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

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

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

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

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

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

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

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

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

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

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

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    Design and Implementation of a Software System for Full-Process Animal Experiment Ethical Review
    XU Xiao, WU Jiemin, LIANG Yi, YE Mingxia, ZHOU Shunchang
    Laboratory Animal and Comparative Medicine    2024, 44 (6): 682-690.   DOI: 10.12300/j.issn.1674-5817.2024.086
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    Institutions of higher education undertake a significant number of animal experiment projects with diverse types. To standardize animal experiments and ensure animal welfare, ethical review has become a core task for the Institutional Animal Care and Use Committee (IACUC) in these higher education institutions. A key issue worth exploring is how to improve the efficiency of animal ethical reviews while maintaining systematic quality of supervision. Based on the current status of ethical reviews at the Laboratory Animal Center of Huazhong University of Science and Technology, this paper designs and implements a full-process ethical review software system. This system seeks to offer solutions for the ethical review and supervision of animal welfare in institutions of higher education through information technology. The system employs a full-process supervision philosophy of "pre-approval, in-process supervision, post-review," focusing on strengthening the review and approval of animal research protocols, as well as post-approval monitoring. At the pre-approval stage, the system optimizes the process for designated members involved in the project review, ensuring that all projects comply with ethical standards and legal regulations. At the in-process stage, veterinary verification and consultation are carried out by executive veterinarians, using a method of "offline inspections and online records" for supervision. At the post-review stage, researchers are required to provide retrospective data to evaluate the experimental process. Additionally, this paper provides an in-depth exploration of the software, detailing its functional and non-functional requirements, as well as its security considerations. It also delves into the comprehensive software architecture, process design, and operational details. Furthermore, it illustrates the system's operational efficacy and impact since its launch. This paper discusses the software's applicability and practicality, analyzing existing limitations and challenges, such as the difficulty of balancing experiment urgency with the stringent nature of review processes, and the traceability blind spots in post-approval supervision. Future research will continue to enhance the accuracy and strength of animal welfare ethical reviews and supervision through new technologies and methods.

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

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

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

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

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    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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    Comparison of the Fixation Effects of Six Composite Fixatives on Retinal Tissue of Golden Hamsters
    WU Haifeng, ZHOU Xiaojiang, LI Chenjiang, LI Huaiyin, GAO Ming
    Laboratory Animal and Comparative Medicine    2024, 44 (6): 675-681.   DOI: 10.12300/j.issn.1674-5817.2024.049
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    Objective To compare the effects of six composite fixatives on paraffin sections of golden hamster retinal tissue, and to optimize the fixation methods for retinal tissue paraffin sections of golden hamsters. Methods Eighteen male SPF grade golden hamsters were taken and randomly divided into six groups of three animals each. After each animal was anesthetized by intraperitoneal injection of sodium pentobarbital, cardiac perfusion was performed using 4% paraformaldehyde, Bouin's, Carnoy, Davidson's, Zenker, and Helly fixatives, respectively. The eye tissues of each animal were taken out to make eye cups and put into the corresponding compound fixative solution for fixation, and then taken out for paraffin embedding after 48 h. The embedded blocks were sliced using microtome, and then stained with hematoxylin and eosin (HE). The morphological characteristics of retinal tissue cells were observed under a light microscope and scored in a double-blind method to statistically analyze the fixation effect and staining quality of various composite fixative solutions. Results Retinal sections fixed with Davidson's fixative exhibited intact morphology without breaks, clear structural layers, well-morphosed nuclei, and tight adhesion between the retina, sclera, and uvea. In contrast, sections fixed with 4% paraformaldehyde, Bouin's, and Carnoy fixatives showed varying degrees of retinal breaks and detachment from the sclera. Sections fixed with Zenker and Helly fixatives demonstrated the poorest quality, characterized by pronounced detachment, large fissures, unclear cell layers, and pale staining. Statistical analysis using SPSS 27.0 software revealed significant differences in mean scores among the six fixatives (P<0.001). The fixation quality ranking was as follows: Davidson's fixative > 4% paraformaldehyde > Bouin's fixative > Carnoy fixative > Helly fixative > Zenker fixative. Zenker and Helly fixatives showed significantly lower scores than the other fixatives (P<0.001), while no significant differences were observed among the remaining fixatives (P>0.05). Conclusion Davidson's fixative provides the best fixation and staining results, followed by 4% paraformaldehyde and Bouin's fixative. Carnoy fixative exhibits suboptimal performance, while Zenker and Helly fixatives result in the poorest outcomes. Therefore, Davidson's fixative is recommended as the optimal fixative for golden hamster retinal tissue, with 4% paraformaldehyde, Bouin's and Carnoy fixatives as alternative options.

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

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    Survey Report Analysis on Parasitic and Microbial Quality of Laboratory Animals in Sichuan Province, 2017-2023
    LIU Lida, CHEN Bing, XIE Na, LIU Li, ZHUANG Siqi, ZOU Yixing
    Laboratory Animal and Comparative Medicine    2024, 44 (6): 654-660.   DOI: 10.12300/j.issn.1674-5817.2024.057
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    Objective To provide reference for further strengthening the management of laboratory animals in Sichuan Province by reviewing and analyzing the results of parasitic and microbial quality inspections of laboratory animals from 2017 to 2023. Methods Sichuan Province has 31 licensed laboratory animal production units, with the main species including mice, rats, guinea pigs, rabbits, dogs, monkeys, and pigs. Sampling inspections and reports were conducted for units with laboratory animal production qualifications in accordance with current national and local standards for laboratory animal classification. The quality inspection results for various laboratory animals in Sichuan Province from 2017 to 2023 were analyzed. Results With the exception of 2018, annual quality inspections of laboratory animals were conducted every year between 2017 and 2023. Mice: In 2017, positive results for helminths, flagellates, Salmonella, murine hepatitis virus antibodies, and murine pneumonia virus antibodies were detected, with a pass rate of only 42.9%. In 2019, Staphylococcus aureus and Klebsiella pneumoniae were detected, with a pass rate of 86.7%. In 2021, Sendai virus antibodies were detected, yielding a pass rate of 85.7%.The pass rate in 2020, 2022, and 2023 was 100%. Rats: In 2017, positive results were found for helminths, mycoplasma antibodies, Staphylococcus aureus, Sendai virus antibodies, murine pneumonia virus antibodies, rat parvovirus RV strain antibodies, rat parvovirus (H-1) strain antibodies, and rat coronavirus antibodies, with a pass rate of 40.0%. In 2019, mycoplasma antibodies, Staphylococcus aureus and Klebsiella pneumoniae were detected, with a pass rate of 35.0%. No positive indicators were detected in 2020. In 2021, Sendai virus antibodies and rat parvovirus RV strain antibodies were detected, with a pass rate of 50.0%. In 2022, positive results for rat parvovirus RV strain antibodies were found, yielding a pass rate of 87.5%. In 2023, Pasteurella pneumotropica and Klebsiella pneumoniae were detected, with a pass rate of 85.7%. Dogs: In 2017 and 2019, the antibody titers for rabies virus and canine distemper virus were below the required standard, with pass rates of 33.3% and 20.0%, respectively. In 2020 and 2022, the pass rate was 100%. Guinea pigs (general grade): In 2019, positive results for Toxoplasma antibodies were detected, with a pass rate of 80.0%. In all other years, the pass rate was 100%. Monkeys: In 2019, positive results for Toxoplasma gondii and rhesus herpesvirus type I antibodies were found, with a pass rate of 87.5%.In 2020 and 2022, rhesus herpesvirus type I antibodies were positively detected, yielding pass rates of 93.3% and 97.5%, respectively. The pass rates in 2021 and 2023 were 100%. Clean guinea pigs, rabbits and pigs all passed the inspection each year. Conclusion Issues related to the parasitic and microbial quality of laboratory animals persist in Sichuan Province. Supervision and sampling inspections have proven to be effective in identifying these issues promptly, serving as a critical measure to ensure the quality of laboratory animals. The results of these inspections offer valuable data to support the healthy development of the laboratory animal industry in Sichuan Province.

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