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    Research Progress in Establishment and Evaluation of Common Asthma Animal Models
    LUO Shixiong, ZHANG Sai, CHEN Hui
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 167-175.   DOI: 10.12300/j.issn.1674-5817.2024.120
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    Bronchial asthma (hereinafter referred to as asthma) is a common chronic respiratory disease characterized by airway inflammation, airway hyperresponsiveness, and airway remodeling. Its pathogenesis is highly complex and heterogeneous, involving multiple factors such as genetics, immunity, and environmental exposure. Currently, therapeutic options for asthma remain relatively limited, making it an urgent priority to explore its underlying mechanisms, identify effective treatment strategies, and develop new drugs. In this context, the establishment of animal models for asthma plays an irreplaceable and crucial role. However, to date, no single ideal animal model has been able to fully and accurately replicate all the features of the onset and progression of human asthma. This study systematically reviews the research progress over the past five years in the establishment methods of asthma animal models. It provides a detailed overview of commonly used experimental animals (such as mice, rats, and guinea pigs), frequently used sensitizing agents (including ovalbumin, house dust mite, lipopolysaccharide, and toluene diisocyanate), and the methods for establishing asthma models using these animals and sensitizers. This study also presents an objective evaluation of the advantages, limitations, and applicability of each model. Evaluation criteria for asthma models are summarized across multiple dimensions, including behavioral assessments, pulmonary function, histopathology, immunological indicators, and pharmacodynamics. Although methods for establishing refractory asthma models remain underdeveloped, several strategies for modeling refractory asthma have been summarized through a review of relevant literature, aiming to provide useful references for related research. Based on current scientific and technological advancements, it is anticipated that future research on asthma animal models will focus more on clinical relevance, technological innovation, and multidisciplinary integration. Specifically, future models are expected to adopt multi-sensitizer induction protocols, apply cutting-edge tools such as gene editing, enhance clinical relevance and promote diversification and personalization of models. Furthermore, advanced technologies such as bioimaging and biosensing are anticipated to enable dynamic monitoring of airway inflammation and remodeling. Organ-on-a-chip platforms may also be explored as potential alternatives to traditional animal models. The ultimate goal is to develop multifactorial, composite models that better simulate the complexity and heterogeneity of human asthma.

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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
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    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.

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    Construction and Evaluation of a Rat Model of Abnormal Uterine Bleeding
    LIAN Hui, JIANG Yanling, LIU Jia, ZHANG Yuli, XIE Wei, XUE Xiaoou, LI Jian
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 130-146.   DOI: 10.12300/j.issn.1674-5817.2024.132
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    Objective By simulating the etiology of abnormal uterine bleeding-ovulatory dysfunction (AUB-O) and establishing a rat model of abnormal uterine bleeding (AUB), this study aims to provide an experimental platform for investigating pathological mechanisms and developing therapeutic drugs for AUB. Methods After acclimation, 24 adult (10-week-old) female SD rats were randomly divided into a normal control group (6 rats) and a model group (18 rats). The normal control group was housed in a barrier environment, while the model group underwent bilateral ovariectomy via dorsal approach in the same environment and rested for one week before starting to receive modeling drugs. In the model group, from Days 1 to 3 of modeling, each rat received a daily subcutaneous injection of 0.5 mg estradiol into the dorsal region. From Days 4 to 7, a daily subcutaneous injection of 5.0 mg progesterone was administered. On Day 6, rats received bilateral injections of 0.5 mL soybean oil per uterine cavity (total 1.0 mL) via the same dorsal surgical incision. On Day 8, mifepristone (10 mg/kg) was administered via oral gavage. The estrous cycle stage and its dynamic changes were continuously monitored during modeling. Uterine bleeding was recorded during the 48-hour observation period post-modeling. Serum and uterine tissue samples were collected from the model group at 0, 12, 24, 36, and 48 h after mifepristone administration, while the normal control group was sampled at 36 h. The samples were subjected to HE staining, serum sex hormone ELISA, immunohistochemistry, TUNEL apoptosis staining, Western blotting, transcriptome sequencing, and bioinformatics analysis for comprehensive evaluation of the AUB rat model. Results The AUB rats exhibited uterine bleeding, endometrial detachment and injury, incomplete uterine restoration, inflammatory cell infiltration in the endometrium, enhanced tissue apoptosis, and structural damage of the stroma, glands, and vasculature. Compared with the normal control group, the levels of serum follicle-stimulating hormone (FSH), estradiol, and luteinizing hormone (LH) were significantly increased in the AUB rats (P<0.05). The vascular density of the endometrium was significantly reduced (P<0.05). The expression of vascular endothelial growth factor (VEGF) was qualitatively observed to be markedly enhanced at the site of endometrial detachment but significantly decreased around the stromal blood vessels (P<0.01). Matrix metalloproteinase-9 (MMP-9) expression was qualitatively observed to be strongly upregulated at the site of endometrial injury but significantly reduced in the non-detached stroma and glands (P<0.01). Endometrial stromal cell apoptosis was significantly enhanced (P<0.01). The expression levels of fibroblast growth factor 2 (FGF2) and endothelin-1 (ET-1) in uterine tissues were significantly decreased (P<0.05). After comparing the transcriptome sequencing results of uterine tissues between AUB and normal rats, a total of 4 723 differentially expressed genes were identified, including 2 191 up-regulated genes and 2 532 down-regulated genes. KEGG enrichment analysis revealed that these differentially expressed genes were significantly enriched in pathways related to inflammation, immune apoptosis, cell signal transduction, proliferation and differentiation, and muscle contraction, among others. Conclusion An AUB rat model can be successfully established using a sequential administration protocol of estrogen, progesterone, and mifepristone to simulate the etiology of AUB-O. In this model, endometrial injury is associated with inflammation and apoptosis, with pathological manifestations influenced by abnormal vasoconstriction and impaired endometrial regeneration. This rat model closely recapitulates pathological characteristics of non-structural AUB observed in clinical practice, making it a validated experimental platform for exploring the pathological mechanisms and therapeutic interventions of non-structural AUB.

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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
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    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.

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    Analysis of Animal Models of Myasthenia Gravis Based on Its Clinical Characteristics in Chinese and Western Medicine
    CHEN Yuhan, CHEN Jinling, LI Xin, OU Yanhua, WANG Si, CHEN Jingyi, WANG Xingyi, YUAN Jiali, DUAN Yuanyuan, YANG Zhongshan, NIU Haitao
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 176-186.   DOI: 10.12300/j.issn.1674-5817.2024.139
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    Myasthenia gravis (MG) is an autoimmune disease characterized primarily by skeletal muscle weakness and, in severe cases, respiratory involvement. Western medical treatment predominantly relies on immunosuppressants, but long-term administration often leads to notable side effects. In contrast, traditional Chinese medicine (TCM) offers the advantage of multi-target interventions. However, the pathogenesis of MG has not been fully elucidated, and the establishment of animal models that accurately reflect the clinical characteristics of both Chinese and Western medicine is essential for mechanism research and new drug development. This paper systematically reviews the etiology and pathogenesis, diagnostic criteria, and progress of animal model research for MG from both Chinese and Western medicine perspectives. In Western medicine, the pathogenesis of MG is closely related to genetic susceptibility, environmental factors, and autoantibody-mediated postsynaptic membrane damage. In TCM, MG is classified under the category of "flaccidity syndrome", attributed to congenital deficiencies and acquired malnourishment. Western diagnostic criteria involve a combination of clinical symptoms, fatigue testing, serum antibody assays, and electrophysiological evaluation. In contrast, TCM diagnosis emphasizes the integration of primary and secondary symptoms with tongue and pulse pattern differentiation. Currently available animal models mainly include experimental autoimmune myasthenia gravis (EAMG) and passive transfer myasthenia gravis (PTMG). The Toredo acetylcholine receptor (AChR) induced EAMG model aligns well with Western diagnostic criteria, but poorly matches secondary symptoms in TCM. The synthetic AChR peptide model is widely used, but shows low conformity with TCM syndromes. Models induced by muscle-specific tyrosine kinase (MuSK), low-density lipoprotein receptor-related protein 4 (LRP4), and transgenic models demonstrate high innovation but exhibit low clinical conformity. Evaluation of these models requires integration of behavioral, electrophysiological, and immunological indicators. However, a systematic framework for modelling TCM syndromes is still lacking. Future research should integrate TCM-based etiological modelling methods with the Western pathological mechanisms to construct disease-syndrome combination models. Additionally, it is crucial to establish a TCM syndrome evaluation system based on "validation by prescription", as well as to improve the scientific rigor and practicality of animal models by the incorporation of emerging technologies. This review provides a theoretical foundation for optimizing MG animal model design, advancing the research on the combination of Chinese and Western medicine, and supporting efficacy assessment and mechanism exploration of Chinese herbal prescriptions.

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    A Mouse Model of Polycystic Ovary Syndrome Established Through Subcutaneous Administration of Letrozole Sustained-Release Pellets and Hepatic Transcriptome Analysis
    XU Qiuyu, YAN Guofeng, FU Li, FAN Wenhua, ZHOU Jing, ZHU Lian, QIU Shuwen, ZHANG Jie, WU Ling
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 119-129.   DOI: 10.12300/j.issn.1674-5817.2024.186
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    Objective Prepubertal mice are administered subcutaneously with letrozole sustained-release pellets behind the neck and treated with a high-fat diet to establish a mouse model of polycystic ovary syndrome (PCOS). The liver transcriptomes of the model mice are compared with those of the placebo control mice to investigate the underlying mechanisms of liver involvement in the pathogenesis of PCOS. Methods A customized 2 mg dose of letrozole sustained-release pellets with a 40-day release period was used. The control placebo and letrozole pellets were implanted subcutaneously in the dorsal cervical region of 3-4-week-old C57BL/6J mice (8 mice per group) to establish the control group and letrozole-induced PCOS model group. Both groups were treated with a high-fat diet starting the day after administration. The modeling period lasted for 5 weeks, during which body weight and 24-hour food intake were monitored in each group every week. When samples were collected, liver weight was recorded. Pathological changes in ovarian and hepatic tissues were examined by hematoxylin-eosin (HE) staining, while hepatic lipid deposition was observed by Oil Red O staining. The extent of macrophage infiltration in the liver was evaluated via F4/80 immunohistochemical staining, and hepatic fibrosis levels were observed by Masson's trichrome staining. Transcriptomic sequencing was performed to analyze differentially expressed genes (DEGs) in liver tissues between the control and model groups, followed by enrichment analysis of significant DEGs. Quantitative real-time fluorescent quantitative PCR (qPCR) was subsequently used to validate the expression of significant DEGs in liver tissues of both groups. Results Compared with the control group, the model group which received subcutaneous letrozole sustained-release pellets combined with a high-fat diet exhibited significantly increased body weight (P<0.001), prominent polycystic ovarian morphology, and significantly decreased liver-to-body weight ratio (P<0.05). However, no significant changes were observed in absolute liver weight (P>0.05), hepatic histomorphology, or lipid deposition. Transcriptome sequencing identified 119 upregulated and 217 downregulated DEGs in the liver tissues of letrozole-treated mice, which were predominantly enriched in pathways related to cholesterol and steroid biosynthesis, steroid hormone metabolism, and inflammatory responses. qPCR validation demonstrated that mRNA expression of HSD3B2 and HMGCR was significantly upregulated in liver (P<0.01), while mRNA expression of IL4, CCL2 and COL1A1 was downregulated (P<0.05) in the model group compared with the control group. However, Masson's trichrome staining and F4/80 immunohistochemical analysis showed no significant changes in hepatic fibrosis or macrophage infiltration. Conclusion Subcutaneous administration of letrozole sustained-release pellets combined with a high-fat diet successfully establishes a mouse model of PCOS. The model mice exhibited significant changes in hepatic gene expression. Liver may contribute to PCOS pathogenesis through regulating cholesterol and steroid metabolism.

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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
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    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.

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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
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    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.

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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
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    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.

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    Special Welfare and Ethical Requirements for Infectious Animal Experiments
    MIN Fangui, FU Hongkun, LIU Yonggang, LIU Xiangmei, LIU Zhonghua, LI Yao, TAO Yufeng
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 239-246.   DOI: 10.12300/j.issn.1674-5817.2024.122
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    Infectious disease animal models serve as indispensable tools for understanding the transmission patterns, pathogenesis, and anti-infective medicine. During the preparation and application of infectious animal disease models, situations inevitably arise that violate animal welfare and ethics, such as animal pain, suffering, and distress. Considering the biosafety factors, animal mortality is still used as the experimental endpoint in most experiments on infectious animals, which poses extremely high requirements for animal welfare and ethics. It is imperative to establish guiding principles or norms for the welfare and ethics of infectious animal experiments. Based on the fundamental principles of the welfare and ethics of experimental animals, this paper explores the special welfare and ethical requirements in infectious animal experiments. It emphasizes that infectious animal experiments should fully consider the balance among the scientific objectives of the research plan, animal welfare and ethics, and occupational health and safety of personnel. Based on literature research and comparative analysis of the welfare and ethical requirements of conventional animal experiments, special welfare and ethics requirements for infectious animal experiments are proposed, including personnel requirements, experimental animal selection standards, living environment management and equipment, special care and veterinary care, and humane endpoints. Personnel are required to undergo effective biosafety training, and sufficient authority should be granted to the Institutional Animal Care and Use Committee (IACUC), veterinarians, and veterinary technicians to ensure the implementation of animal welfare and ethics practices. The selection of laboratory animals should fully consider the requirements of research objectives, welfare, ethics, and biosafety, with the susceptibility and body size of laboratory animals being the key concerns in high-level biosafety laboratories. It is also clarified that the humane endpoint is an indispensable element of welfare and ethics in infectious animal experiments. Environmental enrichment and special care are necessary guarantees for achieving animal welfare and ethics. Therefore, this study can serve as a reference for relevant work.

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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
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    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.

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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
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    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.

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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
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    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.

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    Differential Analysis of Oral Microbiota in db/db Mouse Model of Type 2 Diabetes Utilizing 16S rRNA Sequencing
    PAN Qianjia, GE Junyi, HU Nan, HUA Fei, GU Min
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 147-157.   DOI: 10.12300/j.issn.1674-5817.2024.119
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    Objective To investigate the changes in oral microbiota of db/db mice and provide an experimental basis for exploring the relationship between type 2 diabetes mellitus and oral microecology. Methods Eight 10-week-old male db/db mice were designated as the diabetes experimental group (db/db group), while eight 10-week-old male db/m mice were assigned as the normal control group (db/m group). After a 5-day adaptive feeding period, tail venous blood samples were collected on the 6th and 37th days, and fasting blood glucose (FBG) levels and oral glucose tolerance test (OGTT) were performed for both groups to verify the reliability of the diabetes model. On the 15th day of feeding with the same diet, oral microbiota samples were collected from the buccal mucosa, dorsal and ventral tongue surfaces, oral floor mucosa, hard palate mucosa, and the gingival areas of both the upper and lower jaws of the two groups. Genomic DNA from the oral microbiota was extracted, and the V3-V4 regions of the 16S ribosomal RNA (16S rRNA) gene were amplified using a GeneAmp 9700 thermocycler. The composition of the oral microbiota was evaluated through double-labelled amplification and sequencing on the Illumina MiSeq platform, followed by bioinformatics analysis using QIIME software(version 1.6.0). Results The FBG levels and OGTT results on the 6th and 37th days after the start of the experiment indicated that db/db mice exhibited more pronounced symptoms of type 2 diabetes compared to db/m mice. Alpha diversity (α diversity) analysis showed no significant difference in the diversity of oral microbiota between the two groups (P>0.05); however, there was a significant difference in richness (P<0.05). Principal coordinate analysis(PCoA) revealed differences in the oral microbiota composition between the db/db group and db/m group (P<0.05). Species composition analysis and LEfSe analysis demonstrated that the relative abundance of oral microbiota in db/db group mice, predominantly composed of p_Proteobacteria, increased significantly at the phylum level (P<0.05). At the genus level, the relative abundances of g_Proteus and g_Enterococcus showed a significant increase (P<0.001). Conclusion The composition and diversity of oral microbiota in db/db mice with type 2 diabetes mellitus significantly differed from those without the disease.

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    Key Points for Establishing Occupational Health and Safety Management System in Laboratory Animal Institutions
    SHAO Qiming, BIAN Yong, SHI Aimin
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 188-196.   DOI: 10.12300/j.issn.1674-5817.2024.123
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    As one of the modern enterprise management systems, the Occupational Health and Safety Management System (OHSMS) has garnered increasing attention. The OHSMS has undergone continuous refinement and expansion across various fields, emerging as a pivotal indicator of enterprise competitiveness. Currently, both the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) International and the China National Accreditation Service for Conformity Assessment (CNAS) require for establishing occupational health management related to laboratory animal work. However, within the domestic laboratory animal industry, development of OHSMS is relatively lagging behind due to unfamiliarity with laws and regulations and a lack of experienced management personnel. Consequently, the OHSMS in laboratory animal institutions is still in its early stage. Drawing on the authors' practical experience in establishing OHSMS in laboratory animal institutions, this article first outlines domestic and international occupational health laws, regulations, and safety management systems for laboratory animals, as well as common occupational diseases and their associated risk factors in China. Subsequently, this article highlights key elements for the construction of OHSMS in laboratory animal institutions in areas such as establishing occupational health and safety regulations, conducting training, performing occupational health examinations for staff, monitoring on-site occupational hazard factors, implementing the "three simultaneous" system for occupational disease prevention facilities in construction projects, creating and maintaining occupational health records, developing a notification management system for occupational hazards, and formulating emergency response plans for occupational disease hazard accidents. These points are intended as a reference for professionals in the industry.

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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
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    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.

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    Establishment Methods and Application Progress of Rodent Models for Drug Addiction
    WANG Biying, LU Jiashuo, ZAN Guiying, CHEN Ruosong, CHAI Jingrui, LIU Jinggen, WANG Yujun
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 158-166.   DOI: 10.12300/j.issn.1674-5817.2024.080
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    Drug addiction, also referred to as drug dependence or substance use disorder, is a chronic and recurrent brain disease. Its main characteristics are compulsive drug-seeking behavior, continued use of drugs, and a loss of control over intake. Prolonged use of addictive substances can result in both physiological and psychological dependence. When usage is ceased, individuals may experience intense discomfort, including anxiety, insomnia, nausea, vomiting, and a strong craving for the substances. Drug dependence is classified into two types: physical dependence and psychological dependence. Physical dependence describes a pathological state of adaptation that results from the repeated use of addictive substances, leading to severe withdrawal syndrome upon cessation. Psychological dependence involves a mental craving for addictive substances, which is needed to experience the specific euphoria that follows consumption. Regular or continuous use is required to sustain these euphoric effects. The mechanisms of addiction are complex and influenced by genetic, environmental, and various other factors. They involve higher-level neurological activities, such as memory, reward, and decision-making. Currently, effective treatment methods for drug addiction are insufficient. Due to the complexity of drug addiction, laboratory animal research is essential. Using animal behavioral models to simulate human drug addiction can enhance our understanding of the mechanisms of addiction. This research offers a comprehensive overview of various animal experimental models that explore both physical and psychological dependence. It includes detailed descriptions of the methods and procedures used to assess physical dependence, behavioral sensitization, conditioned place preference, drug discrimination, and self-administration experiments. Additionally, the characteristics of each experimental model are compared, and the relevance of these models is discussed, aiming to provide support for the research on addiction mechanisms and the development of therapeutic methods.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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    Implementation of "Three Simultaneities" for Occupational Health Protection Facilities in Laboratory Animal Institution Construction Project
    CAI Mengshi, SU Xing, SHAO Qiming
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 221-228.   DOI: 10.12300/j.issn.1674-5817.2024.127
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    Occupational health is a discipline dedicated to the identification, assessment, prediction, and control of occupational hazards in the workplace and their potential impacts on health. Its core objective is to ensure the safety of workers through targeted preventive strategies and protective mechanisms, thereby minimizing health risks caused by occupational hazards and promoting physical health, mental well-being, and overall social welfare in the workplace. The "three simultaneities" of occupational health protection facilities in construction projects (hereinafter referred to as "three simultaneities") refers to the principle that occupational disease prevention facilities must be designed, constructed, and put into operation simultaneously with the main project. The implementation of "three simultaneities" involves several stages: a preliminary assessment of occupational disease risks, the design of occupational disease protection facilities in accordance with relevant standards, and ensuring these facilities are constructed in parallel with the primary engineering project to maintain integrity and effectiveness. During the construction phase, occupational disease prevention facilities must be constructed simultaneously with the main project to ensure their integrity and effectiveness. Upon project completion, the construction unit is responsible for evaluating the control effects of occupational disease hazards and conducting the acceptance of occupational disease prevention facilities to ensure the occupational health of employees. The implementation of the "three simultaneities" for occupational disease protection facilities in construction projects is a mandatory national regulation, and non-compliance may result in legal penalties such as fines and forced suspension of work. However, for some laboratory animal institutions, the implementation of the "three simultaneities" remains a challenge. This paper aims to provide practical guidance and feasible implementation strategies for such institutions by introducing the development history of the "three simultaneities" in occupational health, explaining its relevance and necessity in the context of laboratory animal facilities, and outlining step-by-step procedures for compliance and implementation.

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    Review on Occupational Health Risk Prevention and Control for Laboratory Animal Practitioners in Chinese General Universities
    ZHANG Qian, DENG Qingxiu, CAI Lin
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 206-213.   DOI: 10.12300/j.issn.1674-5817.2024.126
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    With the rapid development of the laboratory animal industry in China, animal experimentation has become a crucial tool for both scientific research output and classroom teaching in universities. Correspondingly, the workforce engaged in laboratory animal work has expanded significantly. In recent years, while the Chinese government has begun to emphasize the occupational health of laboratory animal practitioners by introducing relevant laws and policies, current prevention and control efforts within universities remain insufficient. This study conducted an in-depth investigation into the occupational health management and protection of laboratory animal practitioners in nine general universities in China, focusing on the management standards within university laboratory animal research centers. The research was carried out through field visits, interviews, and academic exchanges, focusing on standardized management practices within university laboratory animal research centers. The findings highlight five major occupational health risks faced by these practitioners: laboratory animal allergy, zoonotic infectious diseases, mechanical injuries, chemical exposures, and psychological factors. Key challenges identified include underdeveloped management systems, a lack of risk assessment mechanisms, and inadequate protective facilities. To address these issues, the study proposes a three-tiered prevention and control framework. At the university level, institutions should establish comprehensive regulatory systems and increase funding investment. At the laboratory animal center level, biosafety laboratories should be developed, risk source management strengthened, and emergency preparedness and health monitoring enhanced. At the level of individual practitioners, appropriate use of personal protective equipment should be enforced, occupational health training should be provided, standard operating procedures should be strictly followed, and occupational exposure incidents should be reported promptly. Through multi-party collaboration, this study explores feasible paths for constructing a scientific and efficient occupational health protection system in domestic universities, aiming to provide theoretical support and practical references for the sustainable development of the laboratory animal industry in China.

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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
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    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.

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    Occupational Health Risk Management Measures for Personnel Handling Non-Human Primate Laboratory Animals: An Overview
    LI Qian, CHEN Jiaqi, LI Lihong, ZHANG Feiyan, MAO Huaming, LÜ Longbao
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 197-205.   DOI: 10.12300/j.issn.1674-5817.2024.088
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    Owing to their high genetic and physiological similarities to humans, non-human primates (NHPs) have become pivotal animal models in life sciences research and biomedical development. NHP laboratory animals are not only an ideal platform for exploring the mechanisms of neurological diseases and infectious diseases, but they are also widely used in preclinical safety evaluations of macromolecular drugs, which are considered the "gold standard". Nevertheless, this biological similarity increases the risk of zoonotic disease transmission to personnel working with NHP laboratory animals, their tissues, and body fluids. In light of recent domestic and international outbreaks of zoonotic diseases as well as the implementation of the Biosafety Law, this study examines the occupational risk factors encountered by personnel working with NHPs. This includes biological, chemical, and physical factors. This paper also covers common zoonoses, classification of the corresponding pathogens, transmission routes, risk severity levels, and protocols for post-exposure management. A multidimensional prevention and control framework is proposed, which includes the following components. (1) Risk Assessment and Emergency Response: Regularly identify hazards through an Occupational Health and Safety Committee (OHSC) and develop post-exposure emergency protocols. (2) Optimization of Management Systems: Improve facility design, optimize the allocation of personal protective equipment, and enhance health surveillance and vaccination programs. (3) Technical Training and Standardized Operations: Provide specialized training in NHP laboratory animal ethology and biosafety practices. Additionally, implement intelligent monitoring technologies to reduce the occurrence of aggressive incidents. This paper outlines measures designed to enhance health and safety awareness among personnel working with NHP laboratory animals. It emphasizes the need for strengthened guidance on the use of personal protective equipment (PPE) and the standardization of professional operational practices. The goal is to safeguard personnel health and safety, reduce occupational exposure rates, and effectively prevent occupational diseases related to laboratory animals.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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    Establishment of Occupational Health Related Files in Laboratory Animal Institutions
    XU Chao, SUN Qiufang, SHAO Qiming
    Laboratory Animal and Comparative Medicine    2025, 45 (2): 214-220.   DOI: 10.12300/j.issn.1674-5817.2024.125
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    Establishing occupational health related files is a key component of building an occupational health and safety management system in laboratory animal institutions. These files not only serve as a critical means of protecting the legitimate rights and interests of laboratory animal personnel but also provide institutional safeguards. According to relevant laws and regulations, laboratory animal institutions are responsible for establishing and properly maintaining occupational health surveillance files (also known as occupational hygiene files) for both the institution and its personnel. In accordance with the Occupational Health Archives Management Standards, laboratory animal institutions are required to maintain at least seven categories of occupational health management archive folders: the "three simultaneous" occupational health archives for construction projects; occupational health management folders; occupational health publicity and training folders; folders for monitoring and evaluating occupational disease hazard factors; employer occupational health surveillance folders; individual occupational health surveillance folders for employees; and other relevant folders. Building upon this foundation, the Regulations on the Supervision and Administration of Occupational Health in the Workplaces further specify detailed requirements for occupational health files. These include: documents outlining the responsibility system for occupational disease prevention and control; occupational hygiene management systems and standard operating procedures, lists of types of occupational disease hazard factors in workplaces, their distribution by job distribution, and records of work exposure, basic information on occupational disease prevention and emergency rescue facilities, along with records of their allocation, usage, maintenance, inspection and replacement; records concerning the provision, distribution, maintenance and replacement of personal protective equipment (PPE); and reports on occupational disease hazard incidents; and corresponding emergency response records—amounting to twelve specific requirements in total. In short, occupational health management files are a comprehensive collection of data and documentation related to workers' occupational health. Based on the author's practical experience, this paper outlines the main types of occupational health management files, their basic contents, and key points in their development. It also offers a detailed analysis of how to establish such files, aiming to support laboratory animal institutions in successfully developing occupational health management archives and improving their occupational health and safety management systems.

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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
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    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.

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    Dynamic Evaluation of Vinorelbine-Induced Phlebitis of Dorsalis Pedis Vein in a Rat Model
    JIANG Meng, HAO Shulan, TONG Liguo, ZHONG Qiming, GAO Zhenfei, WANG Yonghui, WANG Xixing, JI Haijie
    Laboratory Animal and Comparative Medicine    2025, 45 (3): 251-258.   DOI: 10.12300/j.issn.1674-5817.2024.163
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    Objective To dynamically observe the clinical symptoms and pathological changes in a rat model of vinorelbine-induced phlebitis via injection into the dorsalis pedis vein. Methods Twenty-eight 11-week-old male SPF-grade SD rats were randomly divided into a model group (n=20) and a control group (n=8). The model group received a single injection of 0.1 mL vinorelbine solution (4 mg/mL) via the right hind limb dorsalis pedis vein, while the control group received an equal volume of normal saline via the same method. The occurrence and grading of phlebitis in both groups were observed and recorded daily. The volume of the injured limb was measured by the drainage method to calculate the swelling rate. The weight-bearing ratio of the injured limb was assessed using a bipedal balance pain meter, and the skin temperature of the injured limb was measured by infrared thermal imaging. These measurements were conducted for 9 consecutive days. Starting from day 1, three rats from the model group were euthanized every other day. A 1-cm segment of the vein extending proximally from the injection site was collected. Pathological changes in the vein tissue were examined by hematoxylin-eosin staining, and ultrastructural changes of the vascular endothelium were observed using scanning electron microscopy. Results Compared to the control group, the injected hindlimb of model rats showed redness and swelling on day 1, with the swelling rate peaking at (81.89±15.75) % on day 3 (P<0.001), then gradually alleviating and decreasing to (15.41±0.33) % by day 9 (P<0.01). Pain was observed in the affected limbs of model rats on day 1 and worsened markedly on day 3, with the weight-bearing ratio decreasing to (36.35±4.91)% (P<0.001). Meanwhile, the skin temperature of the lesion site increased, reaching (36.36±0.40) ℃ on day 5 (P<0.001). Both pain and fever returned to near normal levels by day 9. Phlebitis grading in the model group showed that 75.0% of rats were grade Ⅱ on day 1; grade Ⅲ and Ⅳ each accounted for 37.5% on day 3; from days 5 to 9, most rats exhibited cord-like veins, predominantly grade III. Venous tissue showed peripheral edema and inflammatory cell infiltration on day 1, which gradually progressed to intimal rupture, vessel wall thickening, and even lumen narrowing from day 3 to 9. The venous intima exhibited destruction of tight junctions between endothelial cells and adhesion of blood cells, progressing to roughened, wrinkled, and protruding intimal surfaces. Conclusion The vinorelbine-induced phlebitis of dorsal foot vein in rat model is characterized by local redness, swelling, warmth, and pain from days 3 to 5, which largely resolve by day 9, although cord-like veins can still be observed. With disease progression, venous tissue develops edema, vessel wall thickening, and lumen narrowing. The venous intima shows rupture, roughening, and in some cases, complete loss.

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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
    Abstract428)   HTML49)    PDF (1174KB)(225)       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.

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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
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    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.

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