Loading...

Table of Content

    25 October 2021, Volume 41 Issue 5
    40th Anniversary Expert Forum
    A New Strategy for Constructing Mouse Models of Complex Diseases: Semi-cloning Technology Based on Sperm-like Haploid Embryonic Stem Cells
    LAI Suomei, DING Yifu, LI Jinsong
    2021, 41(5):  369-383.  DOI: 10.12300/j.issn.1674-5817.2021.143
    Asbtract ( 622 )   HTML ( 241)   PDF (6213KB) ( 668 )  
    Figures and Tables | References | Related Articles | Metrics
    The development of haploid genetics has motivated studies on genome evolution and function, especially the technological advancements in recent years have prompted the birth of culture techniques for mammalian haploid embryonic stem cells (haESCs). Sperm-like haESCs are novel haESCs derived from mouse parthenogenetic blastocysts. Sperm-like haESCs only contain paternal genetic material, and their sex chromosome is the X chromosome. They can self-renew, differentiate, and proliferate in vitro for a long time. Furthermore, editing single or multiple genes using the CRISPR system is possible for sperm-like haESCs, which can replace sperms to fertilize oocytes. In contrast to traditional methods for constructing mouse models, such as pronuclear injection, cytoplasmic injection, and tetraploid complementation, by injecting sperm-like haESCs after gene editing into oocytes, semi-cloned mice with a definitive genotype can be obtained efficiently and stably without chimerism, and primary mice can be used for research. The mouse disease model based on multiple precisely edited genes obtained from sperm-like haESCs can explain the effect of multiple genes synergistic interaction at the level of biological individuals to completely simulate various pathological characteristics of complex human diseases that may be affected by multiple genes, and this model facilitates the exploration of novel diagnostic and therapeutic methods.
    Progress on the Accreditation for Laboratory Animal Institutions of CNAS
    WU Xiaohuai
    2021, 41(5):  384-391.  DOI: 10.12300/j.issn.1674-5817.2021.141
    Asbtract ( 745 )   HTML ( 276)   PDF (1886KB) ( 876 )  
    Figures and Tables | References | Related Articles | Metrics
    The accreditation for laboratory animal institutions of China National Accreditation Service for Conformity Assessment (CNAS) is a third-party evaluation program that is in line with international practices and China's national conditions, it is complementary to the national laboratory animal license system, and is an important system for the management of laboratory animal institutions. This paper introduces the origin, development, status quo, problems, and challenges of CNAS laboratory animal accreditation, and proposes countermeasures and suggestions for improving the accreditation system of laboratory animal institutions.
    Advantages and Progress of Using Chinese Hamsters to Construct Disease Models
    GAO Jiping, CHEN Zhaoyang, XUAN Ruijing, ZHANG Ruihu, SONG Guohua
    2021, 41(5):  392-398.  DOI: 10.12300/j.issn.1674-5817.2021.009
    Asbtract ( 541 )   HTML ( 48)   PDF (3619KB) ( 496 )  
    References | Related Articles | Metrics
    Chinese hamsters are experimental animals originally from China. They play an important role in experimental research on medicine and biology. They are widely used in life science research, such as genetics, infectious diseases, tissue culture, especially in animal models and mechanism research, such as diabetes and oral cancer. This paper reviews the relevant applied studies of Chinese hamsters in disease animal models since its development, and discusses the advantages and problems associated with the use of these animals in biomedical research.
    40th Anniversary: Development of Laboratory Animal Sciene Across China
    History and Countermeasure Discovered from 40-year Development of Laboratory Animals in Yunnan Province
    HAN Yuanyuan, LI Na, DAI Jiejie
    2021, 41(5):  399-408.  DOI: 10.12300/j.issn.1674-5817.2021.136
    Asbtract ( 466 )   HTML ( 235)   PDF (1769KB) ( 505 )  
    References | Related Articles | Metrics
    Laboratory animals are strategic resources that are indispensable to support scientific and technological progress and innovation. It is one of the core elements required for country to maintain scientific and technological leadership and improve international scientific and technological competitiveness. In this study, the legalization and standardization, major research institutions, scientific research features and academic achievements, and current opportunities and countermeasures over 40 years of progression of laboratory animals in Yunnan Province were summarized to provide a reference for the further development of laboratory animals in China.
    Original Article: Animal Models of Human Diseases
    Dynamic Changes of Tumor-infiltrating Immune Cells in Mice with MC38 Colon Cancer
    ZHANG Boyang, CHEN Huanpeng, YU Wenlan, LIU Zhonghua, HUANG Chaofeng
    2021, 41(5):  409-417.  DOI: 10.12300/j.issn.1674-5817.2020.157
    Asbtract ( 821 )   HTML ( 44)   PDF (7559KB) ( 448 )  
    Figures and Tables | References | Related Articles | Metrics
    Objective To observe the dynamic changes of tumor-infiltrating immune cells in tumor-bearing mice during tumor development. Methods Mouse colon cancer MC38 cells were transplanted subcutaneously into 24 C57BL/6J mice, and the changes in body mass and tumor size of mice were recorded once a week after tumor-bearing, The levels of tumor-infiltrating immune cells, including CD45+ T cells, CD3+ T cells, CD8+ T cells, CD4+ T cells, natural killer (NK) cells, dendritic cells, macrophages, M1-type macrophages, M2-type macrophages, myeloid-derived suppressor cells (MDSCs), mononuclear-like MDSCs (M-MDSCs), and granulocytic MDSCs (G-MDSCs) were detected by flow cytometry once a week for 4 weeks. Results As the colon cancer tumor of the mouse grows, total CD45+ T cellular level decreased, and CD8+ T cellular level steadily decreased to an irreversible level, but CD4+ T cells proportion rapidly increased at the early stage of the tumor-bearing, quantity of infiltrating cells in the unit volume increased by 8 times, and T cells of tumor-infiltration almost were CD4+ phenotype. The nonspecific immune response level represented by NK cells continued to decline, but the dendritic cells continued to accumulate in the tumor. The immunosuppressive action was further enhanced through the activation of arginase-1 (ARG1) and inducible nitric oxide synthetase (iNOS). Macrophage infiltration levels showed no significant change, but the gliding speed for the quantity of infiltrating type-Ⅰ macrophages was obviously higher than that of type-Ⅱ macrophages. The proportion of MDSCs per unit volume showed no decreasing trend, and the cells continued to accumulate in the tumor. G-MDSC infiltration level showed a steady decrease, while M-MDSC infiltration level showed an increasing trend, the proportion of M-MDSCs were higher than that of G-MDSCs. Conclusion After transplanting MC38 colon cancer cells in mice, the level of immune cell infiltration in the tumor tissue declined, tumor immunology gradually became dominat, thus promoting the development of colon cancer. The Th1/Th2 drift occurred in the tumor. Specific and nonspecific immune response levels were significantly decreased on an average, and the infiltrating monocytes and granulocytes were mainly Ly6C+. Tumor-associated dendritic cells and macrophages play an important immunosuppressive role. CD4+T cells and M-MDSCs might be the key for promoting tumor immunity.
    CD137 Signaling Promotes Cardio-cerebral Angiogenesis in Mice Through Regulating Endothelial-mesenchymal Transition
    WENG Jiayi, MA Xuexing, LI Yuan, SUN Kangyun
    2021, 41(5):  418-425.  DOI: 10.12300/j.issn.1674-5817.2020.210
    Asbtract ( 292 )   HTML ( 18)   PDF (29894KB) ( 128 )  
    Figures and Tables | References | Related Articles | Metrics
    Objective To explore whether CD137-CD137L signaling can promote cardio-cerebral angiogenesis in atherosclerotic plaques via endothelium-mesenchymal transition (End-MT). Methods Mouse brain-derived endothelial cells (Bend.3) and aortas from male mice were divided into the following groups: control (10 ng/mL TNF-α), CD137-activated (10 ng/mL TNF-α + 5 mg/L CD137L agonist antibody), and CD137-inhibited (pretreated with 200 nmol/L IWR-1 for 30 min + 10 ng/mL TNF-α + 5 mg/L CD137L agonist antibody) groups. CD137L agonist antibody was used to activate CD137-CD137L signaling. Fluorescent quantitative PCR was used to determine the expression of CD31, vascular endothelial cadherin (VE-cadherin), fibroblast-specific protein-1F (FSP-1), Wnt, and α-smooth muscle actin (α-SMA) at the mRNA level. Western blotting was used to determine the expression of CD31, VE-cadherin, FSP-1, Wnt, and α-SMA at the protein level. A Transwell assay was used to observe the migration ability of endothelial cells. Matrigel tube formation ability of endothelial cells and mouse aortic ring angiogenesis assay were tested to detect the ability of angiogenesis. Results The expressions of CD31 and VE-cadherin at the mRNA levels in endothelial cells were significantly lower in the CD137-activated group than those in the control group, while the expression was significantly downregulated in the CD137-inhibited group. The expressions of FSP-1, Wnt, and α-SMA were opposite (all P < 0.05). The expressions of CD31 and VE-cadherin proteins in endothelial cells were significantly higher in the CD137-activated group than those in the control group, while the expression was significantly downregulated in the CD137 inhibited group. The expressions of FSP-1, Wnt, and α-SMA were opposite (all P < 0.05). Migration cell number was markably higher in the CD137-activated group than that in the control group, while it was significantly lower in the CD137-inhibited group than that in the control group (P < 0.05). The values of the formation of the tube length (P < 0.05) and branch number (P < 0.05) were both significantly higher in the CD137-activated group than those in the control group, while these values were significantly lower in the CD137-inhibited group than those in the control group (P < 0.05). The number of microvessel outgrowths in the aortic rings was dramatically increased in the CD137-activated group than that in the control group, while it was reduced in the CD137-inhibited group (P < 0.05). Conclusion CD137-CD137L signaling can promote cardio-cerebral angiogenesis in atherosclerotic plaques by activating End-MT.
    Effects of Pharyngeal Refreshing Pills on Immune Cell Subsets and NF-κB Signaling Pathway in Chronic Pharyngitis
    TANG Yi, YU Guodong, HE Xingchen, WANG Jia, ZHANG Tian
    2021, 41(5):  426-434.  DOI: 10.12300/j.issn.1674-5817.2021.006
    Asbtract ( 431 )   HTML ( 17)   PDF (43931KB) ( 149 )  
    Figures and Tables | References | Related Articles | Metrics
    Objective To explore the effect and mechanism of pharyngeal refreshing pills on an animal model of chronic pharyngitis. Methods A total of 42 healthy New Zealand white rabbits were randomly divided into seven groups: control, model, manyanshuning granules (9 g/kg), pharyngeal refreshing pills at low- (0.025 g/kg), medium- (0.05 g/kg), high- (0.1 g/kg) doses, and atomization (0.025 g/kg) groups. Animal models of chronic pharyngitis were established by chemical stimulation in all rabbits except the control group. From the second day after successful modeling, the five treatment groups were administered with manyanshuning granules and different doses of pharyngeal refreshing pills every day, while the control and model groups were fed with white sugar (0.025 g/kg) for 7 days. From the third day after successful modeling, the local condition of the pharyngeal wall of each rabbit was observed every day. Blood samples were collected at the end of drug administration and examined. Histopathological changes in the pharyngeal wall mucosa were examined by hematoxylin-eosin staining, and the levels of CD3+, CD4+, and CD8+ were detected by flow cytometry. The levels of interleukin-6 (IL-6), IL-1, tumor necrosis factor-α (TNF-α), C-reactive protein (CRP), and total antioxidant capacity (T-AOC) in serum were detected by enzyme-linked immunosorbent assay (ELISA). Western blotting was used to detect the expression of IL-6, IL-1, TNF-α, nuclear factor-κappa B (NF-κB) p65 and NF-κB inhibitor (IκB-α) at the protein level in the pharyngeal wall mucosa. qRT-PCR was used to detect NF-κB p65 and IκB-α mRNA expression in the pharyngeal wall mucosa. Results Compared with the control group, the pathological tissues of the posterior pharyngeal mucosa of rabbits with chronic pharyngitis were significantly damaged; the levels of CD3+, CD4+, CD4+/CD8+, and T-AOC were significantly decreased, while the levels of CD8+, IL-6, IL-1, TNF-α, CRP, NF-κB p65, and IκB-α were significantly increased (all P < 0.01). Compared with the model group, the pathological injury of pharyngitis could be alleviated to different degrees in the treatment group; the levels of CD3+, CD4+, CD4+/CD8+, and T-AOC in the pharyngeal refreshing pills at high dose and atomization groups were significantly increased, while the levels of CD8+, IL-6, IL-1, TNF-α, CRP, NF-κB p65, and IκB-α were significantly decreased (all P < 0.05). The effect of high-dose pharyngeal refreshing pill administration was better than that of low-dose and middle-dose pharyngeal refreshing pills, and manyanshuning granules. Conclusion The pharyngeal refreshing pills can significantly improve the inflammatory index of chronic pharyngitis, reduce pathological injury, affect the expression of NF-κB signaling pathway-related factors, and the effect of atomization dose is more obvious.
    Relationship Between the Abnormal Expression of PPARs and Insulin Resistance in Uterus of Rats with Polycystic Ovary Syndrome and Insulin Resistance
    WEI Wei, CHEN Yihua, ZHANG Xiuzhi, LENG Yifu, LI Chun, YIN Tianxiao
    2021, 41(5):  435-442.  DOI: 10.12300/j.issn.1674-5817.2020.206
    Asbtract ( 323 )   HTML ( 23)   PDF (28126KB) ( 163 )  
    Figures and Tables | References | Related Articles | Metrics
    Objective To investigate the relationship between abnormal expression of peroxisome proliferator-activated receptors (PPARs) and insulin resistance in the uterine tissues of rats with polycystic ovary syndrome and insulin resistance (PCOS-IR). Methods Forty female SD rats were randomly divided into normal control and PCOS-IR model groups. PCOS-IR models were established by subcutaneous injection of insulin and human chorionic gonadotropin (INS+hCG) method, and rats in the normal control group were injected with equal amounts of normal saline. Fasting insulin (FINS) and fasting plasma glucose (FPG) levels were measured by enzyme-linked immunosorbent assay (ELISA), and the insulin resistance index (HOMA-IR) was calculated. Uterine tissues of rats were stained with hematoxylin-eosin (HE), PPARs were detected by immunohistochemistry, and the expressions of PPARs, insulin receptors (IRS), glucose transporter 4 (GLUT-4), and insulin-like growth factor-1 (IGF-1) were detected by Western blotting. Results FPG and FINS levels and HOMA-IR in the model group were significantly higher than those in the normal control group (P < 0.05). HE staining of uterine tissues showed that the endometrium of the normal control group was normal, and the endometrium of the model group showed different degrees of proliferative changes, the glandular cavity became smaller, the number of glands decreased, and the arrangement was loose. Immunohistochemistry showed that three subtypes of PPARs (PPARα, PPARβ, and PPARγ) were expressed in the rat uterus. The expression of PPARα and PPARγ in the rat uterus was down-regulated in the model group (P < 0.05), and there was no significant difference in the expression of PPARβ between the two groups (P > 0.05). Western blotting results showed that the expression of PPARα, PPARγ, IRS, and GLUT-4 in the rat uterus of the model group was significantly lower than that of the normal control group (P < 0.05), while the expression of IGF-1 was higher than that of the normal control group (P < 0.05). Conclusion The expression of IRS and GLUT-4 is down-regulated and the expression of IGF-1 is up-regulated in the uterine tissue of PCOS-IR rats. The existence of IR is confirmed in the uterine tissue of PCOS-IR rats, and it may be related to the down-regulation of PPARα and PPARγ expression in the uterine tissues.
    Exploration and Practice: Biosafety
    Prevention and Control of Pathogenic Microorganisms for the Management and Biosafety Control in Laboratory Animal Facilities
    LUO Yinzhu, Min Fangui, WANG Jing, HE Lifang, PAN Jinchun
    2021, 41(5):  443-449.  DOI: 10.12300/j.issn.1674-5817.2020.219
    Asbtract ( 393 )   HTML ( 27)   PDF (1188KB) ( 651 )  
    Figures and Tables | References | Related Articles | Metrics
    Appropriate biosafety management is a critical component of the successful operation of laboratory animal facilities. Standardizing the management of laboratory animal facilities, protecting staff, and strengthening the construction of laboratory animal health monitoring system are inevitable needs for the benign development of the laboratory animal industry. The control of microbial pathogens is a core component of laboratory biosafety management, and it is an important index for the evaluation of laboratory operation and management. The potential biosafety risk of animal laboratories is high owing to the experimentations with live animals, or pathogenic microorganisms. The control of microbial pathogens is an effective measure to reduce the risk of occupational exposure and acquired infections in laboratories. This study discusses microbial pathogen control strategies in laboratory animal facilities for promoting laboratory animal facility management and biosafety control.
    Practice and Exploration of Laboratory Animal and Laboratory Safety Training for Medical Freshmen
    SONG Guoying, XU Yan, ZHU Meilin, ZHANG Hongwei, LI Pei
    2021, 41(5):  450-454.  DOI: 10.12300/j.issn.1674-5817.2021.010
    Asbtract ( 339 )   HTML ( 23)   PDF (1170KB) ( 484 )  
    Figures and Tables | References | Related Articles | Metrics
    Laboratory animal and laboratory safety management in medical colleges and universities is the basic guarantee for the safe operation of teaching laboratories and serves as an important link for the successful completion of teaching tasks in laboratories. Aiming at the safety status of common experimental animals and laboratories, the Experimental Teaching Center of Basic Medical College of Zhengzhou University trained medical freshmen on the characteristics of experimental animals and laboratory safety knowledge. PowerPoint courses, short video lectures, and flipped classrooms were used for offline training. The training contents included the correct use of experimental animals, safety management and laboratory safety knowledge. After training, an online examination was conducted, which 1 061 freshmen passed, and the passing rate was 99.90%. According to the analysis of the answers of the freshmen in the examination, the error rates of knowledge on microorganisms and dangerous chemicals were 20.35% and 14.89%, respectively. This suggests that training should be strengthened in these two aspects in the future. Through the training, the new medical students mastered the correct use of experimental animals, and their knowledge of laboratory safety increased. This will help to prevent future laboratory accidents.
    Design of Two-dimensional Code Information Management System for Animal Corpses in Medical Experimental Teaching
    JIN You, WANG Chenchen, YU Shangbin, LUO Junyu, ZHOU Shunchang
    2021, 41(5):  455-458.  DOI: 10.12300/j.issn.1674-5817.2020.211
    Asbtract ( 289 )   HTML ( 17)   PDF (2118KB) ( 401 )  
    Figures and Tables | References | Related Articles | Metrics
    Experimental animals serve as an important tools for experimental teaching in medical colleges. This study systematically expounds the necessity of establishing a two-dimensional code information management system for the disposal of experimental animal corpses. According to the characteristics of experimental teaching animals, a two-dimensional code information management system for experimental animal corpses based on WeChat applet was designed. Through the seamless connection of participants, the dynamic real-time supervision of the production, collection, transportation, freezing, and treatment of corpses of experimental teaching animals was implemented. In order to effectively promote the standardization of experimental teaching, eliminate the phenomenon of random disposal of experimental animal corpses, and maintain the biological safety of the campus.
    Exploration and Practice: Laboratory Animal Management
    Design and Application of an Online Centralized Procurement System for Laboratory Animals in Universities: Capital Medical University as an Example
    LIU Xiaonan, ZHANG Wei, MENG Xia, LU Jing
    2021, 41(5):  459-465.  DOI: 10.12300/j.issn.1674-5817.2021.089
    Asbtract ( 308 )   HTML ( 18)   PDF (4470KB) ( 410 )  
    Figures and Tables | References | Related Articles | Metrics
    The establishment of a network information platform for a centralized procurement system for laboratory animals in colleges and universities has made the inventory supply and settlement management of laboratory animals centers more systematic, convenient, and scientific. The application of this system in the Laboratory Animals Department of Capital Medical University was examined, and the results showed that the establishment of a clear centralized procurement management module could greatly save labor and time, improve the work efficiency, and standardize procurement and supply personnel. The findings strongly suggested that the network informatization of the centralized procurement system of laboratory animals in colleges and universities is necessary.