI: 10.12300/j.issn.1674-5817.2024.187 Abstract （1435）   HTML （28）    PDF （1104KB）（1003）       Knowledge map    Save Objective To establish background data for a 90-day feeding trial of SD rats to ensure the reliability of research data. Methods Background data from six independent 90-day feeding trials of SD rats conducted by the National Center for Safety Evaluation of Drugs from 2020 to 2023 were summarized. These studies involved a blank control group of 120 SPF-grade 4-week-old SD rats, with an equal number of males and females, which were only given standard full-nutrient pelleted rat feed. After the quarantine period, the animals were observed for an additional 90 days, followed by intraperitoneal injection of Zoletil (50 mg/mL) for anesthesia, blood sampling, euthanasia, and necropsy. By analyzing the data from the blank control group, a relevant background database for SD rats was established. Results Both male and female rats exhibited steady weight gain, with a more pronounced increase in male rats. Within 90 days, the average body weight of male and female rats increased to over 500 g and 300 g, respectively. Three weeks later, the average daily food intake of male rats stabilized at approximately 25~28 g per rat, while that of female rats remained stable at approximately 16~19 g per rat. The food utilization rate of all animals gradually decreased from the first week of the experiment. In the white blood cell (WBC) differential count results, significant differences were observed in the counts of WBCs, neutrophils (Neut), lymphocytes (Lymph), and monocytes (Mono) between males and females (P<0.001). However, there were no significant differences in the percentages of neutrophil (%Neut), lymphocyte (%Lymph), and monocyte (%Mono) between the sexes (P>0.05). The average red blood cell count (RBC), hemoglobin concentration (HGB), hematocrit (HCT), platelet count (PLT), prothrombin time (PT), and activated partial thromboplastin time (APTT) were higher in male animals than in female animals (P<0.05). The average values of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine phosphokinase (CK), lactate dehydrogenase (LDH), glucose (GLU), and triglyceride (TG) in male rats were higher than those in female rats (P<0.05). The urinary pH range for male animals was 5.0 to 8.5, while for female animals it was 6.5 to 9.0. The majority of male animals had a urinary specific gravity lower than 1.020, and the majority of female animals had a urinary specific gravity lower than 1.015. The weights of various organs (excluding the adrenal glands and reproductive organs) in male animals were heavier than those in female animals (P<0.001), while the organ/body weight ratios (excluding the kidneys and reproductive organs) of female animals were higher than those of male animals (P<0.001). Conclusion This study summarizes the background reference ranges for body weight, food intake, hematology, and serum biochemistry indicators in SPF-grade SD rats in the untreated control group from six 90-day feeding trials conducted by the National Center for Safety Evaluation of Drugs. It provides important reference data for related research. By summarizing the background and spontaneous histopathological changes in rats, this study aids in the standardization and normalization of subsequent research, as well as in the evaluation and analysis of abnormal results. Table and Figures | Reference | Related Articles | Metrics

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Construction and Functional Validation of GTKO/hCD55 Gene-Edited Xenotransplant Donor Pigs WANG Jiaoxiang, ZHANG Lu, CHEN Shuhan, JIAO Deling, ZHAO Heng, WEI Taiyun, GUO Jianxiong, XU Kaixiang, WEI Hongjiang Laboratory Animal and Comparative Medicine    2025, 45 (4): 379-392.   DOI: 10.12300/j.issn.1674-5817.2025.024 Abstract （1013）   HTML （62）    PDF （3294KB）（1884）       Knowledge map    Save Objective To develop GTKO (α-1,3-galactosyltransferase gene-knockout, GTKO)/hCD55 (human CD55) gene-edited xenotransplant donor pigs and verify their function. Methods In this study, CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated nuclease 9), PiggyBac transposon technology and somatic cell nuclear transfer technology were used to construct GTKO/hCD55 gene-edited Diannan miniature pigs. The phenotype and function of GTKO/hCD55 pigs were analyzed by Sanger sequencing, real-time fluorescence quantitative PCR, flow cytometry, immunofluorescence, bisulfite sequencing, antigen-antibody binding assays, and complement-dependent cytotoxicity assays. Results After transfection of PX458 and PiggyBac gene editing vectors into wild-type fetal pig fibroblasts, 48 single-cell colonies were obtained through puromycin drug screening. Two single-cell colonies were selected for somatic cell nuclear transfer, resulting in two fetal pigs at 33 days of gestation. The GGTA1(α-1,3-galactosyltransferase) genotypes of fetal pig F01 were -17 bp and wild type (WT), while the GGTA1 genotypes of fetal pig F02 were -26 bp/+2 bp and -3 bp. The hCD55 mRNA expression levels of both fetal pigs were significantly higher than those of WT pigs (P＜0.01). The fetal pig F02 was selected as the donor cell source for recloning, 11 surviving piglets were obtained, all identified as GTKO/hCD55 gene-edited pigs. These pigs showed absence of α-Gal antigen expression, but weak or no expression of hCD55 was observed. Methylation analysis of the hCD55 gene's CpG island showed hypermethylation in kidney tissue lacking hCD55 expression, whereas it was not methylated or partially methylated in kidney tissue expressing hCD55. Moreover, codon optimization of the CpG island of the hCD55 gene to reduce CG content could achieve stable expression of the hCD55 gene. In addition, antigen-antibody binding experiment showed that the amount of human IgM binding to GTKO/hCD55 gene-edited pig fibroblasts was significantly lower than that of WT pigs (P＜0.01). Complement-dependent cytotoxicity experiment showed that the survival rate of fibroblasts in GTKO/hCD55 pigs was significantly higher than that in WT pigs (P＜0.01). Conclusion This study demonstrates the successful generation of GTKO/hCD55 gene-edited xenotransplant donor pigs. Methylation-induced gene silencing of the hCD55 gene can be effectively avoided by reducing the CG content of the CpG island through codon optimization. This study provides a reference for the development of xenotransplant donor pigs and guides subsequent research on xenotransplantation. Table and Figures | Reference | Related Articles | Metrics

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Main Biological Characteristics of Hermetia illucens L. and Its Potential Applications as a Model Organism LIU Song, MO Qianru, WANG Jin, CUI Ying, TIAN Ling Laboratory Animal and Comparative Medicine    2025, 45 (6): 803-809.   DOI: 10.12300/j.issn.1674-5817.2025.141 Abstract （950）   HTML （17）    PDF （5674KB）（1971）       Knowledge map    Save The black soldier fly (Hermetia illucens L.), a globally distributed resource insect native to the tropical regions of the Americas, has become stably established in several regions of China, such as Guangdong and Hainan Provinces, through both natural dispersal and human-mediated introduction. In recent years, owing to its strong environmental adaptability, high resource conversion efficiency, and prominent value in ecology and biological resources, as well as its remarkable advantages in organic waste conversion and high-value bioproduct development, it has emerged as a research hotspot in the field of biological resources. This paper first outlines the biological characteristics and geographical distribution of Hermetia illucens L., clarifying its research basis and resource potential. Subsequently, its efficient and stable breeding system and rearing substrate optimization techniques are introduced, which provide technical support for standardized breeding and growth performance improvement. Further, this paper systematically reviews the medical and health application potential of bioactive components derived from Hermetia illucens L., with a focus on active substances extracted from its larvae and pupae, such as antimicrobial peptides, functional fatty acids, chitosan, and protein hydrolysates, which exhibit broad-spectrum antibacterial, antioxidant, anti-inflammatory, and good biocompatibility. It comprehensively elaborates on the application research progress on the application of these bioactive components of Hermetia illucens L. in fields such as prevention and treatment of multidrug-resistant bacterial infections, anti-inflammatory and antioxidant effects, cancer chemoprevention, tissue repair, and cosmeceuticals for acne care. Moreover, this paper elucidates the advantages of Hermetia illucens L. as a novel invertebrate laboratory animal model, highlighting its high productivity, controllable growth patterns, and suitability for in-depth individual research. It also explores its unique value in fundamental medical research, such as host-microbe interactions and endogenous virus evolution, providing an interdisciplinary research platform that bridges evolutionary biology, microbial ecology, immunology, and preventive medicine. Finally, the paper analyzes the technical bottlenecks faced in the current research and industrialization process of Hermetia illucens L., and reviews and prospects product development strategies, interdisciplinary integration, and clinical translation directions, aiming to offer a systematic reference for the in-depth development and efficient utilization of this resource. Table and Figures | Reference | Related Articles | Metrics

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Applications and Advances of Drosophila in Research of Obesity and Its Related Metabolic Diseases CHEN Haotian, LIU Jingnan Laboratory Animal and Comparative Medicine    2025, 45 (6): 688-704.   DOI: 10.12300/j.issn.1674-5817.2025.104 Abstract （945）   HTML （21）    PDF （7984KB）（2202）       Knowledge map    Save Obesity is a chronic metabolic disease caused by long-term imbalance between energy intake and expenditure, which can significantly increase risks of cardiovascular diseases, type 2 diabetes, various cancers, and premature aging. The latest WHO statistics show that global overweight and obesity rates among adults continue to rise, making obesity a public health problem requiring urgent precise prevention and control. This review systematically summarizes the unique value and application advances of Drosophila melanogaster as a model organism in obesity and metabolic disease research. Due to its short life cycle, low rearing cost, high homology with human disease-related genes, conserved organ functions, and well-developed genetic tools, Drosophila has emerged as an efficient model organism for analyzing obesity and metabolic disorders. Feeding high-sugar/high-fat diets to Drosophila can stably replicate typical obesity-related phenotypes such as lipid accumulation, insulin resistance, impaired cardiometabolic function, and shortened lifespan, and can be combined with tissue- or cell type–specific genetic manipulations for target discovery and mechanism verification. At the organ level, the fat body functions as a central hub for energy storage, metabolic sensing, and endocrine regulation; oenocytes participate in lipid, sterol, and very-long-chain fatty acid metabolism and starvation response; the midgut integrates signals from nutrients and gut microbiota through regionalized absorption and enteroendocrine functions; the malpighian tubules, besides excretion, also regulate reabsorption and influence body size through energy sensing pathways and glucose transporter membrane localization. Muscles are the primary energy-consuming organs in Drosophila with flight muscles exhibiting the highest energy demand. They primarily utilize trehalose and glucose from the hemolymph for energy supply. Additionally, they can mobilize glycogen and fatty acids to participate in energy metabolism. Additionally, circadian rhythm and feeding time (such as time-restricted feeding) can reshape peripheral clock-metabolism coupling, alleviating diet-induced metabolic disorders. Moreover, they regulate systemic metabolic homeostasis through myogenic factors.In cross-organ endocrine regulatory network, brain insulin-like peptide producing cells secrete insulin-like peptides to lower blood sugar and promote metabolic balance, whereas the corpora cardiaca release adipokinetic hormone to increase blood sugar and stimulate lipolysis. These two endocrine systems functionally antagonize each other, together forming a key metabolic homeostasis regulatory axis in Drosophila. Fat body and intestine regulate insulin-like peptide secretion according to nutritional status by releasing Unpaired 2, Limostatin, and adiponectin-like factors, forming a multi-level "gut-fat body-brain-peripheral organ" feedback loop. In summary, Drosophila, with its highly conserved organ functions and metabolic pathways and its powerful genetic tools, provides an efficient and scalable experimental platform for analyzing obesity etiology, elucidating cross-tissue signaling networks, discovering potential translational targets, and evaluating nutritional/pharmacological intervention strategies. Table and Figures | Reference | Related Articles | Metrics

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Discussion on AI-Based Digital Upgrade and Application Practice of Laboratory Animal Centers WANG Tingjun, LUO Hao, CHEN Qi Laboratory Animal and Comparative Medicine    2025, 45 (4): 473-482.   DOI: 10.12300/j.issn.1674-5817.2024.181 Abstract （935）   HTML （45）    PDF （1285KB）（2993）       Knowledge map    Save Objective In traditional laboratory animal centers, there are issues such as low efficiency in cage scheduling, insufficient supervision of personnel behavior, and difficulty in upgrading aging equipment. This study aims to upgrade the information system of existing laboratory animal centers by applying multimodal large language model technology. This upgrade intends to achieve real-time perception of the status of animal cages, intelligent supervision of experimental personnel behavior, and automated processing of business workflows, thereby improving management efficiency and precision. Methods An AI-based approach for upgrading laboratory animal center informatization was proposed by the First Affiliated Hospital of Zhejiang University School of Medicine,compatible with different breeding equipments. The system architecture, from the bottom up, consisted of three layers: hardware layer, core algorithm layer, and application layer. The hardware layer was equipped with cameras and high-speed network transmission devices for collecting information on cages and personnel. The core algorithm layer utilized multi-stage image preprocessing technology and multimodal large language model recognition technology to extract and identify image information. The application layer integrated the recognition results with the existing information of the animal center to generate real-time cage occupancy heatmaps, which visually and clearly showed the density distribution of cage usage in the laboratory animal center. Results The AI-based management system achieved a cage recognition accuracy of 98.5% and a correct wearing identification rate of laboratory coats of 98.8%. The average image processing time was 3.7 seconds per image, the effective utilization rate of cages increased by 23%, and the turnover efficiency improved by 35%. In addition, the management system could track and warn against non-compliant behaviors in real time. After intelligent recognition, the system detected more violations, with the violation detection rate increasing by 90.6%. After continuous use for three months, the weekly average number of violations decreased by 54.0% compared to the baseline period. Conclusion This study applies multimodal large language model to the field of laboratory animal management, achieving real-time monitoring and automated management of cage identification, thereby improving management efficiency and precision. The system integrates multi-source data such as visual recognition and behavior analysis, establishing a comprehensive intelligent supervision system for experimental personnel. It provides research institutions with efficient, accurate, and cost-effective management tools, promoting the intelligent development of laboratory animal management. Table and Figures | Reference | Related Articles | Metrics

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Advances in the Application of Zebrafish in the Research of Inflammatory Bowel Disease Mechanisms and Drug Development ZHAO Xin, WANG Chenxi, SHI Wenqing, LOU Yuefen Laboratory Animal and Comparative Medicine    2025, 45 (4): 422-431.   DOI: 10.12300/j.issn.1674-5817.2024.170 Abstract （918）   HTML （34）    PDF （831KB）（3932）       Knowledge map    Save Inflammatory bowel disease (IBD) is a chronic, relapsing intestinal disorder driven by multiple factors including genetics, immunity, and environment, and is clinically classified into ulcerative colitis and Crohn's disease. Currently, mice and zebrafish are the primary experimental animals used in IBD research, among which zebrafish have emerged as an ideal model due to their unique advantages. Compared with rodent models, zebrafish serve as an effective and convenient model, offering advantages such as a short life cycle, robust reproductive capacity, small size, and transparent embryos. These characteristics make zebrafish highly suitable for dynamic tracking of continuous pathological progression and high-throughput drug screening. Zebrafish share over 70% genetic homology with humans, and their intestinal cellular composition and ontogeny closely resemble those of humans. Moreover, the structure and characteristics of their gut microbiota are similar to the human intestinal microbiome, providing a solid foundation for studying the relationship between gut microbiota and IBD. With advances in biotechnology, zebrafish IBD models generated by chemical induction or genetic engineering can accurately simulate the core pathological features of human IBD, such as intestinal wall thickening, inflammatory cell infiltration, and elevated expression of pro-inflammatory factors. These models have played a significant role in revealing the pathogenesis of IBD as well as the development of targeted therapeutic drugs. This article first outlines the intestinal characteristics of zebrafish and features of zebrafish IBD models, then provides an in-depth analysis of their application in IBD pathogenesis research from multiple aspects, including genetics, immunity, environment and diet, and infection. It also reviews research progress on the application of zebrafish in the development of anti-inflammatory drugs, probiotics, and traditional Chinese medicine therapies, aiming to provide researchers with references for the rational use of zebrafish models at all stages of preclinical research, to advance fundamental IBD research and accelerate breakthroughs in this field. Table and Figures | Reference | Related Articles | Metrics

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A Mouse Model and Mechanism Study of Premature Ovarian Insufficiency Induced by Different Concentrations of Cyclophosphamide GONG Leilei, WANG Xiaoxia, FENG Xuewei, LI Xinlei, ZHAO Han, ZHANG Xueyan, FENG Xin Laboratory Animal and Comparative Medicine    2025, 45 (4): 403-410.   DOI: 10.12300/j.issn.1674-5817.2024.194 Abstract （903）   HTML （27）    PDF （1198KB）（1144）       Knowledge map    Save Objective To observe and compare the effects of different concentrations of cyclophosphamide (CTX) in inducing premature ovarian insufficiency (POI) model in mice and investigate the mechanism of injury. Methods Thirty-two 6~8-week-old female C57BL/6J mice were randomly divided into four groups (n=8 per group) using a weight-based block randomization method. The POI model was established via a single intraperitoneal injection of 75 mg/kg cyclophosphamide (CTX), 120 mg/kg CTX, 120 mg/kg CTX + 12 mg/kg Busulfan, or an equivalent volume of normal saline (control). Ovarian coefficients, serum estradiol (E2) and follicle-stimulating hormone (FSH) levels were measured. Western blotting was performed to assess changes in ovarian expression levels of NAD-dependent deacetylase sirtuin-5 (SIRT5) and forkhead box O3a (FOXO3a) under different modeling conditions. After determining the optimal CTX concentration for modeling, an additional forty 6~8-week-old femal C57BL/6J mice were randomly divided into five groups (n=8 per group) using a weight-based block randomization method: saline control, 120 mg/kg CTX sampling at 1, 2, 7, or 14 days after modeling. Western blotting was used to evaluate temporal changes of ovarian SIRT5 and FOXO3a protein expression. Results Compared with the saline control, all concentrations of CTX (75 mg/kg CTX, 120 mg/kg CTX) and 120 mg/kg CTX + 12 mg/kg Busulfan induced POI injury in mice. The 120 mg/kg CTX group exhibited smaller changes in ovarian coefficients (P<0.001) and E2 levels (P<0.05), whereas the 120 mg/kg CTX + 12 mg/kg Busulfan group showed rough and reduced luster fur, sluggish response and was in the worst state. Compared with the saline control group, FOXO3a expression was significantly down-regulated (P<0.05), while SIRT5 remained unchanged in the 75 mg/kg CTX group (P>0.05). In contrast, both SIRT5 (P<0.05) and FOXO3a (P<0.05) were significantly down-regulated in the 120 mg/kg CTX group. Further analysis revealed that on day 2 and 7 after 120 mg/kg CTX modeling, the expressions of SIRT5 (P<0.01) and FOXO3a (P<0.001) were significantly down-regulated, with the largest decrease observed on day 7 (SIRT5, P<0.000 1; FOXO3a, P<0.000 1). Conclusion Ovarian injury in the POI model induced by 120 mg/kg CTX is milder than that in the POI model induced by 75 mg/kg CTX. Moreover, the expression changes of SIRT5 and FOXO3a are most significant on day 7 after modeling induced by 120 mg/kg CTX, which may be related to the inhibition of the SIRT5-FOXO3a signaling pathway. Table and Figures | Reference | Related Articles | Metrics

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Advances in Mouse Models of Amyotrophic Lateral Sclerosis LUO Lianlian, YUAN Yanchun, WANG Junling, SHI Guangsen Laboratory Animal and Comparative Medicine    2025, 45 (3): 290-299.   DOI: 10.12300/j.issn.1674-5817.2024.161 Abstract （901）   HTML （30）    PDF （816KB）（1677）       Knowledge map    Save Amyotrophic lateral sclerosis (ALS) is an irreversible, fatal neurodegenerative disorder whose incidence is positively correlated with the aging population. ALS is characterized by the progressive loss of motor neurons, leading to muscle weakness, atrophy, and ultimately respiratory failure. The pathogenesis of ALS involves multiple factors, including genetic and environmental influences, with genetic factors playing a particularly significant role. To date, several causative genes have been identified in ALS, such as the Cu/Zn superoxide dismutase 1 (Cu/Zn SOD1, also known as SOD1) gene, transactive response DNA-binding protein 43 (TDP-43) gene, fused in sarcoma (FUS) gene, and chromosome open reading frame 72 (C9orf72). Mutations in these genes have been found not only in familial ALS but also in sporadic ALS. Based on the identified ALS risk genes, various ALS animal models have been established through multiple approaches, including transgenic models, gene knockout/knock-in models, and adeno-associated virus-mediated overexpression models. These models simulate some typical pathological features of human ALS, such as motor neuron loss, ubiquitinated inclusions, and neuromuscular junction degeneration. However, these models still have limitations: (1) single-gene mutation models are insufficient to fully replicate the complex multi-factorial pathogenesis of sporadic ALS; (2) significant differences in microenvironmental regulation mechanisms and the rate of neurodegeneration between model organisms and humans may affect the accurate reproduction of disease phenotypes and the reliable evaluation of drug efficacy. To better understand the pathogenesis of ALS and promote the development of effective therapies, constructing and optimizing ALS animal models is crucial. This review aims to summarize commonly used ALS gene mutation mouse models, analyze their phenotypes and pathological characteristics, including transgenic mouse models, gene knockout/knock-in mouse models, and adeno-associated virus-mediated overexpression mouse models, and further discuss their specific applications in ALS pathogenesis research and drug development by comparing the advantages and limitations of each model. Table and Figures | Reference | Related Articles | Metrics

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An Overview of Strategies for Constructing Animal Models of Traditional Chinese Medicine Syndromes WANG Xiaoming, MENG Chenchen, FAN Lu, LI Yanyang, ZHANG Junping, LÜ Shichao Laboratory Animal and Comparative Medicine    2025, 45 (5): 596-610.   DOI: 10.12300/j.issn.1674-5817.2025.038 Abstract （896）   HTML （24）    PDF （1042KB）（793）       Knowledge map    Save This study aims to explore different construction methods for animal models of traditional Chinese medicine (TCM) syndromes and their advantages and disadvantages, to propose optimization strategies for existing problems in current construction methods, and to provide reference for constructing animal models of TCM syndromes that both preserve the essence of TCM syndromes and conform to modern scientific research standards. Using "traditional Chinese medicine", "syndrome", and "animal model" as key words, articles related to animal models of TCM syndromes from CNKI, Wanfang, and VIP databases are searched and reviewed. Then the theoretical basis, technical characteristics, and existing problems of the main construction methods of current TCM syndrome animal models are systematically sorted out, and corresponding optimization measures are proposed for the existing problems. The construction methods of TCM syndrome animal models include TCM etiology and pathogenesis construction, modern medical etiology and pathology construction, and integration of TCM and Western medicine for diseases and syndromes. The TCM etiology and pathogenesis construction method is guided by a holistic perspective, constructing syndrome models by simulating external factors such as six pathogenic factors and emotional disorders. Although it conforms to TCM theoretical connotation and has simple operation and strong controllability, this method has problems such as low modeling success rate and poor etiology-syndrome fit. The modern medical etiology and pathology construction method is based on microscopic pathological mechanisms, adopting highly controllable technical means such as drug intervention and surgical modeling. Although it has the characteristics of clear objective indicators and excellent reproducibility, this method has defects such as deviation from the essence of TCM "syndrome" and insufficient safety. The integrated TCM-Western medicine disease-syndrome method shows significant complementarity in syndrome essence restoration degree and technical feasibility, achieves systematic integration of TCM basic theories and clinical syndrome differentiation thinking in methodology, and integrates the objective evaluation system of modern medicine, improving the clinical consistency between Western medicine pathological mechanisms and TCM syndrome evolution patterns. However, this method still faces common challenges such as ambiguous syndrome identification standards and distortion of disease progression simulation. The construction of TCM syndrome animal models faces challenges such as poor theoretical adaptability and poor technical standardization, but has irreplaceable value in verifying the efficacy of prescriptions and promoting the internationalization of TCM. In the future, the construction of TCM syndrome animal models should be optimized through measures such as optimizing animal selection, improving the theoretical basis of preparation methods, standardizing the setting of modeling factors, and clarifying the standard for modeling success. Table and Figures | Reference | Related Articles | Metrics

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Optimization of Surgical Procedure and Efficacy Evaluation of Aortic Calcification Model in Rats with Chronic Kidney Disease PAN Yicong, JIANG Wenhong, HU Ming, QIN Xiao Laboratory Animal and Comparative Medicine    2025, 45 (3): 279-289.   DOI: 10.12300/j.issn.1674-5817.2024.128 Abstract （868）   HTML （27）    PDF （2015KB）（2252）       Knowledge map    Save Objective To establish a chronic kidney disease-associated aortic calcification model in SD rats using different nephrectomy surgical methods, and to compare and evaluate surgical duration and survival time to explore a more optimized modeling method. Methods According to different surgical methods, the SD rats were divided into four groups: Group A: intraperitoneal resection of 2/3 of the left kidney followed by right total nephrectomy in the second stage; Group B: intraperitoneal resection of 2/3 of the left kidney and simultaneous right total nephrectomy; Group C: dorsal approach right total nephrectomy followed by resection of 2/3 of the left kidney in the second stage; Group D: dorsal approach resection of 2/3 of the left kidney followed by right total nephrectomy in the second stage. After comparing survival curves of SD rats undergoing intraperitoneal versus dorsal approaches, and staged versus single-stage nephrectomy, the optimal nephrectomy surgical method was determined. Then, twenty-four 8-week-old SPF-grade male SD rats were selected for nephrectomy combined with calcitriol-induced calcification. Experimental group (12 rats): the dorsal approach left 2/3 nephrectomy followed by right total nephrectomy, with intraperitoneal injection of 1 μg/kg calcitriol administered one week later to induce aortic calcification. Control group (12 rats): the intraperitoneal injection of 250 μL/kg physiological saline containing 1% DMSO one week after sham surgery. After intraperitoneal injection of drugs for 3 months, the survival status of rats in each group was observed. Under anesthesia, blood samples were collected from each group to measure serum phosphorus and calcium ion concentrations, as well as serum urea nitrogen and creatinine levels. After euthanizing the rats, a post-mortem examination was performed to observe the residual kidney morphology, and HE staining was used to observe the pathological changes in the coronal section of the kidney. Additionally, the entire aorta of each group was taken, and the degree of aortic calcification was observed by staining with Alizarin red S and von Kossa. Real-time fluorescence quantitative PCR was used to detect the gene expression of smooth muscle actin-associated protein alpha (Sm22), Runt-related transcription factor 2 (Runx2), and osteopontin (OPN) in rat aortic tissue to evaluate the effectiveness of the model. Results The exploratory optimization experiment of different surgical procedures found that the survival rate of group D rats,which underwent 2/3 left kidney resection followed by right whole kidney resection via the dorsal approach, was the highest, indicating that this surgical procedure was the best method for establishing a chronic kidney disease model with renal dysfunction. The experimental group rats treated with this surgical procedure combined with high-dose calcitriol injection had significantly lower serum calcium ion concentration than those in the sham-operated control group (P<0.05), while serum phosphorus ion concentration, serum creatinine, and serum urea nitrogen levels were significantly higher than those of the control group (P<0.05). HE staining of the kidneys showed significant organic changes in the kidneys of the experimental group rats, with a significant decrease in glomerular count compared to that of the control group (P<0.05), indicating the successful establishment of a renal failure model. Alizarin red S staining showed significant pigment deposition in the aortic media of the experimental group rats, while von Kossa staining showed significant silver nitrate deposition in the aortic media of the experimental group rats, which was consistent with the manifestation of aortic calcification in renal failure. Real-time fluorescence quantitative PCR showed that the expression level of Sm22 in the aortic tissue of the experimental group rats decreased (P<0.05), while the expression levels of OPN and Runx2 increased (P<0.05), indicating a transition of aortic smooth muscle cells from smooth muscle phenotype to bone-like phenotype and successful induction of an aortic calcification model. Conclusion The method of establishing an aortic calcification model of chronic kidney disease in SD rats by first removing two-thirds of the left kidney via the dorsal approach followed by right total nephrectomy, combined with high-dose calcitriol administration, shortens the surgical time, improves the success rate of modeling, and increases the animal survival rate. Table and Figures | Reference | Related Articles | Metrics

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Prospects for 3D Bioprinting Research and Transdisciplinary Application to Preclinical Animal Models HU Min, DONG Lexuan, GAO Yi, XI Ziqi, SHEN Zihao, TANG Ruiyang, LUAN Xin, TANG Min, ZHANG Weidong Laboratory Animal and Comparative Medicine    2025, 45 (3): 318-330.   DOI: 10.12300/j.issn.1674-5817.2024.193 Abstract （867）   HTML （12）    PDF （1108KB）（2204）       Knowledge map    Save Animal experiments are widely used in biomedical research for safety assessment, toxicological analysis, efficacy evaluation, and mechanism exploration. In recent years, the ethical review system has become more stringent, and awareness of animal welfare has continuously increased. To promote more efficient and cost-effective drug research and development, the United States passed the Food and Drug Administration (FDA) Modernization Act 2.0 in September 2022, which removed the federal mandate requiring animal testing in preclinical drug research. In April 2025, the FDA further proposed to adopt a series of "new alternative methods" in the research and development of drugs such as monoclonal antibodies, which included artificial intelligence computing models, organoid toxicity tests, and 3D micro-physiological systems, thereby gradually phasing out traditional animal experiment models. Among these cutting-edge technologies, 3D bioprinting models are a significant alternative and complement to animal models, owing to their high biomimetic properties, reproducibility, and scalability. This review provides a comprehensive overview of advancements and applications of 3D bioprinting technology in the fields of biomedical and pharmaceutical research. It starts by detailing the essential elements of 3D bioprinting, including the selection and functional design of biomaterials, along with an explanation of the principles and characteristics of various printing strategies, highlighting the advantages in constructing complex multicellular spatial structures, regulating microenvironments, and guiding cell fate. It then discusses the typical applications of 3D bioprinting in drug research and development,including high-throughput screening of drug efficacy by constructing disease models such as tumors, infectious diseases, and rare diseases, as well as conducting drug toxicology research by building organ-specific models such as those of liver and heart. Additionally,the review examines the role of 3D bioprinting in tissue engineering, discussing its contributions to the construction of functional tissues such as bone, cartilage, skin, and blood vessels, as well as the latest progress in regeneration and replacement. Furthermore, this review analyzes the complementary advantages of 3D bioprinting models and animal models in the research of disease progression, drug mechanisms, precision medicine, drug development, and tissue regeneration, and discusses the potential and challenges of their integration in improving model accuracy and physiological relevance. In conclusion, as a cutting-edge in vitro modeling and manufacturing technology, 3D bioprinting is gradually establishing a comprehensive application system covering disease modeling, drug screening, toxicity prediction, and tissue regeneration. Table and Figures | Reference | Related Articles | Metrics

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Recommendations for Standardized Reporting of Systematic Reviews and Meta-Analysis of Animal Experiments ZHENG Qingyong, YANG Donghua, MA Zhichao, ZHOU Ziyu, LU Yang, WANG Jingyu, XING Lina, KANG Yingying, DU Li, ZHAO Chunxiang, DI Baoshan, TIAN Jinhui Laboratory Animal and Comparative Medicine    2025, 45 (4): 496-507.   DOI: 10.12300/j.issn.1674-5817.2025.017 Abstract （789）   HTML （28）    PDF （1130KB）（2157）       Knowledge map    Save Animal experiments are an essential component of life sciences and medical research. However, the external validity and reliability of individual animal studies are frequently challenged by inherent limitations such as small sample sizes, high design heterogeneity, and poor reproducibility, which impede the effective translation of research findings into clinical practice. Systematic reviews and meta-analysis represent a key methodology for integrating existing evidence and enhancing the robustness of conclusions. Currently, however, the application of systematic reviews and meta-analysis in the field of animal experiments lacks standardized guidelines for their conduct and reporting, resulting in inconsistent quality and, to some extent, diminishing their evidence value. To address this issue, this paper aims to systematically delineate the reporting process for systematic reviews and meta-analysis of animal experiments and to propose a set of standardized recommendations that are both scientific and practical. The article's scope encompasses the entire process, from the preliminary preparatory phase [including formulating the population， intervention， comparison and outcome （PICO） question, assessing feasibility, and protocol pre-registration] to the key writing points for each section of the main report. In the core methods section, the paper elaborates on how to implement literature searches, establish eligibility criteria, perform data extraction, and assess the risk of bias, based on the Preferred Reporting Items for Systematic Reviews and Meta-analysis （PRISMA) statement, in conjunction with relevant guidelines and tools such as Animal Research： Reporting of in Vivo Experiments （ARRIVE) and a risk of bias assessment tool developed by the Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE). For the presentation of results, strategies are proposed for clear and transparent display using flow diagrams and tables of characteristics. The discussion section places particular emphasis on how to scientifically interpret pooled effects, thoroughly analyze sources of heterogeneity, evaluate the impact of publication bias, and cautiously discuss the validity and limitations of extrapolating findings from animal studies to clinical settings. Furthermore, this paper recommends adopting the Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology to comprehensively grade the quality of evidence. Through a modular analysis of the entire reporting process, this paper aims to provide researchers in the field with a clear and practical guide, thereby promoting the standardized development of systematic reviews and meta-analysis of animal experiments and enhancing their application value in scientific decision-making and translational medicine. Table and Figures | Reference | Related Articles | Metrics

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Comparison of Histopathological and Molecular Pathological Phenotypes in Mouse Models of Intrauterine Adhesions Induced by Two Concentrations of Ethanol Perfusion JIANG Juan, SONG Ning, LIAN Wenbo, SHAO Congcong, GU Wenwen, SHI Yan Laboratory Animal and Comparative Medicine    2025, 45 (4): 393-402.   DOI: 10.12300/j.issn.1674-5817.2024.183 Abstract （781）   HTML （30）    PDF （2290KB）（1348）       Knowledge map    Save Objective To construct intrauterine adhesion (IUA) mouse models induced by two different concentrations of ethanol injury, compare the phenotypes, and optimize a more stable IUA modeling method. Methods Twenty 8-week-old female C57BL/6N mice were randomly divided into two groups: the 95% ethanol injury group and the 50% ethanol injury group. Using a self-control method, the left uterine horn was infused with ethanol to establish the IUA model, while the right uterine horn was infused with saline as the sham operation. Five mice from each group were euthanized on day 7 and 15 after modeling, and uterine tissues were collected. Hematoxylin-eosin (HE) staining was used to observe the endometrial pathology, and Masson staining was used to assess the degree of endometrial fibrosis. Quantitative real-time PCR was employed to detect the expression levels of fibrosis markers and pro-inflammatory factors in the uterine tissues. Results Compared to the sham operation, these two ethanol injury led to a significant reduction in elasticity of the uterus, an increase in inflammatory infiltration, and a marked increase in the degree of fibrosis on day 7 after modeling (P<0.05). The 95% ethanol injury group showed a significant decrease in endometrial thickness (P<0.05), whereas no significant change was observed in the 50% ethanol injury group when compared to the sham operation (P>0.05). The expression levels of fibrotic marker molecules collagen type Ⅳ alpha 1 chain (Col4A1), α-smooth muscle actin (α-SMA), transforming growth factor-β (TGF-β), and pro-inflammatory factors tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were significantly elevated in the 50% ethanol injury group when compared to the sham operation (P<0.05), although there was an increasing trend of the same markers in the 95% ethanol injury group, the differences were not statistically significant (P>0.05). On day 15 after modeling, the histopathological changes in both ethanol injury groups were not significant when compared to the sham operation, the expression levels of Col4A1, TGF-β, TNF-α and IL-1β remained significantly higher in the 50% ethanol injury group (P<0.05), while only IL-1β was significantly elevated in the 95% ethanol injury group (P<0.05). Conclusion Uterine infusion with 95% ethanol results in more marked histopathological changes in the IUA mouse model compared to the 50% ethanol injury group. The 95% ethanol injury model is suitable for histopathological studies. However, the 50% ethanol injury group shows higher expression levels of fibrosis markers and pro-inflammatory factors compared to the 95% ethanol injury group, suggesting that the 50% ethanol injury model is more suitable for molecular pathological study. Table and Figures | Reference | Related Articles | Metrics

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Reshaping Intercellular Interactions: Empowering the Exploration of Disease Mechanisms and Therapies Using Organoid Co-Culture Models TAN Dengxu, MA Yifan, LIU Ke, ZHANG Yanying, SHI Changhong Laboratory Animal and Comparative Medicine    2025, 45 (3): 309-317.   DOI: 10.12300/j.issn.1674-5817.2024.164 Abstract （778）   HTML （11）    PDF （1242KB）（1878）       Knowledge map    Save The organoid co-culture model, as a novel tool for recreating a three-dimensional microenvironment to study cell-cell interactions, has demonstrated significant application potential in biomedical research in recent years. By simulating the in vivo tissue microenvironment, this model provides a more precise experimental platform for investigating complex cellular interactions, particularly in areas such as tumor immune evasion mechanisms, drug sensitivity testing, and the pathological characterization of neurodegenerative diseases, where it has demonstrated significant value. However, the organoid co-culture model still faces several challenges in terms of standardized procedures, large-scale cultivation, ethical guidelines, and future development. In particular, in the field of laboratory animal science, how to effectively combine organoids with traditional animal models, and how to select the most appropriate model for different research needs while exploring its potential for replacement, remain pressing issues. In the context of ethical approval and the replacement of animal experiments, the organoid co-culture model offers an experimental approach that better aligns with the "3R" principle (Replacement, Reduction, Refinement), potentially becoming an important tool for replacing traditional animal models. To this end, this paper reviews the latest advances and key challenges in this field, providing a detailed description of the construction methods for organoid co-culture models and discussing their applications in disease mechanism research and drug screening. The paper also systematically compares the organoid co-culture models with traditional animal models, exploring the criteria for selecting the appropriate model for specific applications. Furthermore, this paper discusses the potential value of organoid co-culture models as alternatives to animal experiments and anticipates future development trends of this technology. Through these discussions, the paper aims to promote the innovation and development of organoid co-culture technology and provide new perspectives and scientific evidence for future research. Table and Figures | Reference | Related Articles | Metrics

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Progress and Evaluation of Animal Model of Heart Qi-Yin Deficiency Syndrome LIU Yayi, JIA Yunfeng, ZUO Yiming, ZHANG Junping, LÜ Shichao Laboratory Animal and Comparative Medicine    2025, 45 (4): 411-421.   DOI: 10.12300/j.issn.1674-5817.2024.176 Abstract （773）   HTML （23）    PDF （921KB）（2245）       Knowledge map    Save Animal models combining disease and syndrome are important research tools to explore the nature of traditional Chinese medicine (TCM) syndromes. At present, the construction and evaluation methods of animal models have preliminarily established the foundation for standardized development. Qi-yin deficiency syndrome is a common type of TCM syndrome in cardiovascular diseases. It is an important pathogenic factor causing the onset, pathological damage, and chronic nature of cardiovascular diseases, as well as triggering other illnesses. The establishment of an animal model of cardiovascular disease with the characteristics of Qi-yin deficiency, along with an objective and standardized evaluation system, has become an important part of modern cardiovascular disease research. In recent years, research on the construction and evaluation of animal models of heart Qi-yin deficiency syndrome has increased, but the construction methods and evaluation criteria vary. Compared with other animal models, the literature is limited, lacking statistics and overall analysis. Therefore, based on the scientific connotation of heart Qi-yin deficiency syndrome, this article systematically reviews the evaluation system of its animal model, covering multidimensional methods such as macroscopic characterization assessment, physicochemical indicators and objective evaluation, and syndrome differentiation based on prescriptions. The specific model construction strategies are described, including single-factor induction methods (sleep deprivation, chronic intermittent hypoxia, arterial occlusion, high-salt feeding) and the compound-factor induction methods (sleep deprivation combined with drug method, chronic intermittent hypoxia combined with drug method, exhaustive swimming combined with drug method). Meanwhile, application examples of each model in the research are listed, the existing problems in the current model construction and evaluation are analyzed, and optimization directions are proposed, such as promoting the compound factor induction strategy and improving the objectivity of the evaluation criteria. This article aims to provide theoretical references for constructing an animal model of heart Qi-yin deficiency syndrome that conforms to TCM characteristics, and thereby laying a scientific foundation for the prevention and treatment of cardiovascular diseases with TCM. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Characteristics Analysis of Gut Microbiota and Its Sex Differences in Laboratory Animals SHEN Huangyi, HUANG Yufei, YANG Yunpeng Laboratory Animal and Comparative Medicine    2025, 45 (3): 349-359.   DOI: 10.12300/j.issn.1674-5817.2024.124 Abstract （772）   HTML （18）    PDF （824KB）（5333）       Knowledge map    Save Laboratory animals serve as the cornerstone in life science research, acting as surrogate models for human physiology, pathology, and disease treatment. They play an irreplaceable role in basic research, drug development, and translational medicine. Gut microbiota, a complex microbial community comprising bacteria, fungi, viruses, and unicellular organisms, colonizes the host's intestinal tract and is closely associated with the maintenance of normal physiological metabolism and overall health. Studies have shown that dysbiosis of the gut microbiota can lead to various diseases, including obesity, diabetes, hypertension, inflammatory bowel disease, and Alzheimer's disease. Therefore, conducting characteristic analyses of the gut microbial composition of laboratory animals can not only enhance the reliability of experimental outcomes but also facilitate their translational application. Sex differences represent a critical variable in biological research, significantly influencing the physiological functions, metabolic traits, and gut microbial composition of laboratory animals. However, a pronounced sex bias has been widely observed in many biological studies, thereby limiting the generalizability of results. This study focused on ten commonly used laboratory animals in life sciences, including mice, rats, guinea pigs, hamsters, rabbits, dogs, cats, non-human primates, miniature pigs, and chickens. Their gut microbial composition was summarized and related sex-specific differences of certain species were analyzed. Furthermore, by comparing the gut microbiota of laboratory animals with that of humans, this study offers novel perspectives for comparative medical research. In summary, this study not only deepens researchers' understanding of gut microbiota characteristics and sex-dependent variations across laboratory animal species but also provides practical guidance for selecting appropriate laboratory animals, constructing sex-specific disease models, and interpreting experimental results in scientific studies. Reference | Related Articles | Metrics

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Guidelines for Selecting Animal Models in Preclinical Research of Intervertebral Disc Degeneration (2025 Edition) Expert Committee on Medical Animal Experiments, Chinese Research Hospital Association, Professional Committee on Neural Regeneration and Tissue–Organ Injury Repair, Chinese Research Hospital Association, Section of Engineering Anatomy, Chinese Society for Anatomical Sciences Laboratory Animal and Comparative Medicine    2025, 45 (5): 524-541.   DOI: 10.12300/j.issn.1674-5817.2025.048 Abstract （752）   HTML （74）    PDF （1174KB）（468）       Knowledge map    Save Intervertebral disc herniation is a highly prevalent orthopedic disorder, and intervertebral disc degeneration (IDD), the key pathological basis, is a complex pathological process characterized by progressive degradation of extracellular matrix, structural failure, and loss of biomechanical function, which not only shows higher prevalence in the population, but is also the primary cause of chronic low back pain and dysfunction worldwide, causing a huge socioeconomic burden. Although constructing IDD animal models is important for exploring the pathological mechanisms and promoting translational research of this disease, the etiology and pathophysiological mechanisms of IDD have not been fully elucidated. There are significant differences between humans and common laboratory animals in spinal anatomy, biomechanics, and degenerative course, coupled with the diversity and lack of unified standards of existing IDD animal models. This guide systematically reviews IDD animal models of rodents, non-human primates, as well as different species such as rabbits, goats/sheep, pigs, and dogs, focusing on the modeling principles of three main types of models: inducible models (such as annulus fibrosus/nucleus pulposus/endplate injury and mechanical injury) are suitable for simulating acute injury and rapid screening of therapies due to their high controllability and short cycle; spontaneous models can better simulate the age-related natural degeneration process in humans; genetically modified models provide powerful tools for analyzing specific molecular pathways. The guideline deeply analyzes the key technical points, reproducibility, and clinical relevance of these models. It also compares their advantages, limitations, and applicable research scenarios to guide researchers to conduct "scientific question-driven" precise model selection. Meanwhile, to improve the depth and comparability of research results, this guideline proposes a multidimensional endpoint evaluation system for IDD animal model experiments covering imaging, histology, biochemistry/molecular biology, biomechanics, and pain-related behavior, with recommended observation time windows. It also clarifies the "3Rs (replacement, reduction, and refinement)" ethical principles and animal welfare requirements throughout the experiment. In addition, the guideline outlines future research directions such as integrating single-cell omics, multiscale mechanical analysis, and strengthening pain-related phenotype assessment. This guideline aims to provide researchers with a systematic and standardized methodological framework for the rational selection and application of IDD animal models under specific scientific questions and resource constraints, in order to reduce inter-study heterogeneity, enhance the translation efficiency of preclinical findings, promote high-quality development in the field, and ultimately provide a solid scientific foundation for developing innovative therapies to delay or even reverse IDD. Table and Figures | Reference | Related Articles | Metrics

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Cartilage Protection and Anti-Inflammatory Effects of Fraxetin on Monosodium Iodoacetate-Induced Rat Model of Osteoarthritis LIU Zhiwei, YANG Ran, LIAN Hao, ZHANG Yu, JIN Lilun Laboratory Animal and Comparative Medicine    2025, 45 (3): 259-268.   DOI: 10.12300/j.issn.1674-5817.2024.165 Abstract （742）   HTML （46）    PDF （2277KB）（2423）       Knowledge map    Save Objective To establish a rat model of osteoarthritis and study the anti-inflammatory effects and mechanisms of fraxetin. Methods Eighteen 8-week-old male SPF-grade SD rats were randomly divided into three groups: Rats in the blank group received a right articular cavity injection of 50 μL of normal saline for 1 week; the model and intervention groups were injected with monosodium iodoacetate (MIA) into the right joint cavity to induce osteoarthritis, while the intervention group subsequently received fraxetin (5 mg·kg-1·d-1) for 1 week. Four weeks after drug intervention, abdominal aortic blood was collected. The animals were then euthanized, and knee joint cartilage were collected. The cartilage samples were stained with hematoxylin-eosin, safranin O-fast green, and toluidine blue for histopathological examination and scoring using the Mankin and OARSI scoring systems. The trabecular bone volume/total volume (Tb.BV/TV), trabecular bone surface density/total volume (Tb.BS/TV), and trabecular number (Tb.N) of each group were compared and analyzed using a micro-CT scanning system. The expression levels of various inflammatory factors [tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6)], and cartilage oligomeric matrix protein (COMP) were measured using enzyme-linked immunosorbent assay (ELISA). The expression levels of mitogen-activated protein kinase p38 (p38 MAPK), phosphorylation-p38 MAPK (p-p38 MAPK), c-Jun N-terminal kinase (JNK), and phosphorylation-JNK (p-JNK) were measured by western blotting. Results The staining of cartilage sections of rat knee joints showed that the articular surface defects in the model group were severe, while the cartilage destruction in the intervention group was relatively reduced. Micro-CT results showed that Tb.BV/TV, Tb.BS/TV and Tb.N in the intervention group were significantly higher than those in the model group (P < 0.05); the Mankin score in the model group was significantly higher than that in the blank group (P < 0.05), the Mankin score in the intervention group was significantly lower than that in the model group (P < 0.05); while the OARSI score in the intervention group was significantly lower than that in the model group (P < 0.05). The results of the enzyme-linked immunosorbent assay showed that the serum levels of TNF-α, IL-1β, IL-6, and COMP in the model group were significantly higher than those in the blank group (all P < 0.05), while those in the intervention group were significantly lower than in the model group (P < 0.05). Western blot results showed that the expression levels of p-p38 MAPK and p-JNK in the knee cartilage tissue were significantly lower in the intervention group than in the model group (both P < 0.05), and significantly higher in the model group than in the blank group (both P < 0.05). Conclusion Fraxetin may play a therapeutic role in a monosodium iodoacetate-induced rat model of osteoarthritis through the p38 MAPK pathway. Table and Figures | Reference | Related Articles | Metrics

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Identification and Analysis of MHCⅡ Genes in Wuzhishan Pigs LIU Yuanyuan, XIN Wenshui, CHAO Zhe, CAO Zongxi, CAI Yifei, LI Qiang, LI Lingwei, LIU Guangliang Laboratory Animal and Comparative Medicine    2025, 45 (3): 340-348.   DOI: 10.12300/j.issn.1674-5817.2024.135 Abstract （733）   HTML （13）    PDF （1751KB）（1738）       Knowledge map    Save Objective To obtain the gene sequences of major histocompatibility complex (MHC ) Ⅱgenes of Wuzhishan pigs, analyze their genetic information, and explore the biological functions of their MHC system. Methods Spleen samples were collected from 3 adult male Wuzhishan pigs. Primers were designed according to MHCⅡ gene sequences, and the coding sequences of Wuzhishan pig MHCⅡ genes were amplified by RT-PCR. Sanger sequencing was performed to determine the full-length sequences. Bioinformatics tools were employed to analyze the physicochemical properties, phylogenetic relationships, conserved motifs, structural domains, chromosomal localization, and syntenic relationships of these genes. Results Eight MHCⅡ genes were identified in Wuzhishan pigs, designated as SLA-DRA, SLA-DQA, SLA-DQB, SLA-DRB, SLA-DOB, SLA-DMB, SLA-DMA and SLA-DOA. The full-length sequences of these genes were determined by Sanger sequencing and subsequently deposited in GenBank under accession numbers PQ182796, PQ182797, PQ182798, PQ182799, PQ182800, PQ182801, PQ182802, and PQ164779. Phylogenetic analysis showed that the six MHCⅡ genes of Wuzhishan pigs clustered separately from their counterparts in Duroc, Meishan, Large White, and Bama pigs, indicating distinct evolutionary trajectories. Bioinformatics analysis demonstrated that most MHC Ⅱ proteins were hydrophobic, with molecular weights ranging from 27 700 to 30 000 Da. Genes within the same subregion shared conserved motifs. Specifically, four MHCⅡ proteins encoded by SLA-DQB, SLA-DRB, SLA-DOB, and SLA-DMB contained the MHCⅡβ conserved domain, while those encoded by the genes SLA-DRA, SLA-DQA, SLA-DMA, and SLA-DOA contained the MHCⅡα conserved domain. The eight MHCⅡ genes were scattered along the long arm of chromosome 7 in the Wuzhishan pigs, exhibiting syntenic relationships with three human genes and five Duroc pig genes. Conclusion The MHCⅡ genes of Wuzhishan pigs may possess a unique evolutionary origin. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Animal Models of Gastric Ulcer of Spleen-Stomach Deficiency Cold Type LIU Ziqi, LI Yunying, LI Qin, LI Yuanhan, HE Fangyan, WEN Weibo Laboratory Animal and Comparative Medicine    2025, 45 (5): 574-585.   DOI: 10.12300/j.issn.1674-5817.2025.015 Abstract （680）   HTML （24）    PDF （919KB）（4168）       Knowledge map    Save Gastric ulcer (GU) is one of the common, frequently-occurring and intractable diseases of the digestive system. Spleen-stomach deficiency cold type is the most common and hard-to-cure syndrome pattern of GU, and is both a focus and a challenge in medical research. Therefore, constructing a scientific, reasonable, and clinically practical animal model of GU with spleen-stomach deficiency cold type and formulating objective and effective evaluation criteria are of great significance for in-depth research on the pathogenesis and treatment of GU. In this paper, the methods for constructing GU animal models of spleen-stomach deficiency cold type are comprehensively introduced by systematically reviewing the relevant literature. Firstly, the construction methods of pathological models of GU in Western medicine are introduced, including pyloric ligation method, water immersion-restraint stress method, ethanol-induced method, acetic acid-induced method, etc. This paper expounds the establishment methods for spleen-stomach deficiency cold syndrome type model in traditional Chinese medicine (TCM), including diet disorder method, bitter cold diarrhea method, excessive fatigue method, Qi consumption and Qi impairment method, and overeating sour-flavor method. This paper focuses on the construction methods for disease-syndrome combination GU models of spleen-stomach deficiency cold type, including two-factor modeling method and three-factor modeling method. Meanwhile, the evaluation indices of GU animal models of spleen-stomach deficiency cold type were summarized from various aspects, including animal physical signs ( appearance symptoms, animal behavior, and metabolic indices), as well as tissue morphology and molecular biology-related indicators ( gastric function, oxidative stress, inflammatory factors, other cytokines, four coagulation parameters, intestinal flora detection ), for constructing a comprehensive evaluation system. From the perspective of prescription-based verification, this paper further analyzes the drug composition and pharmacological effects to infer the syndrome type of the treated animal model, so as to verify whether the target animal model is successfully constructed. This review aims to provide a valuable reference for establishing a syndrome-specific GU animal model that closely aligns with clinical reality and embodies the principles of Chinese medicine. This will further advance research on TCM-pattern GU syndromes and deepen the exploration of herbal medicine-based treatments for GU, ultimately promoting the clinical integration and advancement of Chinese medicine in GU therapy. Table and Figures | Reference | Related Articles | Metrics

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Diagnosis of an Outbreak of Canine Distemper in Cynomolgus Monkeys in an Experimental Monkey Farm in 2019 WANG Chenjuan, YANG Lingyan, WANG Lipeng, SUN Xueping, LI Jingwen, GUO Lianxiang, RONG Rong, SHI Changjun Laboratory Animal and Comparative Medicine    2025, 45 (3): 360-367.   DOI: 10.12300/j.issn.1674-5817.2024.160 Abstract （669）   HTML （21）    PDF （1701KB）（2185）       Knowledge map    Save Objective To report the diagnosis of a canine distemper virus outbreak among a colony of cynomolgus monkeys at an experimental monkey farm in 2019. Methods A total of 46 samples were collected from 21 diseased cynomolgus monkeys (exhibiting symptoms such as facial rash, skin scurf, runny nose, and diarrhea) and from one deceased monkey at an experimental monkey breeding farm in South China in late 2019, including serum, skin rash swabs, and anticoagulated whole blood, liver, lung, and skin tissues were submitted for testing. All submitted samples were tested for canine distemper virus gene fragments using real-time quantitative PCR, while immunohistochemical staining was performed to detect canine distemper virus nucleoprotein in lung tissues. The skin tissue of the deceased monkey was ground and sieved. The filtrate was inoculated into a monolayer MDCK cell line for virus isolation. Then, whole-genome sequencing was performed to identify the isolated virus. The Clustal Omega tool was used to align and analyze the homology of different Asian canine distemper virus isolates. A phylogenetic tree was constructed, followed by genetic evolutionary analysis. Results Clinical retrospective analysis revealed that the diseased cynomolgus monkeys exhibited symptoms similar to those observed in cynomolgus monkeys infected with measles virus. Necropsy findings showed red lesions in the lungs and significant hemorrhage in the colonic mucosa. Real-time quantitative PCR detected canine distemper virus nucleic acid in the serum, skin rash swabs of the infected monkeys, and various tissue samples of the deceased monkey, all of which tested positive. Calculation based on the standard curve formula indicated the viral load was highest in the skin tissue. Immunohistochemical staining of the deceased monkey's lung tissue demonstrated aggregation of CDV nucleoprotein in alveolar epithelial cells, bronchi, and bronchioles. A CDV strain was isolated from the skin tissue of the deceased monkey. Phylogenetic analysis indicated that this strain shares the closest relationship (98.86%) with the Asian-1 type canine distemper virus strain CDV/dog/HCM/33/140816, previously identified in dogs in Vietnam. Conclusion Based on comprehensive analysis of clinical symptoms, nucleic acid detection, viral protein immunohistochemistry, and whole-genome sequencing results, the diagnosis confirms that the cynomolgus monkeys in this facility are infected with canine distemper virus. It is recommended to include canine distemper virus as a routine surveillance target in captive monkey populations. Additionally, this study provides a foundation for further research on the molecular biological characteristics of canine distemper virus. Table and Figures | Reference | Related Articles | Metrics

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Key Challenges and Mitigation Strategies for Animal Pregnancy in Non-clinical Reproductive Toxicity Testing of Drugs LIU Kun, LAN Qing, YI Bing, XIE Xiaojie Laboratory Animal and Comparative Medicine    2025, 45 (4): 449-456.   DOI: 10.12300/j.issn.1674-5817.2024.177 Abstract （668）   HTML （13）    PDF （994KB）（3061）       Knowledge map    Save Non-clinical reproductive toxicity studies typically employ mammals like rats, rabbits, and cynomolgus monkeys, with animal pregnancy being a key challenge in such testing. This article focuses on the difficulties encountered in the animal pregnancy process and potential countermeasures. Rats can be used for fertility and early embryonic development toxicity studies (Segment Ⅰ), embryo-fetal development toxicity studies (Segment Ⅱ), and perinatal toxicity studies (Segment Ⅲ). The estrous cycle of female rats can be determined by vaginal smear, and mating behavior is confirmed through copulatory plug checks the following day after pairing one female with one male in the same cage. Rabbits are commonly used in embryo-fetal development toxicity studies (Segment Ⅱ). Mating behavior between male rabbits and estrous females is observed to determine the time of conception. However, challenges such as atypical estrus of female rabbits, large variations in estrus between batches, and mating failure often occur in reproductive toxicity testing, which may be addressed through prolonged light exposure, increased protein supplementation, optimized mating strategies, and environmental modifications like female and male rabbits are raised adjacent to each other. Non-human primates (NHPs) are typically employed in perinatal toxicity studies (Segment Ⅲ), where one of the key challenges lies in accurately determining sexual maturity in males - a critical factor for reproductive toxicity testing, which can be assessed through comprehensive evaluation of age, body weight, and testicular volume. Generally, male macaques are considered sexually mature when they meet the following criteria: age >4.5 years, body weight >4.5 kg, single testis volume >10 mL, and combined testicular volume >20 mL. For pregnancy confirmation, ultrasound examination demonstrating visible gestational sacs is required, though this necessitates experienced veterinary clinicians to establish standardized ultrasound examination protocols. In conclusion, reproductive toxicity studies should employ species-appropriate detection methods and evaluation criteria based on anatomical characteristics of the reproductive system to ensure successful mating and proper study execution. Table and Figures | Reference | Related Articles | Metrics

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Analysis of Common Causes of Out-of-Specification Results in the Test for Depressor Substances TONG Xiyang, QUE Changtian, ZHANG Feng, ZHAO Lu, WANG Hongping Laboratory Animal and Comparative Medicine    2025, 45 (3): 331-339.   DOI: 10.12300/j.issn.1674-5817.2024.143 Abstract （653）   HTML （13）    PDF （1049KB）（3643）       Knowledge map    Save According to General Chapter 1145 of Division IV in the Chinese Pharmacopoeia (2020 Edition), the test for depressor substances is a common method for drug testing. It determines whether the level of depressor substances in a test sample complies with the specified standards by comparing the extent of blood pressure reduction in anesthetized cats induced by the histamine reference substance and the test sample. If an out-of-specification (OOS) result occurs in the test for depressor substances, it may be caused by inherent quality issues of the drug or errors in the testing process. Therefore, analyzing the causes of OOS is particularly important for confirming the test results and evaluating drug quality. Cats are used as experimental animals in the test for depressor substances. Compared with conventional laboratory animals, they are less stable, surgery procedures are more challenging, and the testing process is more complex. These factors make it more difficult to investigate the causes of OOS in this test. Based on a review of the literature and practical work experience, this article analyzes the causes of OOS in the test for depressor substances from the following five aspects: (1) an analysis of the impact of drug standards on OOS from three aspects: standard determination, standard content, and standard drafting; (2) personnel qualifications, including pre-employment training, compliance with standard operating procedures during experimental operations, and the ability to operate instruments; (3) factors related to cats, used as experimental animals in the test for depressor substances, including physiological characteristics, genetic background, and abnormal conditions during the experiment; (4) reference substances, reagents, test samples, and key instruments such as the multi-channel physiological signal instrument; (5) experimental operations including animal anesthesia, arterial and venous catheterization, drug administration, and data processing. This article aims to provide reference approaches for professionals engaged in the testing of pharmaceuticals and biological products when analyzing the causes of OOS in the test for depressor substances. Table and Figures | Reference | Related Articles | Metrics

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Establishment of a Rat Model of Alzheimer's Disease by Introducing Human Triple Mutant APP Gene into Hippocampus via Brain Stereotactic Technology XIAO Linlin, YANG Yixuan, LI Shanshan, LUO Lanshiyu, YIN Siwei, SUN Juming, SHI Wei, OUYANG Yiqiang, LI Xiyi Laboratory Animal and Comparative Medicine    2025, 45 (3): 269-278.   DOI: 10.12300/j.issn.1674-5817.2025.030 Abstract （649）   HTML （31）    PDF （2291KB）（1926）       Knowledge map    Save Objective To establish a rat model of Alzheimer's disease (AD) expressing human triple mutant amyloid precursor protein (APP) in the hippocampus, and to provide a model for the study of disease mechanisms and drug development. Methods Twenty-four 12-week-old SPF-grade female SD rats were randomly divided into a blank control group, a virus control group and an experimental group, with eight rats in each group; among them, the experimental group received a stereotaxic injection of adeno-associated virus (AAV) carrying the human triple mutant APP and NanoLuc luciferase genes into the hippocampus. In vivo imaging was used to observe viral expression in the brains of rats in each group, the novel object recognition test was used to assess the recognition memory of the rats in each group, real-time fluorescent quantitative PCR was used to detect the expression level of the APP gene, HE staining was used to examine the brain histopathology, Nissl staining was used to assess the hippocampal lesions, and immunohistochemistry was used to detect the deposition of amyloid β-protein (Aβ). Results In vivo imaging showed that reporter fluorescence was detected in the brains of rats in both experimental and virus control groups. Fluorescence quantitative PCR showed that the expression level of the APP gene was significantly increased in the brains of rats in the experimental group (P<0.01). Novel object recognition test revealed that the recognition memory of rats in the experimental group was significantly reduced compared with that of the blank control group (P<0.01). Six months after recombinant AAV virus infection, HE staining and Nissl staining of brain tissues showed that the number of neurons and Nissl bodies in the CA1 region of the hippocampus in the experimental group was reduced and disorganized; immuno-histochemistry testing of the CA1 region of the hippocampus and the pyramidal cell layer of the experimental group revealed prominent brown deposits, indicating Aβ protein deposition. Conclusion The rat model successfully established by stereotaxic injection and AAV-mediated delivery of human triple mutant APP gene exhibits typical AD features, providing a valuable animal model for studying AD pathology and developing drug therapies targeting Aβ protein deposition. Table and Figures | Reference | Related Articles | Metrics

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Research Advances and Challenges of Gene Drive Technology in Mosquito-Borne Disease Control YUN Jiaqi, MA Qin, WANG Guandong, SUN Peilu, WANG Yiguan, WANG Sibao Laboratory Animal and Comparative Medicine    2025, 45 (6): 773-783.   DOI: 10.12300/j.issn.1674-5817.2025.138 Abstract （647）   HTML （11）    PDF （2182KB）（504）       Knowledge map    Save Mosquito-borne diseases (such as malaria, dengue fever, Zika virus disease, and Chikungunya) pose major threats to global public health, while traditional control methods based on chemical pesticides face severe challenges including enhanced drug resistance in vector mosquitoes and environmental pollution. Genetic control strategies have become high-potential alternative solutions for mosquito control due to their species specificity and environmental friendliness. Gene drive technology uses gene editing tools such as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) to enable specific genes to efficiently spread in target mosquito populations through "super-Mendelian inheritance", offering a revolutionary strategy for the prevention and control of mosquito-borne diseases. This review systematically summarizes key advances, core challenges, and response strategies of gene drive technology in this field. Research advances: (1) In Anopheles malaria vectors, population suppression drives targeting sex determination genes or female reproductive genes can cause female sterility or skewed sex ratios to achieve population suppression. Population replacement gene drive strategies targeting host genes associated with Plasmodium infection or delivering anti-Plasmodium effector molecules in Anopheles can effectively block pathogen transmission. (2) In Aedes mosquito vectors of arboviruses, targeting female flight-essential genes achieves population suppression, and coupling of antiviral effector systems with drive elements is explored. Optimized split gene drive strategies demonstrate high cutting and recombination efficiency, and models predict safe and controllable spread of disease-resistance traits. (3) In Culex mosquitoes transmitting lymphatic filariasis, homology drive elements are integrated into two genes involved in the eye pigment synthesis pathway, allowing clear visualization of gene drive efficiency through eye color. Core Challenges: technological challenges include low homologous recombination repair efficiency, non-homologous end joining repair causing resistance allele generation, CRISPR/Cas9 off-target effects, and species adaptation differences. Ecological and safety challenges involve gene pool pollution caused by accidental spread of drive elements, potential ecological balance impacts, and long-term irreversible risks. Response strategies and prospects: employing multiplex guide RNA (gRNA) targeting strategies to enhance drive stability and combat potential resistance. Developing reversible designs such as synthetic resistance, reversal drives, and immunizing reversal drives as "genetic brakes". Establishing long-term ecological monitoring systems and mathematical modeling for risk assessment. Exploring "environmentally responsive drives" to enhance controllability. Future research should continuously optimize drive efficiency and specificity, deepen ecological risk evaluation, strengthen international cooperation, and promote ethical consensus and regulatory framework construction, with the aim of making gene drive technology a sustainable prevention and control strategy to address the global health challenge of mosquito-borne diseases under the premise of safety and controllability. Table and Figures | Reference | Related Articles | Metrics

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Drosophila melanogaster Transposons: Characterization, Regulation, and Their Role in Genome Evolution WANG Ye, WANG Lu Laboratory Animal and Comparative Medicine    2025, 45 (6): 676-687.   DOI: 10.12300/j.issn.1674-5817.2025.112 Abstract （630）   HTML （77）    PDF （1107KB）（1565）       Knowledge map    Save Transposable elements (TEs) are mobile DNA sequences in genomes that play key roles in species evolution, genome stability, and gene regulation. Drosophila melanogaster, as a classic model animal with TEs accounting for approximately 20% of its genome, is an ideal model for studying biological characteristics, host defense mechanisms, and functional evolution of TEs, and also provides an important paradigm for understanding mechanisms of TE-related diseases in higher organisms and even humans. This review systematically elucidates the classification and distribution characteristics of TEs in D. melanogaster and their dynamic interactions with host genome, focusing on the host defense system centered on PIWI-interacting RNA (piRNA) pathway. Then, the biological characteristics of key TE families (such as Gypsy, Copia, P-element, and I-element) in D. melanogaster and their dual roles in genomic evolution are analyzed in detail. On the one hand, TE insertions can cause genomic instability, heterozygous sterility, and aging phenotypes, providing a model basis for studying related human diseases (e.g., neurodegenerative diseases, genomic instability syndromes, etc.). On the other hand, their sequences can be co-opted by the host to create novel regulatory elements or functional genes, thereby driving adaptive innovation. Finally, the future research directions of TEs are proposed, including regulation of TE activity by environmental stress, interaction between piRNA pathways and other small RNA systems, as well as regulatory effects of TEs in the occurrence and development of aging and neurodegenerative diseases. The research on TEs in D. melanogaster not only deepens understanding of TE biology, but also provides a key theoretical basis and important inspiration for studying human diseases using experimental animal models, as well as for developing gene therapy and gene editing technologies. Table and Figures | Reference | Related Articles | Metrics

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Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs KONG Zhihao, WEI Xiaofeng, YU Lingzhi, FENG Liping, ZHU Qi, SHI Guojun, WANG Chen Laboratory Animal and Comparative Medicine    2025, 45 (3): 368-375.   DOI: 10.12300/j.issn.1674-5817.2024.166 Abstract （626）   HTML （20）    PDF （1627KB）（1181）       Knowledge map    Save Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. Methods Skin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10? CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10? CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice. Table and Figures | Reference | Related Articles | Metrics

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Aging Inhibits Memory Immune Response of CD8 + T Cells in Lungs of C57BL/6J Mice Against Influenza A (H1N1) Virus WANG Chao, LI Shun, REN Xiaonan, YANG Hua, CHEN Lixiang, XU Chunhua, ZHOU Xiaohui Laboratory Animal and Comparative Medicine    2025, 45 (5): 515-523.   DOI: 10.12300/j.issn.1674-5817.2025.066 Abstract （618）   HTML （124）    PDF （1456KB）（1099）       Knowledge map    Save Objective To compare functional differences of CD8+ T cells in lung tissues between young and aged C57BL/6J mice during the contraction phase and memory immune response phase after infection with influenza A (H1N1) virus. Methods Lung tissues from young (3-month-old) and aged (24-month-old) C57BL/6J female mice without influenza virus infection were collected to prepare single-cell suspensions, which were stimulated with phorbol 12-myristate 13-acetate (PMA)/ionomycin or cluster of differentiation (CD) 3/CD28 antibodies (T-cell antigen receptor/co-stimulatory signals) respectively (non-specific antigens stimulation). Flow cytometry intracellular cytokine staining (ICS) was performed on lung CD8+ T cells to detect their secretion capacity of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). Young and aged C57BL/6J mice were infected intranasally with 490 PFU PR8 influenza virus, and reinfected with homologous influenza virus 28 days later. Lung tissues were isolated on day 28 (the contraction phase) and day 32 (the memory immune response phase) after primary infection. Influenza virus-specific MHC-Ⅰ tetramer staining was used to detect the proportion of virus-specific CD8+ T cells in lung tissue CD8+ T cells, and ICS was used to analyze TNF-α, IFN-γ, and granzyme B expression in CD8+CD44high T cell subset. Results After non-specific antigen stimulation, TNF-α and IFN-γ secretion capacity in lung tissue CD8+ T cells of aged group mice was significantly higher than that of young group (P<0.05). After virus-specific antigen stimulation, there were no statistically significant differences in the proportion of virus-specific CD8+ T cells and the expression levels of TNF-α, IFN-γ, and granzyme B between the two groups of mice during the contraction phase (P>0.05), while during the memory immune response phase, the proportion of virus-specific CD8+ T cells and the expression levels of TNF-α, IFN-γ, and granzyme B in the aged group mice were significantly lower than those in the young group (P<0.05). Conclusion CD8+ T cells in aged mice maintain normal immune-related factor expression function under non-specific antigen stimulation, but show impaired immune-related factor expression function during antigen-specific memory immune response phase, suggesting that aging leads to defects in the formation or maintenance of CD8+ T cell immune memory. Table and Figures | Reference | Related Articles | Metrics

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Approaches and Application Examples for Studying Mitochondrial Morphology and Function in Caenorhabditis elegans SONG Mengjiao, SHEN Yidong Laboratory Animal and Comparative Medicine    2025, 45 (6): 726-737.   DOI: 10.12300/j.issn.1674-5817.2025.119 Abstract （610）   HTML （18）    PDF （4852KB）（1295）       Knowledge map    Save Mitochondria, as the energy metabolism center of cells, have abnormal dynamic morphology and oxidative phosphorylation function, which are directly related to aging and various diseases. Caenorhabditis elegans (C. elegans) is a widely used model animal. This article systematically summarizes the experimental methods for analyzing mitochondrial morphology and function at multiple scales in the laboratory using C. elegans as a model, mainly including: (1) morphological qualitative analysis, which uses a single-blind manual classification method (such as point, rod, and mesh), although it relies on subjective experience, it is easy to operate and suitable for preliminary phenotype screening; (2) morphological quantitative analysis, which relies on the Fiji/ImageJ platform for automated parameter extraction of mitochondria in specific tissues (such as epidermis and body wall muscles), quantifies network connectivity (number of branch points, network length) through skeletonization algorithm, and calculates fragmentation indices (such as area/perimeter ratio, fragment count) with binarization analysis, achieving objective phenotype comparison; (3) high-throughput image processing, which performs batch processing through macro commands, integrates morphological filtering, threshold segmentation, and parameter export workflow, significantly improving research efficiency for large sample sizes; (4) metabolic function monitoring, which uses the Seahorse XF analyzer to measure in vivo respiratory metabolism in C. elegans. By sequential injection of ATP synthase inhibitor N, N'-dicyclohexylcarbodiimide (DCCD), uncoupling agent carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP) and respiratory chain inhibitor sodium azide (NaN3), the basal oxygen consumption rate (OCR), ATP coupled respiratory efficiency, and maximum respiratory capacity (respiratory potential) are accurately analyzed to reveal energy metabolism patterns. This article integrates a dual dimensional approach of morphology and function, which not only provides empirical techniques for mitochondrial research on C. elegans, but also extends its framework to mammalian cell and organoid models, promoting the basic research and drug development targeting mitochondria. Table and Figures | Reference | Related Articles | Metrics

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Common Environmental Problems and Testing Experiences in Laboratory Animal Facilities in Sichuan Province LIU Wentao, LUO Yanhong, LONG Yongxia, LUO Qihui, CHEN Zhengli, LIU Lida Laboratory Animal and Comparative Medicine    2025, 45 (4): 483-489.   DOI: 10.12300/j.issn.1674-5817.2024.142 Abstract （601）   HTML （27）    PDF （724KB）（642）       Knowledge map    Save Laboratory animals are the "living" tools of medical research. Through animal experiments, people can gain continuous insights into the laws of life, reveal the essence of diseases, develop vaccines and drugs for prevention and treatment, and play an important role in the technological development of fields related to human health. The environmental conditions of laboratory animals have a direct impact on their health, quality, and the results of animal experiments. The higher the degree of environmental control, the more reliable the experimental results are in terms of quality. Therefore, environmental control of laboratory animal facilities is important for ensuring that laboratory animals live under required conditions, which is a key factor for conducting effective animal experiments. This article analyzes the current status of environmental testing of laboratory animal facilities in Sichuan Province, briefly summarizing their number, area, and other basic information, and provides detailed statistics on the ability of institutions to conduct environmental testing for laboratory animal facilities in Sichuan Province. It also summarizes the testing requirements for laboratory animal facility environments based on national standards, regulatory requirements, and the quality control needs of facility users. In the analysis of testing indicators for laboratory animal facilities, based on testing data from 40 laboratory animal facilities in Sichuan Province, it was found that static pressure difference is the indicator most prone to non-compliance, followed by illumination and air exchange rate. Using barrier environments as examples, common problems in the process of environmental testing for laboratory animal facilities are summarized in six aspects: testing personnel, instruments, methods, technical materials, testing environment, and reports, and targeted suggestions are proposed. These suggestions help improve environmental control in laboratory animal facilities, and provide practical reference and guidance for relevant testing institutions, as well as laboratory animal producers and users in the industry. Table and Figures | Reference | Related Articles | Metrics

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Exosomes Treat Ischemic Stroke by Regulation of Ferroptosis Through the NRF2/SLC7A11/GPX4 Pathway in Mice XU Yingtao, WANG Mengmeng, LIN Ping, CHI Haitao, WANG Yi, BAI Ying Laboratory Animal and Comparative Medicine    2026, 46 (1): 20-31.   DOI: 10.12300/j.issn.1674-5817.2025.034 Abstract （573）   HTML （31）    PDF （18736KB）（1203）       Knowledge map    Save Objective A middle cerebral artery occlusion (MCAO) mouse model is established by electrocoagulation of the middle cerebral artery. The study examines the mechanism by which exosomes (EXO) derived from human amniotic mesenchymal stem cells (hAMSCs) improve ischemic stroke and regulate neural ferroptosis-related injury. Methods Thirty-two SPF-grade male C57BL/6J mice aged 6 - 8 weeks were randomly divided into four groups (n=8 per group): sham group (Sham), model group (MCAO), MCAO plus normal saline group (MCAO+NaCl), and MCAO plus exosome group (MCAO+EXO). The mouse MCAO model was established by electrocoagulation of the middle cerebral artery. Mice in the Sham group underwent exposure of the middle cerebral artery without electrocoagulation. Twenty-four hours before MCAO induction, mice in the MCAO+EXO group received a tail vein injection of 100 μL of exosomes derived from the culture supernatant of hAMSCs at a concentration of 9.5×1011 particles/mL. Mice in the MCAO+NaCl group were injected with an equal volume of normal saline via the tail vein. Twenty-four hours after model establishment, neurological deficits were evaluated using the Longa neurological deficit scoring system. Cerebral infarct volume was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Hematoxylin and eosin (HE) staining was performed to evaluate morphological changes of neurons in the ischemic brain regions. The contents of ferrous iron (Fe2+), malondialdehyde (MDA), total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) in the infarct core and peri-infarct regions were determined using microcolorimetric assays to evaluate differences among groups. The mRNA expression levels of ferroptosis-related factors, including nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in the infarct core and peri-infarct regions were measured by real-time quantitative PCR. Protein expression levels of NRF2, SLC7A11, and GPX4 in the infarct and peri-infarct regions of each group were analyzed by Western blotting. Results Compared with the MCAO group, the Longa neurological deficit score was significantly reduced in the MCAO+EXO group (P<0.01). Prominent cerebral infarction was observed in the MCAO group, whereas the infarct volume ratio was markedly decreased in the MCAO+EXO group compared with the MCAO group (P<0.001). Histopathological analysis revealed that mice in the MCAO group exhibited obvious neuronal damage, including cytoplasmic vacuolar degeneration, nuclear pyknosis and fragmentation, unclear nuclear structure, and disorganized neuronal arrangement, compared with the Sham group. In contrast, neurons in the MCAO+EXO group showed relatively preserved morphology, with intact cellular structures and large, regular nuclei located centrally within the cells. Biochemical analysis demonstrated that Fe2+ and MDA levels in the infarct core and peri-infarct regions were significantly increased in the MCAO group compared with the Sham group (P<0.001). These levels were significantly reduced in the MCAO+EXO group compared with the MCAO group (P<0.01). In addition, total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) levels were markedly decreased in the MCAO group relative to the Sham group (P<0.01). Compared with the MCAO group, the MCAO+EXO group exhibited significantly increased levels of total GSH and GSH (P<0.001), while no significant change was observed in GSSG levels (P>0.05). Furthermore, both mRNA and protein expression levels of nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) were significantly downregulated in the MCAO group compared with the Sham group (P<0.01, P<0.001). In contrast, both mRNA and protein expression levels of NRF2, SLC7A11, and GPX4 were significantly upregulated in the MCAO+EXO group compared with the MCAO group (P<0.05). Conclusion In the mouse MCAO model, tail vein injection of exosomes derived from hAMSCs can improve motor function, reduce infarct area, protect neuronal cell morphology, and reduce the degree of nerve injury. Exosomes may exert a protective effect by activating the NRF2/SLC7A11/GPX4 pathway and reducing ferroptosis in neuronal cells of MCAO model mice. Table and Figures | Reference | Related Articles | Metrics

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Research Advances on Periplaneta americana as an Experimental Animal Model ZHOU Guanyu, ZHU Shiming, LIU Fangfang Laboratory Animal and Comparative Medicine    2025, 45 (6): 794-802.   DOI: 10.12300/j.issn.1674-5817.2025.110 Abstract （571）   HTML （15）    PDF （986KB）（1395）       Knowledge map    Save The American cockroach (Periplaneta americana), as a representative species of Blattodea and one of the most ancient extant insect groups, is gradually developing from a public-health pest into an emerging experimental animal model in developmental biology and regenerative medicine, due to its remarkable environmental adaptability, fecundity, developmental plasticity, and high sensitivity to RNA interference (RNAi). This review aims to systematically integrate the latest research advances in biological characteristics, genomic resources, developmental regulatory mechanisms, and application value of P. americana, to expand its application potential in basic research and translational medicine, elucidating following aspects: (1) Biological traits of P. americana and its standardized laboratory rearing protocols are outlined, laying a foundation for its use as a stable experimental model. (2) Analysis results of high-quality genome sequencing are elaborated, revealing evolutionary characteristics of repeat sequence-driven expansion and association between significant expansion of metabolism and stress response gene families and their strong environmental adaptation. (3) In developmental biology, the review summarizes how dynamic balance between 20-hydroxyecdysone (20E) and juvenile hormone (JH) governs molting and metamorphic plasticity, how insulin/insulin-like growth factor signaling (IIS) pathway and target of rapamycin complex 1 (TORC1) pathway integrate nutritional signals to regulate developmental rate, and how JH precisely coordinates efficient reproduction regulatory network through multiple mechanisms. (4) The exceptional limb regeneration capacity of P. americana and synergistic effects of conserved pathways and novel regulatory factors behind it are detailed. (5) Concerning environmental adaptation mechanisms, the significant expansion of detoxification-related gene families and innate immune pathways, and JH-mediated sexually dimorphic stress response characteristics are highlighted. (6) The application value of P. americana in traditional drug development (e.g., Kangfuxin Liquid) and precise allergen diagnostics is discussed, and future research directions are prospected. Although technical challenges such as insufficient genetic manipulation tools exist, through deep integration of multi-omics technologies and development of gene-editing tools, P. americana model is expected to play a more significant role in revealing developmental plasticity, regeneration mechanisms, and new targets for pest control. Table and Figures | Reference | Related Articles | Metrics

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Establishment and Multidimensional Pathological Evaluations of a Cigarette Smoke Exposure-Induced Chronic Obstructive Pulmonary Disease Mouse Model HE Jiaqi, ZHOU Yuanyuan, NIE Yongqiang, WANG Zhaoxia, XU Wangjie Laboratory Animal and Comparative Medicine    2026, 46 (1): 11-19.   DOI: 10.12300/j.issn.1674-5817.2025.076 Abstract （571）   HTML （23）    PDF （13520KB）（1051）       Knowledge map    Save Objective To establish a reliable chronic obstructive pulmonary disease (COPD) mouse model based on a self-developed multichannel automatic control system for long-term continuous cigarette smoke exposure in small animals using a novel continuous cigarette smoke exposure method, and to conduct phenotypic evaluation and analysis, thereby providing an animal experimental basis for investigating COPD pathogenesis and prevention strategies. Methods Twenty male C57BL/6J mice aged 6 weeks were randomly and equally divided into a control group and a model group. The model group (n=10) underwent 6 h of continuous cigarette smoke exposure daily (6 cigarettes per day for 12 consecutive weeks), while the control group (n=10) received no intervention. Body weight was monitored biweekly. Post-exposure, in vivo micro-CT imaging was performed. After euthanasia, serum and bronchoalveolar lavage fluid (BALF) levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were quantified by ELISA. Lung tissues underwent H&E and Masson's trichrome staining to observe changes in lung morphology and inflammatory cell infiltration, and the mean linear intercept (MLI) was calculated, thereby comprehensively evaluating the clinical features of COPD in the mouse model. Results Compared with the control group, the model group showed significantly reduced body weight (P<0.01) from the fourth week. Compared with the control group, IL-6 level in the serum and BALF of the model group increased by 27.2% and 140.0%, respectively (P<0.01). TNF-α level in the serum and bronchoalveolar lavage fluid of the model group increased by 16.7% (P<0.01) and 19.3% (P<0.05), respectively. Histopathological examination revealed alveolar wall thinning, septal rupture, emphysematous bullae formation, reduced alveolar count, bronchial wall thickening with lumen narrowing, and inflammatory cell infiltration. MLI was significantly elevated (P<0.01). Masson's staining confirmed collagen deposition and bronchial remodeling. Micro-CT demonstrated localized high-density shadows exhibiting typical features of chronic bronchitis. Conclusion The self-developed device enables long-term continuous smoke exposure, and the successfully established COPD mouse model exhibits pathological features highly consistent with clinical manifestations, offering an efficient and reliable tool for COPD research. Table and Figures | Reference | Related Articles | Metrics

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Evaluation of the Safety and Efficacy of Bone Cement in Experimental Pigs Using Vertebroplasty LIN Zhenhua, CHU Xiangyu, WEI Zhenxi, DONG Chuanjun, ZHAO Zenglin, SUN Xiaoxia, LI Qingyu, ZHANG Qi Laboratory Animal and Comparative Medicine    2025, 45 (4): 466-472.   DOI: 10.12300/j.issn.1674-5817.2025.006 Abstract （566）   HTML （13）    PDF （1400KB）（1077）       Knowledge map    Save Objective The full name of vertebroplasty is percutaneous vertebroplasty (PVP). It is a clinical technique that injects bone cement into the diseased vertebral body to achieve strengthening of the vertebra. The research on the safety and efficacy of bone cement is the basis for clinical application. In this study, vertebroplasty is used to evaluate and compare the safety and efficacy of Tecres and radiopaque bone cement in experimental pigs, and to determine the puncture method suitable for pigs and the pre-clinical evaluation method for the safety and efficacy of bone cement. Methods Twenty-four experimental pigs (with a body weight of 60-80 kg) were randomly divided into an experimental group (Group A) and a control group (Group B). Group A was the Tecres bone cement group, and Group B was the radiopaque bone cement group, with 12 pigs in each group. Under the monitoring of a C-arm X-ray machine, the materials were implanted into the 1st lumbar vertebra (L1) and 4th lumbar vertebra (L4) of the pigs via percutaneous puncture using the unilateral pedicle approach. The animals were euthanized at 4 weeks and 26 weeks after the operation, respectively. The L4 vertebrae were taken for compressive strength testing, and the L1 vertebrae were taken for hard tissue pathological examination to observe the inflammatory response, bone necrosis, and degree of osseointegration at the implantation site. Results The test results of compressive strength between groups A and B showed no significant difference at 4 weeks and 26 weeks after bone cement implantation (P > 0.05). Observation under an optical microscope (×100) revealed that at 4 weeks postoperatively, both groups A and B showed that the bone cement was surrounded by proliferative fibrous tissue, with lymphocyte infiltration around it. The bone cement was combined with bone tissue, the trabecular arrangement was disordered, and osteoblasts and a small amount of osteoid were formed. At 26 weeks postoperatively, bone cement was visible in both groups A and B. The new bone tissue was mineralized, the trabeculae were fused, the trabecular structure was regular and dense with good continuity, and no obvious inflammatory reaction was observed. Conclusion In experimental pig vertebrae, there were no significant differences observed in the compressive strength, inflammation response, bone destruction, and integration with the bone between Tecres and non-radiopaque bone cement. Both exhibited good biocompatibility and osteogenic properties. It indicates that using vertebroplasty to evaluate the safety and efficacy of bone cement in pigs is scientifically sound. Table and Figures | Reference | Related Articles | Metrics

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Observation of Digestive Tract Tissue Morphology in Mice Using Probe-Based Confocal Laser Endomicroscopy LIU Yueqin, XUE Weiguo, WANG Shuyou, SHEN Yaohua, JIA Shuyong, WANG Guangjun, SONG Xiaojing Laboratory Animal and Comparative Medicine    2025, 45 (4): 457-465.   DOI: 10.12300/j.issn.1674-5817.2025.035 Abstract （562）   HTML （12）    PDF （2246KB）（1648）       Knowledge map    Save Objective To explore the application value of probe-based confocal laser endomicroscopy (pCLE) in rapidly detecting and evaluating the morphological characteristics of digestive tract tissues in mice. Methods Twelve male SPF Kunming mice aged 6 weeks were randomly divided into two groups. Six mice were subjected to gastric gavage with 52% Red Star Erguotou to establish the model, and six were given saline by gastric gavage as a control. After 28 days of modeling, 3 mice were randomly selected from each group. After deep anesthesia induced by inhalation of 3% isoflurane, the mice were sacrificed by cervical dislocation. The stomach, duodenum, jejunum, and rectum tissues were excised and immersed in 1% fluorescein sodium solution for staining. The microstructure of the mucosal surface of each tissue was observed using pCLE. The remaining mice in the model group and the control group were deeply anesthetized by inhaling 3% isoflurane, then cardiac perfusion was performed successively with saline and 4% paraformaldehyde. The stomach, duodenum, jejunum, and rectum tissues were excised for dehydration, section and hematoxylin-eosin (HE) staining, and the morphological changes of the tissues were observed under a microscope. Results Under pCLE imaging, fluorescence staining on the surface of the gastrointestinal mucosa was uniform in the control group; the morphology of gastric pits, intestinal villi, and intestinal crypts was intact, arranged compactly, and had distinct boundaries. In the model group, the gastrointestinal mucosa exhibited mucosal swelling and deformation, with uneven fluorescence staining and fluorescein leakage. Furthermore, some tissues showed defects or cell shedding, and the boundaries between adjacent characteristic structures (e.g., gastric pits, intestinal crypts) were blurred. HE staining showed that the gastrointestinal tissue structure of the control group mice was normal and well-organized, with no structural defects. Moreover, submucosal glands were uniform in size, with no hyperplasia observed, and no obvious inflammatory cell infiltration. In the model group, some gastrointestinal mucosal structures were defective and sparsely arranged; submucosal glands showed atrophy, accompanied by obvious inflammatory cell infiltration. The histological characteristics detected by pCLE were consistent with those of HE staining. Conclusion pCLE can be used to obtain rapid, real-time, large-scale, and high-resolution microscopic imaging of the gastrointestinal mucosa, realistically and comprehensively displaying its physiological and microstructural characteristics. It shows promising prospects and practical utility in the histological evaluation of digestive system injuries in small animals. Table and Figures | Reference | Related Articles | Metrics

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Construction and Preliminary Application of Animal Disease Model Digital Atlas Database Platform LI Huiping, GAO Hongbin, WEN Jinyin, YANG Jinchun Laboratory Animal and Comparative Medicine    2025, 45 (3): 300-308.   DOI: 10.12300/j.issn.1674-5817.2024.158 Abstract （557）   HTML （37）    PDF （1848KB）（1273）       Knowledge map    Save Objective Domestic research institutions and researchers have established a wide variety of animal disease models and accumulated a wealth of specialized, distinctive, and targeted atlas data during the model development process. These atlas data are of great value for development and application. Therefore, it is necessary to develop a professional and complete digital atlas database platform for animal models, which can achieve the open sharing of animal model atlas data and the integration and optimization of atlas resources related to disease animal models held by relevant domestic institutions. Methods Based on the B/S architecture, the authors' institution built a digital atlas database of animal models, using Java as the main development language and Oracle database system along with related auxiliary tools. The database platform ran in a Linux environment and could be accessed by users through a web browser. At present, the data on this platform mainly came from the atlas resources submitted by animal model resource units within Guangdong Province. Results In August 2024, a digital atlas database platform for animal models was constructed based on the classification structure of three dimensions: systemic diseases, animal species, and resource units. This platform provided functions such as collection, management, retrieval, and viewing of atlas data. As of January 2025, four resource units had submitted 61 atlas data entries of animal models to the platform, totalling 610 data items. Conclusion The animal model digital atlas database platform has been constructed and put into preliminary use. Although the amount of data on the platform is still limited, it is capable of integrating and openly sharing animal model atlas data. It is believed that with the continuous enrichment of atlas data in the future, this platform is expected to provide important data support for the development of laboratory animal science and comparative medicine research, thereby promoting the efficient utilization of scientific research resources. Table and Figures | Reference | Related Articles | Metrics

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Evolutionary Conservation of Organ Development between Drosophila and Mammals GAO Yinghao, WU Wei Laboratory Animal and Comparative Medicine    2025, 45 (6): 719-725.   DOI: 10.12300/j.issn.1674-5817.2025.126 Abstract （554）   HTML （15）    PDF （2630KB）（345）       Knowledge map    Save Drosophila， as a classic model organism， exhibits high evolutionary conservation with mammals in molecular regulatory networks， signal pathway transduction， and cell fate determination mechanisms involved in its organ development. Focusing on major organs of Drosophila including dorsal vessel （homologous to the mammalian heart）， fat body and oenocytes （liver）， Malpighian tubules and nephrocytes （kidney）， and tracheal system （lung）， this study deeply analyzes conservative characteristics in core regulatory pathways， key gene functions， and cellular behavioral patterns during organogenesis between Drosophila and mammals through multi-dimensional comparative analysis， clarifying the homologous basis of their developmental mechanisms. Integrating specific research examples， it further elaborates the application value of Drosophila in constructing models for human cardiovascular diseases， metabolic disorders， kidney diseases， and respiratory diseases， as well as its unique role in high-throughput screening of drug targets， analysis of disease pathogenic mechanisms， and improvement of the theoretical system of evolutionary developmental biology. The evolutionary conservation of core developmental regulatory mechanisms between Drosophila and mammals has built a key bridge from basic developmental research to human disease translation， providing important theoretical support and a technical platform for in-depth analysis of the molecular mechanisms of complex human diseases and development of novel therapeutic strategies， and holding great significance for promoting interdisciplinary integration of developmental biology and translational medicine. Table and Figures | Reference | Related Articles | Metrics

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Research on Spatiotemporal Gene Expression Profiles and Repair Mechanisms of Spinal Cord Compression and Hemisection Spinal Cord Injury Mouse Models XU Bo, CHEN Tairen, FANG Qian, WU Ji Laboratory Animal and Comparative Medicine    2026, 46 (1): 32-45.   DOI: 10.12300/j.issn.1674-5817.2025.052 Abstract （554）   HTML （28）    PDF （9666KB）（213）       Knowledge map    Save Objective To investigate the gene expression sequence and molecular mechanisms in the local microenvironment during the subacute to chronic phases (1-28 days) in mouse models of spinal cord compression injury and hemisection spinal cord injury, thereby revealing the molecular characteristics of spinal cord repair and providing a theoretical basis for selecting therapeutic targets for spinal cord injury. Methods Thirty-six 8-9-week-old SPF-grade ICR mice were randomly divided into three groups (n=12 per group): sham-operated control (CTR) group, hemisection spinal cord injury (HSCI) group, and spinal cord compression injury (SCC) group. Mice in the CTR group underwent the same surgical preparation and anesthesia, followed by a dorsal midline incision at the T9-T10 segment. After layer-by-layer dissection and removal of the corresponding lamina, the spinal cord dura mater was fully exposed and kept intact. The cord was exposed to air for 10 minutes (matching the duration of the compression injury group), during which any instrument contact with the cord was avoided. The incision was then irrigated and sutured. The HSCI group underwent a 70% transection of the T9 spinal cord segment using micro-instruments to establish a hemisection spinal cord injury model. The SCC group underwent sustained compression of the T10 spinal cord segment for 10 minutes using a self-made compressor (a 30 g solid small iron bar) to establish a spinal cord compression injury model. Motor function recovery was assessed using the modified Basso-Beattie-Bresnahan (BBB) score on postoperative days 1, 3, 7, 14, 21, and 28. On days 7 and 14 post-operation, mice were anesthetized, and the injured spinal cord segments were harvested. The evolution of specific molecular networks in the spinal cord injury mouse models was analyzed via RNA sequencing (RNA-Seq) and enrichment analysis, and the expression of key genes was verified using real time fluorogenic quantitative PCR. Results BBB scores indicated that motor function recovery in the SCC group was significantly better than that in the HSCI group, with BBB scores showing a continuously increasing trend and remaining higher than those in the HSCI group over the 4-week period (P <0.001). Gene ontology （GO)and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses based on RNA-Seq differentially expressed genes revealed that, compared to the CTR group, genes related to the extracellular matrix were significantly up-regulated (P<0.05), while genes related to axon guidance were significantly down-regulated (P <0.05) in the SCC group on day 7 post-operation. On day 21, genes involved in immune regulation and the retinol signaling pathway were significantly activated in the SCC group (P<0.05). In contrast, in the HSCI group, genes associated with inflammation and immune response were significantly up-regulated (P<0.001), while genes related to neuronal differentiation and synapse formation were significantly down-regulated (P <0.001) on day 7. On day 21, genes related to cell-matrix junctions and N-methyl-D-aspartate receptors were significantly up-regulated (P<0.001) in the HSCI group. Furthermore, compared to the SCC group, the HSCI group exhibited different pathway enrichment characteristics in GO and KEGG analyses on days 7 and 21 post-injury. On day 7, genes involved in the NOD-like receptor signaling pathway and the complement and coagulation cascades were significantly up-regulated in the HSCI group (P<0.001). On day 21, genes related to the extracellular matrix-receptor interaction and the neuroactive ligand-receptor interaction pathways were significantly activated (P<0.001). Finally, real time fluorogenic quantitative PCR validation results were highly consistent with the RNA-Seq results, further confirming the differential expression trends of key genes between the SCC and HSCI groups. Conclusion The SCC and HSCI injury models may drive distinct repair pathways: the preservation of some axons in the SCC model predisposes it toward tissue repair, whereas the HSCI model requires the coordination of more complex molecular networks to achieve a new equilibrium. This finding further deepens the understanding of the heterogeneous regulatory mechanisms underlying spinal cord injury. Table and Figures | Reference | Related Articles | Metrics

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Dynamic Monitoring and Analysis of Ammonia Concentration in Laboratory Animal Facilities Under Suspension of Heating Ventilation and Air Conditioning System JIAO Qingzhen, WU Guihua, TANG Wen, FAN Fan, FENG Kai, YANG Chunxiang, QIAO Jian, DENG Sufang Laboratory Animal and Comparative Medicine    2025, 45 (4): 490-495.   DOI: 10.12300/j.issn.1674-5817.2024.130 Abstract （553）   HTML （27）    PDF （668KB）（412）       Knowledge map    Save Objective To monitor the real-time changes in ammonia concentration in the laboratory animal facility environment before, during, and after the air conditioning system stops supplying air, so as to provide a basis and reference for developing emergency plans for the shutdown of the air conditioning system. Methods The laboratory animal facilities of the Wuhan Institute of Biological Products were used as the research object. Ammonia concentration detectors were used to monitor ammonia concentration continuously in the environment of conventional rabbit production facility, SPF hamster production facility, and SPF guinea pig experimental facility before and after the passive shutdown due to repairs and active maintenance shutdown of the air conditioning system, as well as the time for the ammonia concentration to return to daily levels after resuming air supply. Results Under both shutdown modes of the air conditioning system, the trend of ammonia concentration changes in different laboratory animal facilities was consistent, showing a rapid increase after shutdown and a rapid decrease after resuming air supply. Under active maintenance shutdown, the maximum ammonia concentrations in the conventional rabbit production facilities, SPF hamster production facilities, and SPF guinea pig experimental facilities were 9.81 mg/m3, 14.27 mg/m3, and 6.98 mg/m3, respectively. Within 12 minutes after resuming air supply, ammonia concentration could return to normal daily levels. Under passive long-term shutdown, ammonia concentration value was positively correlated with the duration of air supply suspension. As the shutdown duration increased, ammonia concentration continued to increase. The maximum ammonia concentration values in the three facilities occurred at 88 minutes (38.06 mg/m3), 40 minutes (18.43 mg/m3), and 34 minutes (15.61 mg/m3) after air supply suspension, respectively.Within 11 minutes after resuming air supply, ammonia concentration could return to normal daily levels. Conclusion Shutdown of the air conditioning system causes a rapid increase in ammonia concentration in laboratory animal facilities, and the rise in ammonia concentration is positively correlated with the duration of air supply suspension. Therefore, when an emergency shutdown of the air-conditioning system is required due to maintenance or other reasons, backup fans should be provided in accordance with the requirements of GB 50447-2008 "Architectural and Technical Code for Laboratory Animal Facilities". Older facilities should make adequate preparations and develop a scientifically sound emergency plan. Table and Figures | Reference | Related Articles | Metrics