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    History, Current Status, Challenges and Opportunities of Laboratory Monkey Industry in China
    SUN Qiang
    Laboratory Animal and Comparative Medicine    2024, 44 (4): 343-356.   DOI: 10.12300/j.issn.1674-5817.2024.112
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    Laboratory animals play a crucial role in foundational scientific research and clinical medicine. Non-human primates (NHP), particularly Macaca mulatta and Macaca fascicularis, have long been highly valued due to their close resemblance to humans. After more than half a century of development, China's NHP laboratory animal industry has gradually transitioned from its early stage of rapid and unregulated growth to a mature stage of standardization and refinement. However, there has been a dramatic surge in global biopharmaceutical research in recent years, leading to a sharp increase in demand for NHP laboratory animals. This surge, coupled with the lack of long-term strategic planning among breeding enterprises, has resulted in severe aging of breeding populations and a significant decline in reproductive capabilities, further widening the supply gap. Under the dual pressures of rising demand and declining supply, the prices of NHP laboratory animals have surged. Although the cyclical downturn in the biopharmaceutical industry in recent years has lowered the demand for NHP laboratory animals to some extent, leading to significant price drops, the prices remain high. At the same time, against the backdrop of high prices, issues such as the accelerating aging of breeding populations, the lower standards for microbial quality control, insufficient genetic quality control, and blind investment in facility construction have emerged within the NHP laboratory animal industry. This report provides a comprehensive review of the history and current status of China's NHP laboratory animal industry, with a focus on laboratory monkeys. It explores the factors shaping the current industry landscape and identifies potential challenges and opportunities facing the industry. It aims to offer insights and references for the future development of China's NHP laboratory animal industry.

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    Three Dimensions of Animal Experiment Ethics: Analysis Based on Value of Life, Animal Welfare, and Risk Prevention
    ZHAO Yong
    Laboratory Animal and Comparative Medicine    2024, 44 (4): 445-454.   DOI: 10.12300/j.issn.1674-5817.2024.108
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    Ethical review of animal experiments is based on factors such as the necessity of the experiment, its scientific validity, the professional competence of the experimenters, and research conditions, to determine the ethical permissibility of an animal experiment. Attitudes towards laboratory animals and animal experiments vary significantly due to differences in cultural traditions, religious beliefs, personality traits, and roles within the experimentation process. How animal experiment ethics can advance in tandem with the advancements of life sciences, while consistently providing forward-looking guidance and safeguarding fundamental ethics, is a subject that requires continuous exploration, practice, and research. Ensuring the welfare of laboratory animals has now become a widely accepted ethical consensus. However, attitudes and principles towards different types of animal experiments, and the methods to genuinely and effectively ensure the welfare quality of animals during live animal experiments, should be central to animal experiment ethics. Based on the deep integration of biotechnology in the field of laboratory animals, this paper considers the contribution of animal experimentation to the development of scientific ethical concepts. It systematically elucidates the special value of life in laboratory animals and their relationship with biosafety and ecological safety. It also outlines welfare quality assessment methods for different species of laboratory animals, demonstrating that the philosophical ideology of the value of life is the core of animal experiment ethics. The quality and significance of animal experiments determine the ethical level at which the value of life in laboratory animals is realized, and animal welfare technologies provide a robust ethical guarantee for animal experiments. The close integration of ethical theories with life sciences in laboratory animals is an objective requirement for animal experiment ethics. The value of life, animal welfare, and risk prevention collectively form the core elements of ethical review in animal experiments, serving as fundamental factors in improving the quality of ethical reviews and avoiding ethical deviations.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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

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    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
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    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.

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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
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    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.

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    Application Analysis of Animal Models for Pelvic Inflammatory Disease Based on Data Mining
    ZHENG Yiqing, DENG Yasheng, FAN Yanping, LIANG Tianwei, HUANG Hui, LIU Yonghui, NI Zhaobing, LIN Jiang
    Laboratory Animal and Comparative Medicine    2024, 44 (4): 405-418.   DOI: 10.12300/j.issn.1674-5817.2024.012
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    Objective To investigate the key elements for model establishment and determine the evaluation indicators of animal models for pelvic inflammatory disease (PID), providing a reference for improving modelling methods and optimizing the application of PID animal models. Methods The search query "Pelvic Inflammatory Disease" AND "Animal Model" OR "Rat" OR "Mouse" OR" Guinea Pig" OR "Rabbit" OR "Dog" OR "Pig" was used to retrieve relevant literature on PID animal models published from 2013 to 2023 in China Knowledge Network Infrastructure (CNKI), Wanfang, and PubMed databases. The studies were analyzed and categorized based on experimental animal types, modelling methods, modelling cycles, detection indicators, positive control drugs, and administration duration. A database was established for statistical analysis. Results A total of 214 research articles on PID animal models meeting the inclusion criteria were identified. The most commonly used model animals are Sprague Dawley (SD) rats, followed by Wistar rats. The most frequently employed modelling method is a combination of mechanical injury and bacterial infection, followed by the phenol mucilage method. The most common modelling cycles for acute pelvic inflammatory disease (APID) and chronic pelvic inflammatory disease (CPID)/sequelae of pelvic inflammatory disease (SPID) are 8 to 14 days, while for PID models without specific staging, the cycles are 7 days. High-frequency detection methods and indicators include histopathological observation using hematoxylin-eosin staining, enzyme-linked immunosorbent assay (ELISA) for serum-related indicators, morphological changes of tissues observed with the naked eye, and immunohistochemical detection of related protein expression in uterine tissues, and pathological scoring. The most frequently used positive control drugs are Fuke Qianjin Tablets, followed by Jingangteng Capsules. The most common administration duration for APID is 7 days, and for CPID/SPID models, it ranges from 15 to 21 days. Conclusion Currently, SD rats and Wistar rats are commonly used as experimental animals for PID models. The dual modelling method of mechanical injury combined with mixed bacterial infection aligns closely with clinical pathogenesis and can be used to establish a PID model that simulates postoperative uterine cavity infection. Depending on the research objectives, different positive drugs and detection indicators should be selected for comprehensive evaluation. Most existing PID animal model studies are based on western medical diagnosis, with fewer studies focusing on Traditional Chinese Medicine (TCM) syndromes. There is a need to integrate TCM theories of etiology and pathogenesis to construct PID animal models that are more in line with TCM clinical symptoms.

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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
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    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.

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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
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    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.

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    Characteristics Evaluation and Application Analysis on Animal Models of Recurrent Spontaneous Abortion
    DING Tiansong, XIE Jinghong, YANG Bin, LI Heqiao, QIAO Yizhuo, CHEN Xinru, TIAN Wenfan, LI Jiapei, ZHANG Wanyi, LI Fanxuan
    Laboratory Animal and Comparative Medicine    2024, 44 (4): 393-404.   DOI: 10.12300/j.issn.1674-5817.2024.013
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    Objective To summarize and evaluate the characteristics of current recurrent spontaneous abortion (RSA) animal models at home and abroad, and to provide reference and guidance for the standardized preparation of RSA models. Methods "Recurrent spontaneous abortion" and "animal model" were used as co-keywords in CNKI, Wanfang, VIP, PubMed and Web of Science databases to search the RSA animal experimental literature, covering the period up to January 20, 2024, and a total of 1 411 articles were collected. The analysis focused on construction methods and essential elements of RSA animal models, the modeling process and result evaluation, as well as the application of these models in pharmacological and pharmacodynamic research. An Excel table was established for systematic analysis and discussion. Results A total of 138 experimental studies were obtained after screening. In constructing RSA animal models, immunological models were the most widely used in Western medicine (96.92%), with the Clark model being the main one (92.31%). In traditional Chinese medicine (TCM) models, 70.00% were kidney deficiency-luteal inhibition-syndrome combination models, 20.00% were kidney deficiency and blood stasis models, and 10.00% were deficiency-heat syndrome models. Most animals were selected at 6-8 weeks (33.86%) and 8 weeks (32.28%) of age. The majority of animals were paired for mating at 18:00 on the day of cage pairing. In 81.03% of literatures, vaginal plugs were checked once the following morning, with 8:00 being the most common time (17.02%). The most commonly used drug administration cycle was 14 days of continuous gavage after pregnancy. Among the tested drugs, Western drugs were mainly protein-based (29.17%), while TCM drugs were mainly TCM decoction (81.11%). The most frequently used methods for detecting indicators included visual observation of embryos (22.54%), western blot (15.96%), PCR (13.58%), ELISA (12.91%), HE staining (10.80%) and immunohistochemistry (9.39%). Conclusion The etiology of RSA is complex, and corresponding animal models should be established based on different etiologies. Clark model is commonly used in the construction of Western medicine model, while the kidney deficiency-luteal inhibition-syndrome combination model is predominant in TCM. RSA animal model is widely used in related research, but systematic evaluation needs to be strengthened.

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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
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    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.

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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
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    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.

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    Integrative Analysis of Omics Data in Animal Models of Coronavirus Infection
    WU Yue, LI Lu, ZHANG Yang, WANG Jue, FENG Tingting, LI Yitong, WANG Kai, KONG Qi
    Laboratory Animal and Comparative Medicine    2024, 44 (4): 357-373.   DOI: 10.12300/j.issn.1674-5817.2024.008
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    Objective This study analyzes the omics data resources in human-infecting coronavirus animal models collected from various public databases, focusing on data distribution, dataset quantity, data types, species, strains, and research content. It aims to enhance our understanding of biological characteristics and pathogenic mechanisms of coronaviruses, thereby providing a solid foundation for devising effective therapeutic strategies and preventive measures. Methods Query strategies, including specific virus names, time ranges, and inclusion and exclusion criteria, were defined to retrieve data from major public omics databases such as GEO and ArrayExpress. Secondary filtering was performed based on different field types to obtain a more accurate data list. An omics data text database was established for bibliometric analysis. Co-occurrence networks were constructed for the analysis of the correlation strengths between different research themes, technical methods, and involved species. The cell types, organs, and biological pathways involved in studies were examined to further elucidate the pathogenic interplay between pathogens and hosts. Results About twenty public databases containing coronavirus-related omics data were identified, with a primary focus on novel coronavirus infection. Commonly used species include humans, mice, hamsters, and monkeys, while the commonly used virus strains are Wuhan-Hu-1 and USA-WA1/2020. Lung tissues are primarily used in animal models such as mice, macaques, and ferrets, while airway epithelial cells and Calu-3 cells are predominantly employed in human-related studies. Expression profiling data indicate that gene pathways involved in inflammation, cytokine response, complement pathway, cell damage, proliferation, and differentiation are significantly upregulated after infection. Proteomics studies reveal significant changes in phosphoproteome, ubiquitinome, and total proteome of patient samples at different infection stages. Specific protein categories, including viral receptors and proteases, transcription factors, cytokines, proteins associated with coagulation system, angiogenesis-related proteins, and fibrosis markers, show alterations after coronavirus infection. In addition, metabolomics data suggest that phosphocholine, phosphoethanolamine, arachidonic acid, and oleic acid could serve as potential metabolic markers. Epigenomics research indicates m6A methylation plays a role in SARS-CoV-2 replication, infection, and transmission, affecting host cell-virus interactions. Among these, N, S, and non-structural proteins 2 and 3 exhibit the most significant ubiquitination. Trends in microbiomics research suggest that microbial communities in the gut and wastewater are emerging as new research focuses. Conclusion The data types of coronavirus omics are diverse, with a wide variety of models and cell types used. The selection of species and technical methods for modelling varies based on the characteristics of different viruses. Multi-omics data from animal models of coronavirus infection can reveal key interactions between hosts and pathogens, identifying biomarkers and potential therapeutic targets, and provide valuable information for a deeper understanding of biological characteristics and infection mechanisms of coronaviruses.

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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
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    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.

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    Analysis of Institutional Characteristics and Implementation of Guangdong Province Laboratory Animals Ordinance in the Past Decade
    DENG Shaochang, LIN Danrong, LIANG Chujun, LEI Weiqiao, YANG Jinchun, ZHAO Weibo
    Laboratory Animal and Comparative Medicine    2024, 44 (4): 455-462.   DOI: 10.12300/j.issn.1674-5817.2024.030
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    The Guangdong Province Laboratory Animals Ordinance (the GPLAO), officially enacted and implemented in October 2010, marked a new phase of legal management in the administration of laboratory animals in Guangdong Province. The GPLAO clearly authorizes the laboratory animal monitoring institutes in Guangdong Province to conduct laboratory animal quality monitoring and assume corresponding legal responsibilities. It also specifies the standards for the production and use of laboratory animals, while bringing attention to laboratory animal welfare and ethics. Through the formulation of supporting documents, the active promotion of the standardization and regulation of administrative law enforcement, and the implementation of measures to delegate power, streamline administration, and optimize government services, significant improvements have been made in the standardized management of laboratory animals in Guangdong Province and in supporting the innovative development of biomedicine. Moreover, notable achievements have been made in areas such as the construction of experimental animal technology platforms, standardization work, the preservation and development of unique resources, and talent pool development. However, with the advancements in science and technology, social economy, and legislative work, some aspects in the GPLAO have been identified for improvement. These include the need for clearer provisions on laboratory animal welfare, more specific guidelines for the management of genetically engineered animals, improved regulations on the safety and welfare of practitioners, further refined implementation of the "one-vote veto system", continued promotion of laboratory animal standardization, and the exploration of effective mechanisms for resource sharing. This paper reviews and summarizes the institutional characteristics, post-legislation implementation, and effectiveness of the GPLAO, as well as the issues and countermeasures identified during the implementation. It concludes the laboratory animals management practices in Guangdong Province during the past 10 years, aiming to provide a reference for laboratory animal legislation.

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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
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    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.

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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
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    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.

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    Screening of Hypoxia-Sensitive and Hypoxia-Tolerant Wistar Rats and Preliminary Exploration of Hypoxia Sensitivity in Their G 1 Generation
    XIAO Pan, WANG Hongyi, LU Lu, ZHANG Mei, CHEN Keming, SHEN Dongshuai, NIU Tingxian
    Laboratory Animal and Comparative Medicine    2024, 44 (4): 374-383.   DOI: 10.12300/j.issn.1674-5817.2024.017
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    Objective By screening and passaging G0 generation Wistar rats, we obtained hypoxia-sensitive and hypoxia-tolerant G1 generation rats, and then the differences in hypoxia sensitivity among these rats were preliminarily explored. Methods 200 Wistar rats (half male and half female) were selected as G0 generation and placed in a controlled oxygen concentration system. The hypoxia tolerance time, which refers to the time from placement to near death, was recorded for the G0 generation rats at an oxygen volume fraction of 3%. 30 rats (half male and half female) with the shortest hypoxia tolerance time were selected for mating and passage to obtain G1 generation hypoxia-sensitive rats. Similarly, 30 rats (half male and half female) with the longest hypoxia tolerance time were selected for mating and passage to obtain G1 generation hypoxia-tolerant rats. An additional 24 standard Wistar rats were randomly divided into two groups: a control group and a model group, with 12 rats in each group (half male and half female). The control group was kept in a normoxic environment, while the model group, along with the G1 generation hypoxia-sensitive rats (G1 sensitive group) and G1 generation hypoxia-tolerant rats (G1 tolerant group), were placed in a hypobaric hypoxia chamber (simulating an altitude of 5 000 m). After 12 hours, various indicators, including blood gas, complete blood count, blood biochemistry, pathological sections, and hypoxia-related genes were detected or observed to compare the differences in hypoxia sensitivity among the 4 groups. Results Compared with the G0 generation standard rats, the hypoxia tolerance time of G1 generation rats was significantly prolonged (P<0.01). Compared with the model group, the oxygen saturation (SatO2) in G1 tolerant group was significantly higher (P<0.05). In the G1 sensitive group, the levels of white blood cell (WBC) count, neutrophil (NEUT) count, hemoglobin (HGB) concentration, hematocrit (HCT), red blood cell distribution width (RDW), platelet (PLT), and creatinine (Cr) significantly increased (P<0.05 or P<0.01), while actual bicarbonate (AB) content significantly decreased (P<0.05), and the brain and lung coefficients were significantly elevated (P<0.05). In addition, pathological section results showed that the brain and lung tissues in the model group, G1 sensitive group, and G1 tolerant group all suffered from significant damage, with no evident differences in the gene expression levels of hypoxia-inducible factor-1 α (HIF-1α) and vascular endothelial growth factor A (VEGFA) in brain tissues among the three groups (P>0.05). Conclusion Compared with standard rats, G1 generation hypoxia-sensitive/tolerant rats exhibit good signs of hypoxia sensitivity/tolerance traits, but further screening and passage are still needed to purify them.

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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
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    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.

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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
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    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.

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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
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    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.

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    Preliminary Study on Construction Method of Type 1 Diabetes Model in Banna Miniature Pig
    LIU Kai, HU Yuqi, GENG Yatian, CHENG Wenjie, WANG Jing, WEI Taiyun, ZHAO Hongfang, LI Jiayu, JIAO Deling, ZHAO Hongye, WEI Hongjiang
    Laboratory Animal and Comparative Medicine    2024, 44 (4): 384-392.   DOI: 10.12300/j.issn.1674-5817.2024.060
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    Objective To construct a Type 1 diabetes model in miniature pigs and explore postoperative care strategies for effectively prolonging the survival time of the model pigs. Methods Seven Banna miniature pigs were selected for pancreatectomy. Glucose, vitamins, and antibiotics were administered for 3-5 days after surgery to aid recovery. Blood glucose and urine glucose levels were measured twice a day in the morning and evening to adjust insulin supplementation accordingly. The model pigs were observed daily and records were kept, including orexis, psychosis, weakness, skin ulcer, and feces and urine. Body weight was measured weekly until the death of the model animals. Based on the model pigs' condition, glucose injection and Ringer's lactate solution were administered to supplement nutrition and correct electrolyte imbalances. Results All seven Banna miniature pigs showed typical symptoms of diabetes: random blood glucose levels higher than 11.1 mmol/L after pancreatectomy, far exceeding the average blood glucose level of 6.0 mmol/L in normal pigs; positive urine glucose; and progressive weight loss. These features indicated the successful construction of Type 1 diabetes model. Additionally, Type 1 diabetic pigs that survived more than 8 weeks showed progressive hair loss and skin ulceration. Euthanasia was performed on model pigs when they were unable to stand or even eat independently, and pathological examination and HE staining were conducted on tissues collected from affected organs such as the liver, kidneys, and skin. Pathological sections revealed liver congestion, massive glycogen accumulation, ballooning degeneration of hepatocytes, and progressive liver fibrosis, along with glomerular congestion, vacuolar degeneration in renal tubular epithelial cells, proteinuria, dermal congestion, thinning of vascular walls, and varying degrees of parakeratosis and dyskeratosis in the liver, kidneys, and skin tissues due to prolonged hyperglycemia. The average survival time of the constructed Banna miniature pig diabetes model was 44 d, with a maximum survival time of 121 d. Conclusion Type 1 diabetes model can be constructed successfully in Banna miniature pigs through pancreatectomy. With meticulous postoperative care, a long-term Type 1 diabetes model with significant complications can be achieved, providing a stable large-animal model for Type 1 diabetes treatment strategies.

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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
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    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.

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    Analysis of Breeding and Application Data for Laboratory Cats
    WU Xiansheng, HUANG Wei, LIANG Yongfen, DENG Hui, ZHAI Yonghuan, YANG Jiajun, HUANG Ganquan, WANG Gang
    Laboratory Animal and Comparative Medicine    2024, 44 (4): 428-435.   DOI: 10.12300/j.issn.1674-5817.2024.033
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    Objective To cultivate and breed laboratory cats in conventional laboratory animal facilities, collect background data on laboratory cats, and compare them with purchased domestic cats to assess the feasibility of breeding laboratory cats. Methods Indigenous cat breeds were introduced for reproduction and population expansion under conventional laboratory environment, with recording of kitten survival rates and growth curves. Indicators of 20 laboratory cats of F1 generation (half male and half female), including complete blood count, blood biochemistry, organ mass, organ coefficient, heart rate, and blood pressure, were detected and comparisons between sexes were made. Blood pressure values and sensitivity to histamine of these cats were measured using depressor substance detection method in the Pharmacopoeia of the People's Republic of China-Four Parts: 2020, and were compared with the data from 173 concurrently purchased domestic cats. Results Laboratory cats adapted well to the environment of conventional laboratory facilities, with a survival rate of 77.08% of kittens at 8 weeks of age. Red blood cell count, hemoglobin content, mean corpuscular hemoglobin concentration, and hematocrit in male laboratory cats were significantly higher than those in females (P<0.01), while the mean corpuscular volume in males was significantly lower than that in females (P<0.01). The levels of serum alanine aminotransferase, total bilirubin, creatinine, triacylglycerol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol in male laboratory cats were significantly higher than those in females (P<0.05 or P<0.01), while cholesterol, globulin, total protein, and the albumin-globulin ratio were significantly lower in males (P<0.01). The liver coefficient in male laboratory cats was significantly lower than that in female cats (P<0.05), while the kidney coefficient was significantly higher (P<0.05). The spleen-brain and kidney-brain ratios were significantly higher in males compared to females(P<0.05 or P<0.01). No significant differences were found in heart rate, systolic pressure, diastolic pressure, mean blood pressure, or sensitivity to histamine between male and female laboratory cats (P>0.05). Compared to laboratory cats, purchased domestic cats had significantly higher heart rate, systolic pressure, and mean blood pressure (P<0.01), and the magnitude of blood pressure changes induced by medium and high doses of histamine was significantly reduced (P<0.05 or P<0.01). Conclusion It is feasible to breed laboratory cats in conventional laboratory animal facilities. The accuracy of experimental results can be improved by using laboratory cats with clear and standardized background data.

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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
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    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.

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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
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    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.

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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
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    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.

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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
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    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.

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

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    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
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    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.

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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
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    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.

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    Pathological Diagnosis of Systemic Amyloidosis in a New Zealand White Rabbit
    LIN Qingqing, DAI Jinlong, CHEN Zhisen, GUO Jianmin, YANG Wei
    Laboratory Animal and Comparative Medicine    2024, 44 (6): 695-699.   DOI: 10.12300/j.issn.1674-5817.2024.092
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    Objective To provide a reference for the diagnosis of amyloidosis in experimental animals through the pathological diagnosis of systemic amyloidosis in a case of a New Zealand white rabbit. Methods In a 6-month repeated ocular toxicity study, an abnormal finding was noted during the routine gross anatomical examination of one New Zealand white rabbit. Its organs were prepared as paraffin sections and stained with hematoxylin-eosin (HE) staining and Congo red staining. The histopathological features were observed under optical and polarized light microscopy. Results Gross anatomical examination of the animal revealed an enlarged spleen and changes in the color and texture of the lung. HE staining showed that the splenic tissue structure was destroyed, the white pulp of the spleen was surrounded by dense amyloid deposition in the form of nodular rings, along with pressure atrophy of the white pulp. Amyloid deposits were also observed in the submandibular lymph nodes, mesenteric lymph nodes, ileum, sacculus rotundus, vermiform appendix, jejunum, cecum, and rectum. Congo red staining showed that the amyloid deposition in the affected organs appeared salmon-pink, and exhibited characteristic apple green birefringence under polarized light microscopy.Conclusion The histo-pathological features of the New Zealand white rabbit are consistent with the diagnostic characteristics of systemic amyloidosis.

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

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