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Revision of Standards for Microbiological and Parasitological Grades in Laboratory Animals and Its Comparison to Foreign Standards Lianxiang GUO Laboratory Animal and Comparative Medicine    2023, 43 (4): 339-346.   DOI: 10.12300/j.issn.1674-5817.2023.088 Abstract （395）   HTML （74）    PDF （859KB）（664）       Knowledge map    Save The national standard, GB 14922-2022 on "Laboratory Animal Microbiological and Parasitical standards and monitoring " was implemented on July 1st, 2023. This article is compiled according to the speech of the 16th East China Laboratory Annual meeting, explores and critically analyzes the developments made to the revised standard and examines how this framework compares with quality control programs of other established international institutions. The key aspects of establishing quality monitoring programs for animal-associated microorganisms in laboratory animal facilities are briefly discussed. Table and Figures | Reference | Related Articles | Metrics

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Evolution and Prospects of Laboratory Animal Management: A Case Study of Shanghai's Development in the Past Decade Yong ZHAO Laboratory Animal and Comparative Medicine    2023, 43 (5): 492-503.   DOI: 10.12300/j.issn.1674-5817.2023.134 Abstract （343）   HTML （160）    PDF （1101KB）（480）       Knowledge map    Save There are differences in historical and cultural beliefs, development history, and levels of technological development among different countries and regions around the world. However, they have all established corresponding laboratory animal management systems that are suitable for their national conditions. In 2001, the Ministry of Science and Technology, together with six other ministries, jointly issued the administrative licensing system for experimental animals, which was an innovative measure in China's specialized management system for experimental animals.The State Administration for Market Regulation and the National Standards Committee, based on the welfare of experimental animals and the needs of scientific research, have formulated a series of national standards for laboratory animals, and the local experimental animal management institutions, experimental animal quality testing unit and professional training base have also been established, which provide a strong guarantee for the rapid and healthy development of experimental animal science. This paper reviews the development of experimental animal management in Shanghai in the past ten years, reflects the evolution of national experimental animal management in recent years, points out the weak links in the development process, and puts forward suggestions for the innovation and development of experimental animal work. Table and Figures | Reference | Related Articles | Metrics

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Statistical Analysis of the Leakage Situation in the Automated Watering System for Mice in Tsinghua University Laboratory Animal Resources Center Qianqian TANG, Xiuli ZHANG, Zai CHANG Laboratory Animal and Comparative Medicine    2024, 44 (1): 85-91.   DOI: 10.12300/j.issn.1674-5817.2023.132 Abstract （341）   HTML （39）    PDF （1306KB）（244）       Knowledge map    Save Objective To calculate the leakage rate of the automated watering system in Tsinghua University Laboratory Animal Resources Center, to evaluate the safety of the system, and provide references for selection, maintenance, and management of automated watering systems in animal facilities. Methods This study investigated the automated watering system installed in South and North Barriers of Tsinghua University Laboratory Animal Resources Center (Phase II). Water leakage monitoring was conducted over two periods, one over a period of 3 years and the other over 1.5 years. The occurrence of water leakage events at the two barriers during the monitoring period was statistically analyzed, classifying the causes into four categories: mishandling by personnel, animal behavior, obstruction by foreign objects, and deformation of fittings. The total daily leakage rate due to these causes and the daily leakage rate caused by quality issues, i.e. obstruction by foreign objects and deformation of fittings were calculated. Further analysis and discussion focused on the causes of water leakage and its impact on the facilities. At the same time, the number of caretakers at the end of the monitoring period in the Phase I facility without automated watering system and the Phase II facility with automated watering system were counted. Finally the difference in the number of cages per capita under the two watering systems was calculated. Results A total of 52 water leakage incidents occurred in both areas during the monitoring period, with a total daily leakage rate of 0.000 13%. Among them, 31 were caused by personnel mishandling, accounting for approximately 60% of total leakages. Enhanced training, supervision, inspection, and effective reminder measures could reduce leakage caused by personnel mishandling. There were 2 cases of water leakages caused by animal behavior, 0 leakage due to obstruction by foreign objects, and 19 leakages due to system quality issues, with a daily leakage rate of 0.000 07%. According to the operation data of Tsinghua University Laboratory Animal Resources Center, the average number of cages managed per person in facilities equipped with the automated watering system was 908, compared to 570 cages in facilities without the automated watering system. This represents an approximate 59% increase in the number of cages managed per person with the adoption of the automated watering system. Conclusion The daily leakage rate of the automated watering system in the Tsinghua University Laboratory Animal Resources Center is significantly lower than the theoretical design rate of 0.003%, which demonstrates the system's safety and effectiveness. Additionally, the adoption of an automated watering system can signi?cantly enhance caretaking ef?ciency. While initial investments in the system are required, the subsequent increase in ef?ciency leads to a continuous decrease in labor costs, thereby reducing the total operational expenses of the facility. In the context of modernizing animal facility construction, automated watering systems are becoming an essential consideration in facility design and operation. Table and Figures | Reference | Related Articles | Metrics

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Implications on the Development of Animal Disease Models from FDA Modernization Act 2.0 Yinghan WAN, Yexin GU, Yunong YUAN, Min TANG, Li LU Laboratory Animal and Comparative Medicine    2023, 43 (5): 472-481.   DOI: 10.12300/j.issn.1674-5817.2023.083 Abstract （317）   HTML （568）    PDF （1127KB）（411）       Knowledge map    Save Laboratory animals are the foundational conditions and indispensable technical support in life science research and biomedical industry development. The scientific development of animal models of diseases is of great significance to biomedical research and industrial development. In light of the booming development of multiple emerging in vitro modelling technologies over the past decade, in 2022, the U.S. Senate unanimously passed the bill FDA Modernization Act 2.0. This bill rescinded the requirement for animal testing in investigating the safety and effectiveness of a drug—a federal mandate since 1938, and highlighted the potential of various invitro disease modeling approaches in future biomedical fields. This paper provides a comprehensive review of the latest advances and applications of in vitro disease modeling approaches in academia and industry followed by an interpretation of the FDA bill, namely cell culture, organoid, organ-on-a-chip, 3D bio-printing model and computer-based model. The paper next introduces the crossed applications of various disease models and discusses the advantages and disadvantages of each system, thereby providing insights into future trends in the use of animal disease models in China. Reference | Related Articles | Metrics

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Microbiological Monitoring Analysis of Laboratory Rats and Mice from Vendors: Department of Laboratory Animal Science of Fudan University as an Example Ying HUANG, Siyu WEI, Li CAI, Sujing QIANG, Dongting LI, Yuqiang DING Laboratory Animal and Comparative Medicine    2023, 43 (4): 347-354.   DOI: 10.12300/j.issn.1674-5817.2023.060 Abstract （313）   HTML （35）    PDF （1153KB）（406）       Knowledge map    Save Objective Conduct routine microbiological monitoring of laboratory rats and mice from vendors to provide an important basis for the scientific management of laboratory animal facility and ensure the reliability of relevant experimental data obtained from laboratory animals. Methods Taking the Department of Laboratory Animal Science of Fudan University as an example, between April 2021 and April 2023, rats and mice purchased from 7 vendors were sampled for microbiological quality according to the principle of simple random sampling on the arrival days of animal delivery. Then, surveillance tests were conducted to examine the microbiological contaminations according to the national standards of SPF laboratory animals. Results The total qualified rate was 80.36%, with 52.63% in SD rat, 82.76% in inbred mice, 86.67% in outbred mice and 86.36% in immunodeficient mice in details. The most frequent bacteria isolated were Staphylococcus aureus, Pseudomonas aeruginosa, Klebsilla pneumoniae and Rodentibacter heylii, and their detection rates were 10.76%, 3.16%, 2.53% and 0.63%, respectively. Serological assays demonstrated the highest prevalence for virus was Sendai virus, and the detection rate was 2.53%. In addition to the pathogens those must be excluded from SPF rodents, Entamoeba muris and Enterobacter spp. were also detected in inbred mice, and Klebsiella oxytoca was detected in immunodeficient mice, with the detection rates of 1.15%, 2.30% and 4.55%, respectively. Conclusion There are certain incidences of pathogen infections in laboratory rats and mice from vendors, and an efficient microbiological monitoring of laboratory animals should be implemented in animal facilities, in order to eliminate pathogen infections in laboratory animals, which is required for improving the accuracy of research results and protecting the occupational health of laboratory animal practitioners as well. Table and Figures | Reference | Related Articles | Metrics

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Research Progress Report on Microtus fortis as a New Resource of Laboratory Animal Jianyun XIE Laboratory Animal and Comparative Medicine    2023, 43 (5): 482-491.   DOI: 10.12300/j.issn.1674-5817.2023.114 Abstract （293）   HTML （124）    PDF （1243KB）（253）       Knowledge map    Save Microtus fortis (reed vole) is the only mammal known to have natural resistance to Schistosomiasis japonica. Originating from schistosomiasis endemic and non-endemic areas, as well as laboratory bred voles have the same resistance to Schistosoma japonicum. After more than 30 years of laboratory cultivation of wild reed vole, a series of progress have been made in laboratory animalization. A detailed study was conducted on biological traits including growth and development, reproductive physiology, serum biochemistry, hematological indicators and tissue anatomy. At the same time, the anti-schistosomiasis characteristics and anti-schistosomiasis mechanisms of Microtus fortis were studied. The closed Dongtinghu population of Microtus fortis (S: DTMF) cultivated by Shanghai Laboratory Animal Research Center was recognized as a Chinese laboratory animal resource by the Experimental Animal Resources and Evaluation Working Committee of the Chinese Association for Laboratory Animal Sciences in 2021. This review focuses on summarizing the research progress in the biological characteristics, standardization research, genome and anti-schistosomiasis mechanism of reed vole in the past decade, especially in the implementation of the key project in the National Science and Technology Pillar Program. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on Establishing and Evaluation of Acne Animal Models Rui ZHANG, Meiyu LÜ, Jianjun ZHANG, Jinlian LIU, Yan CHEN, Zhiqiang HUANG, Yao LIU, Lanhua ZHOU Laboratory Animal and Comparative Medicine    2023, 43 (4): 398-405.   DOI: 10.12300/j.issn.1674-5817.2023.021 Abstract （288）   HTML （19）    PDF （808KB）（504）       Knowledge map    Save According to understanding of the pathogenesis of acne, scholars have established animal models of acne inflammation, animal models of grafting human skin acne, and natural acne animal models. The acne inflammation model is mainly induced by bacterial infection, chemical drug application, and foreign matter injection. Natural acne animal models include animals that some are sensitivity to hormones and some have clinical symptoms of acne. It is necessary to select appropriate model animals and replicate model methods for the development of acne intervention products with different degrees and mechanisms. At present, there are only human evaluation standards of acne health functions in China, but no animal evaluation standards, which has affected the in-depth study of the pathogenesis of acne as well as the research and development progress of acne products. This article summarizes the conditions for the occurrence of acne, the characteristics of human skin, the bidirectional effect of Cutibacterium acnes on human skin, acne animal models, and commonly used observation and evaluation indicators, providing the reference for studying the pathogenesis of acne, promoting acne treatment and health care, and developing treatment products. Table and Figures | Reference | Related Articles | Metrics

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Establishment of PCR Identification Method for Pig Blood Type Jiaoxiang WANG, Yan WANG, Ke HU, Kaixiang XU, Taiyun WEI, Deling JIAO, Heng ZHAO, Hongye ZHAO, HongJiang WEI Laboratory Animal and Comparative Medicine    2023, 43 (6): 585-594.   DOI: 10.12300/j.issn.1674-5817.2023.065 Abstract （276）   HTML （427）    PDF （2710KB）（278）       Knowledge map    Save Objective Xenotransplantation is an effective way to address the shortage of human organ donors, but it faces serious immune rejection reactions, including hyperacute rejection caused by blood type differences. Establishing a stable, convenient, and reliable method for pig blood type identification can quickly screen suitable donor pigs for xenograft research. Methods Banna miniature inbred pigs, Diannan small eared pigs, and Bama Xiang pigs were selected as the research objects. DNA was extracted from the blood, oral buccal mucosa, and fetal fibroblasts of the three strains of pigs using DNA extraction kits. The target fragment of the ABO homologous gene EAA intron 7 in pigs was amplified using PCR method. Blood agglutination reaction was used to detect hemolysis in pig anterior vena cava whole blood after adding anti A and B antibodies. Immunohistochemical method was used to detect the expression level of A antigen in pig heart, liver, spleen, lung, and kidney tissues. Immunofluorescence method was used to detect the expression level of A antigen in pig oral mucosa. By comparing the results of different methods for determining pig blood types, the stability and reliability of the PCR method were verified, and a convenient PCR based pig blood type identification method was established. Results Firstly, the blood PCR results of 69 inbred strains of Banna miniature pigs, 7 Diannan small eared pigs, and 34 Bama Xiang pigs showed 20 AO blood types, 66 AA blood types, and 24 O blood types. The PCR results of fetal fibroblasts from 47 Diannan small eared pigs showed that all 47 fetuses were O blood type. Among them, the oral mucosal PCR results of 8 gene edited cloned pigs were consistent with those of donor fetal fibroblasts, all of which were O blood type. The oral mucosal PCR results of 8 wild-type pigs (2 inbred lines of Banna miniature pigs, 4 Diannan small eared pigs, and 2 Bama Xiang pigs) were consistent with the blood PCR identification results. Then, 11 inbred lines of Banna miniature pigs, 4 Diannan small eared pigs, and 2 Bama Xiang pigs were randomly selected for blood agglutination reaction validation, and the results were consistent with the PCR identification results of both blood samples and oral mucosa samples. Moreover, immuno-histochemical analysis was performed on the heart, liver, lung, kidney, and spleen tissues of one Banna miniature pig inbred line and two Bama Xiang pigs, and the results were consistent with blood PCR identification and blood agglutination reaction results. Finally, oral mucosal samples were collected from 2 inbred strains of Banna miniature pigs and 1 Bama Xiang pig for immunofluorescence detection, and the results were consistent with the blood PCR identification results. Conclusion By collecting fetal cells and oral mucosal samples from live pigs for PCR detection, the blood type of pigs can be accurately and efficiently identified, providing a convenient method for blood type screening of xenograft donor pigs. Table and Figures | Reference | Related Articles | Metrics

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Explanation and Elaboration for the ARRIVE Guidelines 2.0—Reporting Animal Research and In Vivo Experiments (Ⅲ) Xiaoyu LIU, Xuancheng LU, Xiaomeng SHI, Yuzhou ZHANG, Chao LÜ, Guoyuan CHEN, Xiao LU, Yu BAI, Jing GAO, Yao LI, Yonggang LIU, Yufeng TAO, Wanyong PANG Laboratory Animal and Comparative Medicine    2023, 43 (4): 446-456.   DOI: 10.12300/j.issn.1674-5817.2023.039 Abstract （262）   HTML （27）    PDF （1578KB）（678）       Knowledge map    Save Improving the reproducibility of biomedical research results is a major challenge.Researchers reporting their research process transparently and accurately can help readers 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 applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as the reliability, repeatability, and clinical translatability of animal experimental results. The use of 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 integrity of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is a Chinese translation based on the best practices of international journals following the ARRIVE 2.0 guidelines in international journals, specifically for the complete interpretation of the ARRIVE 2.0 guidelines published in the PLoS Biology journal in 2020 (original text can be found at https://arriveguidelines.org ). The third part of the article includes the items 8-10 of ARRIVE 2.0 Essential 10, which covers "experimental animals" "experimental procedures" and "results". Its aim is to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhance the standardization of experimental animal research and reporting, and promote the high-quality development of experimental animal technology and comparative medicine research in China. Table and Figures | Reference | Related Articles | Metrics

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Animal Models of Pulmonary Arterial Hypertension and Their Application in Drug Research Jiahui YU, Qian GONG, Lenan ZHUANG Laboratory Animal and Comparative Medicine    2023, 43 (4): 381-397.   DOI: 10.12300/j.issn.1674-5817.2023.048 Abstract （227）   HTML （21）    PDF （1028KB）（394）       Knowledge map    Save Pulmonary arterial hypertension is a clinical syndrome characterized by pulmonary vascular remodeling causing increased vascular resistance, which will lead to right heart failure and even death if left untreated. The pathogenesis of pulmonary arterial hypertension has not yet been clarified, and clinical treatments have not been effective in improving prognosis or reducing mortality. To investigate the pathogenesis of pulmonary arterial hypertension and to develop and evaluate more effective and safer drug treatments, establishing related animal disease models is very important. This paper outlines the pathological characteristics of pulmonary arterial hypertension and summarizes the various types of animal models of pulmonary arterial hypertension, as well as describes the progress of the application of these models in three therapeutic pathways and related drug research in the past five years, with a view to providing a reference for the selection of animal models of pulmonary arterial hypertension and research applications. 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 （224）   HTML （207）    PDF （1252KB）（226）       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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Explanation and Elaboration for the ARRIVE Guidelines 2.0—Reporting Animal Research and In Vivo Experiments (Ⅳ) Xiaying LI, Yonglu TIAN, Xiaoyu LIU, Xuancheng LU, Guoyuan CHEN, Xiao LU, Yu BAI, Jing GAO, Yao LI, Yusheng WEI, Wanyong PANG, Yufeng TAO Laboratory Animal and Comparative Medicine    2023, 43 (6): 659-668.   DOI: 10.12300/j.issn.1674-5817.2023.142 Abstract （221）   HTML （33）    PDF （1188KB）（662）       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 applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as the reliability, repeatability, and clinical translatability of animal experimental results. The use of 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 integrity of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is a Chinese translation based on the best practices of international journals following the ARRIVE 2.0 guidelines in international journals, specifically for the complete interpretation of the ARRIVE 2.0 guidelines published in the PLoS Biology journal in 2020 (original text can be found at https://arriveguidelines.org ). The fourth part of the article includes the items 1-5 of ARRIVE 2.0 Recommended 11 section, which covers "Abstract" "Background" "Objectives" "Ethical statement" and "Housing and husbandry". Its aim is to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhance the standardization of experimental animal research and reporting, and promote the high-quality development of experimental animal technology and comparative medicine research in China. Table and Figures | Reference | Related Articles | Metrics

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Analysis of Common Types and Construction Elements of Diabetic Mouse Models Xue WANG, Yonghe HU Laboratory Animal and Comparative Medicine    2023, 43 (4): 415-421.   DOI: 10.12300/j.issn.1674-5817.2023.031 Abstract （212）   HTML （15）    PDF （740KB）（220）       Knowledge map    Save Diabetes mellitus is a disease characterized by absolute or relative lack of insulin, which leads to hyperglycemia, and its high mobidity and complications have a great impact on the lives of patients. Animal models are widely used to study the pathogenesis and treatment of diabetes and its complications. Different types of diabetes, with different pathogenesis and pathognomonic features, have different treatment options. In animal experimental, in addition to considering the genetic factors and physiological characteristics of the animal (such as sex and age), it is also necessary to consider the experimental protocol and various response options, which have a great impact on the experimental data, the reproducibility and stability of the experimental results. Therefore, it is necessary to select suitable animal models for experiments in the study of diabetes. Type 1 diabetes is characterized by absolute insulin deficiency, and existing mouse models of type 1 diabetes include chemically (STZ-induced) induced and spontaneous diabetes model (NOD mice), etc. Type 2 diabetes, characterized by insulin resistance and impaired glucose tolerance, is established in both obese and non-obese animal models, including diet-induced (high-fat diet induced), spontaneous diabetes (including monogenic and polygenic obese mice) models, and genetically modified mouse models. In this review, we discussed the common types of diabetic mouse models and analyzed the elements of their construction, the key factors that should be considered in the selection of diabetic mouse models, and explore the impact of these factors on the research of diabetes. Table and Figures | Reference | Related Articles | Metrics

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Construction and Evaluation of End-to-side Anastomosis Model of Autologous Arteriovenous Fistula in Mice Xin LIU, Shaobo SHI, Cui ZHANG, Bo YANG, Chuan QU Laboratory Animal and Comparative Medicine    2023, 43 (6): 595-603.   DOI: 10.12300/j.issn.1674-5817.2023.093 Abstract （210）   HTML （499）    PDF （1787KB）（280）       Knowledge map    Save Objective To establish an animal model of autologous arteriovenous fistula in mice and evaluate its effect. Methods The left external jugular vein and common carotid artery of 10 8-week-old male C57BL/6 mice were separated by end-to-side anastomosis of external jugular vein and common carotid artery after anesthesia, and the right jugular vein was exposed without suture as a control, so as to establish an animal model of internal arteriovenous fistula. Doppler ultrasound, HE and Masson staining and immunohistochemical staining were used to observe the hemodynamics, intimal hyperplasia and protein expression of smooth muscle cell proliferation in the outflow vein of the internal arteriovenous fistula and the contralateral control vein, and to evaluate the effect of model construction. Results A total of 10 mice were selected for this study, and 9 mice were successfully modeled, with a success rate of 90%. Ultrasound examinations were performed on the day of surgery, 7 and 14 days after surgery, respectively. The results showed that the flow velocity near the anastomosis was linearly correlated with the diameter of the tube. The higher the flow velocity, the larger the diameter of the tube. There was a positive correlation between peak velocity and lumen diameter (P=0.000 6, R2=0.831 7). After surgery 14 days, HE staining results showed that after autologous arteriovenous fistula molding, the average lumen area of outflow segment vein was significantly decreased (P < 0.000 1), the intima area was significantly increased (P < 0.000 1), the intimal area was significantly increased (P < 0.000 1). On the surgical side of arteriovenous fistula, collagen deposition was significantly increased, and the proportion of Masson-positive regions was significantly increased (P < 0.000 1). Immunohistochemical staining showed that the proportion of collagen 1 positive areas on the surgical side of arteriovenous fistula was significantly upregulated (P < 0.000 1), and α-smooth muscle actin (α-SMA) , proliferating cell nuclear antigen (PCNA) positive cells increased significantly (P < 0.000 1), indicating an increase in local cell proliferation level. Conclusion The established mouse autologous arteriovenous fistula model has the advantages of high success rate, good stability and low cost. The model provides a good carrier for exploring the biological mechanism of intimal hyperplasia in arteriovenous fistulas. Table and Figures | Reference | Related Articles | Metrics

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Ginkgolide B Promotes Neural Function Recovery of Ischemic Stroke Mice by Regulating Characteristics of Brain T Cells and Their Interactions with Glial Cells Jia LIU, Yanrong YE, Yun SHEN, Qiying TANG, Meiqing CHEN, Kehui YI, Shaozhuang CHEN Laboratory Animal and Comparative Medicine    2024, 44 (2): 139-148.   DOI: 10.12300/j.issn.1674-5817.2023.121 Abstract （208）   HTML （27）    PDF （1754KB）（177）       Knowledge map    Save Objective To investigate the regulatory effects of Ginkgolide B on the biological characteristics of brain T cells and their interactions with glial cells during the recovery phase of ischemic stroke in mice. Methods 36 adult C57BL/6 mice were randomly assigned to three groups: sham-operated group (Sham group), control group (PBS group), and Ginkgolide B treatment group (GB group). The Sham group underwent only sham surgeries, whereas the PBS and GB groups were subjected to a middle cerebral artery occlusion (MCAO) model using the filament method, followed by intranasal administration of an equivalent volume of either PBS or Ginkgolide B solution for 14 days post-injury. Neurological function changes were evaluated in all three groups using the rotarod test and a neurological scoring system. On day 15, single-cell sequencing was performed on fresh tissues from the brain injury areas, surrounding cortex, corpus callosum, and striatum of mice in the PBS and GB group to assess the biological characteristics of T cells and their subpopulations, and further explore the interactions and mechanisms among T cells, microglia, and oligodendrocytes. Results Compared with the Sham group, both PBS and GB group exhibited significant improvements in neurological scores and reduced pre-fall motor durations (P < 0.001). Compared with the PBS group, the GB group showed a downward trend in neurological scores and an upward trend in pre-fall motor durations on days 5, 10, and 15 post-ischemic brain injury, with a significant increase in pre-fall motor duration on day 15 (P < 0.05). Compared with the PBS group, the GB group exhibited a significant increase in T cell proliferative activity in the brain 15 days post brain injury (P < 0.05). The number of proliferative T cells and the levels of lipid metabolism were significantly elevated (P < 0.05), and there was a significant increase in extracellular matrix remodeling in all T cells (P < 0.05). Additionally, the interactions between T cells and both microglia and oligodendrocytes, as well as among the microglia themselves and between microglia and oligodendrocytes, were significantly enhanced in the GB group. This was primarily evident in the strengthened interactions between CD74 and macrophage migration inhibitory factor (MIF), as well as colony stimulating factor 1 receptor (CSF1R) and colony stimulating factor 1 (CSF1) (P < 0.05). However, the inflammatory levels of T cells showed no significant differences compared with the PBS group. Conclusion A mouse model of ischemic stroke can be successfully established by MCAO operation. Ginkgolide B may promote neurological recovery post-brain injury in mice by modulating the biological characteristics of T cells within the brain and their interactions with glial cells. Table and Figures | Reference | Related Articles | Metrics

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Fine Structure of the Trunk Kidney and Distribution of Its Secreted Exosomes in the Adult Zebrafish Jinxing LIN, Xindong WANG, Xuebing BAI, Liping FENG, Shuwu XIE, Qiusheng CHEN Laboratory Animal and Comparative Medicine    2023, 43 (5): 531-540.   DOI: 10.12300/j.issn.1674-5817.2023.070 Abstract （202）   HTML （123）    PDF （3115KB）（441）       Knowledge map    Save Objective To observe the fine structure of the trunk kidney in zebrafish, and to identify its secreted exosomes. Methods The microstructure and ultrastructure of the trunk kidney in zebrafish were observed by light microscopy and electron microscopy, and the particle size of exosomes was detected by nanoparticle tracking analysis (NTA). Results The trunk kidney was close and parallel to the spine in adult zebrafish. The nephron consisted of renal tubules and renal corpuscles. The renal tubules could be further divided into three types: proximal convoluted tubules, distal convoluted tubules, and cervical segments. The renal corpuscles were composed of glomerulus and renal capsules. The periodic acid-Schiff (PAS) staining results revealed that there were abundant glycogen granules in the proximal convoluted tubules, with brush-like outline in the apical surface of epithelial cells. Under transmission electron microscopy (TEM), there were exosomes distributed in the lumen of renal tubules, with numerous late endosomes and few number of multivesicular bodies (MVBs) in the cytoplasm of the epithelial cells concentrating on the apical side. Meanwhile, MVBs were also distributed in the apical regions of the renal tubules and the podocytes of the renal glomeruli. Immunohistochemical staining results showed that CD9, CD63 and TSG101 were strongly expressed in the lumen surface of the renal tubules, but weakly expressed in the corpuscles and lumen. NTA and TEM results showed that the exosomes isolated from zebrafish trunk kidney were saucer-like outline, and the particle size mode was 144.4 nm, which was consistent with the characteristics of morphological futures of exosome. Conclusion The zebrafish somatic kidney has the typical structure of the mammalian kidney and is the urinary organ in the body. The renal tubules have the ability to secrete exosomes, and their formation is a process of releasing poly-vesicles to the free surface of epithelial cells into the extracellular space. This study laid a morphological foundation for further study of exosomes in urinary function in aquatic experimental animals as well as the development and application of related models. Table and Figures | Reference | Related Articles | Metrics

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Progress in Establishment and Application of Laboratory Animal Models Related to Development of Male Infertility Drugs Shuwu XIE, Ruling SHEN, Jinxing LIN, Chun FAN Laboratory Animal and Comparative Medicine    2023, 43 (5): 504-511.   DOI: 10.12300/j.issn.1674-5817.2023.120 Abstract （199）   HTML （144）    PDF （853KB）（307）       Knowledge map    Save As the incidence of male infertility has been increasing during recent years, it is urgent to reveal the pathogenesis of male infertility, as well as to develop the new drugs for treatment of male infertility, in order to solve the declining birth rate and aging problems. The construction and application of male infertile animal models is critical for drug development, which plays an important role in accurately evaluating the efficacy and mechanism of infertility treatment. A suitable infertility model not only can reduce the repeated drug efficacy evaluations, reduce animal usage and the cost of new drug development, but also has important reference value for subsequent clinical trial research. Male infertility laboratory animal models can be constructed through chemical, physical, endocrine, environmental estrogen, gene modification, and immune methods. This article mainly introduces the existing male infertility animal models available for drug development, and briefly introduces the application progress of each model to provide reference for the male infertility drug researchers. Reference | Related Articles | Metrics

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Establishment of Fluorescence qPCR Method for Detection of Staphylococcus Aureus and Its Application in Feces Detection of Rats and Mice Lingzhi YU, Jianyun XIE, Liping FENG, Xiaofeng WEI Laboratory Animal and Comparative Medicine    2023, 43 (5): 566-573.   DOI: 10.12300/j.issn.1674-5817.2023.022 Abstract （196）   HTML （104）    PDF （1461KB）（244）       Knowledge map    Save Objective To establish a method for rapid and sensitive detection of Staphylococcus aureus. Methods The specific gene nuc of Staphylococcus aureus was selected as the target gene. A pair of specific primers and a TaqMan probe were designed and synthesized according to the published sequence of the nuc gene. Establish a nucleic acid detection method for nuc gene using fluorescence quantitative PCR technology, and apply it clinically in the detection of fecal samples from rats and mice. Results The DNA extracted from Staphylococcus aureus and other non-Staphylococcus aureus strains was detected by qPCR. The results showed that Staphylococcus aureus had a specific amplification curve, while other non-Staphylococcus aureus did not, indicating that the designed primers and probes were specific for Staphylococcus aureus. The sensitivity of this method was determined by diluting the DNA of Staphylococcus aureus by 10 times. The results showed that the detection limit of this method was 10 fg DNA, which was 2 orders of magnitude higher than that of ordinary PCR method. A total of 91 clinical samples were detected in this study, of which 4 rat samples from the same facility had a typical S-curve. The PCR products were sequenced and BLAST compared. The gene sequence of this sample was 100% similar to that of Staphylococcus aureus, indicating that the sample was positive for the nucleic acid of Staphylococcus aureusnuc gene, with a positive rate of 4.40%. The result was consistent with that obtained by bacterial culture method. The nucleic acid extraction adopted a full-automatic nucleic acid purification instrument, and the time required from nucleic acid extraction to detection result determination was less than 1.5 h. Conclusion The qPCR method established in this study to identify Staphylococcus aureus with nuc gene as the target gene has the advantages of fast, high sensitivity and specificity, and can be used for the detection of Staphylococcus aureus in feces of rats and mice. Table and Figures | Reference | Related Articles | Metrics

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Analysis of the Birthing Behaviour of Cynomolgus Macaques Xinyan BIAN, Yong LU, Yan WANG, Qiang SUN Laboratory Animal and Comparative Medicine    2023, 43 (4): 355-362.   DOI: 10.12300/j.issn.1674-5817.2023.086 Abstract （196）   HTML （20）    PDF （1036KB）（187）       Knowledge map    Save Objective Observing and analyzing the delivery behaviour of cynomolgus macaques, to establish the criteria for determining the occurrence of delivery in cynomolgus macaques, and then combining with veterinary assistance in order to improve the live birth rate of cynomolgus macaques. Methods By backtracking and analyzing the surveillance videos of 112 perinatal cynomolgus macaques with a gestation period of 140 d or more from 2017 to 2021, we observed and recorded the main behavioural manifestations of the cynomolgus macaques during labour, including Valsalva's maneuver, touching and licking the birth canal, lying on their backs or stomachs, and rolling and tumbling of the body. On this basis, we established the weights of delivery-related behavioural indicators and exhaustively analysed the perinatal behavioural performances of 30 cynomolgus macaques for delivery determination. Results The perinatal behavioural validation results of the 30 cynomolgus macaques showed that the behavioural indicators of Valsalva's maneuver, touching and licking the birth canal, lying on the stomach or on the floor, body rolling and tumbling occurred with different frequencies, among which Valsalva's maneuver and lying on the stomach or on the back were the most important, with weight values of 35.5% and 27.2%, respectively. These two behaviours can be used to accurately determine the onset of parturition in cynomolgus macaques. The average live birth rate of the monkeys that were accurately determined to have given birth and were assisted by veterinarians reached 87.1%, which was significantly higher than that of the monkeys that had unassisted spontaneous deliveries, which was 63.5%, and there was a significant difference between these two rates (P<0.05). Conclusion The combination of accurate birth determination and veterinary assisted delivery can significantly increase the live birth rate of experimental cynomolgus macaques. Table and Figures | Reference | Supplementary Material | Related Articles | Metrics

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Application of Allograft Endometriosis Rat Model in Pharmaco-dynamic Evaluation of GnRH Agonists Ruihua ZHONG, Guoting LI, Wenjie YANG, Xiangjie GUO, Jieyun ZHOU, Yingyi HU, Qicheng NI, Ye YANG, Min ZHANG, Yan ZHU Laboratory Animal and Comparative Medicine    2024, 44 (2): 127-138.   DOI: 10.12300/j.issn.1674-5817.2023.150 Abstract （190）   HTML （34）    PDF （2085KB）（207）       Knowledge map    Save Objective To establish an allogeneic rat model of endometriosis and to evaluate the effects of gonadotropin-releasing hormone (GnRH) agonist GenSci006 on experimental rat endometriosis. Methods Endometrium from SPF grade donor female SD rats were transplanted onto the abdominal wall of recipient female rats to construct an allogeneic endometriosis model. The rats undergoing sham surgery were divided into the sham group. Three weeks later, the length, width and height of the ectopic endometrium were measured, and the volume of the endometrium (V1) was calculated before drug administration. The modeling rats were randomly divided into four groups: model group, triptorelin group (0.25 mg/kg), GenSci006-1 group (0.125 mg/kg) and GenSci006-2 group (0.25 mg/kg). Each group had 16 rats and received a single dose of the corresponding drug. The sham group and model group were administered an equal volume of solvent. Three weeks after administration, ectopic endometrium was measured to calculate the volume V2 and inhibition rate. The effect of GenSci006 on rat uterus and ovarian tissues was assessed by comparing organ coefficients and changes in pathological sections. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of serum estradiol (E2), progesterone (P4), follicle stimulating hormone (FSH), and luteinizing hormone (LH). Real-time fluorescent quantitative PCR was used to detect the expression of GnRH receptor (GnRHR) mRNA in the hypothalamus and pituitary. Western blot was used to detect the expression of estradiol receptor alpha (ERα), beta (ERβ) and progesterone receptor (PR) in ectopic endometrium. Results Three weeks after administration, compared with the model group, the body weight of rats in the triptorelin and GenSci006-2 groups significantly increased (P < 0.05), while the volume of ectopic endometrium significantly decreased (P < 0.05). Compared with the sham group, the model group showed no significant changes in uterine and ovarian organ coefficients or endometrial thickness (P > 0.05). Compared with the model group, the uterine organ coefficients and endometrial thickness were significantly reduced in the triptorelin and GenSci006-2 groups (P < 0.05). Compared with the sham group, the serum levels of E2, P4, FSH and LH in the model group showed no significant changes (P > 0.05). Compared with the model group, the ovarian organ coefficient and serum P4 levels of rats in the Triptorelin, GenSci006-1, and GenSci006-2 groups were significantly reduced (P < 0.05), while the serum LH levels of rats in the GenSci006-1 group were significantly increased (P < 0.05). However, there were no significant changes in serum E2 and FSH levels in each group (P > 0.05). Compared with the model group, the expression levels of GnRHR mRNA in the pituitary tissue of rats in the triptorelin and GenSci006-2 groups were significantly downregulated (P < 0.05), with no significantly changes in the hypothalamus (P > 0.05). There were no significant changes in the expression level of GnRHR mRNA in the hypothalamus or the protein levels of ERα, ERβ and PR in the ectopic endometrial tissue in any group (P > 0.05). Conclusion The allogeneic endometriosis rat model is a suitable animal model for screening and evaluating drugs for treating endometriosis. The volume of ectopic endometrium, inhibition rate, uterine and ovarian organ coefficients, and serum E2 levels may serve as indicators for detecting drug efficacy. Table and Figures | Reference | Related Articles | Metrics

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Creation and Analysis of Related Genetic Characteristics of BALB/cA.Cg.SHJH hr Mice Xiaoqian TAN, Hao YANG, Huiqing TANG, Wei QU, Liang LI, Zhen QIAN, Jianzhong GU, Ping XU, Junhua XIAO Laboratory Animal and Comparative Medicine    2023, 43 (4): 363-370.   DOI: 10.12300/j.issn.1674-5817.2023.055 Abstract （181）   HTML （33）    PDF （1768KB）（211）       Knowledge map    Save Objective To introduce the Hr gene of spontaneously mutated SHJH hr mice into BALB/cAShjh inbred mice with clear genetic background,and provide a basis for study on the molecular mechanism of Hr gene mutation-induced abnormal phenotype and the application of this model. Methods Using a backcross-intercross breeding method guided by phenotypic monitoring, mutant genes from SHJH hr mice bred by spontaneous mutation were introduced into inbred BALB/cAShjh mice by homozygous mutation introgression, and the mice were bred into BALB/cA.Cg.SHJH hr (abbreviated as C.Cg.SHJH hr ) mice after 10 generations. The genotypes of 90 single nucleotide polymorphism (SNP) detection sites were analyzed in C.Cg.SHJH hr mice by multiplex PCR library construction followed by next generation sequencing. Then 14 biochemical locus marker genes were detected in C.Cg.SHJH hr mice according to the method of GB/T 14927.1-2008. Finally, whole genome exon sequencing was utilized to detect the mutated genes in this mouse. Results From May 2018 to March 2022, a total of 10 generations of backcross-intercross were conducted to complete the construction of the C.Cg.SHJH hr mouse line. Among the 90 SNPs loci detected, except for rs13484115 and rs13484116, all the other loci had the same genotype as the recipient mice BALB/cAShjh. The results of biochemical marker gene detection showed that all the 14 loci of the mouse were the same as those of the recipient mouse. Whole genome exon sequencing found that the mouse had 109 site mutations compared with the recipient mouse strain, including 71 synonymous mutations, 1 stopgain, 37 missense mutations, and 20 genes involved in protein sequence alterations (including the reported Hr gene). Conclusion C.Cg.SHJH hr mice were created. Through exon sequencing and genetic analysis, three Hr mutated genes and associated mutated genes that mainly cause phenotypic variations were identified, which provides a basis for expanding the application of C.Cg.SHJH hr mice in biomedical research. 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 （180）   HTML （31）    PDF （1051KB）（206）       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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Discussion on the Operation, Maintenance and Care Modes of Laboratory Animals Facilities Yue HUANG, Ye DONG, Jiale SHU Laboratory Animal and Comparative Medicine    2024, 44 (1): 92-96.   DOI: 10.12300/j.issn.1674-5817.2023.078 Abstract （180）   HTML （22）    PDF （697KB）（210）       Knowledge map    Save With the rapid development of the biopharmaceutical industry in China, the laboratory animal sector has entered a stage of rapid growth, and the construction of facility operation systems has become increasingly automated and intelligent. Compared to traditional laboratory animal facilities, new facilities require a more specialized technical team for the maintenance of air supply and exhaust systems, air conditioning, automated control, and the entire barrier system. The Lingang Laboratory’s animal facilities accommodate both large and small animals for feeding and experimental purposes. The facility management team has summarized daily maintenance experiences and explored various operational and maintenance modes based on the characteristics of laboratory operation. After analyzing the advantages and disadvantages of three common modes, this paper provides new ideas for the management of these laboratory animal facilities, and offers guidance for peers in choosing the most appropriate professional maintenance mode. Table and Figures | Reference | Related Articles | Metrics

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Generation of 12 Drosophila Transgenic Negative Control Lines Based on Site-specific ΦC31 Integrase and pUASTattB Vector Longmei XU, Ruling SHEN, Chun FAN, Wei WU Laboratory Animal and Comparative Medicine    2023, 43 (5): 541-547.   DOI: 10.12300/j.issn.1674-5817.2023.100 Abstract （177）   HTML （122）    PDF （1374KB）（377）       Knowledge map    Save Objective Construction of a negative control line for the Drosophila transgenic system based on ΦC31 integrase and vector plasmid pUASTattB to provide a more scientific negative control for transgenic Drosophila research experiments. Methods The vector plasmid pUASTattB was microinjected into four different genetic backgrounds Drosophila lines attP-25C6, attP-68A4, attP-75B1 and attP-86F8 embryos carrying ΦC31 integrase. All of the injected embryos were incubuated to get G0 adults, and each of them was crossed with balancer stock ywR13S separately in a single vial (1 adult of the G0 generation and 3 of the ywR13S in each vial). The probability of successful insertion was calculated by observing the colour of the compound eyes of the G1 generation of Drosophila to determine whether there was a mini-White insertion. The G1 generation Drosophila adults successfully inserted into mini-White were then selected to make single-vial crosses (one G1 generation male Drosophila crossed with three virgins of balancer Drosophila line) with each of the three balancer Drosophila strains DB, ywR13S and yw122, respectively, for balanced seed preservation. The genomic DNA of the conserved Drosophila lines was extracted and the vector plasmid pUASTattB was identified for transfer by PCR. Results 12 Drosophila strains were obtained, all of which were red-eyedDrosophila melanogaster carrying the mini-White marker, and were identified by PCR as having the pUASTattB sequence insertion. Conclusion The 12 transgenic Drosophila strains can meet the negative control requirements for the transgenic fly research experiments that constructed with pUASTattB as the vector basically, enriching the Drosophila resources in the National Drosophila Resource Center of China. Table and Figures | Reference | Related Articles | Metrics

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Literature Analysis of the Preparation Elements of Animal Models of Skin Photoaging and the Data of Subjects Yasheng DENG, Jiang LIN, Chiling GAN, Guanfeng ZENG, Jiayin HUANG, Huifang DENG, Yingxian MA, Siyin HAN Laboratory Animal and Comparative Medicine    2023, 43 (4): 406-414.   DOI: 10.12300/j.issn.1674-5817.2023.026 Abstract （173）   HTML （7）    PDF （805KB）（219）       Knowledge map    Save Objective To analyze the modeling elements and subjects of the animal model of skin photoaging, and to provide a reference for the preparation and improvement of the model and a basis for the scientific evaluation of the subject. Methods By searching and collecting relevant literature on the preparation of animal models of skin photoaging from 2010 to 2022 in the China National Knowledge Infrastructure, Wanfang Database, and PubMed database, the model animal species, gender, modeling method, modeling cycle, radiation source and its distance from the modeling site, cumulative radiation volume, detection indicators, and subjects (drugs or treatments) recorded in the literature were collated and summarized, and a database was established for statistical analysis. Results 257 articles that met the inclusion criteria were selected. Among them, the most common animal model was SKH-1 hairless mice, followed by SD rats and KM mice; the gender of animals was mainly female, medium-wave ultraviolet B (UVB) was often used as the radiation source, the distance between the radiation source and the modelling site was mostly 30 cm, and the modelling period was usually 40-60 days. The cumulative dose of long-wave ultraviolet A (UVA) was between 100-150 J/cm2, and the cumulative dose of UVB was between 5-10 J/cm2. The tests used after model establishment were skin histopathological examination, skin tissue homogenization, fibre staining, immunoblotting, etc. Subjects included Chinese herbal medicines, Chinese herbal extracts, Chinese patent medicines, Chinese herbal compound medicines, chemical drugs, biological agents and other treatments, while the animal model of skin photoaging was also used for clinical efficacy studies of external Chinese medicine, physiotherapy and positive control drugs. Conclusion In skin photoaging animal experiments, female SKH-1 hairless mice are often used, and UVB is used as the radiation source. The modeling period is usually 40-60 days, and the minimum erythema dose (MED) is incremented week by week. The cumulative UVB irradiation dose ranges from 0 to 10 J/cm2, which has the advantages of high success rate, good reproducibility and high similarity with clinical disease. Table and Figures | Reference | Related Articles | Metrics

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Establishment of Preeclampsia Model in Goat and Evaluation on Maternal Biological Characteristics Jin LU, Jian WANG, Lian ZHU, Guofeng YAN, Zhengwen MA, Yao LI, Jianjun DAI, Yinqiu ZHU, Jing ZHOU Laboratory Animal and Comparative Medicine    2023, 43 (4): 371-380.   DOI: 10.12300/j.issn.1674-5817.2023.036 Abstract （170）   HTML （22）    PDF （2128KB）（205）       Knowledge map    Save Objective Establish an animal model of preeclampsia in goats, collect data on various physiological indicators and maternal biological characteristics of the disease model to compare with clinical feature of the disease in humans, provide reference for the prevention and treatment of preeclampsia in humans. Methods Twenty-three goats bred in Chongming district were divided into three groups: Control group, no surgical procedure was performed on animals of this group; sham group, ewes in this group underwent the sham operation on the 100±5th day of gestation, and only the uterine artery was exposed and dissociated; surgical group, a silver vascular clamp was clipped on one side of the uterine body artery of the ewe to narrow the inner diameter of the artery at the same gestation period (100±5) days. Heart rate and hindlimb blood pressure were continuously monitored in control and surgical ewes from 100 to 140 d of gestation, and blood flow data within the lateral branches of the uterine arteries of ewes in the sham group were collected using a hemodynamometer in combination with a hemodynamic probe and an animal physiological signal collector, as well as changes in blood flow within the uterine arteries in the lateral branches of the uterine arteries of the surgical group before and after placement of vascular clips in the surgical ewes. At the expected date of delivery, jugular vein blood was taken from ewes for routine blood test, creatinine, urea nitrogen, and blood ion analysis; urine was also collected from ewes in each group for analysis of urinary protein and urinary creatinine. All experimental groups were subjected to cesarean section on the 140±5th day of gestation in ewes, and then liver, kidney, uterus and placenta tissues were taken from ewes in each group and stained with HE for pathological observation. Results After 15 minutes of preeclampsia modeling surgery, blood flow volume remained stable in the vessel stenosis segment and the volume differential was relatively reduced in comparison to the control group and sham group (P＜0.05，P＜0.01). Compared with the control group, the ewes in the surgical group showed prenatal changes such as increased serum osmolality, decreased hemoglobin, increased blood glucose and urea nitrogen values, as well as increased levels of calcium, sodium, and chloride ions (all P<0.05) and proteinuria, with urinary creatinine and urinary protein-creatinine ratios were significantly higher than those in the control group and sham group (all P<0.05). The elastic lamina of the uterine body arteries on the operated side of the animals in the surgical group was thicker than that on the opposite side, but the structure was loose. The placenta on the operated side showed pathological changes such as cell interstitial swelling and inflammatory cell infiltration. The above physiological index characteristics were more consistent with the clinical features of human preeclampsia disease. Conclusion In this experiment, we successfully constructed a goat preeclampsia model and obtained data on relevant physiological indexes of this model, which further verified the correlation between preeclampsia disease and uterine artery lesions. 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 （170）   HTML （23）    PDF （1936KB）（155）       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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Study on the Antibody Production Efficiency in Modified Big-BALB/c Mice Dan WANG, Xiaolu ZHANG, Yan WANG, Bo FU, Wendong WANG, Jing LIU, Suyin ZHANG, Yihe WU, Deguo WU, Xiaoyan DU, Dawei ZHAN, Xiulin ZHANG, Changlong LI Laboratory Animal and Comparative Medicine    2023, 43 (6): 612-618.   DOI: 10.12300/j.issn.1674-5817.2023.035 Abstract （167）   HTML （20）    PDF （1032KB）（186）       Knowledge map    Save Objective To compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c (B-BALB/c) mice, and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies induced in vivo through hybridoma. Methods Individuals weighing more than 5% of the weight of normal animals at 4 weeks of age (the criterion for late selection is more than 10%) were selected from a population of conventionally bred BALB/c mice and bred individually, and a subline of B-BALB/c mice was prepared after 10 generations of selection. A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks, half male and half female, were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin, respectively. Mice ascites containing monoclonal antibodies were obtained by in vivo induction. The antibody titer was tested by indirect ELISA. The mice were grouped based on the sub-strains, gender and inoculation type of hybridoma to analyze the ascites production, antibody titer and antibody production, and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains. Results After 10 generations of breeding, the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3% and 12.8%, respectively, compared with BALB/c mice of the same age. Compared with BALB/c mice, B-BALB/c mice had better tolerance and adaptation to secondary ascites collection. Compared with BALB/c mice, the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased, especially in the hybridoma cell line G23 vaccination group (both P＜0.000 1) . After inoculation with the hybridoma cell lines G23 or Y15, the average antibody production of B-BALB/c mice (14.99×104 U and 33.22×104 U) was higher than that of BALB/c mice (5.33×104 U and 19.31×104 U) (both P＜0.01). After inoculation with hybridoma cell line G23, the average antibody production per unit body weight of B-BALB/c mice (0.55×104 U/g) was higher than that of BALB/c mice (0.23×104 U/g) (P＜0.000 1). And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice (both P＜0.01). Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation, which can achieve the purpose of reducing the number of experimental animals used, lowering the labor cost, and improving the efficiency of antibody preparation. Table and Figures | Reference | Related Articles | Metrics

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Progress in the Application of Animal Disease Models in the Medical Research on Colorectal Cancer Yanjuan CHEN, Ruling SHEN Laboratory Animal and Comparative Medicine    2023, 43 (5): 512-523.   DOI: 10.12300/j.issn.1674-5817.2023.076 Abstract （166）   HTML （135）    PDF （1046KB）（329）       Knowledge map    Save Colorectal cancer (CRC) is the third most common malignant tumor in the world. The latest statistics show that CRC accounts for 10% of all cancer cases worldwide and is the second leading cause of cancer deaths. CRC is a highly heterogeneous disease, the development of which is driven by functional abnormalities or epigenetic changes caused by multiple gene expression mutations, and there are different pathways that lead to tumor formation. Complex factors such as genetics, environment, ethics, and individual differences of patients themselves limit the study of CRC in humans, so the disease animal models have become an indispensable tool for the study of CRC, and play an important role in prevention, treatment, preclinical research and basic research. There are various types of CRC animal models, of which mouse models are the most widely used. According to different model establishing methods, the models are divided into spontaneous, chemically induced, transplanted tumor and genetic-engineering mouse models. Different models have different characteristics and application prospects. In this study, we focus on these mouse models of CRC in detail, and introduce the latest research progress of CRC models in rats, experimental pigs and zebrafish, to provide reference for the selection and application of animal models of CRC. Table and Figures | Reference | Related Articles | Metrics

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Current Status and Reflection on the Study of Welfare for Laboratory Fish Liping FENG, Qi ZHU, Jinxing LIN Laboratory Animal and Comparative Medicine    2023, 43 (5): 524-530.   DOI: 10.12300/j.issn.1674-5817.2023.069 Abstract （165）   HTML （131）    PDF （818KB）（234）       Knowledge map    Save In recent years, with the rapid development of life sciences, the use of laboratory fish in toxicology, genetics, developmental biology and medicine has increased dramatically, and they have gradually become important new model organisms. At the same time, the welfare of laboratory fish has also received increasing attention. Although the research level of experimental fish welfare is still in a relatively early stage compared to terrestrial experimental animals, developed regions such as Europe and America have established corresponding legal frameworks to safeguard the welfare of laboratory fish in research. This article elucidates the current developmental status of laboratory fish welfare, discusses the rationale behind the imperative to prioritize and enhance their welfare, deeply investigates factors influencing their welfare from the feeding stage and experimental stage. Moreover, it explores strategies for augmenting welfare standards, with the overarching aim of propelling the continual improvement of laboratory fish welfare in our country. 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 （164）   HTML （190）    PDF （2172KB）（223）       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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Exploration of Laboratory Animal Science Teaching Practice from Perspectives of Curriculum Ideology and Politics Ya ZHAO, Caiqin ZHANG, Han MENG, Jing QIN, Bing BAI, Yong ZHAO, Xu GE, Changhong SHI Laboratory Animal and Comparative Medicine    2023, 43 (6): 641-646.   DOI: 10.12300/j.issn.1674-5817.2023.109 Abstract （162）   HTML （116）    PDF （803KB）（240）       Knowledge map    Save The ideological and political content of the laboratory animal science degree course with the basic task of "cultivating morality and cultivating people" is organically integrated into the teaching system of laboratory animal science. It can have a subtle influence on students' thoughts and behaviors. Combined with the curriculum design and professional characteristics of laboratory animal science, this article discussed the ideological and political elements contained in this course, proposed the forms and methods of integrating ideological and political elements into the curriculum design in each chapter. Additionally, the typical cases and characteristic practices of the organic connection of ideological and political education in the teaching system of laboratory animal science were summarized. Practice has proved that integrating the ideological and political elements into the teaching system of laboratory animal science can enhance teacher's awareness and ability of politics, thus effectively improving the compre-hensive quality of students and enhancing the effectiveness of ideological and political education in laboratory animal science. Table and Figures | Reference | Related Articles | Metrics

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Construction and Verification of Quality Evaluation Indicator System for Extracorporeal Membrane Oxygenation Animal Experimental Platform Shuo WANG, Yunhui LÜ, Xiaokang WANG, Zhenhao ZHANG, Yongchun CUI Laboratory Animal and Comparative Medicine    2023, 43 (6): 604-611.   DOI: 10.12300/j.issn.1674-5817.2023.107 Abstract （161）   HTML （15）    PDF （1017KB）（260）       Knowledge map    Save Objective To establish a standardized and professional service quality evaluation index system for extracorporeal membrane oxygenation (ECMO) animal experimental platform. Methods The literature research and expert consultation were used to establish a factor set for the quality evaluation of ECMO animal experimental platform. Then, experts used the 1/9-9 scale method to compare and score pair-two indicators. Based on the principles of fuzzy analytic hierarchy process and expert scoring results, the ECMO animal experimental platform quality evaluation system was constructed. In order to verify the actual efficacy of this system, a case study was carried out on the ECMO animal experiment platform of FW Animal Experimental Center (FAEC) laboratory. Results A total of 10 experts were included in this study, the questionnaire recovery rate was 100%, the judgment coefficient (Ca) and familiarity coefficient (Cs) were both greater than 0.50, the expert authority was high (Cr>0.80), the validity test was P<0.01, and the coordination was good. The quality evaluation system of ECMO animal experiment platform includes two levels. There are 4 first-level indicators, with professionalism, safety, functionality, and stability ranked from high to low in terms of their weights. There are 15 second-level indicators, and the top 5 weights are personnel's technical expertise, attractiveness of hardware facility, auditability of data, confidentiality capabilities of data, and professionalism in service process. To facilitate the popularization and application of the system, this study also proposed a "star" system to represent the evaluation results of an ECMO animal experimental platform quality. The quality evaluation system established in this study was used to evaluate the FAEC laboratory as a case study, and the evaluation result was five-star. The actual potency value of FAEC laboratory was 0.910, reaching the five-star level, but the average actual appraisal values of "service continuity" and "sufficiency of project completion" were lower than 0.80, which needs to be improved. Conclusion A standardized and professional ECMO animal experimental platform quality evaluation system was established in this study, which would provide a measurable basis for the demander to select the supplier and a method for the supplier to complete the animal experiment of ECMO research and development with high quality. Table and Figures | Reference | Related Articles | Metrics

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Transcriptome Data and Comparative Medical Analysis of COVID-19 Virus Infection Tingting FENG, Yitong LI, Yue WU, Jue WANG, Qi KONG Laboratory Animal and Comparative Medicine    2024, 44 (1): 62-73.   DOI: 10.12300/j.issn.1674-5817.2023.079 Abstract （160）   HTML （11）    PDF （1282KB）（157）       Knowledge map    Save Objective To provide more basic information of comparative medicine for the study of biological changes and pathogenesis of COVID-19 by systematical sorting and analyzing the transcriptome data. Methods Following a retrieval strategy, using COVID-19 and SARS-CoV-2 as key words, transcriptome datasets related to COVID-19 from January 2020 to May 2023 were collected from GEO, ArrayExpress and GEN Transcriptome databases. The composition, distribution, and research application of COVID-19 transcriptome data resources were analyzed. Data distribution was visually displayed and correlation analysis was performed. The research applications and limitations of existing COVID-19 transcriptome data were analyzed from the perspectives of clinical medicine and comparative medicine, focusing on clinical-related molecular mechanisms, biomarkers and related immune responses, treatment intervention strategies, etc. The research value and application prospects were discussed. Results A total of 975 sets of COVID-19 transcriptome data were included. Among three databases, datasets from humans accounted for the highest proportion, namely 71.9%, 77.9%, and 90%, respectively. Species other than humans, such as mice, were the main sources of data, with the respiratory and nervous systems having the highest distribution of data. Twenty-seven datasets were associated with clinical significance. Analysis revealed that respiratory tract injury and other related molecular mechanisms were obtained through transcriptome data mining. Biomarkers such as cfDNA could be used as therapeutic targets. The severity of COVID-19 infection was associated with cell changes and disorders represented by M1 macrophages. Comparative medical analysis showed that mice, hamsters, and other animals were susceptible to SARS-CoV-2. Rhesus monkeys and cynomolgus monkeys exhibited infection characteristics highly similar to human. Apart from respiratory symptoms, hamsters also exhibited digestive system symptoms. SARS-CoV-2 can replicate in the respiratory organs of various susceptible animals, the intestines of ferrets and the ears of minks, resulting in interstitial pneumonia, diffuse lung injury and other pathological changes of varying degrees. Based on the differences in immune responses, hamsters can be used for neutralizing antibody reaction research. Conclusion Currently there is a wealth of COVID-19 transcriptome data, but there is a lack of comparative transcriptome research. Transcriptomics can be combined with comparative medicine to further explore the differences in comparative medicine of COVID-19. 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 （154）   HTML （20）    PDF （841KB）（176）       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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Research on the Virulence Identification and Preservation Methods of Desert-type Leishmania donovani Strains Lifu LIAO, Yun LUO, Shen SHI, Yimei XU Laboratory Animal and Comparative Medicine    2023, 43 (6): 619-625.   DOI: 10.12300/j.issn.1674-5817.2023.034 Abstract （152）   HTML （8）    PDF （1190KB）（161）       Knowledge map    Save Objective To determine the virulence of desert-type Leishmania donovani strains through animal infection experiments and to explore preservation methods for maintaining their pathogenicity. Methods The isolated strain was cultured in vitro for 7, 30, 36, 44, 60, 90, and 150 days, respectively， and inoculated into Lagurus lagurus (L.lagurus) with the dose of 2.6×105 per animal by intraperitoneal injection. The spleen coefficient, infection rate, and antibody positive rate of the inoculated animals were detected at day 60 after infection. The desert-type Leishmania donovani strain was further inoculated with Cricetulus migratorius (C.migratorius) and L. lagurus, respectively, for passaging and preservation. The survival time of two kinds of animals and pathogenicity change of the stain in their bodies were compared. Results After inoculation of desert-type Leishmania donovani strains cultured in vitro for 7-150 days, the spleen coefficient of inoculated L.lagurus gradually increased from 1% on day 7 to 2.2% on day 30, which was more than 10 times of the normal spleen coefficient. Additionally, on day 60, the spleen coefficient remained 3 times higher than the normal value. The infection rate and antibody positive rate decreased from 80% on day 7 to 0% on day 60. At 90 days, there were no significant differences between the infected groups and the control group, and all the observed indexes were within the normal range. The survival time of L.lagurus infected with the in vivo passage strain ranged from 1 to 13 months, and half of the infected individuals died within 4 months. In contrast, C.migratorius had a survival time ranging from 5 to 31 months, and half of the infected individuals died within an average of 13.7 months. There was a significant difference in the average time of death between the two groups (t=0.000 1, P<0.001), but no significant difference in spleen coefficient (t=0.990, P>0.05). This strain exhibited equal virulence in both animals and remained virulent for up to 4 years after continuous passage. Conclusion With the prolonged culture time, the virulence of the strain decreases gradually. At 90 d, it has no pathogenicity to L. lagurus. Long-term in vitro culture fails to preserve it's pathogenicity to L.lagurus. Only in vivo inoculation can maintain the virulence of this strain. Table and Figures | Reference | Related Articles | Metrics

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Application of Optimized Latex Perfusion Technique in the Establishment of Craniofacial Venous Model in Mice Chengji WANG, Jue WANG, Haijie WANG, Weisheng LU, Yan SHI, Zhengye GU, Mingqiu WAN, Ruling SHEN Laboratory Animal and Comparative Medicine    2023, 43 (5): 574-578.   DOI: 10.12300/j.issn.1674-5817.2023.095 Abstract （151）   HTML （109）    PDF （1145KB）（194）       Knowledge map    Save Objective Optimize the latex perfusion technique and apply it to the construction of a murine craniofacial venous vascular model. Methods A total of nine 8-week-old male C57BL/6 mice weighing (25.0±1.3) g were randomly divided into three groups: 60% latex physiological saline group, 60% latex heparin group, and 30% latex heparin group. After completion of the perfusion, the specimens were immersed in 4 °C formalin fixative for 24 h, followed by dissection, observation, and measurement of the extracranial blood vessel diameters. Results After 200 μL latex perfusion solution was injected into the external jugular vein, the supraorbital vein, infraorbital vein, temporal vein, retrofacial vein, masseter vein and external jugular vein were perfused in each group.After comparing the perfusion degree of the distal branches of blood vessels, sublingual vein and tip venule, it was found that the 30% latex heparin group had the best perfusion effect, followed by the 60% latex heparin group, and the 60% latex saline group had the worst perfusion effect. Conclusion The optimized latex perfusion technique can effectively infuse the veins in the head and face of mice, and this technique can provide a good reference for the study of the direction and morphology of facial veins in mice. Table and Figures | Reference | Related Articles | Metrics