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History, Current Status, Challenges and Opportunities of Laboratory Monkey Industry in China SUN Qiang Laboratory Animal and Comparative Medicine    2024, 44 (4): 343-356.   DOI: 10.12300/j.issn.1674-5817.2024.112 Abstract （1654）   HTML （756）    PDF （1167KB）（3238）       Knowledge map    Save Laboratory animals play a crucial role in foundational scientific research and clinical medicine. Non-human primates (NHP), particularly Macaca mulatta and Macaca fascicularis, have long been highly valued due to their close resemblance to humans. After more than half a century of development, China's NHP laboratory animal industry has gradually transitioned from its early stage of rapid and unregulated growth to a mature stage of standardization and refinement. However, there has been a dramatic surge in global biopharmaceutical research in recent years, leading to a sharp increase in demand for NHP laboratory animals. This surge, coupled with the lack of long-term strategic planning among breeding enterprises, has resulted in severe aging of breeding populations and a significant decline in reproductive capabilities, further widening the supply gap. Under the dual pressures of rising demand and declining supply, the prices of NHP laboratory animals have surged. Although the cyclical downturn in the biopharmaceutical industry in recent years has lowered the demand for NHP laboratory animals to some extent, leading to significant price drops, the prices remain high. At the same time, against the backdrop of high prices, issues such as the accelerating aging of breeding populations, the lower standards for microbial quality control, insufficient genetic quality control, and blind investment in facility construction have emerged within the NHP laboratory animal industry. This report provides a comprehensive review of the history and current status of China's NHP laboratory animal industry, with a focus on laboratory monkeys. It explores the factors shaping the current industry landscape and identifies potential challenges and opportunities facing the industry. It aims to offer insights and references for the future development of China's NHP laboratory animal industry. Table and Figures | Reference | Related Articles | Metrics

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Three Dimensions of Animal Experiment Ethics: Analysis Based on Value of Life, Animal Welfare, and Risk Prevention ZHAO Yong Laboratory Animal and Comparative Medicine    2024, 44 (4): 445-454.   DOI: 10.12300/j.issn.1674-5817.2024.108 Abstract （1310）   HTML （65）    PDF （1033KB）（2890）       Knowledge map    Save Ethical review of animal experiments is based on factors such as the necessity of the experiment, its scientific validity, the professional competence of the experimenters, and research conditions, to determine the ethical permissibility of an animal experiment. Attitudes towards laboratory animals and animal experiments vary significantly due to differences in cultural traditions, religious beliefs, personality traits, and roles within the experimentation process. How animal experiment ethics can advance in tandem with the advancements of life sciences, while consistently providing forward-looking guidance and safeguarding fundamental ethics, is a subject that requires continuous exploration, practice, and research. Ensuring the welfare of laboratory animals has now become a widely accepted ethical consensus. However, attitudes and principles towards different types of animal experiments, and the methods to genuinely and effectively ensure the welfare quality of animals during live animal experiments, should be central to animal experiment ethics. Based on the deep integration of biotechnology in the field of laboratory animals, this paper considers the contribution of animal experimentation to the development of scientific ethical concepts. It systematically elucidates the special value of life in laboratory animals and their relationship with biosafety and ecological safety. It also outlines welfare quality assessment methods for different species of laboratory animals, demonstrating that the philosophical ideology of the value of life is the core of animal experiment ethics. The quality and significance of animal experiments determine the ethical level at which the value of life in laboratory animals is realized, and animal welfare technologies provide a robust ethical guarantee for animal experiments. The close integration of ethical theories with life sciences in laboratory animals is an objective requirement for animal experiment ethics. The value of life, animal welfare, and risk prevention collectively form the core elements of ethical review in animal experiments, serving as fundamental factors in improving the quality of ethical reviews and avoiding ethical deviations. Reference | Related Articles | Metrics

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Advantages of Ciona intestinalis as a Model Organism and Its Applications Ruiqi LI, Han DUAN, Luo GAN, Yuan ZHENG, Wen YANG Laboratory Animal and Comparative Medicine    2024, 44 (2): 162-179.   DOI: 10.12300/j.issn.1674-5817.2023.159 Abstract （937）   HTML （45）    PDF （2131KB）（2840）       Knowledge map    Save With the continuous development of experimental biology, the limitations of commonly utilized model organisms are becoming increasingly apparent. Discrepancies between research conducted on laboratory animals and humans significantly impede the translational application of findings derived from animal experiments. This review introduces ascidian Ciona intestinalis as a novel model organism, an invertebrate that is evolutionarily closest to vertebrates and is a sister group to vertebrates. The review summarizes recent research progress on Ciona intestinalis in various fields to illustrate the significant advantages and promising application prospects of it as a model organism. The research progress outlined in the review mainly encompasses: (1) The whole-genome sequencing of Ciona intestinalis has been determined and numerous related databases have been established. Various embryonic gene editing technologies have been successfully applied, making it an animal model easy to manipulate genetically and study the functions and interactions of target genes visually. (2) In the field of neurobiology, Ciona intestinalis boasts a central nervous system structure similar to that of vertebrates and possesses numerous homologous neuropeptides and hormone molecules. These features grant it an edge in exploring the mechanisms and functional evolution of endocrine and neuroendocrine-related molecules. Additionally, the sensitivity and habituation of its larvae to light stimulation provide an avenue for exploring mechanisms related to behavioral plasticity. (3) In the field of immunology, Ciona intestinalis possesses a mature innate immune system and has evolved precursor genes to the adaptive immune system, with a relatively simple coding of immune-related genes. These features make it an exemplary model organism for immunological studies. (4) In the field of developmental biology, many studies have focused on the notochord development process in Ciona intestinalis and the regulatory mechanisms of gene expression within it, indicating common evolutionary developmental strategies among chordates. Additionally, insights into its heart development also significantly enhance our comprehension on the genetic network of human heart development. (5) In medical research, the ability of Ciona intestinalis to regenerate its neural complex and siphon, as well as the resilience of its heart to recover contractile function from substantial damage, renders it a valuable animal model for the study of regeneration and heart injury. It also has unique advantages as a research model for Alzheimer's disease and new drug development. Furthermore, its brief five-month lifespan facilitates the observation and recording of the entire aging process and the exploration of the effects of various factors on aging. In summary, this review aims to demonstrate that Ciona intestinalis stands out as a model organism with unique attributes and is expected to play a significant role in a wider range of scientific research areas. Table and Figures | Reference | Related Articles | Metrics

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Guidelines for the Selection of Animal Models and Preclinical Drug Trials for Spontaneous Intracerebral Hemorrhage (2024 Edition) Committee of Experts on Medical Animal Experiments, Chinese Research Hospital Association Laboratory Animal and Comparative Medicine    2024, 44 (1): 3-30.   DOI: 10.12300/j.issn.1674-5817.2024.001 Abstract （1203）   HTML （258）    PDF （1252KB）（2823）       Knowledge map    Save Spontaneous intracerebral hemorrhage (sICH), the most prevalent and lethal subtype of stroke, is characterized by spontaneous hemorrhage in the brain parenchyma. Presently, there are no effective methods for preventing and treating sICH. The existing sICH animal models can be broadly categorized into three classes: (1) induced intracerebral hemorrhage models, including autologous blood injection model, collagenase injection model, microballoon inflation model, and hyperglycemia-induced sICH hematoma expansion model; (2) spontaneous hypertensive intracerebral hemorrhage models mainly include stroke-prone spontaneously hypertensive rats (SHRsp) and stroke-prone renovascular hypertensive rats (RHRsp); (3) gene-modified models encompassing transgenic hypertensive intracerebral hemorrhage, transgenic cerebral amyloid angiopathy, arteriovenous malformation-related, cerebral cavernous malformation-related and collagen-related genetically modified animal models for sICH. These models contribute not only to unraveling the pathogenesis of sICH and exploring preventive or therapeutic interventions, but also serve as invaluable tools for conducting preclinical drug trials to advance novel treatments. This guide comprehensively reviews sICH pathogenesis, delineates the superiority and inferiority of different species of modeling animals, explains the modeling principles and techniques for various sICH animal models, elucidates the technical details of animal model production, summarizes the pathophysiological mechanism simulated by the models and their clinical relevance, outlines the neurobehavioral evaluation methodologies for sICH animal models, compares the advantages and disadvantages of various models, and suggests their applicable research areas. Additionally, it underscores critical considerations in the design of preclinical drug trials for sICH. Table and Figures | Reference | Related Articles | Metrics

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Construction and Evaluation of a Rat Model of Abnormal Uterine Bleeding LIAN Hui, JIANG Yanling, LIU Jia, ZHANG Yuli, XIE Wei, XUE Xiaoou, LI Jian Laboratory Animal and Comparative Medicine    2025, 45 (2): 130-146.   DOI: 10.12300/j.issn.1674-5817.2024.132 Abstract （624）   HTML （43）    PDF （4039KB）（2818）       Knowledge map    Save Objective By simulating the etiology of abnormal uterine bleeding-ovulatory dysfunction (AUB-O) and establishing a rat model of abnormal uterine bleeding (AUB), this study aims to provide an experimental platform for investigating pathological mechanisms and developing therapeutic drugs for AUB. Methods After acclimation, 24 adult (10-week-old) female SD rats were randomly divided into a normal control group (6 rats) and a model group (18 rats). The normal control group was housed in a barrier environment, while the model group underwent bilateral ovariectomy via dorsal approach in the same environment and rested for one week before starting to receive modeling drugs. In the model group, from Days 1 to 3 of modeling, each rat received a daily subcutaneous injection of 0.5 mg estradiol into the dorsal region. From Days 4 to 7, a daily subcutaneous injection of 5.0 mg progesterone was administered. On Day 6, rats received bilateral injections of 0.5 mL soybean oil per uterine cavity (total 1.0 mL) via the same dorsal surgical incision. On Day 8, mifepristone (10 mg/kg) was administered via oral gavage. The estrous cycle stage and its dynamic changes were continuously monitored during modeling. Uterine bleeding was recorded during the 48-hour observation period post-modeling. Serum and uterine tissue samples were collected from the model group at 0, 12, 24, 36, and 48 h after mifepristone administration, while the normal control group was sampled at 36 h. The samples were subjected to HE staining, serum sex hormone ELISA, immunohistochemistry, TUNEL apoptosis staining, Western blotting, transcriptome sequencing, and bioinformatics analysis for comprehensive evaluation of the AUB rat model. Results The AUB rats exhibited uterine bleeding, endometrial detachment and injury, incomplete uterine restoration, inflammatory cell infiltration in the endometrium, enhanced tissue apoptosis, and structural damage of the stroma, glands, and vasculature. Compared with the normal control group, the levels of serum follicle-stimulating hormone (FSH), estradiol, and luteinizing hormone (LH) were significantly increased in the AUB rats (P<0.05). The vascular density of the endometrium was significantly reduced (P<0.05). The expression of vascular endothelial growth factor (VEGF) was qualitatively observed to be markedly enhanced at the site of endometrial detachment but significantly decreased around the stromal blood vessels (P<0.01). Matrix metalloproteinase-9 (MMP-9) expression was qualitatively observed to be strongly upregulated at the site of endometrial injury but significantly reduced in the non-detached stroma and glands (P<0.01). Endometrial stromal cell apoptosis was significantly enhanced (P<0.01). The expression levels of fibroblast growth factor 2 (FGF2) and endothelin-1 (ET-1) in uterine tissues were significantly decreased (P<0.05). After comparing the transcriptome sequencing results of uterine tissues between AUB and normal rats, a total of 4 723 differentially expressed genes were identified, including 2 191 up-regulated genes and 2 532 down-regulated genes. KEGG enrichment analysis revealed that these differentially expressed genes were significantly enriched in pathways related to inflammation, immune apoptosis, cell signal transduction, proliferation and differentiation, and muscle contraction, among others. Conclusion An AUB rat model can be successfully established using a sequential administration protocol of estrogen, progesterone, and mifepristone to simulate the etiology of AUB-O. In this model, endometrial injury is associated with inflammation and apoptosis, with pathological manifestations influenced by abnormal vasoconstriction and impaired endometrial regeneration. This rat model closely recapitulates pathological characteristics of non-structural AUB observed in clinical practice, making it a validated experimental platform for exploring the pathological mechanisms and therapeutic interventions of non-structural AUB. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in Establishment and Evaluation of Common Asthma Animal Models LUO Shixiong, ZHANG Sai, CHEN Hui Laboratory Animal and Comparative Medicine    2025, 45 (2): 167-175.   DOI: 10.12300/j.issn.1674-5817.2024.120 Abstract （984）   HTML （47）    PDF （846KB）（2790）       Knowledge map    Save Bronchial asthma (hereinafter referred to as asthma) is a common chronic respiratory disease characterized by airway inflammation, airway hyperresponsiveness, and airway remodeling. Its pathogenesis is highly complex and heterogeneous, involving multiple factors such as genetics, immunity, and environmental exposure. Currently, therapeutic options for asthma remain relatively limited, making it an urgent priority to explore its underlying mechanisms, identify effective treatment strategies, and develop new drugs. In this context, the establishment of animal models for asthma plays an irreplaceable and crucial role. However, to date, no single ideal animal model has been able to fully and accurately replicate all the features of the onset and progression of human asthma. This study systematically reviews the research progress over the past five years in the establishment methods of asthma animal models. It provides a detailed overview of commonly used experimental animals (such as mice, rats, and guinea pigs), frequently used sensitizing agents (including ovalbumin, house dust mite, lipopolysaccharide, and toluene diisocyanate), and the methods for establishing asthma models using these animals and sensitizers. This study also presents an objective evaluation of the advantages, limitations, and applicability of each model. Evaluation criteria for asthma models are summarized across multiple dimensions, including behavioral assessments, pulmonary function, histopathology, immunological indicators, and pharmacodynamics. Although methods for establishing refractory asthma models remain underdeveloped, several strategies for modeling refractory asthma have been summarized through a review of relevant literature, aiming to provide useful references for related research. Based on current scientific and technological advancements, it is anticipated that future research on asthma animal models will focus more on clinical relevance, technological innovation, and multidisciplinary integration. Specifically, future models are expected to adopt multi-sensitizer induction protocols, apply cutting-edge tools such as gene editing, enhance clinical relevance and promote diversification and personalization of models. Furthermore, advanced technologies such as bioimaging and biosensing are anticipated to enable dynamic monitoring of airway inflammation and remodeling. Organ-on-a-chip platforms may also be explored as potential alternatives to traditional animal models. The ultimate goal is to develop multifactorial, composite models that better simulate the complexity and heterogeneity of human asthma. Table and Figures | Reference | Related Articles | Metrics

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

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

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

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Construction of Dmd Gene Mutant Mice and Phenotype Verification in Muscle and Immune Systems Min LIANG, Yang GUO, Jinjin WANG, Mengyan ZHU, Jun CHI, Yanjuan CHEN, Chengji WANG, Zhilan YU, Ruling SHEN Laboratory Animal and Comparative Medicine    2024, 44 (1): 42-51.   DOI: 10.12300/j.issn.1674-5817.2023.089 Abstract （847）   HTML （50）    PDF （1936KB）（2278）       Knowledge map    Save Objective The aim is to utilize CRISPR/Cas9 gene editing technology to construct Dmd gene mutant mice with a point mutation in exon 23 of the Dmd gene. Subsequently, the phenotypic changes of the mice in muscles and immune systems are analyzed and verified, providing an evaluation model for Duchenne muscular dystrophy and other related diseases. Methods Based on the sequence characteristics of exon 23 of the Dmd gene, small guide RNA (sgRNA) was designed and synthesized. Cas9 mRNA, sgRNA fragments, and oligo donor DNA were microinjected into fertilized eggs of C57BL/6J mice. After transferring the fertilized eggs to surrogate mice, F0 generation mice were born. After mating with F0 generation mice, offspring mice were obtained, and Dmd gene positive mutant (DmdMu/+) mice were obtained after genotype identification. Male hemizygous DmdMu/+(DmdMu/Y) mice were selected for phenotype validation. The body weight of live 3- and 9-month-old mice were recorded. Muscle tension was evaluated through the grid test. Hearts and semitendinosus muscles were collected, and the histopathological changes were observed using HE staining. Further, the expression of Dmd protein in muscle tissue of 9-month-old mice was analyzed by Western blotting. An acute inflammation model was established in DmdMu/Y mice using lipopolysaccharide induction. Peripheral blood from the submandibular vein was collected, and the changes in the proportion of neutrophils and monocytes were detected by flow cytometry. Results The results of genome sequencing and Western blotting confirmed the successful construction of Dmd gene point mutant mice (DmdMu/+ mice). Dmd protein expression was not detected in skeletal muscle and myocardium of DmdMu/+ mice, and it was significantly reduced compared to wild-type C57BL/6J mice (P<0.05). Compared with wild-type mice of the same background, DmdMu/Y mice at 3 and 9 months of age showed significant weight loss (P<0.01) and decreased muscle tension (P<0.05). 9-month-old DmdMu/Y mice exhibited significant pathological changes in skeletal muscle and myocardium, including widening of intermuscular space. Under normal condition, compared with wild-type mice, the proportion of neutrophils and monocytes in the peripheral blood of 3-month-old DmdMu/Y mice was significantly lower than that of wild-type mice (P<0.01). After lipopolysaccharide stimulation, the proportion of neutrophils in peripheral blood of 3-month-old DmdMu/Y mice remained significantly lower compared to that of wild-type mice (P<0.01). The proportion of neutrophils in peripheral blood of 9-month-old DmdMu/Y mice significantly decreased after lipopolysaccharide induction (P<0.01), with a trend of change observed in monocytes between groups. Conclusion The successful construction of the Dmd gene mutant mouse model has confirmed the vital function of Dmd gene in maintaining normal muscle tissue morphology and muscle tone. It preliminarily indicated that Dmd gene deletion could significantly reduce the proportion of neutrophils in peripheral blood, offering a new perspective for the study of immune system alterations in Duchenne muscular dystrophy patients. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Animal Models of Gastric Ulcer of Spleen-Stomach Deficiency Cold Type LIU Ziqi, LI Yunying, LI Qin, LI Yuanhan, HE Fangyan, WEN Weibo Laboratory Animal and Comparative Medicine    2025, 45 (5): 574-585.   DOI: 10.12300/j.issn.1674-5817.2025.015 Abstract （315）   HTML （7）    PDF （919KB）（2271）       Knowledge map    Save Gastric ulcer (GU) is one of the common, frequently-occurring and intractable diseases of the digestive system. Spleen-stomach deficiency cold type is the most common and hard-to-cure syndrome pattern of GU, and is both a focus and a challenge in medical research. Therefore, constructing a scientific, reasonable, and clinically practical animal model of GU with spleen-stomach deficiency cold type and formulating objective and effective evaluation criteria are of great significance for in-depth research on the pathogenesis and treatment of GU. In this paper, the methods for constructing GU animal models of spleen-stomach deficiency cold type are comprehensively introduced by systematically reviewing the relevant literature. Firstly, the construction methods of pathological models of GU in Western medicine are introduced, including pyloric ligation method, water immersion-restraint stress method, ethanol-induced method, acetic acid-induced method, etc. This paper expounds the establishment methods for spleen-stomach deficiency cold syndrome type model in traditional Chinese medicine (TCM), including diet disorder method, bitter cold diarrhea method, excessive fatigue method, Qi consumption and Qi impairment method, and overeating sour-flavor method. This paper focuses on the construction methods for disease-syndrome combination GU models of spleen-stomach deficiency cold type, including two-factor modeling method and three-factor modeling method. Meanwhile, the evaluation indices of GU animal models of spleen-stomach deficiency cold type were summarized from various aspects, including animal physical signs ( appearance symptoms, animal behavior, and metabolic indices), as well as tissue morphology and molecular biology-related indicators ( gastric function, oxidative stress, inflammatory factors, other cytokines, four coagulation parameters, intestinal flora detection ), for constructing a comprehensive evaluation system. From the perspective of prescription-based verification, this paper further analyzes the drug composition and pharmacological effects to infer the syndrome type of the treated animal model, so as to verify whether the target animal model is successfully constructed. This review aims to provide a valuable reference for establishing a syndrome-specific GU animal model that closely aligns with clinical reality and embodies the principles of Chinese medicine. This will further advance research on TCM-pattern GU syndromes and deepen the exploration of herbal medicine-based treatments for GU, ultimately promoting the clinical integration and advancement of Chinese medicine in GU therapy. Table and Figures | Reference | Related Articles | Metrics

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Analysis of Animal Models of Myasthenia Gravis Based on Its Clinical Characteristics in Chinese and Western Medicine CHEN Yuhan, CHEN Jinling, LI Xin, OU Yanhua, WANG Si, CHEN Jingyi, WANG Xingyi, YUAN Jiali, DUAN Yuanyuan, YANG Zhongshan, NIU Haitao Laboratory Animal and Comparative Medicine    2025, 45 (2): 176-186.   DOI: 10.12300/j.issn.1674-5817.2024.139 Abstract （582）   HTML （17）    PDF （882KB）（2214）       Knowledge map    Save Myasthenia gravis (MG) is an autoimmune disease characterized primarily by skeletal muscle weakness and, in severe cases, respiratory involvement. Western medical treatment predominantly relies on immunosuppressants, but long-term administration often leads to notable side effects. In contrast, traditional Chinese medicine (TCM) offers the advantage of multi-target interventions. However, the pathogenesis of MG has not been fully elucidated, and the establishment of animal models that accurately reflect the clinical characteristics of both Chinese and Western medicine is essential for mechanism research and new drug development. This paper systematically reviews the etiology and pathogenesis, diagnostic criteria, and progress of animal model research for MG from both Chinese and Western medicine perspectives. In Western medicine, the pathogenesis of MG is closely related to genetic susceptibility, environmental factors, and autoantibody-mediated postsynaptic membrane damage. In TCM, MG is classified under the category of "flaccidity syndrome", attributed to congenital deficiencies and acquired malnourishment. Western diagnostic criteria involve a combination of clinical symptoms, fatigue testing, serum antibody assays, and electrophysiological evaluation. In contrast, TCM diagnosis emphasizes the integration of primary and secondary symptoms with tongue and pulse pattern differentiation. Currently available animal models mainly include experimental autoimmune myasthenia gravis (EAMG) and passive transfer myasthenia gravis (PTMG). The Toredo acetylcholine receptor (AChR) induced EAMG model aligns well with Western diagnostic criteria, but poorly matches secondary symptoms in TCM. The synthetic AChR peptide model is widely used, but shows low conformity with TCM syndromes. Models induced by muscle-specific tyrosine kinase (MuSK), low-density lipoprotein receptor-related protein 4 (LRP4), and transgenic models demonstrate high innovation but exhibit low clinical conformity. Evaluation of these models requires integration of behavioral, electrophysiological, and immunological indicators. However, a systematic framework for modelling TCM syndromes is still lacking. Future research should integrate TCM-based etiological modelling methods with the Western pathological mechanisms to construct disease-syndrome combination models. Additionally, it is crucial to establish a TCM syndrome evaluation system based on "validation by prescription", as well as to improve the scientific rigor and practicality of animal models by the incorporation of emerging technologies. This review provides a theoretical foundation for optimizing MG animal model design, advancing the research on the combination of Chinese and Western medicine, and supporting efficacy assessment and mechanism exploration of Chinese herbal prescriptions. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Animal Models of Long Bone Fractures Guangyuan YAO, Ping DONG, Hao WU, Mei BAI, Ying DANG, Yue WANG, Kai HU Laboratory Animal and Comparative Medicine    2024, 44 (3): 289-296.   DOI: 10.12300/j.issn.1674-5817.2023.183 Abstract （718）   HTML （30）    PDF （777KB）（2197）       Knowledge map    Save Traumatic fractures and stress fractures are common orthopedic diseases, and there is great potential in researching bone turnover, repair, and promotion of fracture healing. Basic medical experiments often use animal models of long bone fractures in limbs to study the mechanisms of various interventions on fracture healing. Fracture healing is a complex process influenced by multiple factors and involves multiple molecules and pathways. Therefore, to explore the mechanisms more deeply, accelerate the translation of results, and improve the clinical efficacy, it is particularly important to choose the appropriate animal fracture modeling methods in experimental research. Based on this, this paper conducts a literature review of animal species and modeling methods commonly used for long bone fracture models in experimental research. It summarizes five methods: bone defect method, physical impact method, mechanical bending method, open osteotomy method, and drilling method. A side-by-side comparison of their advantages, disadvantages, and scope of application is made, aiming to provide suitable fracture models for studying the mechanisms of fracture healing interventions. Table and Figures | Reference | Related Articles | Metrics

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Interpretation and Elaboration for the ARRIVE Guidelines 2.0—Animal Research: Reporting In Vivo Experiments (V) Zhengwen MA, Xiaying LI, Xiaoyu LIU, Yao LI, Jian WANG, Jin LU, Guoyuan CHEN, Xiao LU, Yu BAI, Xuancheng LU, Yonggang LIU, Wanyong PANG, Yufeng TAO Laboratory Animal and Comparative Medicine    2024, 44 (1): 105-114.   DOI: 10.12300/j.issn.1674-5817.2023.146 Abstract （668）   HTML （60）    PDF （1051KB）（2014）       Knowledge map    Save Improving the reproducibility of biomedical research results is a major challenge. Transparent and accurate reporting of the research process enables readers to evaluate the reliability of the research results and further explore the experiment by repeating it or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), provide a checklist that is applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as enhance the reliability, repeatability, and clinical translation of animal experimental results. The use of the ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and completeness of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is based on the best practices following the ARRIVE 2.0 guidelines in international journals, and it interprets, explains, and elaborates in Chinese the fifth part of the comprehensive version of the ARRIVE 2.0 guidelines published in PLoS Biology in 2020 (the original text can be found at https://arriveguidelines.org ). This section includes the items 6-11 of Recommended 11 section, covering "Animal Care and Monitoring", "Interpretation/Scientific Implications", "Generalisability/Translation", "Protocol Registration", "Data Access" and "Declaration of Interests". Its aim is to promote a comprehensive understanding and use of the ARRIVE 2.0 guidelines among domestic researchers, to enhance the standardization of experimental animal research and reporting, and to promote high-quality development of experimental animal sciences and comparative medicine research in China. Table and Figures | Reference | Related Articles | Metrics

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

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Advances in Comparative Medical Research on Anatomy and Histological Structure of Intervertebral Discs in Humans and Other Animals Li ZHANG, Yu KUANG, Lingxia HAN Laboratory Animal and Comparative Medicine    2024, 44 (2): 192-201.   DOI: 10.12300/j.issn.1674-5817.2023.141 Abstract （761）   HTML （47）    PDF （1064KB）（1895）       Knowledge map    Save The 2023 China Health Report on Spine Degeneration noted a significant increase in lumbar surgery among patients under 35 years old in recent years, indicating a trend towards younger onset of cervical and lumbar diseases. Lumbar intervertebral disc herniation has become a major concern, making the study of disc degeneration pathogenesis and treatment methods clinically significant. At present, human intervertebral disc diseases are primarily diagnosed through imaging due to the challenges of obtaining tissue samples from the spine. Therefore, experimental animals have emerged as alternative research subjects because they are cost-effective, have short experimental cycles, and are easily accessible. Given the structural and physiological differences between human and other animal intervertebral discs, comparing their anatomy and histological characteristics forms the foundation of research into human disc degeneration. The purpose of this paper is to collect and review relevant studies on anatomical and histological structures of intervertebral discs in different animals and conduct a comparative analysis from four aspects, namely, intervertebral disc height, lumbar disc geometry, lumbar disc cartilaginous endplate characteristics, and extracellular matrix components. The results show that humans, kangaroos, sheep, pigs, and rats exhibit similar relative heights between the sixth and seventh cervical vertebrae. Mice possess lumbar disc geometries most akin to humans. Compared to other animals, humans have the thickest cartilaginous endplates and the lowest cell densities. The collagen within the fibrous annulus differs most notably in pigs compared to humans, while water content in the nucleus pulposus is consistent across pigs, sheep, rabbits, rats, and humans. Additionally, this paper describes the commonalities and discrepancies in disc degeneration manifestations between humans and animals, and summarizes modeling methods for disc degeneration in different experimental animals. Ultimately, the aims of this paper is to provide fundamental data for selecting suitable experimental animal models for the study of intervertebral disc degeneration. Table and Figures | Reference | Related Articles | Metrics

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Differential Analysis of Oral Microbiota in db/db Mouse Model of Type 2 Diabetes Utilizing 16S rRNA Sequencing PAN Qianjia, GE Junyi, HU Nan, HUA Fei, GU Min Laboratory Animal and Comparative Medicine    2025, 45 (2): 147-157.   DOI: 10.12300/j.issn.1674-5817.2024.119 Abstract （469）   HTML （31）    PDF （1993KB）（1850）       Knowledge map    Save Objective To investigate the changes in oral microbiota of db/db mice and provide an experimental basis for exploring the relationship between type 2 diabetes mellitus and oral microecology. Methods Eight 10-week-old male db/db mice were designated as the diabetes experimental group (db/db group), while eight 10-week-old male db/m mice were assigned as the normal control group (db/m group). After a 5-day adaptive feeding period, tail venous blood samples were collected on the 6th and 37th days, and fasting blood glucose (FBG) levels and oral glucose tolerance test (OGTT) were performed for both groups to verify the reliability of the diabetes model. On the 15th day of feeding with the same diet, oral microbiota samples were collected from the buccal mucosa, dorsal and ventral tongue surfaces, oral floor mucosa, hard palate mucosa, and the gingival areas of both the upper and lower jaws of the two groups. Genomic DNA from the oral microbiota was extracted, and the V3-V4 regions of the 16S ribosomal RNA (16S rRNA) gene were amplified using a GeneAmp 9700 thermocycler. The composition of the oral microbiota was evaluated through double-labelled amplification and sequencing on the Illumina MiSeq platform, followed by bioinformatics analysis using QIIME software(version 1.6.0). Results The FBG levels and OGTT results on the 6th and 37th days after the start of the experiment indicated that db/db mice exhibited more pronounced symptoms of type 2 diabetes compared to db/m mice. Alpha diversity (α diversity) analysis showed no significant difference in the diversity of oral microbiota between the two groups (P>0.05); however, there was a significant difference in richness (P<0.05). Principal coordinate analysis(PCoA) revealed differences in the oral microbiota composition between the db/db group and db/m group (P<0.05). Species composition analysis and LEfSe analysis demonstrated that the relative abundance of oral microbiota in db/db group mice, predominantly composed of p_Proteobacteria, increased significantly at the phylum level (P<0.05). At the genus level, the relative abundances of g_Proteus and g_Enterococcus showed a significant increase (P<0.001). Conclusion The composition and diversity of oral microbiota in db/db mice with type 2 diabetes mellitus significantly differed from those without the disease. Table and Figures | Reference | Related Articles | Metrics

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Preliminary Study on Construction Method of Type 1 Diabetes Model in Banna Miniature Pig LIU Kai, HU Yuqi, GENG Yatian, CHENG Wenjie, WANG Jing, WEI Taiyun, ZHAO Hongfang, LI Jiayu, JIAO Deling, ZHAO Hongye, WEI Hongjiang Laboratory Animal and Comparative Medicine    2024, 44 (4): 384-392.   DOI: 10.12300/j.issn.1674-5817.2024.060 Abstract （584）   HTML （44）    PDF （2116KB）（1776）       Knowledge map    Save Objective To construct a Type 1 diabetes model in miniature pigs and explore postoperative care strategies for effectively prolonging the survival time of the model pigs. Methods Seven Banna miniature pigs were selected for pancreatectomy. Glucose, vitamins, and antibiotics were administered for 3-5 days after surgery to aid recovery. Blood glucose and urine glucose levels were measured twice a day in the morning and evening to adjust insulin supplementation accordingly. The model pigs were observed daily and records were kept, including orexis, psychosis, weakness, skin ulcer, and feces and urine. Body weight was measured weekly until the death of the model animals. Based on the model pigs' condition, glucose injection and Ringer's lactate solution were administered to supplement nutrition and correct electrolyte imbalances. Results All seven Banna miniature pigs showed typical symptoms of diabetes: random blood glucose levels higher than 11.1 mmol/L after pancreatectomy, far exceeding the average blood glucose level of 6.0 mmol/L in normal pigs; positive urine glucose; and progressive weight loss. These features indicated the successful construction of Type 1 diabetes model. Additionally, Type 1 diabetic pigs that survived more than 8 weeks showed progressive hair loss and skin ulceration. Euthanasia was performed on model pigs when they were unable to stand or even eat independently, and pathological examination and HE staining were conducted on tissues collected from affected organs such as the liver, kidneys, and skin. Pathological sections revealed liver congestion, massive glycogen accumulation, ballooning degeneration of hepatocytes, and progressive liver fibrosis, along with glomerular congestion, vacuolar degeneration in renal tubular epithelial cells, proteinuria, dermal congestion, thinning of vascular walls, and varying degrees of parakeratosis and dyskeratosis in the liver, kidneys, and skin tissues due to prolonged hyperglycemia. The average survival time of the constructed Banna miniature pig diabetes model was 44 d, with a maximum survival time of 121 d. Conclusion Type 1 diabetes model can be constructed successfully in Banna miniature pigs through pancreatectomy. With meticulous postoperative care, a long-term Type 1 diabetes model with significant complications can be achieved, providing a stable large-animal model for Type 1 diabetes treatment strategies. Table and Figures | Reference | Related Articles | Metrics

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Establishment and Evaluation of a Rat Model of Non-Puerperal Mastitis YIN Yulian, MA Lina, TU Siyuan, CHEN Ling, YE Meina, CHEN Hongfeng Laboratory Animal and Comparative Medicine    2024, 44 (6): 587-596.   DOI: 10.12300/j.issn.1674-5817.2024.065 Abstract （963）   HTML （255）    PDF （1898KB）（1753）       Knowledge map    Save Objective This study aims to establish a non-puerperal mastitis (NPM) rat model by simulating hyperprolactinemia and immune-inflammatory states, and to evaluate its local inflammatory characteristics in the mammary gland, thereby laying the foundation for research on the diagnosis and treatment of this clinically challenging disease. Methods Twelve SPF-grade Wistar female rats were evenly divided into a control group and a model group. During the experiment, the control group received no experimental treatment or medication. The model group received daily subcutaneous injections of 100 mg/kg metoclopramide hydrochloride for 7 consecutive days. Serum prolactin (PRL) levels were measured using ELISA on the 10th, 20th, and 30th days after the first injection. After 7 days of injections, 200 μL of lactating SD rat milk was mixed with 200 μL of complete Freund's adjuvant to prepare an oil-in-water emulsion, which was administered by multiple subcutaneous injections into the back of the Wistar rats for the initial immunization. Seven days after the initial immunization, the emulsion was injected subcutaneously into the third, fourth, and fifth mammary glands for the final immunization. After the final immunization, the rats were observed for 28 days for changes in mammary gland appearance, and the size of mammary nodules was calculated. On the 3rd, 7th, 14th, and 28th days, hematoxylin-eosin (HE) staining was used to analyze mammary tissue morphology. Immunohistochemistry was employed to detect CD138 expression levels. ELISA was used to measure the levels of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, and inducible nitric oxide synthase (iNOS) in mammary tissue to comprehensively assess the model. Results Rats in the model group exhibited mammary skin ulceration and foul odor at the ulcer sites. Palpation and ultrasound revealed the formation of mammary nodules. HE staining showed that on the 3rd day after the final immunization, normal ductal and lobular structures in the mammary glands disappeared, with significant infiltration of plasma cells. On the 7th day, ductal dilation, epithelial necrosis and detachment, and pronounced periductal plasma cell and lymphocyte (predominantly T-lymphocytes) infiltration were observed. On the 14th day, there was a proliferation of fibrofatty tissue, small blood vessels, and granulation tissue, with scattered plasma cells in the interstitium. By the 28th day, inflammatory cell infiltration and fibrous tissue proliferation were reduced, with granuloma formation. Serum PRL levels in the model group were significantly increased on the 10th day (P<0.05) and the 20th day (P<0.001). IL-6 and TNF-α levels in mammary tissue were higher in the model group compared to the control group on the 3rd, 7th, 14th, and 28th days (P<0.05). IL-1β levels were higher on the 3rd, 7th, and 14th days compared with the control group (P<0.01) but lower than the control group on the 28th day (P>0.05). iNOS levels were significantly higher on the 7th day after the final immunization (P<0.001). Conclusion A successful NPM model was established in rats, which exhibited typical pathological features such as local mammary masses, abscesses, ulcers, ductal dilation and plasma cell infiltration. This model can serve as a foundation for further research into the diagnosis and treatment of this clinically challenging disease. Table and Figures | Reference | Related Articles | Metrics

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

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Advances in Nucleic Acid Drugs and Gene Therapies based on Animal Models of Duchenne Muscular Dystrophy LIU Siyu, LAI Yuezhao, GUO Wenting, CHEN Xuejin Laboratory Animal and Comparative Medicine    2024, 44 (6): 613-625.   DOI: 10.12300/j.issn.1674-5817.2024.168 Abstract （678）   HTML （26）    PDF （1488KB）（1628）       Knowledge map    Save Duchenne muscular dystrophy (DMD) is a severe X-linked recessive genetic disorder caused by mutations in the DMD gene, making it one of the most common forms of hereditary muscular dystrophy. The DMD gene, which encodes dystrophin, is the largest known gene in the human genome. Mutations in the DMD gene are highly diverse, including exon deletions, duplications, point mutations, and small insertions or deletions, posing significant challenges for treatment. Currently, there is no cure for DMD, and existing treatment strategies focus primarily on symptom management, which cannot reverse or halt disease progression. Advances in biotechnology position nucleic acid drugs and gene therapies at the forefront of DMD treatment research. These treatments aim to restore dystrophin expression by repairing or replacing mutated genes, thereby improving muscle function or slowing muscle degeneration. Preclinical studies in animal models and early-phase clinical trials demonstrate promising efficacy and offer new hope for DMD patients. This review briefly outlines the pathological mechanisms and genetic characteristics of DMD before delving into recent progress in therapeutic strategies, with a particular focus on nucleic acid drugs (including antisense oligonucleotides for exon skipping therapy and translation readthrough inducers) and gene therapy approaches (including gene replacement therapy and gene editing). The development and application of these therapies not only provide new treatment options for DMD patients, but also offer valuable insights for addressing other genetic disorders. However, numerous challenges impede the clinical translation of DMD treatments. Future studies must optimize existing therapeutic strategies, improve their efficacy and applicability, and explore innovative approaches to deliver more effective and sustainable treatments for DMD patients. Table and Figures | Reference | Related Articles | Metrics

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

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Key Challenges and Mitigation Strategies for Animal Pregnancy in Non-clinical Reproductive Toxicity Testing of Drugs LIU Kun, LAN Qing, YI Bing, XIE Xiaojie Laboratory Animal and Comparative Medicine    2025, 45 (4): 449-456.   DOI: 10.12300/j.issn.1674-5817.2024.177 Abstract （316）   HTML （7）    PDF （994KB）（1588）       Knowledge map    Save Non-clinical reproductive toxicity studies typically employ mammals like rats, rabbits, and cynomolgus monkeys, with animal pregnancy being a key challenge in such testing. This article focuses on the difficulties encountered in the animal pregnancy process and potential countermeasures. Rats can be used for fertility and early embryonic development toxicity studies (Segment Ⅰ), embryo-fetal development toxicity studies (Segment Ⅱ), and perinatal toxicity studies (Segment Ⅲ). The estrous cycle of female rats can be determined by vaginal smear, and mating behavior is confirmed through copulatory plug checks the following day after pairing one female with one male in the same cage. Rabbits are commonly used in embryo-fetal development toxicity studies (Segment Ⅱ). Mating behavior between male rabbits and estrous females is observed to determine the time of conception. However, challenges such as atypical estrus of female rabbits, large variations in estrus between batches, and mating failure often occur in reproductive toxicity testing, which may be addressed through prolonged light exposure, increased protein supplementation, optimized mating strategies, and environmental modifications like female and male rabbits are raised adjacent to each other. Non-human primates (NHPs) are typically employed in perinatal toxicity studies (Segment Ⅲ), where one of the key challenges lies in accurately determining sexual maturity in males - a critical factor for reproductive toxicity testing, which can be assessed through comprehensive evaluation of age, body weight, and testicular volume. Generally, male macaques are considered sexually mature when they meet the following criteria: age >4.5 years, body weight >4.5 kg, single testis volume >10 mL, and combined testicular volume >20 mL. For pregnancy confirmation, ultrasound examination demonstrating visible gestational sacs is required, though this necessitates experienced veterinary clinicians to establish standardized ultrasound examination protocols. In conclusion, reproductive toxicity studies should employ species-appropriate detection methods and evaluation criteria based on anatomical characteristics of the reproductive system to ensure successful mating and proper study execution. Table and Figures | Reference | Related Articles | Metrics

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Integrative Analysis of Omics Data in Animal Models of Coronavirus Infection WU Yue, LI Lu, ZHANG Yang, WANG Jue, FENG Tingting, LI Yitong, WANG Kai, KONG Qi Laboratory Animal and Comparative Medicine    2024, 44 (4): 357-373.   DOI: 10.12300/j.issn.1674-5817.2024.008 Abstract （580）   HTML （412）    PDF （3355KB）（1586）       Knowledge map    Save Objective This study analyzes the omics data resources in human-infecting coronavirus animal models collected from various public databases, focusing on data distribution, dataset quantity, data types, species, strains, and research content. It aims to enhance our understanding of biological characteristics and pathogenic mechanisms of coronaviruses, thereby providing a solid foundation for devising effective therapeutic strategies and preventive measures. Methods Query strategies, including specific virus names, time ranges, and inclusion and exclusion criteria, were defined to retrieve data from major public omics databases such as GEO and ArrayExpress. Secondary filtering was performed based on different field types to obtain a more accurate data list. An omics data text database was established for bibliometric analysis. Co-occurrence networks were constructed for the analysis of the correlation strengths between different research themes, technical methods, and involved species. The cell types, organs, and biological pathways involved in studies were examined to further elucidate the pathogenic interplay between pathogens and hosts. Results About twenty public databases containing coronavirus-related omics data were identified, with a primary focus on novel coronavirus infection. Commonly used species include humans, mice, hamsters, and monkeys, while the commonly used virus strains are Wuhan-Hu-1 and USA-WA1/2020. Lung tissues are primarily used in animal models such as mice, macaques, and ferrets, while airway epithelial cells and Calu-3 cells are predominantly employed in human-related studies. Expression profiling data indicate that gene pathways involved in inflammation, cytokine response, complement pathway, cell damage, proliferation, and differentiation are significantly upregulated after infection. Proteomics studies reveal significant changes in phosphoproteome, ubiquitinome, and total proteome of patient samples at different infection stages. Specific protein categories, including viral receptors and proteases, transcription factors, cytokines, proteins associated with coagulation system, angiogenesis-related proteins, and fibrosis markers, show alterations after coronavirus infection. In addition, metabolomics data suggest that phosphocholine, phosphoethanolamine, arachidonic acid, and oleic acid could serve as potential metabolic markers. Epigenomics research indicates m6A methylation plays a role in SARS-CoV-2 replication, infection, and transmission, affecting host cell-virus interactions. Among these, N, S, and non-structural proteins 2 and 3 exhibit the most significant ubiquitination. Trends in microbiomics research suggest that microbial communities in the gut and wastewater are emerging as new research focuses. Conclusion The data types of coronavirus omics are diverse, with a wide variety of models and cell types used. The selection of species and technical methods for modelling varies based on the characteristics of different viruses. Multi-omics data from animal models of coronavirus infection can reveal key interactions between hosts and pathogens, identifying biomarkers and potential therapeutic targets, and provide valuable information for a deeper understanding of biological characteristics and infection mechanisms of coronaviruses. Table and Figures | Reference | Related Articles | Metrics

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Application Analysis of Animal Models for Pelvic Inflammatory Disease Based on Data Mining ZHENG Yiqing, DENG Yasheng, FAN Yanping, LIANG Tianwei, HUANG Hui, LIU Yonghui, NI Zhaobing, LIN Jiang Laboratory Animal and Comparative Medicine    2024, 44 (4): 405-418.   DOI: 10.12300/j.issn.1674-5817.2024.012 Abstract （776）   HTML （44）    PDF （974KB）（1559）       Knowledge map    Save Objective To investigate the key elements for model establishment and determine the evaluation indicators of animal models for pelvic inflammatory disease (PID), providing a reference for improving modelling methods and optimizing the application of PID animal models. Methods The search query "Pelvic Inflammatory Disease" AND "Animal Model" OR "Rat" OR "Mouse" OR" Guinea Pig" OR "Rabbit" OR "Dog" OR "Pig" was used to retrieve relevant literature on PID animal models published from 2013 to 2023 in China Knowledge Network Infrastructure (CNKI), Wanfang, and PubMed databases. The studies were analyzed and categorized based on experimental animal types, modelling methods, modelling cycles, detection indicators, positive control drugs, and administration duration. A database was established for statistical analysis. Results A total of 214 research articles on PID animal models meeting the inclusion criteria were identified. The most commonly used model animals are Sprague Dawley (SD) rats, followed by Wistar rats. The most frequently employed modelling method is a combination of mechanical injury and bacterial infection, followed by the phenol mucilage method. The most common modelling cycles for acute pelvic inflammatory disease (APID) and chronic pelvic inflammatory disease (CPID)/sequelae of pelvic inflammatory disease (SPID) are 8 to 14 days, while for PID models without specific staging, the cycles are 7 days. High-frequency detection methods and indicators include histopathological observation using hematoxylin-eosin staining, enzyme-linked immunosorbent assay (ELISA) for serum-related indicators, morphological changes of tissues observed with the naked eye, and immunohistochemical detection of related protein expression in uterine tissues, and pathological scoring. The most frequently used positive control drugs are Fuke Qianjin Tablets, followed by Jingangteng Capsules. The most common administration duration for APID is 7 days, and for CPID/SPID models, it ranges from 15 to 21 days. Conclusion Currently, SD rats and Wistar rats are commonly used as experimental animals for PID models. The dual modelling method of mechanical injury combined with mixed bacterial infection aligns closely with clinical pathogenesis and can be used to establish a PID model that simulates postoperative uterine cavity infection. Depending on the research objectives, different positive drugs and detection indicators should be selected for comprehensive evaluation. Most existing PID animal model studies are based on western medical diagnosis, with fewer studies focusing on Traditional Chinese Medicine (TCM) syndromes. There is a need to integrate TCM theories of etiology and pathogenesis to construct PID animal models that are more in line with TCM clinical symptoms. Table and Figures | Reference | Related Articles | Metrics

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

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

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Research Advances in Mongolian Gerbil Models of Cerebral Ischemia and Auditory Impairment ZHANG Zishan, WU Ying, LI Feiyang, DU Xiaoyan Laboratory Animal and Comparative Medicine    2024, 44 (4): 419-427.   DOI: 10.12300/j.issn.1674-5817.2024.032 Abstract （431）   HTML （33）    PDF （999KB）（1473）       Knowledge map    Save The Mongolian gerbil currently used as laboratory animals worldwide all originates from China. As early as the 1930s, wild Mongolian gerbils were domesticated and introduced into medical research. Today, they have become recognized multifunctional laboratory animals and are extensively used in various fields such as brain nerve studies, parasitology and microbiology, and oncology, etc. Mongolian gerbils possess unique anatomical characteristics in the basal cerebral arteries, such as a congenital absence of the Willis' circle, making it possible to construct cerebral ischemia or cerebral ischemia-reperfusion injury models with simple procedures of unilateral common carotid artery ligation, while also enabling intra-individual control. These anatomical features also increase their sensitivity to cerebral ischemia and make them more prone to cochlear ischemia, therefore playing a crucial role in the preparation of auditory impairment models. The disease progression and pathological manifestations in Mongolian gerbils show many similarities to those observed in human patients. Researchers have successfully used Mongolian gerbils to develop models of cerebral ischemia, cerebral ischemia-reperfusion, cochlear ischemia, cochlear implantation, and sensorineural hearing loss, achieving significant results. This article focuses on the current methods and assessment indicators for constructing Mongolian gerbils models of cerebral ischemia and auditory impairment. It discusses the advantages and disadvantages of various modelling techniques, and explores their application progress, aiming to provide a theoretical basis and reference for the application of Mongolian gerbils in these two important research areas. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Animal Models of Sepsis-Related Organ Injury YANG Jiahao, DING Chunlei, QIAN Fenghua, SUN Qi, JIANG Xusheng, CHEN Wen, SHEN Mengwen Laboratory Animal and Comparative Medicine    2024, 44 (6): 636-644.   DOI: 10.12300/j.issn.1674-5817.2024.087 Abstract （1065）   HTML （31）    PDF （794KB）（1456）       Knowledge map    Save Sepsis is a multi-organ dysfunction syndrome caused by infection and immune dysfunction, with a high mortality rate. It affects multiple important organs such as the heart, lungs, kidneys, liver, and brain. Establishing corresponding animal models of organ dysfunction syndrome is an essential step in clarifying its pathogenesis, researching potential effective drugs, and evaluating the effectiveness and safety of treatment plans. This article first summarizes classic modeling methods for sepsis related organ injury, including the destruction of intestinal barrier tissue integrity and the implantation of pathogens or toxic drugs. The former mainly includes cecal ligation and puncture, ascending colon stent implantation, and cecal ligation incision. The latter is divided into intraperitoneal injection, intravenous injection, and intratracheal administration based on the clinical infection route being simulated. Cecal ligation and puncture and lipopolysaccharide intraperitoneal injection are the most commonly used methods. Secondly, this article summarizes the common modeling methods and evaluation methods for animal models of sepsis-induced cardiomyopathy, acute lung injury, acute kidney injury, acute liver injury, and brain dysfunction. It points out that almost all organ injuries use classic modeling methods, and different organ injury models have additional modifications according to their different pathogenesis. For example, in addition to the classic modeling methods, lipopolysaccharide instillation in the trachea is more effective in modeling acute lung injury as it better simulates lung barrier dysfunction. Cecal ligation and puncture followed by Pseudomonas instillation in the trachea in a secondary challenge model better represents sepsis-induced acute kidney injury. Intraperitoneal injection of galactosamine is a mature modeling method of sepsis-induced acute liver injury. Intracerebral injection of lipopolysaccharide is a feasible model of sepsis-associated encephalopathy. In addition to the different modeling methods, there are differences in the administration time, dosage and experimental time points according to the different experimental purposes. This article reviews the research progress of animal experimental models for sepsis-induced cardiomyopathy, acute lung injury, acute kidney injury, acute liver injury, and brain dysfunction, aiming to provide a reference for the selection of animal experimental models and optimization of experimental design. Table and Figures | Reference | Related Articles | Metrics

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Challenges and Development in Suzhou Laboratory Animal Industry Over the Past Five Decades ZHAO Lijuan, XIAO Chunlan, SHENG Yajie, LU Xi, ZHOU Zhengyu Laboratory Animal and Comparative Medicine    2024, 44 (6): 645-653.   DOI: 10.12300/j.issn.1674-5817.2024.113 Abstract （753）   HTML （57）    PDF （1401KB）（1441）       Knowledge map    Save Since the 1970s, the laboratory animal industry in Suzhou has gone through five stages: its inception, emergence, growth, transformation, and scaling up. It began with the manufacturing of caging equipment for laboratory animals, initially by imitation and later through independent innovation. The industry evolved from sporadic factories to clustered enterprises, gradually growing and opening up the export market for caging equipment. In the 21st century, with industrial upgrading and transformation, purification systems and related products began to develop, and industry organizations emerged. As China has modernized, the rise of automation and intelligent production has led to technological innovation in enterprises and the emergence of various outsourcing services in the laboratory animal industry, driving the large-scale development of the industrial chain. After nearly half a century of growth, the laboratory animal industry in Suzhou has formed a complete industrial chain, including the production of laboratory animals, caging equipment, feed and bedding materials, design and construction of laboratory animal facilities, quality testing of laboratory animals and environments, and animal experimentation services. Laboratory animal breeding equipment, the core of the industry, has reached the level of developed countries, and the industry's scale and influence are unmatched in China. Since the 21st century, biopharmaceuticals have become the "No.1 industry" in the development of Suzhou. With government support, the guidance of the local economy, and the assistance from universities and research institutes, the animal experiment outsourcing industry has begun to cluster in Suzhou. The continuous influx of CROs has driven the construction of large-scale laboratory animal facilities, and key research projects have been initiated, significantly enhancing the industry's R&D capabilities. The Suzhou laboratory animal industry has quickly expanded alongside the "No. 1 industry," creating a unique "Suzhou Path" for laboratory animals. Over nearly fifty years, the laboratory animal industry in Suzhou has been essential to the rapid development of the biopharmaceutical industry in Suzhou and China. Table and Figures | Reference | Related Articles | Metrics

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

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Optimization and Evaluation of Conditions for Orthotopic Nude Mouse Models of Human Liver Tumor Cells MENG Yu, LIANG Dongli, ZHENG Linlin, ZHOU Yuanyuan, WANG Zhaoxia Laboratory Animal and Comparative Medicine    2024, 44 (5): 511-522.   DOI: 10.12300/j.issn.1674-5817.2024.048 Abstract （775）   HTML （27）    PDF （2747KB）（1354）       Knowledge map    Save Objective The study aims to optimize the conditions for constructing orthotopic nude mouse models of liver cancer by injecting human liver tumor cell lines and to explore appropriate timings for drug administration. Methods Human hepatocellular carcinoma Hep3B and hepatoblastoma HepG2 cell lines, which stably expressing the luciferase reporter gene (LUC), were selected. The linear correlation between the luciferase luminescence intensity and the number of liver tumor cells was analyzed using a Small Animal In Vivo Imaging system to verify the luminescent efficiency of the human liver tumor cells. Different concentrations (8×106, 2.4×107, 7.2×107 cells/mL) and resuspension media (PBS, Matrigel) of human liver tumor cell suspensions HepG2-LUC and Hep3B-LUC were orthotopically inoculated into the liver lobes of 5-week-old female BALB/c nude mice (12 groups, 7 mice each) to construct human liver tumor nude mouse orthotopic cancer models. Every 7 days, the weights of mice were recorded, and the growth of orthotopic tumors was monitored using the Small Animal In Vivo Imaging system. On day 35 post-cell inoculation, mouse livers were dissected, and pathological slices were prepared for HE staining to observe histopathological changes in liver tissues. Results The luminescence intensity of human liver tumor cell lines was positively correlated with the number of cells (R2=0.983 1, R2=0.970 5), indicating their suitability for orthotopic model construction. Successful modeling was achieved in the high-concentration groups of HepG2-LUC, the low-, medium-, and high-concentration groups of HepG2-LUC+Matrigel, the medium- and high-concentration groups of Hep3B-LUC, and the low-, medium-, and high-concentration groups of Hep3B-LUC+Matrigel. For both HepG2-LUC+Matrigel and Hep3B-LUC+Matrigel groups, mice in the high-concentration groups exhibited significantly reduced body weight compared to the low- and medium-concentration groups (both with P<0.05). The luminescence intensity of successfully modeled mice increased exponentially over time (R2>0.950 0), and reached a minimum of 1.0×107 p/(s·cm2·sr) by day 14 post-transplantation. Mice in the low- and medium-concentration groups of HepG2-LUC and the low-concentration group of Hep3B-LUC showed no significant pathological changes, while the other groups exhibited evident liver tumors and hepatocyte lesions. Conclusion For the HepG2-LUC cell line, the recommended injection volume is 50 μL with a cell density of 2.4×107 cells/mL, resuspended with Matrigel, followed by drug administration or prognostic measures on day 7 post-modeling. For the Hep3B-LUC cell line, the recommended injection volume is 50 μL with a cell density of 7.2×107 cells/mL, not resuspended with Matrigel, with administration or prognostic measures on day 14 post-modeling. Table and Figures | Reference | Related Articles | Metrics

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Diagnosis of an Outbreak of Canine Distemper in Cynomolgus Monkeys in an Experimental Monkey Farm in 2019 WANG Chenjuan, YANG Lingyan, WANG Lipeng, SUN Xueping, LI Jingwen, GUO Lianxiang, RONG Rong, SHI Changjun Laboratory Animal and Comparative Medicine    2025, 45 (3): 360-367.   DOI: 10.12300/j.issn.1674-5817.2024.160 Abstract （347）   HTML （17）    PDF （1701KB）（1350）       Knowledge map    Save Objective To report the diagnosis of a canine distemper virus outbreak among a colony of cynomolgus monkeys at an experimental monkey farm in 2019. Methods A total of 46 samples were collected from 21 diseased cynomolgus monkeys (exhibiting symptoms such as facial rash, skin scurf, runny nose, and diarrhea) and from one deceased monkey at an experimental monkey breeding farm in South China in late 2019, including serum, skin rash swabs, and anticoagulated whole blood, liver, lung, and skin tissues were submitted for testing. All submitted samples were tested for canine distemper virus gene fragments using real-time quantitative PCR, while immunohistochemical staining was performed to detect canine distemper virus nucleoprotein in lung tissues. The skin tissue of the deceased monkey was ground and sieved. The filtrate was inoculated into a monolayer MDCK cell line for virus isolation. Then, whole-genome sequencing was performed to identify the isolated virus. The Clustal Omega tool was used to align and analyze the homology of different Asian canine distemper virus isolates. A phylogenetic tree was constructed, followed by genetic evolutionary analysis. Results Clinical retrospective analysis revealed that the diseased cynomolgus monkeys exhibited symptoms similar to those observed in cynomolgus monkeys infected with measles virus. Necropsy findings showed red lesions in the lungs and significant hemorrhage in the colonic mucosa. Real-time quantitative PCR detected canine distemper virus nucleic acid in the serum, skin rash swabs of the infected monkeys, and various tissue samples of the deceased monkey, all of which tested positive. Calculation based on the standard curve formula indicated the viral load was highest in the skin tissue. Immunohistochemical staining of the deceased monkey's lung tissue demonstrated aggregation of CDV nucleoprotein in alveolar epithelial cells, bronchi, and bronchioles. A CDV strain was isolated from the skin tissue of the deceased monkey. Phylogenetic analysis indicated that this strain shares the closest relationship (98.86%) with the Asian-1 type canine distemper virus strain CDV/dog/HCM/33/140816, previously identified in dogs in Vietnam. Conclusion Based on comprehensive analysis of clinical symptoms, nucleic acid detection, viral protein immunohistochemistry, and whole-genome sequencing results, the diagnosis confirms that the cynomolgus monkeys in this facility are infected with canine distemper virus. It is recommended to include canine distemper virus as a routine surveillance target in captive monkey populations. Additionally, this study provides a foundation for further research on the molecular biological characteristics of canine distemper virus. Table and Figures | Reference | Related Articles | Metrics

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

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Construction and Evaluation of a Mouse Model with Intestinal Injury by Acute Hypoxic Stress in Plateau Jianhua ZHENG, Yunzhi FA, Qiaoyan DONG, Yefeng QIU, Jingqing CHEN Laboratory Animal and Comparative Medicine    2024, 44 (1): 31-41.   DOI: 10.12300/j.issn.1674-5817.2023.118 Abstract （822）   HTML （223）    PDF （2172KB）（1307）       Knowledge map    Save Objective By simulating acute hypoxic conditions, an experimental model of intestinal stress injury in plateau mice was established to explore the pathogenic mechanism of acute gastrointestinal diseases in plateau, and to lay foundation for preventive and therapeutic measures. Methods Thirty-six SPF-grade adult male BALB/c mice were randomly divided into four groups: normoxic 24 h, normoxic 72 h, hypoxic 24 h, and hypoxic 72 h, based on body weight using a randomized numerical table method, with nine mice in each group. Mice in the normoxic group were kept in a conventional barrier environment, while those in the hypoxic group were placed in a hypoxic chamber within the barrier environment with oxygen concentration set at 10% to simulate plateau conditions. They were subjected to stress for 24 h and 72 h, respectively, in order to establish a model of intestinal injury induced by acute hypoxia. After modeling, the mice were weighed, anesthetized with 1% pentobarbital sodium, and then euthanized by cervical dislocation. Duodenal and colonic tissues were collected. Histopathological morphology of intestinal tissues was observed after HE staining. Western blotting and immunohistochemistry were used to detect the expression levels of tight junction-related proteins in intestinal tissues. Real-time fluorescence quantitative PCR was performed to measure the expression levels of inflammatory cytokines and chemokines. TUNEL staining was used to assess apoptotic activity of intestinal epithelial cells, thus evaluating intestinal injury-related phenotypes in this model. Results Compared with the normoxic groups, mice in the 24 h and 72 h hypoxia groups showed weight loss, shortened duodenal villi, abnormal crypt structure, and decreased villus/crypt ratio. The colonic mucosa was infiltrated with inflammatory cells and irregular crypt structure. Expression levels of Occludin and zonula occludens-1 (ZO-1) were significantly decreased in duodenal and colonic tissues of mice in the 24 h and 72 h hypoxia groups (P<0.05). The expression of pro-apoptotic protein Bax was significantly up-regulated while expression of anti-apoptotic protein Bcl-2 was significantly down-regulated in duodenal tissues (P<0.05). Apoptotic activity of intestinal epithelial cells was significantly enhanced (P<0.05). In addition, interleukin (IL)-1β, IL-6, monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor-α (TNF-α) mRNA levels were significantly increased in duodenal tissues after 24 and 72 h of hypoxic stress(P<0.05). After 24 h of hypoxic stress, there was no significant change in the expression levels of inflammatory cytokines in colon tissues (P>0.05), but after 72 h, the expression levels of pro-inflammatory factors IL-1β, TNF-α, IL-6, MCP-1, and anti-inflammatory factor IL-10 mRNAs significantly increased in colon tissues of mice (P<0.05). Conclusion The usage of a hypoxia chamber to simulate an acute hypoxic environment in plateau can lead to abnormal intestinal tissue structure, intestinal barrier dysfunction, and induce intestinal epithelial cell apoptosis, triggering an intestinal inflammatory response in stress mice. These findings indicate the successful construction of a mouse model for an acute hypoxic stress-induced intestinal injury. Table and Figures | Reference | Related Articles | Metrics

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

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Preparation Methods and Evaluation Criteria Analysis of Animal Models for Perimenopausal Syndrome Tianwei LIANG, Yasheng DENG, Hui HUANG, Na RONG, Xin LIU, Yujie WANG, Jiang LIN Laboratory Animal and Comparative Medicine    2024, 44 (1): 74-84.   DOI: 10.12300/j.issn.1674-5817.2023.062 Abstract （1054）   HTML （44）    PDF （841KB）（1227）       Knowledge map    Save Objective To comprehensively analyze the reported preparation methods for animal models of perimenopausal syndrome (PS), to compare the advantages and disadvantages of various preparation elements and detection indexes, so as to provide useful references for the optimization of the relevant animal models as well as the standardization of their application in the efficacy evaluation of new drugs. Methods In this paper, literature research methods were applied using "perimenopausal syndrome" as the subject term. The publication period of the literature was limited to January 2016 to February 2023. Relevant literature on the preparation of PS animal models was retrieved from databases such as China National Knowledge Infrastructure, Wanfang database, and PubMed. After screening the experimental literature that met the inclusion and exclusion criteria, detailed information on experimental animal strains, modeling methods, duration of drug administration, positive drugs, detection indexes and other relevant information were collected. After the above information was standardized, the PS animal model database was established using Excel 2010 software. The model preparation elements and evaluation indexes were summarized systematically, and the statistical results were processed and analyzed using Excel 2010 software. Results A total of 247 articles were screened. SD rats (164 times, 65.86%) and Wistar rats (35 times, 14.06%) were often used to prepare PS animal models. Bilateral ovariectomy (139 times, 53.87%) and natural aging (43 times, 16.80%) were chosen as modeling methods. The ages of rats used for modeling ranged from 7 weeks to 18 months, with 3-month-old rats (22 times, 21.78%) being the most common. The detection indexes were comprehensively evaluated from multiple perspectives, including serum biochemistry, vaginal exfoliated cell smear, histomorphology, general observation, behavioral observation, and organ tissue protein immunoblotting. Western medical evaluation indexes were commonly used to test the successful preparation of models, with vaginal exfoliated cell smears being the most frequently used method (125 times, 85.04%). A model was considered successfully prepared when estrous cycle disorder or irregularity was observed. Some literature also determined modeling success by detecting a significant decrease in serum estradiol levels (5 times, 3.04%). Traditional Chinese medicine (TCM) syndrome evaluation often used a combination of Chinese and Western medical evaluation indexes for comprehensive evaluation, with researchers determining the TCM syndrome through vaginal exfoliated cell smears supplemented by general observation (3 times, 2.04%). Conclusion There are many methods for preparing PS animal models, but there are still significant differences in the selection of animal species, age, criteria for successful modeling, and TCM syndrome evaluation in the related literature. Table and Figures | Reference | Related Articles | Metrics

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Glycyrrhizic Acid Showed Therapeutic Effects on Severe Pulmonary Damages in Mice Induced by Pneumonia Virus of Mice Infection Yun LIU, Tingting FENG, Wei TONG, Zhi GUO, Xia LI, Qi KONG, Zhiguang XIANG Laboratory Animal and Comparative Medicine    2024, 44 (3): 251-258.   DOI: 10.12300/j.issn.1674-5817.2024.014 Abstract （577）   HTML （41）    PDF （1718KB）（1215）       Knowledge map    Save Objective In this study, inbred BALB/c mice infected with the pneumonia virus of mice (PVM) were used to establish an animal model of viral pneumonia, and the changes in the pro-inflammatory alarmin molecule, high mobility group box 1 protein (HMGB1), during PVM infection were observed, as well as the in vivo intervention effects of the HMGB1 inhibitor, glycyrrhizic acid (GA), on PVM-induced lung injury. Methods Three-week-old female BALB/c mice were randomly divided into three groups, each consisting of 6 mice. One group, uninfected by PVM, served as the control group (Control). The other two groups were inoculated intranasally with PVM at a dose of 1×104 50% tissue culture infective dose (TCID50)/25 μL, and subsequently treated with GA saline solution (GA group) or plain saline solution (normal saline, NS group) via gavage for 15 consecutive days. During this period, changes in body weight and appearance were monitored in each group. At the end of the experiment, lung tissue samples were collected from all groups. The distribution of PVM and HMGB1 proteins in the lung tissues was analyzed using hematoxylin-eosin staining and immunohistochemistry. The expression levels of HMGB1 and its Toll-like receptor 4 (TLR-4), advanced glycosylation end-product-specific receptor (AGER), and inflammatory cytokines such as interleukin (IL)-1β, IL-2, and tumor necrosis factor-α (TNF-α) in lung tissues of mice were measured using real time fluorescence quantitative PCR. Results Compared with the Control group, the NS group showed a significant weight loss after 6 days (P<0.05). Histopathological tests revealed pronounced inflammatory lesions in their lungs. Immunohistochemistry results showed that HMGB1 was released from the nucleus to the cytoplasm, and real time fluorescence quantitative PCR results indicated that the expression levels of HMGB1, IL-1β, and IL-2 were significantly upregulated (P<0.05). In the GA group, there was no significant change in the clinical symptoms or body weight. However, compared with the NS group, the pathological damages of lung tissues in the GA group were significantly reduced, and the expression levels of HMGB1, IL-1β, IL-2, and interferon-γ (IFN-γ) in lung tissues were also significantly decreased (P<0.05), although the expression level of AGER was significantly increased (P<0.05). Conclusion PVM infection can cause significant inflammatory pathological lung damages in mice, and GA can effectively alleviate the damages. Its therapeutic effect may be related to the activation of HMGB1 signaling pathway. Table and Figures | Reference | Related Articles | Metrics