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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 Abstract （984）   HTML （47）    PDF （846KB）（2790）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Polymorphism and Tissue Expression Analysis of TYR and MC1 R Genes in Guinea Pigs with Different Coat-Color Phenotypes TANG Yingen, FENG Yaxian, ZHONG Min, WEI Zhen, WANG Lie, LIU Diwen Laboratory Animal and Comparative Medicine    2025, 45 (1): 21-29.   DOI: 10.12300/j.issn.1674-5817.2024.105 Abstract （688）   HTML （21）    PDF （1784KB）（974）       Knowledge map    Save Objective To explore the polymorphism of tyrosinase (TYR) and melanocortin 1 receptor (MC1R) genes and their mRNA expression levels in relation to coat-color phenotypes in guinea pigs, providing genetic markers for locating dominant traits in guinea pigs. Methods A total of 57 self-bred ordinary-level guinea pigs were selected and divided into three groups based on coat color: white (n=22), variegated (n=22) and black (n=13). The guinea pigs were euthanized with an overdose of pentobarbital sodium via intraperitoneal injection. DNA was then extracted from the dorsal skin tissue. Polymorphism in the coding sequence (CDS) of the exons of the TYR and MC1R genes in each group was detected by cloning and sequencing. The mRNA expression of the two genes in skin tissues was detected by real-time fluorescent quantitative PCR to investigate the relationship between these genes and guinea pig coat color. Results A single nucleotide polymorphism (SNP) site was found in the CDS region of TYR exon Ⅰ, where the base A was replaced by G. All white guinea pigs had the G/G genotype for TYR, while no deep-colored (variegated and black) guinea pigs exhibited the G/G genotype for TYR. Most deep-colored guinea pigs had the A/A genotype, and a few had A/G genotype. The A/A genotype frequency in black guinea pigs was higher than in variegated guinea pigs. A 2 760 bp sequence deletion was identified in the exon of the MC1R gene, marked as the - gene, with non-deleted samples marked as N gene. Most white guinea pigs had the -/- genotype for MC1R, variegated guinea pigs mainly had the -/N genotype, and black guinea pigs mainly had the N/N genotype, with a few showing the -/N. The TYR gene expression level was higher in white guinea pigs, lower in variegated guinea pigs, and intermediate in black guinea pigs, but there was no significant difference among the three groups (P>0.05). The MC1R gene expression level in white guinea pigs was extremely low, while both variegated and black guinea pigs showed significantly higher levels than white guinea pigs (P<0.01). Black guinea pigs showed significantly higher levels than variegated guinea pigs (P<0.05). Conclusion The TYR and MC1R genes synergistically regulate coat color of guinea pigs. The G-site mutation in the TYR gene may lead to albinism, and the change of N-site in the MC1R gene affects the depth of the coat color. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Animal Models for Hernia Diseases and New Hernia Repair Materials FEI Bin, GUO Wenke, GUO Jianping Laboratory Animal and Comparative Medicine    2025, 45 (1): 55-66.   DOI: 10.12300/j.issn.1674-5817.2024.121 Abstract （671）   HTML （19）    PDF （889KB）（2655）       Knowledge map    Save Hernia is a common and frequently occurring condition in general surgery, referring to the displacement of an organ or part of an organ from its normal anatomical position through a congenital or acquired weak point, defect, or space into another area. Its pathogenesis is complex, involving multiple factors such as abdominal wall weakness or increased intra-abdominal pressure. The clinical manifestations of hernia vary depending on its type, location, and severity. As the aging of the population continues to advance, the incidence of hernia has been increasing annually. Animal models serve as an important tool in hernia research. They enable the evaluation of the safety and efficacy of new repair materials and techniques, as well as assisting clinicians in developing new surgical methods and investigating the mechanisms and novel therapies for certain hernia diseases and their complications. Given the significant differences in the pathophysiological mechanisms of different types of hernia diseases, the methods and evaluation criteria for establishing animal models are highly diverse. Furthermore, the methods for establishing animal models are closely related to experimental objectives, and different experimental goals require different animal models. Therefore, selecting appropriate animal models based on experimental objectives is crucial for ensuring the smooth progress of research and obtaining reliable results. To this end, this review summarizes effective methods for establishing animal models for external abdominal hernias (including incisional hernia, inguinal hernia, umbilical hernia, parastomal hernia, incarcerated hernia, and pelvic floor hernia), congenital diaphragmatic hernia, hiatal hernia, and cerebral hernia. It provides a detailed analysis of the advantages, disadvantages, and evaluation criteria of these models. Additionally, this review summarizes recent preclinical applications of new hernia repair materials, aiming to provide references for animal experimental research in the field of hernia studies. Table and Figures | Reference | Related Articles | Metrics

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Improving the Certainty of Evidence in Animal Experiment Systematic Review/Meta-Analysis: An Empirical Study of the GRADE Method LI Tengfei, ZHENG Qingyong, XU Jianguo, LI Yiyi, ZHOU Yongjia, XU Caihua, ZHANG Mingyue, TIAN Jiexiang, WANG Gang, TIAN Jinhui Laboratory Animal and Comparative Medicine    2025, 45 (1): 101-111.   DOI: 10.12300/j.issn.1674-5817.2024.109 Abstract （644）   HTML （20）    PDF （1114KB）（852）       Knowledge map    Save Animal experiments are essential tools in biomedical research, serving as a bridge between basic research and clinical trials. Systematic reviews and meta-analyses (SRs/MAs) of animal experiments are crucial methods for integrating evidence from animal experiment, which can facilitate the translation of findings into clinical research, reduce translational risks, and promote resource integration in basic research. With the continuous development of the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) methodology, its application in SRs/MAs of animal experiments has gained increasing attention. This article first outlines the principles and specific applications of the GRADE methodology in SRs/MAs of animal experiments, including qualitative descriptive systematic reviews, meta-analyses, and network meta-analyses. It then deeply analyzes the misuse of the GRADE methodology in practice, including incorrect evidence grading, improper classification of evidence, misapplication in qualitative systematic reviews, inconsistencies between the documentation of the upgrading and downgrading process and results, and inappropriate use for making recommendations. Furthermore, this article comprehensively discusses the factors influencing the grading of evidence certainty in SRs/MAs of animal experiments, including the impact of bias risk, indirectness, inconsistency, imprecision, and publication bias on evidence downgrading, as well as the role of large effect sizes and cross-species consistency in evidence upgrading. Finally, in response to the issues discussed, improvement strategies are proposed, including further research and optimization of the GRADE methodology for SRs/MAs of animal experiments, the development of reporting guidelines tailored to the characteristics of SRs/MAs in animal experiment research, and enhanced professional training for researchers in the GRADE methodology. This article aims to improve the quality of evidence in SRs/MAs of animal experiments, strengthen their reliability in clinical decision-making, and promote the more efficient translation of findings from animal experiment research into clinical practice. Table and Figures | Reference | Related Articles | Metrics

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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 Abstract （624）   HTML （43）    PDF （4039KB）（2818）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Analysis of Kidney Differential Metabolites and Hypoxia Adaptation Mechanism of Plateau Pikas Based on UHPLC-QE-MS HE Yuxin, BAI Zhenzhong, XUE Hua, GUO Zixu, CAO Xuefeng Laboratory Animal and Comparative Medicine    2025, 45 (1): 3-12.   DOI: 10.12300/j.issn.1674-5817.2024.095 Abstract （621）   HTML （98）    PDF （1785KB）（1526）       Knowledge map    Save Objective To explore the potential mechanisms of hypoxic adaptive metabolic changes in the kidneys of plateau pikas at different altitudes using non-targeted metabolomics analysis via ultra-high-performance liquid chromatography coupled with quadrupole electrostatic field orbital trap-mass spectrometry (UHPLC-QE-MS). Methods 10 plateau pikas were captured at an altitude of 4 360 m in Xingxiuhai area, Maduo County, Guoluo Tibetan Autonomous Prefecture, Qinghai Province (MD group), and 10 plateau pikas were captured at an altitude of 2 900 m in Menyuan area, Haibei Tibetan Autonomous Prefecture, Qinghai Province (MY group). After anesthesia, serum samples were collected, and kidney samples were collected after euthanasia. General physiological and biochemical indicators were measured and metabolomics analysis was performed. Part of the serum samples was used for hematology analysis, another part for blood gas analysis, and the remaining part for biochemical indicator detection. Metabolites were extracted from the kidney tissue samples and then analyzed using UHPLC-QE-MS. Differential metabolites were analyzed using metabolomics principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA), with screening criteria set as variable importance in projection (VIP)>1.5 and fold change (FC)>1.5, or VIP>1.5 and FC<1/1.5. Correlation analysis heatmaps, significance analysis volcano plots, signaling pathway recognition bubble charts, and rectangular graphs were used for the analysis of differential metabolites and related signaling pathways. Results The red blood cell count, glucose, urea nitrogen, uric acid, and homocysteine levels in the MD group plateau pikas were higher than those in the MY group, while hemoglobin, hematocrit, creatinine, and carbon dioxide combining power were lower than those in the MY group. This indicated a significant difference in the blood oxygen-carrying capacity of plateau pikas at different altitudes. The principal component pattern recognition analyses, and OPLS-DA permutation test showed that the kidney metabolites of the MD and MY groups of plateau pikas had distinct clustering distributions (R2Y=0.930, Q2=0.655). According to the screening criteria and database comparison, 46 differential metabolites were identified in the kidneys of plateau pikas at different altitudes. In the MD group of plateau pikas, the expression levels of bufadienolide, adenosine, adenine, diosgenin, berberine chloride, carnosol, and astaxanthin were significantly increased (VIP>1.5, P<0.05), while the levels of arachidonic acid, histamine, and coumarin were significantly decreased (VIP>1.5, P<0.05). The analysis of related signaling pathways showed that the biosynthetic pathways of valine, leucine, and isoleucine had the largest impact factors (P<0.05), while the biosynthetic pathways of pantothenate and coenzyme A showed the most significant enrichment (P<0.05). Conclusion The differential metabolites of amino acids, pantothenate, and coenzyme A pathways in the kidneys of plateau pikas at different altitudes may be involved in the metabolic mechanisms of plateau pikas' hypoxia adaptation in high-altitude environments. Table and Figures | Reference | Related Articles | Metrics

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Dynamic Monitoring and Correlation Analysis of General Body Indicators, Blood Glucose, and Blood Lipid in Obese Cynomolgus Monkeys WEI Yanye, SHEN Guo, ZHANG Pengfei, SHI Songping, HU Jiahao, ZHANG Xuzhe, HUA Huiyuan, HUA Guanyang, LU Hongzheng, ZENG Yong, JI Feng, WEI Zhumei Laboratory Animal and Comparative Medicine    2025, 45 (1): 30-36.   DOI: 10.12300/j.issn.1674-5817.2024.091 Abstract （603）   HTML （19）    PDF （908KB）（1210）       Knowledge map    Save Objective This study aims to investigate the dynamic changes in general body parameters, blood glucose, and blood lipid profiles in obese cynomolgus monkeys, exploring the correlations among these parameters and providing a reference for research on the obese cynomolgus monkey model. Methods 30 normal male cynomolgus monkeys aged 5 - 17 years old (with body mass index < 35 kg/m2 and glycated hemoglobin content < 4.50%) and 99 spontaneously obese male cynomolgus monkeys (with body mass index ≥35 kg/m2 and glycated hemoglobin content < 4.50%) were selected. Over a period of three years, their abdominal circumference, skinfold thickness, body weight, body mass index, fasting blood glucose, glycated hemoglobin, and four blood lipid indicators were monitored. The correlations between each indicator were analyzed using repeated measurement ANOVA, simple linear regression, and multiple linear regression correlation analysis method. Results Compared to the control group, the obese group exhibited significantly higher levels of abdominal circumference, skinfold thickness, body weight, body mass index, and triglyceride (P＜0.05). In the control group, skinfold thickness increased annually, while other indicators remained stable. Compared with the first year, the obese group showed significantly increased abdominal circumference, skinfold thickness, body weight, body mass index, triglyceride, and fasting blood glucose in the second year(P＜0.05), with this increasing trend persisting in the third year (P＜0.05). In the control group, the obesity incidence rates in the second and third years were 16.67% and 23.33%, respectively, while the prevalence of diabetes remained at 16.67%. In the obese group, the diabetes incidence rates were 29.29% and 44.44% in years 2 and 3, respectively. Among the 11-13 year age group, the incidence rates were 36.36% and 44.68%, while for the group older than 13 years, the rates were 28.13% and 51.35%. Correlation analysis revealed significant associations (P＜0.05) between fasting blood glucose and age, abdominal circumference, skinfold thickness, body weight, and triglyceride in the diabetic monkeys. Conclusion Long-term obesity can lead to the increases in general physical indicators and fasting blood glucose levels in cynomolgus monkeys, and an increase in the incidence of diabetes. In diabetic cynomolgus monkeys caused by obesity, there is a high correlation between their fasting blood glucose and age, weight, abdominal circumference, skinfold thickness, and triglyceride levels, which is of some significance for predicting the occurrence of spontaneous diabetes. Table and Figures | Reference | Related Articles | Metrics

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Study on the 90-day Feeding Experimental Background Data of SD Rats for Drug Safety Evaluation QIN Chao, LI Shuangxing, ZHAO Tingting, JIANG Chenchen, ZHAO Jing, YANG Yanwei, LIN Zhi, WANG Sanlong, WEN Hairuo Laboratory Animal and Comparative Medicine    2025, 45 (4): 439-448.   DOI: 10.12300/j.issn.1674-5817.2024.187 Abstract （599）   HTML （20）    PDF （1104KB）（524）       Knowledge map    Save Objective To establish background data for a 90-day feeding trial of SD rats to ensure the reliability of research data. Methods Background data from six independent 90-day feeding trials of SD rats conducted by the National Center for Safety Evaluation of Drugs from 2020 to 2023 were summarized. These studies involved a blank control group of 120 SPF-grade 4-week-old SD rats, with an equal number of males and females, which were only given standard full-nutrient pelleted rat feed. After the quarantine period, the animals were observed for an additional 90 days, followed by intraperitoneal injection of Zoletil (50 mg/mL) for anesthesia, blood sampling, euthanasia, and necropsy. By analyzing the data from the blank control group, a relevant background database for SD rats was established. Results Both male and female rats exhibited steady weight gain, with a more pronounced increase in male rats. Within 90 days, the average body weight of male and female rats increased to over 500 g and 300 g, respectively. Three weeks later, the average daily food intake of male rats stabilized at approximately 25~28 g per rat, while that of female rats remained stable at approximately 16~19 g per rat. The food utilization rate of all animals gradually decreased from the first week of the experiment. In the white blood cell (WBC) differential count results, significant differences were observed in the counts of WBCs, neutrophils (Neut), lymphocytes (Lymph), and monocytes (Mono) between males and females (P<0.001). However, there were no significant differences in the percentages of neutrophil (%Neut), lymphocyte (%Lymph), and monocyte (%Mono) between the sexes (P>0.05). The average red blood cell count (RBC), hemoglobin concentration (HGB), hematocrit (HCT), platelet count (PLT), prothrombin time (PT), and activated partial thromboplastin time (APTT) were higher in male animals than in female animals (P<0.05). The average values of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine phosphokinase (CK), lactate dehydrogenase (LDH), glucose (GLU), and triglyceride (TG) in male rats were higher than those in female rats (P<0.05). The urinary pH range for male animals was 5.0 to 8.5, while for female animals it was 6.5 to 9.0. The majority of male animals had a urinary specific gravity lower than 1.020, and the majority of female animals had a urinary specific gravity lower than 1.015. The weights of various organs (excluding the adrenal glands and reproductive organs) in male animals were heavier than those in female animals (P<0.001), while the organ/body weight ratios (excluding the kidneys and reproductive organs) of female animals were higher than those of male animals (P<0.001). Conclusion This study summarizes the background reference ranges for body weight, food intake, hematology, and serum biochemistry indicators in SPF-grade SD rats in the untreated control group from six 90-day feeding trials conducted by the National Center for Safety Evaluation of Drugs. It provides important reference data for related research. By summarizing the background and spontaneous histopathological changes in rats, this study aids in the standardization and normalization of subsequent research, as well as in the evaluation and analysis of abnormal results. Table and Figures | Reference | Related Articles | Metrics

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Study on Cardiac Aging Phenotypes of SHJH hr Mice LIU Rongle, CHENG Hao, SHANG Fusheng, CHANG Shufu, XU Ping Laboratory Animal and Comparative Medicine    2025, 45 (1): 13-20.   DOI: 10.12300/j.issn.1674-5817.2024.100 Abstract （584）   HTML （90）    PDF （1909KB）（739）       Knowledge map    Save Objective To investigate the spontaneous premature cardiac aging in SHJH hr mice. Methods A comparative study was conducted between SHJH hr mice (SHJH hr group) and wild-type ICR mice (WT group) at different ages (10 and 24 weeks). Cardiac function was analyzed using a small animal in vivo ultrasound imaging system. After euthanasia, organs were collected and weighed to assess the extent of cardiac atrophy. Cardiac pathological damage was observed using hematoxylin-eosin (HE) staining. Cardiac fibrosis was analyzed using Masson staining. Myocardial cell area was analyzed after wheat germ agglutinin (WGA) staining. The activities of oxidative damage indicators in myocardial tissue, including superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT), as well as the content of 8-hydroxy-2'-deoxyguanosine (8-OHdG), were measured using enzyme-linked immunosorbent assay. Real-time fluorescence quantitative PCR was used to measure the mRNA expression levels of factors associated with inflammation, fibrosis, and oxidative stress. Colorimetric assay was used to measure malondialdehyde (MDA) levels. Results Compared to WT group mice of the same age, 10-week-old mice in the SHJH hr group showed no significant differences in stroke volume (SV), ejection fraction (EF), fractional shortening (FS), or heart and lung weights. However, at 24 weeks of age, mice in the SHJH hr group had significantly lower SV, EF, and FS values compared to mice of the same age in the WT group (P<0.05), with no significant change in lung weight but a significant reduction in heart weight (P<0.05). Histological analysis of heart tissue from 24-week-old mice revealed no significant difference in cardiac fibrosis levels between SHJH hr and WT groups, but WGA staining showed a significant reduction in myocardial cell area in mice in the SHJH hr group (P<0.05). PCR analysis revealed a significant downregulation of mRNA levels of oxidative stress factors Sod2, Gpx1, and Cat genes (P<0.05). Biochemical assays indicated significantly reduced activities of oxidative damage-related enzymes SOD, GPX, and CAT in myocardial tissue (P<0.05), while the levels of oxidative damage markers 8-OHdG and MDA significantly increased (P<0.05). Conclusion Mice in the SHJH hr group exhibit premature cardiac aging, which may be associated with oxidative stress damage in myocardial tissue. Table and Figures | Reference | Related Articles | Metrics

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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 Abstract （582）   HTML （17）    PDF （882KB）（2214）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Optimization of Surgical Procedure and Efficacy Evaluation of Aortic Calcification Model in Rats with Chronic Kidney Disease PAN Yicong, JIANG Wenhong, HU Ming, QIN Xiao Laboratory Animal and Comparative Medicine    2025, 45 (3): 279-289.   DOI: 10.12300/j.issn.1674-5817.2024.128 Abstract （568）   HTML （27）    PDF （2015KB）（1311）       Knowledge map    Save Objective To establish a chronic kidney disease-associated aortic calcification model in SD rats using different nephrectomy surgical methods, and to compare and evaluate surgical duration and survival time to explore a more optimized modeling method. Methods According to different surgical methods, the SD rats were divided into four groups: Group A: intraperitoneal resection of 2/3 of the left kidney followed by right total nephrectomy in the second stage; Group B: intraperitoneal resection of 2/3 of the left kidney and simultaneous right total nephrectomy; Group C: dorsal approach right total nephrectomy followed by resection of 2/3 of the left kidney in the second stage; Group D: dorsal approach resection of 2/3 of the left kidney followed by right total nephrectomy in the second stage. After comparing survival curves of SD rats undergoing intraperitoneal versus dorsal approaches, and staged versus single-stage nephrectomy, the optimal nephrectomy surgical method was determined. Then, twenty-four 8-week-old SPF-grade male SD rats were selected for nephrectomy combined with calcitriol-induced calcification. Experimental group (12 rats): the dorsal approach left 2/3 nephrectomy followed by right total nephrectomy, with intraperitoneal injection of 1 μg/kg calcitriol administered one week later to induce aortic calcification. Control group (12 rats): the intraperitoneal injection of 250 μL/kg physiological saline containing 1% DMSO one week after sham surgery. After intraperitoneal injection of drugs for 3 months, the survival status of rats in each group was observed. Under anesthesia, blood samples were collected from each group to measure serum phosphorus and calcium ion concentrations, as well as serum urea nitrogen and creatinine levels. After euthanizing the rats, a post-mortem examination was performed to observe the residual kidney morphology, and HE staining was used to observe the pathological changes in the coronal section of the kidney. Additionally, the entire aorta of each group was taken, and the degree of aortic calcification was observed by staining with Alizarin red S and von Kossa. Real-time fluorescence quantitative PCR was used to detect the gene expression of smooth muscle actin-associated protein alpha (Sm22), Runt-related transcription factor 2 (Runx2), and osteopontin (OPN) in rat aortic tissue to evaluate the effectiveness of the model. Results The exploratory optimization experiment of different surgical procedures found that the survival rate of group D rats,which underwent 2/3 left kidney resection followed by right whole kidney resection via the dorsal approach, was the highest, indicating that this surgical procedure was the best method for establishing a chronic kidney disease model with renal dysfunction. The experimental group rats treated with this surgical procedure combined with high-dose calcitriol injection had significantly lower serum calcium ion concentration than those in the sham-operated control group (P<0.05), while serum phosphorus ion concentration, serum creatinine, and serum urea nitrogen levels were significantly higher than those of the control group (P<0.05). HE staining of the kidneys showed significant organic changes in the kidneys of the experimental group rats, with a significant decrease in glomerular count compared to that of the control group (P<0.05), indicating the successful establishment of a renal failure model. Alizarin red S staining showed significant pigment deposition in the aortic media of the experimental group rats, while von Kossa staining showed significant silver nitrate deposition in the aortic media of the experimental group rats, which was consistent with the manifestation of aortic calcification in renal failure. Real-time fluorescence quantitative PCR showed that the expression level of Sm22 in the aortic tissue of the experimental group rats decreased (P<0.05), while the expression levels of OPN and Runx2 increased (P<0.05), indicating a transition of aortic smooth muscle cells from smooth muscle phenotype to bone-like phenotype and successful induction of an aortic calcification model. Conclusion The method of establishing an aortic calcification model of chronic kidney disease in SD rats by first removing two-thirds of the left kidney via the dorsal approach followed by right total nephrectomy, combined with high-dose calcitriol administration, shortens the surgical time, improves the success rate of modeling, and increases the animal survival rate. Table and Figures | Reference | Related Articles | Metrics

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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 Abstract （560）   HTML （47）    PDF （4184KB）（650）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Construction and Functional Validation of GTKO/hCD55 Gene-Edited Xenotransplant Donor Pigs WANG Jiaoxiang, ZHANG Lu, CHEN Shuhan, JIAO Deling, ZHAO Heng, WEI Taiyun, GUO Jianxiong, XU Kaixiang, WEI Hongjiang Laboratory Animal and Comparative Medicine    2025, 45 (4): 379-392.   DOI: 10.12300/j.issn.1674-5817.2025.024 Abstract （543）   HTML （51）    PDF （3294KB）（1293）       Knowledge map    Save Objective To develop GTKO (α-1,3-galactosyltransferase gene-knockout, GTKO)/hCD55 (human CD55) gene-edited xenotransplant donor pigs and verify their function. Methods In this study, CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated nuclease 9), PiggyBac transposon technology and somatic cell nuclear transfer technology were used to construct GTKO/hCD55 gene-edited Diannan miniature pigs. The phenotype and function of GTKO/hCD55 pigs were analyzed by Sanger sequencing, real-time fluorescence quantitative PCR, flow cytometry, immunofluorescence, bisulfite sequencing, antigen-antibody binding assays, and complement-dependent cytotoxicity assays. Results After transfection of PX458 and PiggyBac gene editing vectors into wild-type fetal pig fibroblasts, 48 single-cell colonies were obtained through puromycin drug screening. Two single-cell colonies were selected for somatic cell nuclear transfer, resulting in two fetal pigs at 33 days of gestation. The GGTA1(α-1,3-galactosyltransferase) genotypes of fetal pig F01 were -17 bp and wild type (WT), while the GGTA1 genotypes of fetal pig F02 were -26 bp/+2 bp and -3 bp. The hCD55 mRNA expression levels of both fetal pigs were significantly higher than those of WT pigs (P＜0.01). The fetal pig F02 was selected as the donor cell source for recloning, 11 surviving piglets were obtained, all identified as GTKO/hCD55 gene-edited pigs. These pigs showed absence of α-Gal antigen expression, but weak or no expression of hCD55 was observed. Methylation analysis of the hCD55 gene's CpG island showed hypermethylation in kidney tissue lacking hCD55 expression, whereas it was not methylated or partially methylated in kidney tissue expressing hCD55. Moreover, codon optimization of the CpG island of the hCD55 gene to reduce CG content could achieve stable expression of the hCD55 gene. In addition, antigen-antibody binding experiment showed that the amount of human IgM binding to GTKO/hCD55 gene-edited pig fibroblasts was significantly lower than that of WT pigs (P＜0.01). Complement-dependent cytotoxicity experiment showed that the survival rate of fibroblasts in GTKO/hCD55 pigs was significantly higher than that in WT pigs (P＜0.01). Conclusion This study demonstrates the successful generation of GTKO/hCD55 gene-edited xenotransplant donor pigs. Methylation-induced gene silencing of the hCD55 gene can be effectively avoided by reducing the CG content of the CpG island through codon optimization. This study provides a reference for the development of xenotransplant donor pigs and guides subsequent research on xenotransplantation. Table and Figures | Reference | Related Articles | Metrics

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Advances in Mouse Models of Amyotrophic Lateral Sclerosis LUO Lianlian, YUAN Yanchun, WANG Junling, SHI Guangsen Laboratory Animal and Comparative Medicine    2025, 45 (3): 290-299.   DOI: 10.12300/j.issn.1674-5817.2024.161 Abstract （524）   HTML （26）    PDF （816KB）（876）       Knowledge map    Save Amyotrophic lateral sclerosis (ALS) is an irreversible, fatal neurodegenerative disorder whose incidence is positively correlated with the aging population. ALS is characterized by the progressive loss of motor neurons, leading to muscle weakness, atrophy, and ultimately respiratory failure. The pathogenesis of ALS involves multiple factors, including genetic and environmental influences, with genetic factors playing a particularly significant role. To date, several causative genes have been identified in ALS, such as the Cu/Zn superoxide dismutase 1 (Cu/Zn SOD1, also known as SOD1) gene, transactive response DNA-binding protein 43 (TDP-43) gene, fused in sarcoma (FUS) gene, and chromosome open reading frame 72 (C9orf72). Mutations in these genes have been found not only in familial ALS but also in sporadic ALS. Based on the identified ALS risk genes, various ALS animal models have been established through multiple approaches, including transgenic models, gene knockout/knock-in models, and adeno-associated virus-mediated overexpression models. These models simulate some typical pathological features of human ALS, such as motor neuron loss, ubiquitinated inclusions, and neuromuscular junction degeneration. However, these models still have limitations: (1) single-gene mutation models are insufficient to fully replicate the complex multi-factorial pathogenesis of sporadic ALS; (2) significant differences in microenvironmental regulation mechanisms and the rate of neurodegeneration between model organisms and humans may affect the accurate reproduction of disease phenotypes and the reliable evaluation of drug efficacy. To better understand the pathogenesis of ALS and promote the development of effective therapies, constructing and optimizing ALS animal models is crucial. This review aims to summarize commonly used ALS gene mutation mouse models, analyze their phenotypes and pathological characteristics, including transgenic mouse models, gene knockout/knock-in mouse models, and adeno-associated virus-mediated overexpression mouse models, and further discuss their specific applications in ALS pathogenesis research and drug development by comparing the advantages and limitations of each model. Table and Figures | Reference | Related Articles | Metrics

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Exploration and Practice of Safe Access System Construction for Barrier Environment Facilities of Laboratory Animals: A Case Study on Xianlin Campus of Nanjing University HOU Dongxia, TIE Zuoxiu, LU Yong, NAN Panpan, BAO Jie Laboratory Animal and Comparative Medicine    2025, 45 (1): 96-100.   DOI: 10.12300/j.issn.1674-5817.2024.106 Abstract （511）   HTML （25）    PDF （905KB）（542）       Knowledge map    Save Laboratory animals are essential in scientific research and experimental teaching in colleges and universities. Disciplines such as life sciences, medicine, pharmacy, chemistry, and biomedical engineering heavily rely on animal experiments. The standardized barrier environmental facility for laboratory animals provides a fundamental platform for stable, scientific, and reliable animal experiment results. Rigorous access management for such facilities is a vital safeguard for maintaining standardized operations of facilities, controlling the quality and stability of laboratory animals, mitigating pathogen contamination risks among animals and laboratory staff, and preventing biosecurity incidents such as zoonotic disease outbreaks. Taking the small-scale barrier facilities for laboratory rats and mice at Nanjing University's Xianlin Campus, operational since 2019, as an example, this study focuses on the safety access management system of these facilities. Based on five years of operational data and accumulated experience in studying and optimizing the access management system, this study, from the perspectives of management system development and the formulation and implementation of standard operating procedures, reviews five aspects of access management: personnel access, animals access, material access, equipment access, and air circulation control. Furthermore, these aspects are systematically analyzed and summarized to serve as a reference for the construction and management of the laboratory animal facilities in universities, while also contributing to scientific research, public health security, and the well-being of experimental personnel. Table and Figures | Reference | Related Articles | Metrics

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Discussion on AI-Based Digital Upgrade and Application Practice of Laboratory Animal Centers WANG Tingjun, LUO Hao, CHEN Qi Laboratory Animal and Comparative Medicine    2025, 45 (4): 473-482.   DOI: 10.12300/j.issn.1674-5817.2024.181 Abstract （511）   HTML （35）    PDF （1285KB）（1603）       Knowledge map    Save Objective In traditional laboratory animal centers, there are issues such as low efficiency in cage scheduling, insufficient supervision of personnel behavior, and difficulty in upgrading aging equipment. This study aims to upgrade the information system of existing laboratory animal centers by applying multimodal large language model technology. This upgrade intends to achieve real-time perception of the status of animal cages, intelligent supervision of experimental personnel behavior, and automated processing of business workflows, thereby improving management efficiency and precision. Methods An AI-based approach for upgrading laboratory animal center informatization was proposed by the First Affiliated Hospital of Zhejiang University School of Medicine,compatible with different breeding equipments. The system architecture, from the bottom up, consisted of three layers: hardware layer, core algorithm layer, and application layer. The hardware layer was equipped with cameras and high-speed network transmission devices for collecting information on cages and personnel. The core algorithm layer utilized multi-stage image preprocessing technology and multimodal large language model recognition technology to extract and identify image information. The application layer integrated the recognition results with the existing information of the animal center to generate real-time cage occupancy heatmaps, which visually and clearly showed the density distribution of cage usage in the laboratory animal center. Results The AI-based management system achieved a cage recognition accuracy of 98.5% and a correct wearing identification rate of laboratory coats of 98.8%. The average image processing time was 3.7 seconds per image, the effective utilization rate of cages increased by 23%, and the turnover efficiency improved by 35%. In addition, the management system could track and warn against non-compliant behaviors in real time. After intelligent recognition, the system detected more violations, with the violation detection rate increasing by 90.6%. After continuous use for three months, the weekly average number of violations decreased by 54.0% compared to the baseline period. Conclusion This study applies multimodal large language model to the field of laboratory animal management, achieving real-time monitoring and automated management of cage identification, thereby improving management efficiency and precision. The system integrates multi-source data such as visual recognition and behavior analysis, establishing a comprehensive intelligent supervision system for experimental personnel. It provides research institutions with efficient, accurate, and cost-effective management tools, promoting the intelligent development of laboratory animal management. Table and Figures | Reference | Related Articles | Metrics

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Establishment of an Intestinal Fibrosis Model Associated with Inflammatory Bowel Disease in VDR -/- Mice Induced by Helicobacter hepaticus Infection and Mechanism Exploration WU Zhihao, CAO Shuyang, ZHOU Zhengyu Laboratory Animal and Comparative Medicine    2025, 45 (1): 37-46.   DOI: 10.12300/j.issn.1674-5817.2024.090 Abstract （500）   HTML （26）    PDF （3011KB）（1485）       Knowledge map    Save Objective To employ Helicobacter hepaticus (H.hepaticus, H.h) to induce intestinal fibrosis in vitamin D receptor deletion (VDR-/-) mice, thereby establishing a model of inflammatory bowel disease to investigate its pathological characteristics and underlying mechanisms. Methods Five male WT and five male VDR-/- mice were orally administered a suspension containing 2×108 CFU of H.hepaticus (referred to as the WT+H.h group and the VDR-/-+H.h group, respectively), with treatments occurring every other day for three administrations. Concurrently, two uninfected control groups were established, consisting of five WT and five VDR-/- mice, which were administered an equivalent volume of PBS. Seven days after the final administration, the infection status of the mice was assessed, and their body weight was recorded weekly. At the 16th week post-infection, the mice were dissected, and the length of the colon tissue was measured, with fecal moisture content analyzed. The colon tissue was partitioned into four parts: one for paraffin embedding for HE, alcian blue-periodic acid Schiff (AB-PAS), Masson's trichrome staining, and immunohistochemical analysis; one for DNA extraction to evaluate the colonization levels of H.hepaticus through real-time fluorescent quantitative polymerase chain reaction (RFQ-PCR), thereby assessing the impact of the infection; one for RNA extraction to analyze cytokine expression via reverse transcription-PCR (RT-PCR); and one for protein extraction to measure the expression levels of alpha smooth muscle actin (α-SMA) and interleukin (IL)-33 using Western blotting. Results All mice in the infected groups successfully were infected with H. hepaticus after three oral gavages. Compared to VDR-/- control group, VDR-/- mice exhibited significant weight loss (P<0.05), intestinal hemorrhage, and higher fecal water content after 16 weeks of H. hepaticus infection than the uninfected control group and the WT+H.h group (P<0.05). Compared to the WT+H.h group, HE staining of the VDR-/-+H.h group showed inflammatory cell infiltration, AB-PAS staining revealed irregular atrophy of intestinal glands and reduced acini, and Masson staining showed increased collagen area. RT-PCR demonstrated that the transcription levels of inflammation and fibrosis-related genes, including IL-6, IL-33, tumor necrosis factor-α (TNF-α), and α-SMA (P < 0.000 1), were significantly upregulated in the colon tissues of VDR-/-+H.h group. Additionally, immunohistochemical analysis and Western blotting showed that the protein expression levels of IL-33 and α-SMA were markedly increased (P<0.001) in the VDR-/-+H.h group. Conclusion VDR-/- mice infected with H.hepaticus exhibit more severe inflammatory responses, including mucosal inflammatory infiltration, impaired mucosal tissue function, and collagen deposition, indicating successful construction of the inflammatory bowel disease model. Further research suggests that VDR deficiency may exacerbate the intestinal fibrosis process associated with inflammatory bowel disease by affecting IL-33 expression. Table and Figures | Reference | Related Articles | Metrics

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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 Abstract （496）   HTML （18）    PDF （915KB）（900）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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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 Abstract （488）   HTML （17）    PDF （802KB）（725）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Design of a Capture Stress-Free Marmoset Monkey Chair Device for Experiments and Its Preliminary Application XU Shengye, HUANG Junfeng, CHEN Yihang, CHANG Liangtang Laboratory Animal and Comparative Medicine    2025, 45 (1): 67-72.   DOI: 10.12300/j.issn.1674-5817.2024.097 Abstract （486）   HTML （20）    PDF （1203KB）（421）       Knowledge map    Save Objective To avoid stress responses in experimental monkeys caused by direct capture, and to improve the adaptability and experimental efficiency of marmosets in behavioral, two-photon imaging, and electrophysiological experiments, a device for immobilizing marmosets without the need for capture is developed. Methods A set of compatible transport cage and monkey chair was produced through 3D graphic design and printing. First, the transport cage was aligned with the feeding outlet of the experimental housing cage, and the marmoset was gently guided into the transport cage. Then, the transport cage was connected to the monkey chair, and the marmoset was gently guided into the chair for immobilization. Subsequent experiments were carried out afterward. The effectiveness was evaluated by observing the efficiency of transport and immobilization, the marmoset cooperation level, and stress responses. Results After testing and improvements, the device successfully completed immobilization of marmosets without the need for capture, significantly improving the fluency and efficiency of the experiment. As the number of operations increased, the marmosets became more cooperative, and the operation speed was significantly enhanced. After using the device, the stress responses were noticeably reduced, with marmosets showing lower stress levels. In particular, compared to traditional capture methods, the use of this device significantly reduced marmoset anxiety and discomfort, increasing their cooperation levels during the experiment. Conclusion The monkey chair device designed allows for restraint of marmosets without the need for capture, ensuring smooth progress of subsequent experiments while also safeguarding animal welfare. This device is easy to operate, highly practical, cost-effective, and has great potential for widespread application. Table and Figures | Reference | Related Articles | Metrics

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Identification and Analysis of MHCⅡ Genes in Wuzhishan Pigs LIU Yuanyuan, XIN Wenshui, CHAO Zhe, CAO Zongxi, CAI Yifei, LI Qiang, LI Lingwei, LIU Guangliang Laboratory Animal and Comparative Medicine    2025, 45 (3): 340-348.   DOI: 10.12300/j.issn.1674-5817.2024.135 Abstract （470）   HTML （12）    PDF （1751KB）（912）       Knowledge map    Save Objective To obtain the gene sequences of major histocompatibility complex (MHC ) Ⅱgenes of Wuzhishan pigs, analyze their genetic information, and explore the biological functions of their MHC system. Methods Spleen samples were collected from 3 adult male Wuzhishan pigs. Primers were designed according to MHCⅡ gene sequences, and the coding sequences of Wuzhishan pig MHCⅡ genes were amplified by RT-PCR. Sanger sequencing was performed to determine the full-length sequences. Bioinformatics tools were employed to analyze the physicochemical properties, phylogenetic relationships, conserved motifs, structural domains, chromosomal localization, and syntenic relationships of these genes. Results Eight MHCⅡ genes were identified in Wuzhishan pigs, designated as SLA-DRA, SLA-DQA, SLA-DQB, SLA-DRB, SLA-DOB, SLA-DMB, SLA-DMA and SLA-DOA. The full-length sequences of these genes were determined by Sanger sequencing and subsequently deposited in GenBank under accession numbers PQ182796, PQ182797, PQ182798, PQ182799, PQ182800, PQ182801, PQ182802, and PQ164779. Phylogenetic analysis showed that the six MHCⅡ genes of Wuzhishan pigs clustered separately from their counterparts in Duroc, Meishan, Large White, and Bama pigs, indicating distinct evolutionary trajectories. Bioinformatics analysis demonstrated that most MHC Ⅱ proteins were hydrophobic, with molecular weights ranging from 27 700 to 30 000 Da. Genes within the same subregion shared conserved motifs. Specifically, four MHCⅡ proteins encoded by SLA-DQB, SLA-DRB, SLA-DOB, and SLA-DMB contained the MHCⅡβ conserved domain, while those encoded by the genes SLA-DRA, SLA-DQA, SLA-DMA, and SLA-DOA contained the MHCⅡα conserved domain. The eight MHCⅡ genes were scattered along the long arm of chromosome 7 in the Wuzhishan pigs, exhibiting syntenic relationships with three human genes and five Duroc pig genes. Conclusion The MHCⅡ genes of Wuzhishan pigs may possess a unique evolutionary origin. Table and Figures | Reference | Related Articles | Metrics

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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 Abstract （469）   HTML （31）    PDF （1993KB）（1850）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Advances in the Application of Zebrafish in the Research of Inflammatory Bowel Disease Mechanisms and Drug Development ZHAO Xin, WANG Chenxi, SHI Wenqing, LOU Yuefen Laboratory Animal and Comparative Medicine    2025, 45 (4): 422-431.   DOI: 10.12300/j.issn.1674-5817.2024.170 Abstract （464）   HTML （23）    PDF （831KB）（1656）       Knowledge map    Save Inflammatory bowel disease (IBD) is a chronic, relapsing intestinal disorder driven by multiple factors including genetics, immunity, and environment, and is clinically classified into ulcerative colitis and Crohn's disease. Currently, mice and zebrafish are the primary experimental animals used in IBD research, among which zebrafish have emerged as an ideal model due to their unique advantages. Compared with rodent models, zebrafish serve as an effective and convenient model, offering advantages such as a short life cycle, robust reproductive capacity, small size, and transparent embryos. These characteristics make zebrafish highly suitable for dynamic tracking of continuous pathological progression and high-throughput drug screening. Zebrafish share over 70% genetic homology with humans, and their intestinal cellular composition and ontogeny closely resemble those of humans. Moreover, the structure and characteristics of their gut microbiota are similar to the human intestinal microbiome, providing a solid foundation for studying the relationship between gut microbiota and IBD. With advances in biotechnology, zebrafish IBD models generated by chemical induction or genetic engineering can accurately simulate the core pathological features of human IBD, such as intestinal wall thickening, inflammatory cell infiltration, and elevated expression of pro-inflammatory factors. These models have played a significant role in revealing the pathogenesis of IBD as well as the development of targeted therapeutic drugs. This article first outlines the intestinal characteristics of zebrafish and features of zebrafish IBD models, then provides an in-depth analysis of their application in IBD pathogenesis research from multiple aspects, including genetics, immunity, environment and diet, and infection. It also reviews research progress on the application of zebrafish in the development of anti-inflammatory drugs, probiotics, and traditional Chinese medicine therapies, aiming to provide researchers with references for the rational use of zebrafish models at all stages of preclinical research, to advance fundamental IBD research and accelerate breakthroughs in this field. Table and Figures | Reference | Related Articles | Metrics

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Cartilage Protection and Anti-Inflammatory Effects of Fraxetin on Monosodium Iodoacetate-Induced Rat Model of Osteoarthritis LIU Zhiwei, YANG Ran, LIAN Hao, ZHANG Yu, JIN Lilun Laboratory Animal and Comparative Medicine    2025, 45 (3): 259-268.   DOI: 10.12300/j.issn.1674-5817.2024.165 Abstract （459）   HTML （42）    PDF （2277KB）（791）       Knowledge map    Save Objective To establish a rat model of osteoarthritis and study the anti-inflammatory effects and mechanisms of fraxetin. Methods Eighteen 8-week-old male SPF-grade SD rats were randomly divided into three groups: Rats in the blank group received a right articular cavity injection of 50 μL of normal saline for 1 week; the model and intervention groups were injected with monosodium iodoacetate (MIA) into the right joint cavity to induce osteoarthritis, while the intervention group subsequently received fraxetin (5 mg·kg-1·d-1) for 1 week. Four weeks after drug intervention, abdominal aortic blood was collected. The animals were then euthanized, and knee joint cartilage were collected. The cartilage samples were stained with hematoxylin-eosin, safranin O-fast green, and toluidine blue for histopathological examination and scoring using the Mankin and OARSI scoring systems. The trabecular bone volume/total volume (Tb.BV/TV), trabecular bone surface density/total volume (Tb.BS/TV), and trabecular number (Tb.N) of each group were compared and analyzed using a micro-CT scanning system. The expression levels of various inflammatory factors [tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6)], and cartilage oligomeric matrix protein (COMP) were measured using enzyme-linked immunosorbent assay (ELISA). The expression levels of mitogen-activated protein kinase p38 (p38 MAPK), phosphorylation-p38 MAPK (p-p38 MAPK), c-Jun N-terminal kinase (JNK), and phosphorylation-JNK (p-JNK) were measured by western blotting. Results The staining of cartilage sections of rat knee joints showed that the articular surface defects in the model group were severe, while the cartilage destruction in the intervention group was relatively reduced. Micro-CT results showed that Tb.BV/TV, Tb.BS/TV and Tb.N in the intervention group were significantly higher than those in the model group (P < 0.05); the Mankin score in the model group was significantly higher than that in the blank group (P < 0.05), the Mankin score in the intervention group was significantly lower than that in the model group (P < 0.05); while the OARSI score in the intervention group was significantly lower than that in the model group (P < 0.05). The results of the enzyme-linked immunosorbent assay showed that the serum levels of TNF-α, IL-1β, IL-6, and COMP in the model group were significantly higher than those in the blank group (all P < 0.05), while those in the intervention group were significantly lower than in the model group (P < 0.05). Western blot results showed that the expression levels of p-p38 MAPK and p-JNK in the knee cartilage tissue were significantly lower in the intervention group than in the model group (both P < 0.05), and significantly higher in the model group than in the blank group (both P < 0.05). Conclusion Fraxetin may play a therapeutic role in a monosodium iodoacetate-induced rat model of osteoarthritis through the p38 MAPK pathway. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Characteristics Analysis of Gut Microbiota and Its Sex Differences in Laboratory Animals SHEN Huangyi, HUANG Yufei, YANG Yunpeng Laboratory Animal and Comparative Medicine    2025, 45 (3): 349-359.   DOI: 10.12300/j.issn.1674-5817.2024.124 Abstract （457）   HTML （13）    PDF （824KB）（2462）       Knowledge map    Save Laboratory animals serve as the cornerstone in life science research, acting as surrogate models for human physiology, pathology, and disease treatment. They play an irreplaceable role in basic research, drug development, and translational medicine. Gut microbiota, a complex microbial community comprising bacteria, fungi, viruses, and unicellular organisms, colonizes the host's intestinal tract and is closely associated with the maintenance of normal physiological metabolism and overall health. Studies have shown that dysbiosis of the gut microbiota can lead to various diseases, including obesity, diabetes, hypertension, inflammatory bowel disease, and Alzheimer's disease. Therefore, conducting characteristic analyses of the gut microbial composition of laboratory animals can not only enhance the reliability of experimental outcomes but also facilitate their translational application. Sex differences represent a critical variable in biological research, significantly influencing the physiological functions, metabolic traits, and gut microbial composition of laboratory animals. However, a pronounced sex bias has been widely observed in many biological studies, thereby limiting the generalizability of results. This study focused on ten commonly used laboratory animals in life sciences, including mice, rats, guinea pigs, hamsters, rabbits, dogs, cats, non-human primates, miniature pigs, and chickens. Their gut microbial composition was summarized and related sex-specific differences of certain species were analyzed. Furthermore, by comparing the gut microbiota of laboratory animals with that of humans, this study offers novel perspectives for comparative medical research. In summary, this study not only deepens researchers' understanding of gut microbiota characteristics and sex-dependent variations across laboratory animal species but also provides practical guidance for selecting appropriate laboratory animals, constructing sex-specific disease models, and interpreting experimental results in scientific studies. Reference | Related Articles | Metrics

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Evaluation of Proficiency Validation Results for Air Change Rate Testing in Laboratory Animal Facilities LIU Wei, ZHANG Xinyan, HOU Fengtian, XU Zhongkan, MA Liying Laboratory Animal and Comparative Medicine    2025, 45 (1): 87-95.   DOI: 10.12300/j.issn.1674-5817.2024.101 Abstract （452）   HTML （19）    PDF （1178KB）（681）       Knowledge map    Save Objective By organizing and implementing a laboratory proficiency validation plan for air change rate testing, this study aims to explore proficiency testing approaches in laboratory animal facilities, assess the current status of relevant laboratories regarding standard application and test capabilities, standardize air change rate testing methods, and ensure the accuracy and reliability of test results. Methods From September to November 2023, the National Institutes for Food and Drug Control (NIFDC) organized a laboratory proficiency validation plan for air change rate testing in laboratory animal facilities (Plan Number: NIFDC-PT-417). The proficiency testing was conducted on-site and consisted of two parts: a written test and practical operation. The written test was open-book. True/false questions focused on participants' understanding of specific clauses in relevant standards, while application-based questions assessed their ability to handle data processing in simulated testing scenarios. The practical operation was conducted according to the relevant criteria of the China National Accreditation Service for Conformity Assessment (CNAS). Two laboratory animal rooms were prepared as proficiency testing samples using a sample splitting approach. These rooms underwent uniformity and stability testing according to CNAS requirements and were approved. Participating laboratories were required to conduct three tests on each of the two laboratory animal rooms, complete the testing and calculation of air change rate within the specified timeframe, and submit their test result reports and original records. Results A total of 27 laboratories registered and participated in the proficiency testing. All participating laboratories submitted their results within the designated timeframe, and the outcomes of all tested laboratories were rated as satisfactory. Conclusion This proficiency validation program objectively and scientifically evaluates the air change rate testing capabilities of selected domestic laboratories, effectively promoting the overall improvement of testing capabilities in the industry. It provides technical support for regulatory authorities to standardize testing institutions and offers reliable references for the purchase of testing services. Through this activity, it was identified that some laboratories need to further enhance their calibration of instruments and the utilization of calibration results. Future efforts should focus on refining related standards to improve the accuracy and reliability of testing. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Human Ovarian Aging Using Non-Human Primates as Laboratory Animals XIAO Wenxian, LÜ Longbao Laboratory Animal and Comparative Medicine    2025, 45 (1): 47-54.   DOI: 10.12300/j.issn.1674-5817.2024.114 Abstract （442）   HTML （26）    PDF （793KB）（1357）       Knowledge map    Save The ovary has two main functions: folliculogenesis and hormone secretion, both of which are closely related to female fertility. Ovarian aging is characterized by morphological changes, a reduction in follicle numbers, and fluctuations in hormone levels. It not only leads to a decline in female fertility, but is also considered to be a key driver of multi-organ aging. In addition, the disruption of sex hormone secretion associated with ovarian aging can lead to the occurrence of related diseases and symptoms, such as cardiovascular diseases, sleep disorders, and hot flashes. Due to the influence of social pressures and personal career planning, many modern women are increasingly postponing childbearing. However, ovarian aging does not slow down with advancing age. As a result, many women face issues such as infertility when they are ready to have children, having missed their optimal childbearing age. This leads to growing interest in research on delaying ovarian aging. Non-human primates share the closest evolutionary relationship with humans, with a genomic sequence identity of 93%, which grants them unparalleled advantages over other model animals in studies on physiological metabolism, reproductive endocrinology, and developmental aging. Findings obtained in non-human primates are also more reliably translatable to human medical research. This study begins by discussing the current state of ovarian aging research and treatment strategies, highlighting the advantages of non-human primates as laboratory animals for ovarian aging research. It then reviews research progress in areas such as reproductive endocrine hormone levels, ovarian morphology and function, and other physiological changes associated with ovarian aging. Furthermore, it summarizes existing challenges and future research directions, aiming to provide valuable insights for researchers. Reference | Related Articles | Metrics

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Prospects for 3D Bioprinting Research and Transdisciplinary Application to Preclinical Animal Models HU Min, DONG Lexuan, GAO Yi, XI Ziqi, SHEN Zihao, TANG Ruiyang, LUAN Xin, TANG Min, ZHANG Weidong Laboratory Animal and Comparative Medicine    2025, 45 (3): 318-330.   DOI: 10.12300/j.issn.1674-5817.2024.193 Abstract （427）   HTML （11）    PDF （1108KB）（582）       Knowledge map    Save Animal experiments are widely used in biomedical research for safety assessment, toxicological analysis, efficacy evaluation, and mechanism exploration. In recent years, the ethical review system has become more stringent, and awareness of animal welfare has continuously increased. To promote more efficient and cost-effective drug research and development, the United States passed the Food and Drug Administration (FDA) Modernization Act 2.0 in September 2022, which removed the federal mandate requiring animal testing in preclinical drug research. In April 2025, the FDA further proposed to adopt a series of "new alternative methods" in the research and development of drugs such as monoclonal antibodies, which included artificial intelligence computing models, organoid toxicity tests, and 3D micro-physiological systems, thereby gradually phasing out traditional animal experiment models. Among these cutting-edge technologies, 3D bioprinting models are a significant alternative and complement to animal models, owing to their high biomimetic properties, reproducibility, and scalability. This review provides a comprehensive overview of advancements and applications of 3D bioprinting technology in the fields of biomedical and pharmaceutical research. It starts by detailing the essential elements of 3D bioprinting, including the selection and functional design of biomaterials, along with an explanation of the principles and characteristics of various printing strategies, highlighting the advantages in constructing complex multicellular spatial structures, regulating microenvironments, and guiding cell fate. It then discusses the typical applications of 3D bioprinting in drug research and development,including high-throughput screening of drug efficacy by constructing disease models such as tumors, infectious diseases, and rare diseases, as well as conducting drug toxicology research by building organ-specific models such as those of liver and heart. Additionally,the review examines the role of 3D bioprinting in tissue engineering, discussing its contributions to the construction of functional tissues such as bone, cartilage, skin, and blood vessels, as well as the latest progress in regeneration and replacement. Furthermore, this review analyzes the complementary advantages of 3D bioprinting models and animal models in the research of disease progression, drug mechanisms, precision medicine, drug development, and tissue regeneration, and discusses the potential and challenges of their integration in improving model accuracy and physiological relevance. In conclusion, as a cutting-edge in vitro modeling and manufacturing technology, 3D bioprinting is gradually establishing a comprehensive application system covering disease modeling, drug screening, toxicity prediction, and tissue regeneration. Table and Figures | Reference | Related Articles | Metrics

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An Overview of Strategies for Constructing Animal Models of Traditional Chinese Medicine Syndromes WANG Xiaoming, MENG Chenchen, FAN Lu, LI Yanyang, ZHANG Junping, LÜ Shichao Laboratory Animal and Comparative Medicine    2025, 45 (5): 596-610.   DOI: 10.12300/j.issn.1674-5817.2025.038 Abstract （424）   HTML （6）    PDF （1042KB）（350）       Knowledge map    Save This study aims to explore different construction methods for animal models of traditional Chinese medicine (TCM) syndromes and their advantages and disadvantages, to propose optimization strategies for existing problems in current construction methods, and to provide reference for constructing animal models of TCM syndromes that both preserve the essence of TCM syndromes and conform to modern scientific research standards. Using "traditional Chinese medicine", "syndrome", and "animal model" as key words, articles related to animal models of TCM syndromes from CNKI, Wanfang, and VIP databases are searched and reviewed. Then the theoretical basis, technical characteristics, and existing problems of the main construction methods of current TCM syndrome animal models are systematically sorted out, and corresponding optimization measures are proposed for the existing problems. The construction methods of TCM syndrome animal models include TCM etiology and pathogenesis construction, modern medical etiology and pathology construction, and integration of TCM and Western medicine for diseases and syndromes. The TCM etiology and pathogenesis construction method is guided by a holistic perspective, constructing syndrome models by simulating external factors such as six pathogenic factors and emotional disorders. Although it conforms to TCM theoretical connotation and has simple operation and strong controllability, this method has problems such as low modeling success rate and poor etiology-syndrome fit. The modern medical etiology and pathology construction method is based on microscopic pathological mechanisms, adopting highly controllable technical means such as drug intervention and surgical modeling. Although it has the characteristics of clear objective indicators and excellent reproducibility, this method has defects such as deviation from the essence of TCM "syndrome" and insufficient safety. The integrated TCM-Western medicine disease-syndrome method shows significant complementarity in syndrome essence restoration degree and technical feasibility, achieves systematic integration of TCM basic theories and clinical syndrome differentiation thinking in methodology, and integrates the objective evaluation system of modern medicine, improving the clinical consistency between Western medicine pathological mechanisms and TCM syndrome evolution patterns. However, this method still faces common challenges such as ambiguous syndrome identification standards and distortion of disease progression simulation. The construction of TCM syndrome animal models faces challenges such as poor theoretical adaptability and poor technical standardization, but has irreplaceable value in verifying the efficacy of prescriptions and promoting the internationalization of TCM. In the future, the construction of TCM syndrome animal models should be optimized through measures such as optimizing animal selection, improving the theoretical basis of preparation methods, standardizing the setting of modeling factors, and clarifying the standard for modeling success. Table and Figures | Reference | Related Articles | Metrics

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Effects of Different Durations of Light Exposure on Body Weight and Learning and Memory Abilities of NIH Mice ZHANG Nan, LI Huaiyin, LIAN Xiaodi, WEI Juanpeng, GAO Ming Laboratory Animal and Comparative Medicine    2025, 45 (1): 73-78.   DOI: 10.12300/j.issn.1674-5817.2024.062 Abstract （422）   HTML （18）    PDF （793KB）（943）       Knowledge map    Save Objective This study aims to investigate the effects of varying durations of light exposure on body weight and learning and memory abilities of pubertal NIH mice. Methods Forty pubertal NIH mice, evenly split by gender and with similar initial weights, were subjected to a 12 h light-dark cycle for one week. They were then randomly assigned to groups with daily light exposure durations of 0, 6, 12, 18, and 24 hours, with 8 mice in each group. The experimental period lasted for 7 weeks, with the first 5 weeks as the feeding phase under different light exposure conditions, and the last 2 weeks as the behavioral testing phase. Their body weight was monitored, and learning and memory abilities were assessed using the T-maze, object location test, and eight-arm maze tests. Results During the light exposure period, there were no significant differences in body weight among groups (P>0.05). However, the weight gain of mice in the 24 h group was significantly higher than that of the 0 h group and the 6 h group during the second and third weeks of light exposure (P<0.05). After five weeks of light exposure, in the T-maze test, the latency time of the 0 h light exposure group was significantly longer than that of the 12 h group (P<0.01), and the latency time of the 24 h light exposure group was significantly longer than that of the 12 h group (P<0.05). In the object location test, the mice in 12 h group exhibited a higher discrimination index and spent more time observing the new location compared to the other groups, with significant differences in comparison to the 18 h group (P<0.01) and the 24 h group (P<0.05). In the eight-arm maze test, the time to find food, the reference memory error rate, and the working memory error rate in the 12 h group were all lower than those in the 0 h group, with significant differences (P<0.05). Moreover, the working memory error rate in the 24 h group was higher than that in the 12 h group, with significant differences (P<0.05). Conclusion Continuous 24 h light exposure affects body weight gain, while light exposure durations exceeding 18 h or below 6 h per day weaken the learning and memory abilities of NIH mice. Table and Figures | Reference | Related Articles | Metrics

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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 Abstract （418）   HTML （21）    PDF （835KB）（771）       Knowledge map    Save 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. Reference | Related Articles | Metrics

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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 Abstract （409）   HTML （10）    PDF （791KB）（756）       Knowledge map    Save 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. Reference | Related Articles | Metrics

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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 Abstract （404）   HTML （21）    PDF （984KB）（628）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Progress and Evaluation of Animal Model of Heart Qi-Yin Deficiency Syndrome LIU Yayi, JIA Yunfeng, ZUO Yiming, ZHANG Junping, LÜ Shichao Laboratory Animal and Comparative Medicine    2025, 45 (4): 411-421.   DOI: 10.12300/j.issn.1674-5817.2024.176 Abstract （402）   HTML （17）    PDF （921KB）（714）       Knowledge map    Save Animal models combining disease and syndrome are important research tools to explore the nature of traditional Chinese medicine (TCM) syndromes. At present, the construction and evaluation methods of animal models have preliminarily established the foundation for standardized development. Qi-yin deficiency syndrome is a common type of TCM syndrome in cardiovascular diseases. It is an important pathogenic factor causing the onset, pathological damage, and chronic nature of cardiovascular diseases, as well as triggering other illnesses. The establishment of an animal model of cardiovascular disease with the characteristics of Qi-yin deficiency, along with an objective and standardized evaluation system, has become an important part of modern cardiovascular disease research. In recent years, research on the construction and evaluation of animal models of heart Qi-yin deficiency syndrome has increased, but the construction methods and evaluation criteria vary. Compared with other animal models, the literature is limited, lacking statistics and overall analysis. Therefore, based on the scientific connotation of heart Qi-yin deficiency syndrome, this article systematically reviews the evaluation system of its animal model, covering multidimensional methods such as macroscopic characterization assessment, physicochemical indicators and objective evaluation, and syndrome differentiation based on prescriptions. The specific model construction strategies are described, including single-factor induction methods (sleep deprivation, chronic intermittent hypoxia, arterial occlusion, high-salt feeding) and the compound-factor induction methods (sleep deprivation combined with drug method, chronic intermittent hypoxia combined with drug method, exhaustive swimming combined with drug method). Meanwhile, application examples of each model in the research are listed, the existing problems in the current model construction and evaluation are analyzed, and optimization directions are proposed, such as promoting the compound factor induction strategy and improving the objectivity of the evaluation criteria. This article aims to provide theoretical references for constructing an animal model of heart Qi-yin deficiency syndrome that conforms to TCM characteristics, and thereby laying a scientific foundation for the prevention and treatment of cardiovascular diseases with TCM. Table and Figures | Reference | Related Articles | Metrics

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Recommendations for Standardized Reporting of Systematic Reviews and Meta-Analysis of Animal Experiments ZHENG Qingyong, YANG Donghua, MA Zhichao, ZHOU Ziyu, LU Yang, WANG Jingyu, XING Lina, KANG Yingying, DU Li, ZHAO Chunxiang, DI Baoshan, TIAN Jinhui Laboratory Animal and Comparative Medicine    2025, 45 (4): 496-507.   DOI: 10.12300/j.issn.1674-5817.2025.017 Abstract （395）   HTML （21）    PDF （1130KB）（1121）       Knowledge map    Save Animal experiments are an essential component of life sciences and medical research. However, the external validity and reliability of individual animal studies are frequently challenged by inherent limitations such as small sample sizes, high design heterogeneity, and poor reproducibility, which impede the effective translation of research findings into clinical practice. Systematic reviews and meta-analysis represent a key methodology for integrating existing evidence and enhancing the robustness of conclusions. Currently, however, the application of systematic reviews and meta-analysis in the field of animal experiments lacks standardized guidelines for their conduct and reporting, resulting in inconsistent quality and, to some extent, diminishing their evidence value. To address this issue, this paper aims to systematically delineate the reporting process for systematic reviews and meta-analysis of animal experiments and to propose a set of standardized recommendations that are both scientific and practical. The article's scope encompasses the entire process, from the preliminary preparatory phase [including formulating the population， intervention， comparison and outcome （PICO） question, assessing feasibility, and protocol pre-registration] to the key writing points for each section of the main report. In the core methods section, the paper elaborates on how to implement literature searches, establish eligibility criteria, perform data extraction, and assess the risk of bias, based on the Preferred Reporting Items for Systematic Reviews and Meta-analysis （PRISMA) statement, in conjunction with relevant guidelines and tools such as Animal Research： Reporting of in Vivo Experiments （ARRIVE) and a risk of bias assessment tool developed by the Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE). For the presentation of results, strategies are proposed for clear and transparent display using flow diagrams and tables of characteristics. The discussion section places particular emphasis on how to scientifically interpret pooled effects, thoroughly analyze sources of heterogeneity, evaluate the impact of publication bias, and cautiously discuss the validity and limitations of extrapolating findings from animal studies to clinical settings. Furthermore, this paper recommends adopting the Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology to comprehensively grade the quality of evidence. Through a modular analysis of the entire reporting process, this paper aims to provide researchers in the field with a clear and practical guide, thereby promoting the standardized development of systematic reviews and meta-analysis of animal experiments and enhancing their application value in scientific decision-making and translational medicine. Table and Figures | Reference | Related Articles | Metrics

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Comparison of Histopathological and Molecular Pathological Phenotypes in Mouse Models of Intrauterine Adhesions Induced by Two Concentrations of Ethanol Perfusion JIANG Juan, SONG Ning, LIAN Wenbo, SHAO Congcong, GU Wenwen, SHI Yan Laboratory Animal and Comparative Medicine    2025, 45 (4): 393-402.   DOI: 10.12300/j.issn.1674-5817.2024.183 Abstract （388）   HTML （23）    PDF （2290KB）（771）       Knowledge map    Save Objective To construct intrauterine adhesion (IUA) mouse models induced by two different concentrations of ethanol injury, compare the phenotypes, and optimize a more stable IUA modeling method. Methods Twenty 8-week-old female C57BL/6N mice were randomly divided into two groups: the 95% ethanol injury group and the 50% ethanol injury group. Using a self-control method, the left uterine horn was infused with ethanol to establish the IUA model, while the right uterine horn was infused with saline as the sham operation. Five mice from each group were euthanized on day 7 and 15 after modeling, and uterine tissues were collected. Hematoxylin-eosin (HE) staining was used to observe the endometrial pathology, and Masson staining was used to assess the degree of endometrial fibrosis. Quantitative real-time PCR was employed to detect the expression levels of fibrosis markers and pro-inflammatory factors in the uterine tissues. Results Compared to the sham operation, these two ethanol injury led to a significant reduction in elasticity of the uterus, an increase in inflammatory infiltration, and a marked increase in the degree of fibrosis on day 7 after modeling (P<0.05). The 95% ethanol injury group showed a significant decrease in endometrial thickness (P<0.05), whereas no significant change was observed in the 50% ethanol injury group when compared to the sham operation (P>0.05). The expression levels of fibrotic marker molecules collagen type Ⅳ alpha 1 chain (Col4A1), α-smooth muscle actin (α-SMA), transforming growth factor-β (TGF-β), and pro-inflammatory factors tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were significantly elevated in the 50% ethanol injury group when compared to the sham operation (P<0.05), although there was an increasing trend of the same markers in the 95% ethanol injury group, the differences were not statistically significant (P>0.05). On day 15 after modeling, the histopathological changes in both ethanol injury groups were not significant when compared to the sham operation, the expression levels of Col4A1, TGF-β, TNF-α and IL-1β remained significantly higher in the 50% ethanol injury group (P<0.05), while only IL-1β was significantly elevated in the 95% ethanol injury group (P<0.05). Conclusion Uterine infusion with 95% ethanol results in more marked histopathological changes in the IUA mouse model compared to the 50% ethanol injury group. The 95% ethanol injury model is suitable for histopathological studies. However, the 50% ethanol injury group shows higher expression levels of fibrosis markers and pro-inflammatory factors compared to the 95% ethanol injury group, suggesting that the 50% ethanol injury model is more suitable for molecular pathological study. Table and Figures | Reference | Related Articles | Metrics

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Analysis of Common Causes of Out-of-Specification Results in the Test for Depressor Substances TONG Xiyang, QUE Changtian, ZHANG Feng, ZHAO Lu, WANG Hongping Laboratory Animal and Comparative Medicine    2025, 45 (3): 331-339.   DOI: 10.12300/j.issn.1674-5817.2024.143 Abstract （377）   HTML （13）    PDF （1049KB）（2300）       Knowledge map    Save According to General Chapter 1145 of Division IV in the Chinese Pharmacopoeia (2020 Edition), the test for depressor substances is a common method for drug testing. It determines whether the level of depressor substances in a test sample complies with the specified standards by comparing the extent of blood pressure reduction in anesthetized cats induced by the histamine reference substance and the test sample. If an out-of-specification (OOS) result occurs in the test for depressor substances, it may be caused by inherent quality issues of the drug or errors in the testing process. Therefore, analyzing the causes of OOS is particularly important for confirming the test results and evaluating drug quality. Cats are used as experimental animals in the test for depressor substances. Compared with conventional laboratory animals, they are less stable, surgery procedures are more challenging, and the testing process is more complex. These factors make it more difficult to investigate the causes of OOS in this test. Based on a review of the literature and practical work experience, this article analyzes the causes of OOS in the test for depressor substances from the following five aspects: (1) an analysis of the impact of drug standards on OOS from three aspects: standard determination, standard content, and standard drafting; (2) personnel qualifications, including pre-employment training, compliance with standard operating procedures during experimental operations, and the ability to operate instruments; (3) factors related to cats, used as experimental animals in the test for depressor substances, including physiological characteristics, genetic background, and abnormal conditions during the experiment; (4) reference substances, reagents, test samples, and key instruments such as the multi-channel physiological signal instrument; (5) experimental operations including animal anesthesia, arterial and venous catheterization, drug administration, and data processing. This article aims to provide reference approaches for professionals engaged in the testing of pharmaceuticals and biological products when analyzing the causes of OOS in the test for depressor substances. Table and Figures | Reference | Related Articles | Metrics

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A Mouse Model and Mechanism Study of Premature Ovarian Insufficiency Induced by Different Concentrations of Cyclophosphamide GONG Leilei, WANG Xiaoxia, FENG Xuewei, LI Xinlei, ZHAO Han, ZHANG Xueyan, FENG Xin Laboratory Animal and Comparative Medicine    2025, 45 (4): 403-410.   DOI: 10.12300/j.issn.1674-5817.2024.194 Abstract （374）   HTML （17）    PDF （1198KB）（555）       Knowledge map    Save Objective To observe and compare the effects of different concentrations of cyclophosphamide (CTX) in inducing premature ovarian insufficiency (POI) model in mice and investigate the mechanism of injury. Methods Thirty-two 6~8-week-old female C57BL/6J mice were randomly divided into four groups (n=8 per group) using a weight-based block randomization method. The POI model was established via a single intraperitoneal injection of 75 mg/kg cyclophosphamide (CTX), 120 mg/kg CTX, 120 mg/kg CTX + 12 mg/kg Busulfan, or an equivalent volume of normal saline (control). Ovarian coefficients, serum estradiol (E2) and follicle-stimulating hormone (FSH) levels were measured. Western blotting was performed to assess changes in ovarian expression levels of NAD-dependent deacetylase sirtuin-5 (SIRT5) and forkhead box O3a (FOXO3a) under different modeling conditions. After determining the optimal CTX concentration for modeling, an additional forty 6~8-week-old femal C57BL/6J mice were randomly divided into five groups (n=8 per group) using a weight-based block randomization method: saline control, 120 mg/kg CTX sampling at 1, 2, 7, or 14 days after modeling. Western blotting was used to evaluate temporal changes of ovarian SIRT5 and FOXO3a protein expression. Results Compared with the saline control, all concentrations of CTX (75 mg/kg CTX, 120 mg/kg CTX) and 120 mg/kg CTX + 12 mg/kg Busulfan induced POI injury in mice. The 120 mg/kg CTX group exhibited smaller changes in ovarian coefficients (P<0.001) and E2 levels (P<0.05), whereas the 120 mg/kg CTX + 12 mg/kg Busulfan group showed rough and reduced luster fur, sluggish response and was in the worst state. Compared with the saline control group, FOXO3a expression was significantly down-regulated (P<0.05), while SIRT5 remained unchanged in the 75 mg/kg CTX group (P>0.05). In contrast, both SIRT5 (P<0.05) and FOXO3a (P<0.05) were significantly down-regulated in the 120 mg/kg CTX group. Further analysis revealed that on day 2 and 7 after 120 mg/kg CTX modeling, the expressions of SIRT5 (P<0.01) and FOXO3a (P<0.001) were significantly down-regulated, with the largest decrease observed on day 7 (SIRT5, P<0.000 1; FOXO3a, P<0.000 1). Conclusion Ovarian injury in the POI model induced by 120 mg/kg CTX is milder than that in the POI model induced by 75 mg/kg CTX. Moreover, the expression changes of SIRT5 and FOXO3a are most significant on day 7 after modeling induced by 120 mg/kg CTX, which may be related to the inhibition of the SIRT5-FOXO3a signaling pathway. Table and Figures | Reference | Related Articles | Metrics

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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 Abstract （373）   HTML （8）    PDF （742KB）（382）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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A Case Study of Using Assisted Reproductive Technology to Rescue Genetically Modified Mice with Reproductive Disorder Phenotypes WANG Qianqian, TAO Sijue, WEI Zhen, JIN Huihui, LIU Ping, WANG Lie Laboratory Animal and Comparative Medicine    2025, 45 (1): 79-86.   DOI: 10.12300/j.issn.1674-5817.2024.107 Abstract （370）   HTML （10）    PDF （2017KB）（1927）       Knowledge map    Save Objective The utilization of assisted reproductive technology to rescue genetically modified mouse strains with reproductive disorders provides a reference for improving techniques to preserve valuable experimental mouse strains. Methods In vitro fertilization-embryo transfer (IVF-ET) technology was performed on 28 strains of infertile male mice aged 9-18 months. Several indicators such as sperm density and sperm motility in infertile male mice were assessed to select the most viable sperm for IVF-ET experiments. Fertility rate, abnormal egg rate, and birth rate were recorded after the birth of the pups. An optimized ovarian transplantation procedure was applied to 12 strains of infertile female mice aged 8-18 months. 6-week-old female mice with the same genetic background were selected as recipients. One intact ovary was removed from each recipient mouse, and the contralateral oviduct was ligated. An ovary from a donor mouse was isolated and transplanted orthotopically into the side where the ovary had been removed in the recipient mouse. Twenty-one days post-surgery, recipient mice were co-housed with 8-week-old wild type male mice of the same genetic background for breeding. Data such as the pregnancy rate and live birth rate of the recipients were recorded after the birth of the pups. Results IVF-ET successfully rescued 28 mouse strains, with the oldest male mice being 18 months old. The success rate of the first round of IVF-ET experiments was 89.29% (25/28). The average fertility rate of IVF in infertile male mice was (51.01±14.97)%, the abnormal egg rate was (9.03±5.28)%, and the birth rate of offspring mice was (18.60±7.03)%. 39 out of 40 ovarian transplant recipient mice survived, with a pregnancy rate of 33.33% (13/39) for ovarian transplant recipients, and a live birth rate of 17.95% (7/39). Four mouse strains were successfully rescued using optimized ovarian transplantation technology, with the oldest female mice being 18 months old. 8 strains were not rescued as they failed to produce offspring that survived to sexual maturity. Conclusion IVF-ET is an effective approach for rescuing mice with reproductive disorders caused by different reasons, especially for those beyond the optimal breeding age. Ovarian transplantation technology can also be used as an alternative for aged female mice. But its success rate is relatively lower than that of IVF-ET, and carries a higher experimental risk. Table and Figures | Reference | Related Articles | Metrics