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Explanation and Elaboration of the ARRIVE Guidelines 2.0—Reporting Animal Research and In Vivo Experiments (Ⅱ) Guoyuan CHEN, Xiao LU, Yu BAI, Lingzhi YU, Ying QIAO, Jian WANG, Jin LU, Xiaoyu LIU, Xuancheng LU, Jing GAO, Yao LI, Wanyong PANG Laboratory Animal and Comparative Medicine    2023, 43 (3): 323-331.   DOI: 10.12300/j.issn.1674-5817.2023.042 Abstract （305）   HTML （30）    PDF （1187KB）（311）       Knowledge map    Save Improving the reproducibility of biomedical research results remains a major challenge. Transparent and accurate reporting of progress can help readers evaluate the reliability of research results and further explore an experiment by repeating 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 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 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 the second part of the Chinese translation of the complete interpretation of the ARRIVE 2.0 guidelines published in PLoS Biology in 2020 (original text can be found at https://arriveguidelines.org ) and based on the best practices for following the ARRIVE 2.0 guidelines in international journals. This part includes Items 4-7 of "ARRIVE Essential 10" in the ARRIVE 2.0 guidelines: "Randomization", "Blinding", "Outcome Measurement", and "Statistical Methods". Our Chinese translated version aims to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhancing the standardization of experimental animal research and reporting, and promoting the high-quality development of experimental animal technology and comparative medicine research in China. Reference | Related Articles | Metrics

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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 （284）   HTML （57）    PDF （859KB）（374）       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 （283）   HTML （158）    PDF （1101KB）（273）       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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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 （243）   HTML （562）    PDF （1127KB）（190）       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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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 （233）   HTML （124）    PDF （1243KB）（116）       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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H1 Linker Histone Gene Regulates Lifespan via Dietary Restriction Pathways in Caenorhabditis elegans Hui CHENG, Fei FANG, Jiahao SHI, Hua YANG, Mengjie ZHANG, Ping YANG, Jian FEI Laboratory Animal and Comparative Medicine    2023, 43 (3): 271-281.   DOI: 10.12300/j.issn.1674-5817.2022.183 Abstract （227）   HTML （12）    PDF （1521KB）（168）       Knowledge map    Save Objective To reveal the physiological function of H1 linker histone gene (hil-1) and its molecular mechanism for regulating the lifespan in Caenorhabditis elegans (C. elegans). Methods C. elegans was used as a model organism and hil-1 gene was knock-down, knock-out and over-expressed via RNA interference technology, hil-1(gk229) mutants backcross purification and microinjection technology. Then the survival and oviposition of C. elegans were observed. Physiological tests including heat shock test, paraquat stress test and heavy metal Cr6+ stress test were conducted to evaluate the stress resistance of hil-1 mutants. After constructing a dual mutant nematode, real-time fluorescence quantitative PCR (RT-qPCR) was used to further identify the signaling pathways and target sites associated with hil-1 gene regulatory lifespan. Results Compared with wild-type N2 worms, the lifespan of C. elegans of RNA interference and hil-1(gk229) mutants were significantly shortened (P<0.001), while overexpression of hil-1 in the whole body increased lifespan (P<0.05). The tolerance of hil-1(gk229) mutants to heat stress and oxidative stress was significantly decreased (P<0.001, P<0.05), but the tolerance to heavy metals was not different compared to wild-type N2 worms (P>0.05). In addition, the developmental cycle of hil-1(gk229) mutants was shortened and the time of oviposition was advanced (P<0.001), but there was no significant change in total number of oviposition (P>0.05). After feeding hil-1 RNA interference bacteria to eat-2(ad465) mutants, the down-regulation of hil-1 expression did not affect the lifespan of eat-2(ad465) mutants (P>0.05). Compared with wild-type N2 worms, the expression level of daf-16 in hil-1(gk229) mutants was significantly down-regulated (P<0.001), and the expressions of downstream genes, mtl-1 and ctl-1, were also down-regulated (P<0.05, P<0.001). Compared with daf-2(e1370) mutants, the lifespan of daf-2 (e1370); hil-1(gk229) mutants did not shortened (P>0.05). Compared with daf-16(mu86) mutants, the lifespan of daf-16(mu86); hil-1(gk229) mutants was significantly shortened (P<0.001). The knockdown of hil-1 via RNA interference technology, specifically in epidermis and intestine, was sufficient for lifespan reduction (P<0.001). Conclusion The deletion of hil-1 gene significantly shortened the lifespan of C. elegans and decreased the tolerance to heat and oxidative stress. The hil-1 gene regulates the lifespan of C. elegans via dietary restriction pathway and acts mostly in epidermis and intestine. Table and Figures | 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 （224）   HTML （34）    PDF （1153KB）（206）       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 | Supplementary Material | Related Articles | Metrics

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Injurious Effect of Cisplatin on the Function of Hypothalamus-pituitary-adrenal/gonadal Axis in Mice and the Intervention Effect of Dehydroepiandrosterone Zhiqiang PAN, Zixin NONG, Haina XIE, Peike PENG Laboratory Animal and Comparative Medicine    2023, 43 (3): 229-242.   DOI: 10.12300/j.issn.1674-5817.2022.182 Abstract （195）   HTML （38）    PDF （3170KB）（449）       Knowledge map    Save Objective To study the pathway of cis-dichlorodiamineplatinum (DDP) inhibiting the synthesis of steroid hormones in mice, and to observe the intervention effect of dehydroepiandrosterone (DHEA). Methods Sixty adult ICR mice were randomly divided into three groups: control group, DDP modeling group, and DHEA group, with 10 male and 10 female mice in each group. The DDP modeling group mice were intraperitoneally injected with DDP solution at a dose of 2.5 mg·kg-1·d-1, once every 3 days, a total of 7 times. On the same day of modeling, the control group mice were injected with an equal amount of physiological saline intraperitoneally. The DEHA treatment group mice were treated with DDP and given a dose of 8.3 mg·kg-1·d -1 of DHEA by gavage for 21 consecutive days. The changes of fatigue indexes of mice were observed by open field, grip and rod rotation tests. The morphology changes of adrenal gland, testicular and ovarian tissue were observed by pathological section and HE staining. The levels of serum steroid hormones were detected by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The mRNA and protein expression levels of the related genes of the hypothalamus, hypophysis, adrenal, testis and ovary were tested by real-time fluorescent quantitative PCR (RT-qPCR) and Western blotting. Results Compared with control group, both male and female mice in DDP modeling group were significantly losing weight (P＜0.05), their abilities in horizontal movement and vertical movement decreased (all P＜0.05), and the stay time and grip also significantly decreased (all P＜0.05) in female mice. Indexes of fatigue were improved after DHEA supplement (all P＜0.05). In the DDP modeling group, the arrangement of spermatogenic cells at all levels in the testicular tissue was disordered and the testicular interstitial edema was observed, and a large number of primordial follicles in the ovarian tissue were activated, the number of atresia follicles increased, and the number of granulosa cells in the follicles decreased; while in the DHEA group, the damaged phenotype of testicles and ovaries was significantly improved. Compared with control group, the levels of serum testosterone and dihydrotestosterone in both male and female DDP modeling mice significantly decreased (P＜0.01), the pregnenolone was down-regulated but corticosterone was up-regulated significantly (P＜0.05) in male mice, the corticosterone was down-regulated significantly (P＜0.05) in female mice. Compared with the DDP group, after DHEA supplement, the pregnenolone in male mice and the progesterone in female mice increased significantly (P＜0.05), but the pregnenolone in female mice and the progesterone in male mice decreased significantly (P＜0.05). Compared with control group, the expression levels of Cyp21a1 and Cyp11a1 genes in the adrenal gland and Gnrh gene in the hypothalamus of male and female mice in the DDP modeling group significantly decreased (all P＜0.05); the expression levels of Hsd3b2 gene in the adrenal gland, Star, Cyp11a1, and Lhr genes in the ovaries, Crh, Pomc, and Lhb genes in the hypothalamus, pituitary, and pituitary of female mice significantly decreased (all P＜0.05); the expression levels of Star gene and StAR protein in the testicles of male mice, as well as Fshb and Lhb genes in the pituitary gland, were significantly down-regulated (all P＜0.05). After DHEA supplement, compared with the DDP modeling group, the mRNA expression levels of Cyp17a1 in the adrenal gland of male mice and Cyp17a1, Lhr and Fshr genes in testis were down-regulated significantly (P＜0.05); the expression level of Cyp11a1 gene in the adrenal gland of female mice was also decreased (P＜0.05); while the expression levels of Hsd3b2 gene in the adrenal gland, Star, Cyp11a1, Hsd3b2 and Lhr gene in the ovary, and Lhb gene in the pituitary gland were all up-regulated ( P＜0.05). Conclusion The function of hypothalamus-pituitary-adrenal/gonadal axis was inhibited by DDP intermittent injection, especially in female. Supplementation of DHEA can help regulate the homeostasis of steroid hormone levels. 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 （194）   HTML （23）    PDF （1578KB）（265）       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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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 （191）   HTML （16）    PDF （808KB）（213）       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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An Optimized Experimental Zebrafish Breeding Scheme for Significantly Enhancing Reproductive Efficiency and Service Life Shirong JIN, Ye HUA, Huaxing ZI, Xufei DU, Jiwen BU Laboratory Animal and Comparative Medicine    2023, 43 (3): 297-306.   DOI: 10.12300/j.issn.1674-5817.2023.004 Abstract （182）   HTML （29）    PDF （1580KB）（187）       Knowledge map    Save Objective To solve the problems of delayed growth and development and insufficient spawning of experimental Zebrafish, so as to improve the reproductive efficiency and service life of experimental Zebrafish. Methods The zebrafish at the age of 2 months after fertilization were divided into two groups. The experimental group was fed with dry commercial diets specifically designed for ornamental fish or frozen adult brine shrimp, while control group was fed with live laval brine shrimp. Within a period of 70 days, the growth performance of the zebrafish was evaluated by measuring body length and weight, and the reproductive performance was assessed by measuring the fecundity and spawning rate. Zebrafish with apparent goiter disease were fed with dry commercial diets, and the inhibitory effect of the pellets on this disease was evaluated by measuring the diameter of the thyroid enlargement lesion. The three feeding methods were combined, and the feeding plan was optimized. The actual effects of the plan on zebrafish rearing were validated through reproductive performance tests. Results Starting from 60 days post-fertilization (dpf) until 111 dpf, the body length and weight of the dry commercial diets feed group gradually surpassed those of control group (all P<0.000 1). From 60 dpf to 96 dpf, the growth trend in body length of the adult brine shrimp group was similar to that of control group, but the female fish in the adult brine shrimp group had significantly higher body weight than the female fish in control group at 75-82 dpf (P<0.000 1). Compared to control group, there was a significant difference in body color between males and females in the adult brine shrimp group, and at 75 dpf, gender could be accurately distinguished by body color differences. Furthermore, the spawning rate of the zebrafish in the adult brine shrimp group at 3 months of age was significantly higher than that of control group (94.44% vs. 27.78%, P<0.05). Additionally, after feeding with the dry commercial diets for 130 days, all thyroid enlargement lesions in the experimental zebrafish disappeared. Based on the above results, the three feeding methods were combined and the feeding plan for zebrafish older than 2 months of age was optimized as follows: feed live brine shrimp in the morning, and alternate between dry commercial diets and adult brine shrimp in the afternoon. This feeding plan lasts until the age of 12 months. The spawning rate of Zebrafish can maintan 70%, and the spawning amount can reach (233.6±3.95) eggs. The fertilization rate and hatching rate were 97.47% and 90.24%, respectively, both significantly higher than those of control group (P<0.001, or P=0.01). Conclusion Compared to live brine shrimp feed, the dry commercial diets feed significantly improves the growth performance of zebrafish and has a therapeutic effect on thyroid enlargement disease. On the other hand, adult brine shrimp feed significantly enhances the early reproductive performance of zebrafish. The optimized feeding plan successfully improves the spawning efficiency of laboratory zebrafish, prolonging their reproductive lifespan and better supporting relevant scientific research. 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 （180）   HTML （417）    PDF （2710KB）（127）       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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Repairing Effects of Ginsenoside Rg1 on Traumatic Brain Injury in Mice Wenwen GUO, Ya ZHAO, Yinghua WANG, Ke LIU, Xu GE, Yanying ZHANG, Yongfeng WANG, Changhong SHI Laboratory Animal and Comparative Medicine    2023, 43 (3): 243-252.   DOI: 10.12300/j.issn.1674-5817.2022.187 Abstract （176）   HTML （20）    PDF （2002KB）（146）       Knowledge map    Save Objective To explore the effects of ginsenoside Rg1 on blood-brain barrier, neuroinflammation and behavioral function of traumatic brain injury (TBI) mouse model. Methods The experiment was divided into two parts. In the first part, 27 SPF male BALB/c mice were randomly divided into blank group, sham operation group and TBI model group, with 9 mice in each group. TBI model group was made by controlled cortical impact (CCI) after craniotomy, while sham operation group was only performed craniotomy without any treatment, and the blank group was not treated at all. The effect of modeling was evaluated after operation. In the second part, 50 male BALB/c mice were randomly divided into sham operation group, three different drug dosage groups and solvent (DMSO) control group, with 8 mice in each group. The drug treatment groups were injected with ginsenoside Rg1 at the doses of 10, 20 and 40 mg/kg respectively 6 hours after TBI model had been successfully established, while the DMSO control group was given the same amount of 1% DMSO for one week, twice a day. Modified neurological severity scores (mNSS) were performed on the 1st, 3rd, 7th and 14th day after modeling, and the blood-brain barrier leakage was detected by Western blotting on the 3rd day after modeling. On the 14th and 16th day, the elevated cross maze test and water maze test were used to detect the neurobehavioral function. On the 28th day after anesthesia and perfusion, the brains were taken out, and the neuroinflammation such as activation of microglia and astrocytes was observed by immunofluorescence staining. Results The expression level of MMP-9, a marker of blood-brain barrier, decreased in ginsenoside Rg1 treatment group (P<0.01). The number of microglia （Iba-1 positive) and astrocyte (GFAP positive) cells decreased significantly (P<0.05), which indicated that neuroinflammation was inhibited, and the best effect was achieved at the dosage of 20 mg/kg (P<0.01). The mNSS of mice in ginsenoside Rg1 treatment group were significantly lower than those in DMSO control group (P < 0.01), and the proportion of times they entered the open arm was significantly higher than that in DMSO control group (P < 0.05). The time ratio in the quadrant where the water maze experimental platform was located and the times of crossing the platform were significantly higher than those in control group (P < 0.05), and the dosage of 20 mg/kg had the best effect. Conclusion The TBI mouse model was successfully constructed and applied to the study of ginsenoside Rg1 repair of mouse traumatic brain injury. Ginsenoside Rg1 can significantly improve blood-brain barrier, alleviate neuroinflammation and improve neurobehavioral function in TBI model mice, and the effect is the most significant at the dose of 20 mg/kg. Table and Figures | Reference | Related Articles | Metrics

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Downregulation of Micall2a Gene Expression Inhibited Vascular Development in Zebrafish Jinxian YANG, Shujuan WANG, Jinyun ZHAI, Shunxing ZHU Laboratory Animal and Comparative Medicine    2023, 43 (3): 282-287.   DOI: 10.12300/j.issn.1674-5817.2022.166 Abstract （155）   HTML （16）    PDF （1075KB）（142）       Knowledge map    Save Objective To explore the expression pattern of Micall2a gene during the early development of zebrafish embryos and the effect of this gene on zebrafish vascular development. Methods Whole embryo in situ hybridization was used to detect Micall2a expression levels at different stages of early embryo development of Tg (fli:GFP) transgenic (labeled with green fluorescent protein) and wild type zebrafish (AB). Micall2a gene expression was downregulated by microinjection of a morpholine antisense oligonucleotide, and real-time fluorescent quantitative PCR was used to detect mRNA expression of the gene at different developmental stages of zebrafish embryos. Laser confocal microscopy was used to observe and analyze vascular phenotypic changes in zebrafish after the downregulation of Micall2a. Results Micall2a was expressed in the brain, heart, and vascular system of zebrafish embryos at the 24th, 36th, and 48th hours post fertilization. The mRNA level of Micall2a increased after microinjection of morpholine antisense oligonucleotides, inhibiting vascular development in zebrafish embryos, resulting in internode angiogenesis defects in zebrafish. Conclusion Downregulation of Micall2a expression inhibits the development of blood vessels in zebrafish. Table and Figures | Reference | Related Articles | Metrics

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Effects of Pogostemon cablin on Serum Metabolomiceof Guizhou Miniature Pigs and It's mechanism Taofeng LU, Hui ZHANG, Jie ZHOU, Qian LI, Shuguang WU, Yanjun WU Laboratory Animal and Comparative Medicine    2023, 43 (3): 253-261.   DOI: 10.12300/j.issn.1674-5817.2022.186 Abstract （148）   HTML （16）    PDF （1940KB）（154）       Knowledge map    Save Objective Based on the liquid chromatography-tandem mass spectormetry (LC-MS/MS), to study the effects of Pogostemon cablin on serum metabolism of Guizhou miniature pigs， and to explore its pharmacological mechanism. Methods Nine healthy Guizhou miniature pigs were divided into two groups, namely Pogostemon cablin drug group (n=5) and control group (n=4). The pigs in Pogostemon cablin drug group were orally fed with traditional Chinese medicine formula granules, each 0.5 g per day, for consecutive 8 days, while those in control group were given normal feeding without additional treatment. After the feeding experiment, serum samples were collected and analyzed using the LC-MS/MS technology. The metabolomics data was annotated and compared with KEGG, HMDB and Lipidmaps databases. Bioinformatics analysis methods including partial least squares discriminant analysis (PLS-DA), intergroup clustering, differential metabolite analysis and functional enrichment were used to screen differential metabolic biomarkers and their possible metabolic pathways. Results Forty-four differential metabolites (P<0.05) were screened out from the 443 metabolites, eight differential metabolites were significantly up-regulated (P<0.01), namely cinnamoylglycine, N-benzyl-N-isopropyl-N'-[4-(trifluoromethoxy) phenyl]urea, hypotaurine, D-glucose 6-phosphate, cis-2-decenoic acid, 11(Z),14(Z)-eicosadienoic acid, prostaglandin A2 and 10-hydroxydecanoic acid, and three differential metabolites were significantly down-regulated (P<0.01), namely lysophosphatidyl choline 22:5, lysophosphatidic acid 22:6 and lysophosphatidic acid 22:5. The differential metabolites were mainly enriched in the metabolic pathways of alanine, aspartate and glutamate metabolism (MapID: map00250) and taurine and hypotaurine metabolism (MapID: map00430). Conclusion Pogostemon cablin can significantly affect the metabolism of lysophosphatidic acids in porcine, and relieve the disorder of amino acids metabolism and regulate the occurrence of inflammation by affecting the metabolic pathways of alanine, aspartate and glutamate metabolism and taurine and hypotaurine metabolism. Table and Figures | Reference | Supplementary Material | 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 （147）   HTML （10）    PDF （740KB）（106）       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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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 （142）   HTML （30）    PDF （1768KB）（103）       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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Advances and Applications in Animal Models of Neuroblastoma Zhigang TAN, Jinxin LIU, Chuya ZHENG, Wenfeng LIAO, Luping FENG, Hongli PENG, Xiu YAN, Zhenjian ZHUO Laboratory Animal and Comparative Medicine    2023, 43 (3): 288-296.   DOI: 10.12300/j.issn.1674-5817.2022.194 Abstract （142）   HTML （6）    PDF （1067KB）（227）       Knowledge map    Save Neuroblastoma (NB) is one of the most common malignant solid tumors in children, ranks fourth in the incidence of pediatric tumors, and accounts for 15% of pediatric tumor deaths in children in China. Despite the development of new treatment options, the prognosis for high-risk patients is still poor. An animal model that can replicate the tumorigenesis of NB is an important tool for the prevention and treatment of NB. However, there are currently no animal models that can simulate all features of human NB. To provide a reference for the construction of animal models and treatment of NB, this article introduced several animal models of NB that have been extensively researched: the mouse, chick embryo chorioallantoic membrane, and zebrafish models. At the same time, it elaborated on the species, construction methods, characteristics, advantages and disadvantages, and research progress in NB. 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 （141）   HTML （14）    PDF （1028KB）（164）       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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Influence of Corneal Staining in Rabbits on the Evaluation of Eye Irritation Test Results Honghua XU, Tian JIN, Hai WANG, Mengying SHEN, Rui WANG, Yijia ZHOU, Ying TAN Laboratory Animal and Comparative Medicine    2023, 43 (3): 307-313.   DOI: 10.12300/j.issn.1674-5817.2022.169 Abstract （139）   HTML （16）    PDF （1439KB）（184）       Knowledge map    Save Objective To observe the influence of the staining phenomenon after fluorescein sodium staining on eye irritation in normal rabbits. Methods In the experimental rabbit eye irritation test conducted with sodium chloride eye drops, Siwei Zhenceng Bingpeng eye drops, sodium hyaluronate eye drops, sodium cromoglycate eye drops, and compound aspartate eye drops (4 in each group, half male and half female), the left eyes of rabbits were administered normal saline (self-negative control) and the right eyes were administered the experimental medicine; the eyes were stained with 1% sodium fluorescein, and eye irritation was observed and scored using slit lamp microscope for 31 days. Morphological changes of corneal epithelial staining were recorded and the incidence of staining was calculated. After the observation, the eyeballs and Hasselblad glands were examined histopathologically, and the staining rate of the left eye was compared with that of the right eye which was administered the corresponding medicine. Results Neither eye had any irritation symptoms; the scores were 0, and the total incidences of corneal staining were 3% (left) and 1% (right), respectively. There was no significant difference between the two groups (P > 0.05). Corneal epithelial staining showed single-spot staining, scattered dot, localized, or large areas of fusion staining. No histopathological changes were found in the eyeballs or Hasselblad glands, and the results were evaluated as non-irritative. Conclusion The irregularity of corneal epithelial staining in rabbits did not influence the results of the ocular irritation test. Table and Figures | Reference | Related Articles | Metrics

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Quantification of Uric Acid of Rat Serum by Liquid Chromatography-ultraviolet Detection and Its Comparison Study Ziyin XIA, Yuanyuan CHAI, Yunxia XU, Qinwei YU, Xin HUANG, Luyong ZHANG, Zhenzhou JIANG Laboratory Animal and Comparative Medicine    2023, 43 (3): 314-322.   DOI: 10.12300/j.issn.1674-5817.2022.189 Abstract （138）   HTML （15）    PDF （1203KB）（101）       Knowledge map    Save Objective To establish a more accurate and sensitive liquid chromatography-ultraviolet (LC-UV) method for the determination of uric acid in rat serum, and compare the results with those of commercial kits, providing a new method for the accurate determination of uric acid in the rat hyperuricemia model induced by potassium oxonate. Methods A hyperuricemia model was established by intraperitoneal injection of potassium oxonate (300 mg/kg) into SPF-grade male SD rats, and the control group was administered an equal amount of 0.5% sodium carboxymethylcellulose solution. Blood samples were collected from the posterior orbital venous plexus and centrifuged to obtain serum samples. After precipitation with 0.1% trifluoroacetic acid-acetonitrile (containing the internal standard 3,4-dihydroxybenzylamine hydrobromide), the supernatant was injected for analysis. Uric acid was separated on a Waters XBridge HILIC column (150 mm×4.6 mm, 3.5 μm) using acetonitrile (containing 0.5% formic acid and 2 mmol/mL ammonium formate) as the organic phase and methanol solution (methanol∶water=1∶1, containing 0.5% formic acid with 2 mmol/L ammonium formate) as the aqueous phase for isocratic elution and detection at 290 nm. Serum samples treated with activated carbon were used as substitute matrices for the methodological verification. Serum uric acid levels in rats with potassium oxonate-induced hyperuricemia were measured using the established LC-UV method and commercially available kits (uricase and phosphotungstic acid methods), and the accuracies of the three methods were compared. Results Serum uric acid showed a good linear relationship (R>0.999) at mass concentration of 10–200 μg/mL in rats, the lower limit of quantification was 10 μg/mL, the accuracy ranged from -2.17% to 2.21%, the intra-batch precision ranged from 0.52% to 1.95%, the inter-batch precision ranged from 3.04% to 4.90%, and the extraction recovery ranged from 83.12% to 89.91%. In the rat model, the results obtained using the commercially available phosphotungstic acid method kit were significantly higher than those of the LC-UV method, and those obtained using the commercially available uricase method kit were significantly lower than those of the LC-UV method, but the LC-UV method showed the best recovery of the spiked sample (95.90%–99.96%). Conclusion The LC-UV method developed in this study can determine the concentration of uric acid in rat serum with higher accuracy than commercially available kits and is recommended for the determination of serum uric acid in the rat model of hyperuricemia induced by potassium oxonate. 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 （137）   HTML （18）    PDF （1036KB）（112）       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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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 （137）   HTML （121）    PDF （1374KB）（149）       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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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 （135）   HTML （26）    PDF （1188KB）（213）       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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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 （132）   HTML （143）    PDF （853KB）（146）       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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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 （129）   HTML （121）    PDF （3115KB）（102）       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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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 （128）   HTML （98）    PDF （1461KB）（99）       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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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 （128）   HTML （493）    PDF （1787KB）（107）       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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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 （118）   HTML （9）    PDF （1017KB）（77）       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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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 （116）   HTML （133）    PDF （1046KB）（133）       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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Whole-brain Transcriptomic Analysis of Weight Gain Mice induced by Olanzapine Yuan ZHANG, Han LI, Chengfang ZHANG Laboratory Animal and Comparative Medicine    2023, 43 (3): 262-270.   DOI: 10.12300/j.issn.1674-5817.2023.006 Abstract （116）   HTML （9）    PDF （1961KB）（107）       Knowledge map    Save Objective The transcriptome sequencing results of brain tissues of olanzapine-treated mice were analyzed to screen out differentially-expressed genes and explore potential targets of atypical antipsychotics leading to body weight gain. Methods Twenty female C57BL/6 mice were randomly divided into control group (Ctrl) and Olanzapine administration group (Olz), which were given saline and Olanzapine solution by gavage, respectively. The whole brain tissues were collected 8 weeks later for Transcriptome sequencing (RNA-Seq). The possible targets of olanzapine-induced body weight gain were identified by the Gene Ontology (GO) functional annotation analysis, the Kyoto Encyclopedia of Genes and Gnomes (KEGG) pathway enrichment analysis, and protein-protein interaction (PPI) network analysis. Differential expression levels of mRNAs were further verified by real-time quantitative fluorescence PCR (RT-qPCR). Results Compared with Ctrl group, 591 differentially expressed genes were screened in Olz group, including 251 up-regulated genes and 340 down-regulated genes. GO analysis showed that differential genes were widely involved in transcriptional process, among which the expression of genes related to the regulation of digestive system and cold-induced thermogenesis were significantly enriched. KEGG analysis showed that differential genes were widely involved in the interaction between neuroactive ligands and receptors, and the differential genes were significantly enriched in oxytocin signaling, fat digestion and absorption, and cholesterol metabolism pathways. RT-qPCR were performed to verify the expression levels of genes enriched in feeding regulation, gastric kinesis, thermogenesis, fat metabolism and other processes (Oxt, Trpv1, Adipoq, Phox2b, Abcg5, Mogat2, Dbh, Plac8 and Neurog1) as well as hub genes in PPI network (Fos, Dusp1 and Egr2), and the results were consistent with the trend of RNA-Seq. Conclusion Olanzapine administration resulted in changes in central feeding regulation, gastrointestinal motility, thermogenesis and other physiological processes in mice, which might be involved in body weight gain induced by olanzapine. 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 （115）   HTML （14）    PDF （2128KB）（102）       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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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 （110）   HTML （15）    PDF （1032KB）（69）       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