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Establishment and Evaluation of a Moderate-to-Severe Knee Osteoarthritis Model in Rats Induced by Surgery SUN Xiaorong, SU Dan, GUI Wenjuan, CHEN Yue Laboratory Animal and Comparative Medicine    2024, 44 (6): 597-604.   DOI: 10.12300/j.issn.1674-5817.2024.066 Abstract （1047）   HTML （233）    PDF （1789KB）（618）       Knowledge map    Save Objective To establish a rat model of moderate-to-severe knee osteoarthritis, laying the foundation for studying the pathogenesis of moderate-to-severe knee osteoarthritis and its prevention and treatment methods. Methods Thirty male SD rats were randomly divided into three groups: a sham surgery group, an 8-week model group, and a 20-week model group, with 10 rats in each group. Rats in the 8-week and 20-week model groups underwent surgery to cut the anterior and posterior cruciate ligaments and medial collateral ligament of the right knee joint, and remove the medial and lateral menisci. After surgery, the rats were allowed to move freely. The rats in the sham surgery group had only skin incisions to expose the joint without any surgical treatment. At 8 and 20 weeks post-surgery, Micro-CT scans were performed to analyze the femoral osteoporosis in the rats. After euthanizing the rats, gross observations of the knee joints were made, and the cartilage of the joint surface was scored using the Pelletier scoring system. The knee joints were collected for hematoxylin and eosin (HE) staining and safranin O-fast green staining to observe changes in cartilage morphology. The modified Mankin's scoring system was used to assess the tissue pathology of the joint surface. Immunohistochemical staining was used to detect the expression of type II collagen and matrix metalloproteinase 13 (MMP13), reflecting the anabolic and catabolic metabolism of the knee joint cartilage. Results The knee joint cartilage in the 8-week and 20-week model groups was severely damaged, with Pelletier and modified Mankin's scores significantly higher than those in the sham surgery group (both P<0.01). The Pelletier and modified Mankin's scores in the 20-week model group were significantly higher than those in the 8-week model group (P<0.01). Micro-CT observations revealed irregular joint surfaces, osteophyte formation, and signs of osteoporosis in both the 8-week and 20-week model groups, with the 20-week model group showing more loose bodies around the knee joints. Immunohistochemical staining showed increased expression of MMP13 and decreased expression of type II collagen in the knee joint tissues of the model groups, indicating that the balance of anabolic and catabolic metabolism in the joint cartilage was disrupted. MMP13 increased while type II collagen decreased. Conclusion The surgical method of cutting the anterior and posterior cruciate ligaments and medial collateral ligament and removing the medial and lateral menisci successfully creates a moderate-to-severe knee osteoarthritis model in rats. Imaging examinations reveal osteophytes, osteoporosis, and loose bodies in the knee joints, while pathological observations show a reduction or even disappearance of joint cartilage, with a disruption in the balance of cartilage anabolic and catabolic metabolism. Table and Figures | Reference | Related Articles | Metrics

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Genetic Characteristics and Research Progress of Feline Coronavirus TAO Lingyun Laboratory Animal and Comparative Medicine    2024, 44 (6): 661-666.   DOI: 10.12300/j.issn.1674-5817.2024.069 Abstract （1040）   HTML （26）    PDF （734KB）（648）       Knowledge map    Save Feline coronavirus (FCoV) is classified into two biotypes: feline infectious peritonitis virus (FIPV) and feline enteric coronavirus (FECV). FIPV and FECV might evolve and mutate via genetic recombination and mutation, leading to novel subtypes and variants. This study examined the genomic structure and biological subtyping of FCoV, analyzed the infection characteristics of FIPV and FECV, and investigated the mechanisms of FECV transforming into FIPV. The findings revealed that while their genome structures were fundamentally similar, differences in their ability to efficiently infect monocytes/macrophages significantly influenced their pathogenicity and transmission characteristics, with FIPV exhibiting higher virulence. Moreover, the analysis of the open reading frames (ORF)3/7 as well as the N/S sequences of FIPV indicated that its non-structural proteins were associated with modulation of the host immune system. These proteins enabled immune evasion, leading to more severe disease. The genomic variability of FCoV constitutes an important foundation for studying the pathogenicity and epidemiology of FIPV and FECV, and offers references for virus detection and drug development. Reference | Related Articles | Metrics

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Advances and Challenges in the Research of Integration Methods of Animal Experimental Evidence ZHENG Qingyong, LI Tengfei, XU Jianguo, ZHOU Yongjia, MA Zhichao, WANG Na, LI Molan, YANG Wenjing, WU Peirun, WANG Haidong, TIAN Jinhui Laboratory Animal and Comparative Medicine    2024, 44 (5): 567-576.   DOI: 10.12300/j.issn.1674-5817.2024.079 Abstract （1022）   HTML （29）    PDF （1365KB）（1094）       Knowledge map    Save Integrating evidence from animal experiments is a critical component of biomedical research, providing essential prior information for in-depth investigations of disease mechanisms and new drug development. Animal models have played an irreplaceable role in simulating human diseases. However, the integration of evidence from animal experiments has faced numerous challenges, including insufficient emphasis, significant heterogeneity in study designs, high publication bias, and discrepancies with clinical research practices. This paper first identifies existing issues in the original research evidence from animal experiments, such as the selection and applicability of animal models, considerations in the design of experimental studies, and factors influencing the translation of animal experimental evidence. It then discusses various methods for integrating this evidence, including systematic review and meta-analysis, overview of systematic review/umbrella review, scoping review, and evidence mapping, while highlighting recent advancements in their application. Finally, the paper addresses the main challenges currently encountered in the integration of evidence from animal experiments and proposes targeted improvement strategies aimed at enhancing the efficiency of translating research outcomes into clinical practice and promoting the advancement of evidence-based medicine. By continuously optimizing original experimental research protocols and evidence integration practices, this work aims to establish a more efficient and scientific environment for the synthesis of evidence from animal experiments, ultimately contributing to clinical trials and human health. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Animal Models of Sepsis-Related Organ Injury YANG Jiahao, DING Chunlei, QIAN Fenghua, SUN Qi, JIANG Xusheng, CHEN Wen, SHEN Mengwen Laboratory Animal and Comparative Medicine    2024, 44 (6): 636-644.   DOI: 10.12300/j.issn.1674-5817.2024.087 Abstract （832）   HTML （29）    PDF （794KB）（999）       Knowledge map    Save Sepsis is a multi-organ dysfunction syndrome caused by infection and immune dysfunction, with a high mortality rate. It affects multiple important organs such as the heart, lungs, kidneys, liver, and brain. Establishing corresponding animal models of organ dysfunction syndrome is an essential step in clarifying its pathogenesis, researching potential effective drugs, and evaluating the effectiveness and safety of treatment plans. This article first summarizes classic modeling methods for sepsis related organ injury, including the destruction of intestinal barrier tissue integrity and the implantation of pathogens or toxic drugs. The former mainly includes cecal ligation and puncture, ascending colon stent implantation, and cecal ligation incision. The latter is divided into intraperitoneal injection, intravenous injection, and intratracheal administration based on the clinical infection route being simulated. Cecal ligation and puncture and lipopolysaccharide intraperitoneal injection are the most commonly used methods. Secondly, this article summarizes the common modeling methods and evaluation methods for animal models of sepsis-induced cardiomyopathy, acute lung injury, acute kidney injury, acute liver injury, and brain dysfunction. It points out that almost all organ injuries use classic modeling methods, and different organ injury models have additional modifications according to their different pathogenesis. For example, in addition to the classic modeling methods, lipopolysaccharide instillation in the trachea is more effective in modeling acute lung injury as it better simulates lung barrier dysfunction. Cecal ligation and puncture followed by Pseudomonas instillation in the trachea in a secondary challenge model better represents sepsis-induced acute kidney injury. Intraperitoneal injection of galactosamine is a mature modeling method of sepsis-induced acute liver injury. Intracerebral injection of lipopolysaccharide is a feasible model of sepsis-associated encephalopathy. In addition to the different modeling methods, there are differences in the administration time, dosage and experimental time points according to the different experimental purposes. This article reviews the research progress of animal experimental models for sepsis-induced cardiomyopathy, acute lung injury, acute kidney injury, acute liver injury, and brain dysfunction, aiming to provide a reference for the selection of animal experimental models and optimization of experimental design. Table and Figures | Reference | Related Articles | Metrics

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Establishment and Evaluation of a Rat Model of Non-Puerperal Mastitis YIN Yulian, MA Lina, TU Siyuan, CHEN Ling, YE Meina, CHEN Hongfeng Laboratory Animal and Comparative Medicine    2024, 44 (6): 587-596.   DOI: 10.12300/j.issn.1674-5817.2024.065 Abstract （816）   HTML （252）    PDF （1898KB）（1222）       Knowledge map    Save Objective This study aims to establish a non-puerperal mastitis (NPM) rat model by simulating hyperprolactinemia and immune-inflammatory states, and to evaluate its local inflammatory characteristics in the mammary gland, thereby laying the foundation for research on the diagnosis and treatment of this clinically challenging disease. Methods Twelve SPF-grade Wistar female rats were evenly divided into a control group and a model group. During the experiment, the control group received no experimental treatment or medication. The model group received daily subcutaneous injections of 100 mg/kg metoclopramide hydrochloride for 7 consecutive days. Serum prolactin (PRL) levels were measured using ELISA on the 10th, 20th, and 30th days after the first injection. After 7 days of injections, 200 μL of lactating SD rat milk was mixed with 200 μL of complete Freund's adjuvant to prepare an oil-in-water emulsion, which was administered by multiple subcutaneous injections into the back of the Wistar rats for the initial immunization. Seven days after the initial immunization, the emulsion was injected subcutaneously into the third, fourth, and fifth mammary glands for the final immunization. After the final immunization, the rats were observed for 28 days for changes in mammary gland appearance, and the size of mammary nodules was calculated. On the 3rd, 7th, 14th, and 28th days, hematoxylin-eosin (HE) staining was used to analyze mammary tissue morphology. Immunohistochemistry was employed to detect CD138 expression levels. ELISA was used to measure the levels of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, and inducible nitric oxide synthase (iNOS) in mammary tissue to comprehensively assess the model. Results Rats in the model group exhibited mammary skin ulceration and foul odor at the ulcer sites. Palpation and ultrasound revealed the formation of mammary nodules. HE staining showed that on the 3rd day after the final immunization, normal ductal and lobular structures in the mammary glands disappeared, with significant infiltration of plasma cells. On the 7th day, ductal dilation, epithelial necrosis and detachment, and pronounced periductal plasma cell and lymphocyte (predominantly T-lymphocytes) infiltration were observed. On the 14th day, there was a proliferation of fibrofatty tissue, small blood vessels, and granulation tissue, with scattered plasma cells in the interstitium. By the 28th day, inflammatory cell infiltration and fibrous tissue proliferation were reduced, with granuloma formation. Serum PRL levels in the model group were significantly increased on the 10th day (P<0.05) and the 20th day (P<0.001). IL-6 and TNF-α levels in mammary tissue were higher in the model group compared to the control group on the 3rd, 7th, 14th, and 28th days (P<0.05). IL-1β levels were higher on the 3rd, 7th, and 14th days compared with the control group (P<0.01) but lower than the control group on the 28th day (P>0.05). iNOS levels were significantly higher on the 7th day after the final immunization (P<0.001). Conclusion A successful NPM model was established in rats, which exhibited typical pathological features such as local mammary masses, abscesses, ulcers, ductal dilation and plasma cell infiltration. This model can serve as a foundation for further research into the diagnosis and treatment of this clinically challenging disease. Table and Figures | Reference | Related Articles | Metrics

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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 （746）   HTML （36）    PDF （846KB）（1768）       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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Optimal Adaptation Period for Metabolic Cage Experiments in Mice at Different Developmental Stages TAN He, YANG Xiaohui, ZHANG Daxiu, WANG Guicheng Laboratory Animal and Comparative Medicine    2024, 44 (5): 502-510.   DOI: 10.12300/j.issn.1674-5817.2024.034 Abstract （636）   HTML （16）    PDF （1923KB）（822）       Knowledge map    Save Objective To investigate the optimal adaptation period for mice at different developmental stages during metabolic cage experiments, aiming to provide a reference for conducting metabolic research using mice. Methods A total of 80 male C57BL/6J mice at three developmental stages (weaning period M1, adolescent M2, and adulthood M3) were subjected to a 7-day metabolic cage experiment. Data on food intake, water intake, energy expenditure, respiratory quotient, body weight, and activity levels were recorded every five minutes. The collected data were processed using time series decomposition and comprehensive cluster analysis. Statistical differences were compared using repeated measures ANOVA combined with t-test to determine the optimal adaptation period. Results Significant differences in metabolism were observed among mice in different developmental stages (P<0.01). Compared with adolescent (M2) and adult (M3) mice, weaned mice (M1) exhibited lower activity level (P<0.01) and less distinct circadian rhythm. M1 mice had higher oxygen consumption, carbon dioxide production, and energy expenditure, as well as a lower respiratory quotient (all P<0.001), indicating that they mainly relied on fat as an energy source. Analysis of food intake, water intake, and energy expenditure revealed significant differences between the first light cycle (0-12 h) and the second light cycle (24-36 h) across all developmental stages (all P<0.05) . However, there was no significant difference in daily food intake or water intake after 24 hours (both P>0.05). Comprehensive cluster analysis of multiple indicators showed that the overall indicators of mice during the first 24 hours in the metabolic cages did not cluster with those of the subsequent 6 days, demonstrating significant differences. Conclusion Metabolic cage experiment can be used to detect continuous physiological changes in mice. The results suggest that mice can adapt to new metabolic cages environment within 24 hours, providing a theoretical basis for the design of metabolic experiments using mice. Table and Figures | Reference | Related Articles | Metrics

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Advances in Development of PM 2.5-Exposed Animal Models and Their Application in Reproductive Toxicity Research TIAN Fang, PAN Bin, SHI Jiayi, XU Yanyi, LI Weihua Laboratory Animal and Comparative Medicine    2024, 44 (6): 626-635.   DOI: 10.12300/j.issn.1674-5817.2024.068 Abstract （616）   HTML （29）    PDF （846KB）（647）       Knowledge map    Save Atmospheric fine particulate matter (particulate matter 2.5,PM2.5) is a major component of haze, and its potential hazards to human reproductive health have garnered widespread attention. Establishing appropriate animal models is crucial for in-depth research into the reproductive toxicity of PM2.5 exposure and its underlying mechanisms. This paper, based on recent literature, summarizes current methods for establishing PM2.5-exposed animal models and the evaluation criteria for reproductive toxicity research. The primary modeling methods for PM2.5 exposure include whole-body inhalation exposure and intratracheal instillation exposure. While whole-body inhalation exposure effectively simulates real-life human inhalation environments, it requires sophisticated experimental equipment. Conversely, intratracheal instillation exposure is more cost-effective and easier to operate but faces challenges in accurately mimicking the distribution and deposition of PM2.5 during natural inhalation. Therefore, researchers must carefully weigh these exposure methods to enhance model rigor and achieve the most realistic simulation of human exposure conditions. When summarizing the application evaluation indicators of PM2.5-induced reproductive toxicity, this review finds that the main indicators of male reproductive toxicity include reduced sperm quality, testicular tissue damage, and hormonal imbalances. For female reproductive toxicity, the primary indicators are reduced ovarian reserve, endocrine dysfunction, endometrial damage, and adverse perinatal reactions. Additionally, this review highlights the need for detailed chemical composition analysis of PM2.5, exploring the reproductive toxic targets and mechanisms of particles containing different chemical components, such as heavy metals and polycyclic aromatic hydrocarbons. Long-term studies are also necessary to assess the effects of PM2.5 exposure on reproductive health and transgenerational effects, to predict potential long-term risks for humans. Additionally, interdisciplinary collaboration should be encouraged, involving cooperation between environmental science, toxicology, reproductive medicine, and other disciplines, to comprehensively assess the environmental health risks of PM2.5 and provide scientific support for the development of integrated prevention and control strategies. This review summarizes animal modeling methods, evaluation criteria, and their applications, providing valuable methodological references for future reproductive toxicity research on PM2.5. Table and Figures | Reference | Related Articles | Metrics

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Optimization and Evaluation of Conditions for Orthotopic Nude Mouse Models of Human Liver Tumor Cells MENG Yu, LIANG Dongli, ZHENG Linlin, ZHOU Yuanyuan, WANG Zhaoxia Laboratory Animal and Comparative Medicine    2024, 44 (5): 511-522.   DOI: 10.12300/j.issn.1674-5817.2024.048 Abstract （565）   HTML （26）    PDF （2747KB）（1274）       Knowledge map    Save Objective The study aims to optimize the conditions for constructing orthotopic nude mouse models of liver cancer by injecting human liver tumor cell lines and to explore appropriate timings for drug administration. Methods Human hepatocellular carcinoma Hep3B and hepatoblastoma HepG2 cell lines, which stably expressing the luciferase reporter gene (LUC), were selected. The linear correlation between the luciferase luminescence intensity and the number of liver tumor cells was analyzed using a Small Animal In Vivo Imaging system to verify the luminescent efficiency of the human liver tumor cells. Different concentrations (8×106, 2.4×107, 7.2×107 cells/mL) and resuspension media (PBS, Matrigel) of human liver tumor cell suspensions HepG2-LUC and Hep3B-LUC were orthotopically inoculated into the liver lobes of 5-week-old female BALB/c nude mice (12 groups, 7 mice each) to construct human liver tumor nude mouse orthotopic cancer models. Every 7 days, the weights of mice were recorded, and the growth of orthotopic tumors was monitored using the Small Animal In Vivo Imaging system. On day 35 post-cell inoculation, mouse livers were dissected, and pathological slices were prepared for HE staining to observe histopathological changes in liver tissues. Results The luminescence intensity of human liver tumor cell lines was positively correlated with the number of cells (R2=0.983 1, R2=0.970 5), indicating their suitability for orthotopic model construction. Successful modeling was achieved in the high-concentration groups of HepG2-LUC, the low-, medium-, and high-concentration groups of HepG2-LUC+Matrigel, the medium- and high-concentration groups of Hep3B-LUC, and the low-, medium-, and high-concentration groups of Hep3B-LUC+Matrigel. For both HepG2-LUC+Matrigel and Hep3B-LUC+Matrigel groups, mice in the high-concentration groups exhibited significantly reduced body weight compared to the low- and medium-concentration groups (both with P<0.05). The luminescence intensity of successfully modeled mice increased exponentially over time (R2>0.950 0), and reached a minimum of 1.0×107 p/(s·cm2·sr) by day 14 post-transplantation. Mice in the low- and medium-concentration groups of HepG2-LUC and the low-concentration group of Hep3B-LUC showed no significant pathological changes, while the other groups exhibited evident liver tumors and hepatocyte lesions. Conclusion For the HepG2-LUC cell line, the recommended injection volume is 50 μL with a cell density of 2.4×107 cells/mL, resuspended with Matrigel, followed by drug administration or prognostic measures on day 7 post-modeling. For the Hep3B-LUC cell line, the recommended injection volume is 50 μL with a cell density of 7.2×107 cells/mL, not resuspended with Matrigel, with administration or prognostic measures on day 14 post-modeling. Table and Figures | Reference | Related Articles | Metrics

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Challenges and Development in Suzhou Laboratory Animal Industry Over the Past Five Decades ZHAO Lijuan, XIAO Chunlan, SHENG Yajie, LU Xi, ZHOU Zhengyu Laboratory Animal and Comparative Medicine    2024, 44 (6): 645-653.   DOI: 10.12300/j.issn.1674-5817.2024.113 Abstract （556）   HTML （57）    PDF （1401KB）（1295）       Knowledge map    Save Since the 1970s, the laboratory animal industry in Suzhou has gone through five stages: its inception, emergence, growth, transformation, and scaling up. It began with the manufacturing of caging equipment for laboratory animals, initially by imitation and later through independent innovation. The industry evolved from sporadic factories to clustered enterprises, gradually growing and opening up the export market for caging equipment. In the 21st century, with industrial upgrading and transformation, purification systems and related products began to develop, and industry organizations emerged. As China has modernized, the rise of automation and intelligent production has led to technological innovation in enterprises and the emergence of various outsourcing services in the laboratory animal industry, driving the large-scale development of the industrial chain. After nearly half a century of growth, the laboratory animal industry in Suzhou has formed a complete industrial chain, including the production of laboratory animals, caging equipment, feed and bedding materials, design and construction of laboratory animal facilities, quality testing of laboratory animals and environments, and animal experimentation services. Laboratory animal breeding equipment, the core of the industry, has reached the level of developed countries, and the industry's scale and influence are unmatched in China. Since the 21st century, biopharmaceuticals have become the "No.1 industry" in the development of Suzhou. With government support, the guidance of the local economy, and the assistance from universities and research institutes, the animal experiment outsourcing industry has begun to cluster in Suzhou. The continuous influx of CROs has driven the construction of large-scale laboratory animal facilities, and key research projects have been initiated, significantly enhancing the industry's R&D capabilities. The Suzhou laboratory animal industry has quickly expanded alongside the "No. 1 industry," creating a unique "Suzhou Path" for laboratory animals. Over nearly fifty years, the laboratory animal industry in Suzhou has been essential to the rapid development of the biopharmaceutical industry in Suzhou and China. Table and Figures | Reference | Related Articles | Metrics

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Advances in Nucleic Acid Drugs and Gene Therapies based on Animal Models of Duchenne Muscular Dystrophy LIU Siyu, LAI Yuezhao, GUO Wenting, CHEN Xuejin Laboratory Animal and Comparative Medicine    2024, 44 (6): 613-625.   DOI: 10.12300/j.issn.1674-5817.2024.168 Abstract （524）   HTML （23）    PDF （1488KB）（1525）       Knowledge map    Save Duchenne muscular dystrophy (DMD) is a severe X-linked recessive genetic disorder caused by mutations in the DMD gene, making it one of the most common forms of hereditary muscular dystrophy. The DMD gene, which encodes dystrophin, is the largest known gene in the human genome. Mutations in the DMD gene are highly diverse, including exon deletions, duplications, point mutations, and small insertions or deletions, posing significant challenges for treatment. Currently, there is no cure for DMD, and existing treatment strategies focus primarily on symptom management, which cannot reverse or halt disease progression. Advances in biotechnology position nucleic acid drugs and gene therapies at the forefront of DMD treatment research. These treatments aim to restore dystrophin expression by repairing or replacing mutated genes, thereby improving muscle function or slowing muscle degeneration. Preclinical studies in animal models and early-phase clinical trials demonstrate promising efficacy and offer new hope for DMD patients. This review briefly outlines the pathological mechanisms and genetic characteristics of DMD before delving into recent progress in therapeutic strategies, with a particular focus on nucleic acid drugs (including antisense oligonucleotides for exon skipping therapy and translation readthrough inducers) and gene therapy approaches (including gene replacement therapy and gene editing). The development and application of these therapies not only provide new treatment options for DMD patients, but also offer valuable insights for addressing other genetic disorders. However, numerous challenges impede the clinical translation of DMD treatments. Future studies must optimize existing therapeutic strategies, improve their efficacy and applicability, and explore innovative approaches to deliver more effective and sustainable treatments for DMD patients. 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 （511）   HTML （16）    PDF （889KB）（2061）       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 （501）   HTML （19）    PDF （1114KB）（690）       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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Establishment Methods and Application Evaluation of Animal Models in Reproductive Toxicology Research HUANG Dongyan, WU Jianhui Laboratory Animal and Comparative Medicine    2024, 44 (5): 550-559.   DOI: 10.12300/j.issn.1674-5817.2024.028 Abstract （492）   HTML （17）    PDF （870KB）（772）       Knowledge map    Save Reproductive toxicology is a discipline that uses toxicological methods to study the mechanisms by which foreign substances interfere with the generation of eggs or sperm and their detrimental effects on offspring. Research includes evaluating the damaging effects of test substances on reproductive function of parents and the toxicity evaluation of offspring embryos. People are exposed to a wide range of drugs, chemicals and environmental pollutants on a daily basis, and determining whether these substances have reproductive toxicity is crucial for the health of future generations. Reproductive toxicology research is therefore critical. Given the specificity and importance of reproductive toxicity evaluation, corresponding institutions both domestically and internationally have issued guidelines, national standards, or industry standards, all of which involve animal experiments. In the study of reproductive system diseases, numerous animal models have been developed to investigate key reproductive organs, such as testicles and ovaries. Each model involves the selection of animals, the establishment of methods, and the quantification of evaluation indicators, and all have advantages and limitations. The choice of model should be based on experimental needs and the characteristics of the model. This paper summarizes commonly used animal models for reproductive and development toxicity evaluation in reproductive toxicology research, including rat models for fertility and early embryonic development toxicity, rat models for embryo-fetal development toxicity, rabbit models for embryo-fetal development toxicity evaluation, minipig models for embryo-fetal development toxicity, rat models for perinatal toxicity, zebrafish models for embryonic development toxicity, and models for evaluating ovarian toxicity induced by chemical drugs, radiotherapy, autoimmunity, and ovariectomy, as well as models for evaluating testicular toxicity caused by chemical drugs and environmental factors. The methods for establishing these models, their application scope, and characteristics are reviewed in order to provide references for relevant research applications. 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 （491）   HTML （20）    PDF （1784KB）（773）       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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Establishment of a New Hyperglycemic Obesity Cardiac Dysfunction Mouse Model with Triacsin C ZHAO Xiaona, WANG Peng, YE Maoqing, QU Xinkai Laboratory Animal and Comparative Medicine    2024, 44 (6): 605-612.   DOI: 10.12300/j.issn.1674-5817.2024.078 Abstract （475）   HTML （196）    PDF （1446KB）（709）       Knowledge map    Save Objective This study aims to establish a novel hyperglycemic obesity mouse model by utilizing Triacsin C, an inhibitor of acyl-CoA synthetase long-chain family member 1 (ACSL1), combined with a high-fat diet, to simulate the changes in adipose tissue and cardiac function observed in patients with obesity-related type 2 diabetes. Methods Twenty adult SPF-grade male C57BL/6J mice were randomly divided into two groups: the Control group (injected intraperitoneally with citric acid-sodium citrate buffer, Con group) and the TC group (injected intraperitoneally with Triacsin C, TC group). After four consecutive weeks of intraperitoneal injections, both groups were fed high-fat diets. Body weight and glucose tolerance of the mice were assessed every eight weeks. The models were considered successful if fasting blood glucose exceeded 8 mmol/L or blood glucose was above 15 mmol/L two hours after glucose injection. Cardiac function, including ventricular end-diastolic diameter (LVEDD), left ventricular end systolic diameter (LVESD), end-diastolic interventricular septal thickness (EDIVS), left ventricular ejection fraction (LVEF), and left ventricular short-axis fractional shortening (FS), was measured by echocardiography. HE staining was used to detect the changes in epididymal white adipose tissue (WAT) and brown adipose tissue (BAT). Immunofluorescence technology was used to analyze changes in CD31 and UCP1 in BAT. ACSL1 expression in myocardial tissue was tested by Western blotting. Results The fasting blood glucose levels were (8.14±1.43) mmol/L in the Con group and (8.18±0.85) mmol/L in the TC group (P>0.05) , and the 2-hour postprandial blood glucose levels were (19.8±4.01) mmol/L in the Con group and (22.60±3.97) mmol/L in the TC group (P<0.05). This indicated that both groups of diabetic mouse models were successfully established. Compared to the Con group, the TC group showed poor glucose tolerance; significant decreases in LVEDD, LVEF and FS (P<0.05); significant increases in WAT and BAT areas (P<0.05); significant decreases in CD31 and UCP1 expression (P<0.05); and a significant decrease in the expression of ACSL1 in myocardial tissues (P<0.05). Conclusion Compared with the high-fat diet-induced type 2 diabetes model, the new hyperglycemic obesity and cardiac dysfunction mouse model, created by the combination of Triacsin C and a high-fat diet, is feasible and allows for easier observation of brown adipose tissue whitening, insulin resistance and cardiac dysfunction. 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 （469）   HTML （97）    PDF （1785KB）（1442）       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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Analysis of the Progress in Identification and Evaluation of Laboratory Animal Resources in China DU Xiaoyan, LIU Yunbo Laboratory Animal and Comparative Medicine    2024, 44 (5): 469-474.   DOI: 10.12300/j.issn.1674-5817.2024.050 Abstract （453）   HTML （44）    PDF （873KB）（597）       Knowledge map    Save Laboratory animals are not only a national strategic resource but also an important support for development of science and technology. The Committee of Identification and Evaluation for Laboratory Animal Resources, organized by Chinese Association for Laboratory Animal Sciences and established in May 2019, is currently the only specialized academic agency dedicated to the identification and evaluation of laboratory animal resources in China. This paper first discusses the significance of identifying and evaluating laboratory animal resources, summarizes three new approaches to developing these resources, including the domestication and standardization of laboratory animals (economic, ornamental, agricultural, and wild animals, etc.), the acquisition of new strains (species) through natural mutation and induced mutation, and the creation of new laboratory animal resources through gene editing technology. It then introduces the workflow for resource identification and evaluation, including preliminary review (format review), written review (expert review), joint review or on-site inspection, final review (voting and public announcement)，and the issuance of certificates. The required materials to be submitted include application, summary report, research or identification reports, appendices and other necessary documents. The paper further discusses related requirements for resource identification and evaluation, including population, genetic classification, biological characteristics, genetic stability, and application value. Finally, the current status of newly identified laboratory animal strains (species) and issues in current work practices are analyzed, as well as solutions to these issues. This paper aims to provide a valuable reference for further research in this field. 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 （437）   HTML （87）    PDF （1909KB）（611）       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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Guideline Checklist for Publishing Research Papers on Animal Experimentation and Comparative Medicine in China (2024 Edition) Editorial Board of Laboratory Animal and Comparative Medicine Laboratory Animal and Comparative Medicine    2024, 44 (5): 577-582.   DOI: 10.12300/j.issn.1674-5817.2024.149 Abstract （433）   HTML （47）    PDF （651KB）（374）       Knowledge map    Save The standardization, transparency, and completeness of research papers in animal experimentation and comparative medicine research are crucial for ensuring the credibility, reproducibility, and clinical applicability of research findings. Internationally recognized guidelines, such as the ARRIVE guidelines, serve to standardize animal experimentation and reporting. In China, there is already a comprehensive framework of regulations and standards governing laboratory animal research. However, the practical guidance in these documents remains insufficient to effectively guide Chinese researchers in writing and publishing in vivo animal research papers. To address this gap, the Editorial Board of Laboratory Animals and Comparative Medicine has developed a checklist of publication standards tailored for Chinese scholars, informed by the ARRIVE 2.0 guidelines and Chinese regulations and standards. This checklist is applicable for authors to self-check during manuscript preparation and submission, for peer experts during the review process, for journal editors in pre-publication verification, and for readers to evaluate published works. It aims to effectively promote the standardization and high-quality development of laboratory animal and comparative medicine research in China. 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 （422）   HTML （25）    PDF （2015KB）（598）       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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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 （410）   HTML （17）    PDF （908KB）（909）       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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Perioperative Animal Care for Xenotransplantation from Genetically Edited Pigs to Monkeys ZHU Chan, ZHANG Dongliang, ZHAO Deli, SHI Xueqin, QIAN Lei, ZHANG Xuan, JIN Yan, DUAN Wei, QI Ruocheng, LIU Chaohua, YANG Xuekang, HAN Juntao, PAN Dengke Laboratory Animal and Comparative Medicine    2024, 44 (5): 495-501.   DOI: 10.12300/j.issn.1674-5817.2024.043 Abstract （403）   HTML （19）    PDF （1645KB）（807）       Knowledge map    Save Objective To discuss the perioperative care and wound protection of xenotransplantation from genetically edited pigs to monkeys, with the goal of improving the success rate of such experimental procedures. Methods From October 2022 to October 2023, perioperative care and wound protection were performed on 7 recipient rhesus monkeys undergoing xenotransplantation of genetically edited pig tissues and organs. Customized wound protective garments were designed based on monkeys' size and surgical area to protect the wounds, alongside meticulous perioperative care. This included preoperative preparation and medication, intraoperative monitoring of physiological indicators and anesthesia management, and postoperative care comprising wound protection, observation and monitoring, and nutritional support. Results All seven monkeys successfully underwent xenotransplantation. With the aid of protective garments and detailed care, all surgical wounds healed by first intention, and postoperative recovery was satisfactory. Conclusion Proper care and wound protection during xenotransplantation from genetically edited pigs to monkeys not only promote wound healing, but also alleviate pain and harm to animals. This has significant implications for advancing experimental research in pig-monkey xenotransplantation and enhancing animal welfare. 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 （395）   HTML （32）    PDF （4039KB）（1442）       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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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 （392）   HTML （40）    PDF （4184KB）（499）       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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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 （389）   HTML （17）    PDF （882KB）（1342）       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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Advances and Challenges of Using Experimental Pigs in Da Vinci Surgical Robot Training LIU Yishu, CAI Liping Laboratory Animal and Comparative Medicine    2024, 44 (6): 667-674.   DOI: 10.12300/j.issn.1674-5817.2024.075 Abstract （372）   HTML （24）    PDF （1379KB）（587）       Knowledge map    Save Experimental pigs occupy a crucial position in life sciences research and have been indispensable in advancing the practical application of new clinical technologies and methods. The Da Vinci Surgical Robot System, developed by Intuitive Surgical in the United States, has been widely used across various surgical fields since its approval by the U.S. Food and Drug Administration (FDA) in 2000, and is highly esteemed for its precision and accuracy. With the continuous advancement of surgical robot technology, the skill requirements for medical professionals have also increased. Consequently, surgical skills training has become an essential component of ensuring both surgical safety and effectiveness. This article briefly reviews the current status of Da Vinci surgical robot training, both domestically and internationally, with a focus on the practical application of experimental pigs in domestic Da Vinci surgical robot training. It emphasizes that experimental pigs not only effectively simulate the human surgical environment, enabling trainees to practice in a safe and controlled setting, but also help accelerate the trainees' familiarity with and mastery of the surgical robot. This, in turn, significantly shortens the learning curve, enhances the precision and stability of surgical procedures, and reduces surgical risks. However, the use of experimental animals in surgical robot training also encounters challenges, including limitations caused by the differences between experimental animals and humans, potential ethical concerns, and public opinion pressures. In response to these challenges, the paper proposes suggestions such as improving and enforcing ethical regulations, as well as advancing the development of virtual reality (VR) and augmented reality (AR) technologies. These efforts aim to reduce reliance on experimental animals in surgical training while enhancing training effectiveness, thereby contributing to the innovation and development of Da Vinci surgical robot training models. Table and Figures | Reference | Related Articles | Metrics

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Evaluation of Simulated Weightlessness Model of Hindlimb Unloading Miniature Pigs and Their Tissue Damage TU Yingxin, JI Yilan, WANG Fei, YANG Dongming, WANG Dongdong, SUN Zhixin, DAI Yuexin, WANG Yanji, Guanghan KAN, WU Bin, ZHAO Deming, YANG Lifeng Laboratory Animal and Comparative Medicine    2024, 44 (5): 475-486.   DOI: 10.12300/j.issn.1674-5817.2024.038 Abstract （369）   HTML （20）    PDF （3774KB）（639）       Knowledge map    Save Objective To establish a weightlessness simulation animal model using miniature pigs, leveraging the characteristic of multiple systems’ tissue structures and functions similar to those of humans, and to observe pathophysiological changes, providing a new method for aerospace research. Methods Nine standard-grade miniature pigs were selected and randomly divided into an experimental group (n=7) and a control group (n=2). The experimental group was fixed using customized metal cages, with canvas slings suspending their hind limbs off the ground, and the body positioned at a -20° angle relative to the ground to simulate unloading for 30 days (24 hours a day). Data on body weight, blood volume, and blood biochemistry indicators were collected at different time points for statistical analysis of basic physiological changes. After the experiment, the miniature pigs were euthanized and tissue samples were collected for histopathological observation of the cardiovascular, skeletal and muscle systems HE and Masson staining. Statistical analysis was also conducted on the thickness of arterial vessels and the diameter of skeletal muscle fibers. Additionally, western blotting was employed to detect the expression levels of skeletal muscle atrophy-related proteins, including muscle-specific RING finger protein 1 (MuRf-1) and muscle atrophy F-box (MAFbx, as known as Atrogin-1), while immunohistochemistry was used to detect the expression of glial fibrillary acidic protein (GFAP), an indicator of astrocyte activation in the brain, reflecting the pathophysiological functional changes across systems. Results After hindlimb unloading, the experimental group showed significant decreases in body weight (P<0.001) and blood volume (P<0.01). During the experiment, hemoglobin, hematocrit, and red blood cell count levels significantly decreased (P<0.05) but gradually recovered. The expression levels of alanine aminotransferase and γ-glutamyltransferase initially decreased (P<0.05) before rebounding, while albumin significantly decreased (P<0.001) and globulin significantly increased (P<0.01). Creatinine significantly decreased (P<0.05). The average diameter of gastrocnemius muscle fibers in the experimental group significantly shortened (P<0.05), with a leftward shift in the distribution of muscle fiber diameters and an increase in small-diameter muscle fibers. Simultaneously, Atrogin-1 expression in the gastrocnemius and paravertebral muscles significantly increased (P<0.05). These changes are generally consistent with the effects of weightlessness on humans and animals in space. Furthermore, degenerative changes were observed in some neurons of the cortical parietal lobe, frontal lobe, and hippocampal regions of the experimental group, with a slight reduction in the number of Purkinje cells in the cerebellar region, and a significant enhancement of GFAP-positive signals in the hippocampal area (P<0.05). Conclusion Miniature pigs subjected to a -20° angle hind limb unloading for 30 days maybe serve as a new animal model for simulating weightlessness, applicable to related aerospace research. 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 （357）   HTML （21）    PDF （793KB）（1279）       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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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 （356）   HTML （38）    PDF （3294KB）（602）       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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Transcriptomic Analysis of Menstrual Blood-Derived Stem Cells Transplantation Combined with Exercise Training in Promoting Spinal Cord Injury Recovery in Rats QI Longju, CHEN Shiyuan, LIAO Zehua, SHI Yuanhu, SUN Yuyu, WANG Qinghua Laboratory Animal and Comparative Medicine    2024, 44 (5): 531-542.   DOI: 10.12300/j.issn.1674-5817.2024.031 Abstract （353）   HTML （8）    PDF （2080KB）（794）       Knowledge map    Save Objective To explore the potential therapeutic targets and molecular mechanisms of menstrual blood-derived stem cells (MenSCs) transplantation combined with exercise training in promoting recovery in rats with spinal cord injury (SCI) through transcriptome sequencing analysis. Methods Female SD rats aged two months were selected and a SCI model was established by a hemisection at the tenth thoracic vertebra (T10). The rats were then divided into two groups: the Cell and Treadmill Training (CTMT) group, which received MenSCs transplantation and treadmill training after SCI, and the SCI group (control), with 12 rats in each group. One week after modeling, the CTMT group received a microinjection of 1×105 MenSCs at the injury site, followed by two weeks of weight-supported aerobic exercise training. Spinal cord tissue from the injury site was selected for transcriptome sequencing, and mRNA expression data from both the SCI and CTMT groups were analyzed. Differential gene expression, GO (Gene Ontology) functional enrichment, KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment, and protein-protein interaction (PPI) network analyses were performed. Motor function recovery was assessed using the Basso, Beattie, and Bresnahan (BBB) score, while histopathological changes at the injury site were evaluated through hematoxylin-eosin (HE) staining. Real-time fluorescent quantitative PCR and Western blotting were used to verify the expression of differentially expressed genes. Results Transcriptome sequencing analysis showed 247 upregulated genes and 174 downregulated genes in the CTMT group compared to the SCI group. Notably, genes such as Bdnf, Hmox1, Sd4, Mmp3, and Cd163 were significantly upregulated [|log2(FoldChange)|≥0.66, P<0.05]. KEGG pathway enrichment analysis and GO functional enrichment analysis indicated that these differentially expressed genes were mainly involved in growth and development, metabolic reactions, and immune-inflammatory processes, such as axon growth and the electron transport chain. The Bdnf gene was notably enriched in the PI3K-Akt signaling pathway. The BBB score showed that MenSCs transplantation combined with exercise training significantly improved the motor function of SCI rats. HE staining revealed that pathological changes at the injury site were significantly reduced in the treatment group. Furthermore, real-time quantitative PCR and Western blotting confirmed that brain-derived neurotrophic factor (BDNF) mRNA and protein expression levels in the CTMT group were significantly higher than those in the SCI group (P<0.001). Conclusion The combined exercise training with MenSCs effectively promotes the recovery of motor function in SCI rats by upregulating BDNF expression, providing a novel strategy for SCI treatment. 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 （352）   HTML （18）    PDF （915KB）（631）       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 （349）   HTML （15）    PDF （802KB）（492）       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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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 （348）   HTML （19）    PDF （905KB）（497）       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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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 （342）   HTML （26）    PDF （3011KB）（992）       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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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 （342）   HTML （19）    PDF （1203KB）（338）       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 （341）   HTML （11）    PDF （1751KB）（832）       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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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 （334）   HTML （37）    PDF （2277KB）（517）       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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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 （332）   HTML （18）    PDF （793KB）（809）       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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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 （327）   HTML （19）    PDF （816KB）（425）       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