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    28 October 2021, Volume 38 Issue 5
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    Promotion of the Ethical Review of Laboratory Animal Welfare through Institutional Construction
    2021, 38(5):  1. 
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    Abstract:Although great progress has been made in the ethical review of laboratory animal welfare in China, there are still some problems in the performance of IACUC including the unclear legal status of IACUC, the absence of rules and regulations, weak ability of IACUC and the insufficient attention of researchers. Based on the ethical review method of biomedical research involving human beings, 4 suggestions are put forward. Firstly, the management requirements of the ethical work of laboratory animal welfare are put forward in the “ Statute on the Administration of Laboratory Animal Use” . Secondly, it formulates the national level of laboratory animal welfare ethics review regulation. Thirdly, the ability of IACUC should be improved by training. Finally, the project application and paper publishing process should strictly abide by the animal welfare ethics. It is urgent for China to establish welfare ethical review system, to improve the ethical review of laboratory animal welfare and facilitate the development of science and technology.
    Effects of Heat Stress on Superovulation and Follicular Development in the Female Mice
    TIAN Yonglu, LI Na, TIAN Zhen, ZENG Shenming, ZHU Desheng
    2021, 38(5):  4. 
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    Abstract: Objective To explore the effect of short-term heat stress on superovulation and follicular development in mice. Method Selected the super-ovulated 20 female mice ( ICR) , which should be 7 to 8 weeks old and have approximate weight. These mice were randomly divided into 2 groups, the control group and the heat group ( n = 10) . The control group was fed in the SPF room and the heat group was heated in the 38 ℃ biochemical incubator for 10 hours every day. After 6 days, these mice were super-ovulated and then were killed. The ova and ovarian in the oviduct were draw. Result The experimental parameters of the heat group, such as the superovulation, the total number of follicles and the number of corpus luteum, were significantly higher than the ones of the control group ( P<0. 01) . The rate of follicular atresia of the heat group was also higher than that of the control group, but there was no significant difference between the 2 groups ( P >0. 05) . Conclusion Heat stress severely affects the reproductive function of mice, especially the follicular development process.
    Study on the Role of Caveolin-1 in Liver Fibrosis
    ZHANG Jie, YANG Ziheng, WANG Yan, SUN Quan
    2021, 38(5):  8. 
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    Abstract: Objective In this study, wild-type and Caveolin-1 ( Caveolin-1) knockout mice were induced by 10% carbon tetrachloride ( CCl4) olive oil solution to establish liver fibrosis model. The effect of Caveolin-1 knockout on liver injury and collagen deposition was analyzed to explore the role of Caveolin-1 in liver fibrosis. Method Wildtype mice and Caveolin-1 knockout mice were intraperitoneally injected with 10% CCl4 olive oil solution at 1 μL / g body weight, twice a week. Five mice were sacrificed on day 1, 3, 7, 14 and 28 after the injection. Serum and liver were collected, and the degree of liver injury was analyzed by HE staining and serum ALT and AST levels. The liver collagen deposition, together with the mRNA and protein expression of α-SMA, CollagenⅠ- and Collagen-Ⅲ were observed by Sirius red staining, Real-time PCR and Western blot to analyze the degree of liver fibrosis. Result On day 3, the contents of ALT and AST in serum of the knockout mice were significantly increased ( P<0. 01) . On day 3, 7 and 14, the necrotic area of liver of knockout mice was significantly increased ( P<0. 01 or P<0. 05) . On day 7, 14 and 28, the collagen staining area in Caveoin-1 knockout mice increased significantly ( P<0. 01 or P<0. 05) . Hepatic stellate cell activation marker, α-SMA mRNA significantly increased on day 3 and day 14 ( P<0. 05) . mRNA expression of Collagen- Ⅰ and Collagen-Ⅲ rise significantly on day 14 ( P <0. 01) . The protein expression levels of α-SMA, Collagen- Ⅰ and Collagen-Ⅲ were significantly higher in Caveolin-1 knockout mice ( P<0. 01 or P<0. 05) . Conclusion Caveolin-1 inhibited the liver inflammatory injury and fibrosis in mice induced by CCl4.
    Percutaneous Pulmonary Artery Denervation Cryoablation was Performed in Therapeutic Effect Evaluation of Canine Pulmonary Hypertension
    CHEN Hao , LIN Xianliang, WEN Fuli, XUE Laien, CHEN Jing, LI Linrui, WU Xiaojuan, LIN Yaze, YANG Shen
    2021, 38(5):  15. 
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    Abstract: Objective To observe the changes of mean pulmonary artery pressure ( mPAP ), pulmonary capillary wedge pressure (PCWP) and endothelin receptor 1(ET-1) after 6 months of pulmonary artery denervation (PADN) cryoablation in canine model of pulmonary artenal hypertension induced by monocyrinine, and to evaluate the safety and efficacy of cryoablation denervation of pulmonary artery for beagle dogs. Method Fifteen beagles were randomly divided into group A ( the cryoablation operation group) , group B ( the sham operation group) and group C ( the blank control group) , and each group with five dogs. The changes of mPAP, PCWP and ET-1 levels were observed after operation in beagles. Result Six months after surgery, the levels of mPAP, PCWP and ET-1 in groups A, B and C were lower than the preoperative level and the difference was statistically significant ( P<0. 05 ) . Conclusion Percutaneous pulmonary artery denervation cryoablation of canine pulmonary hypertension is safe and effective. The postoperative result showed a continuous reduction of pulmonary artery pressure.
    Analysis of Random Testing Results of Laboratory Animals in Beijing Area from 2017 to 2019
    WEI Jie, HUANG Jian, LIU Wenju, FU Rui, XING Jin, WANG Hong, WANG Ji, LI Xiaobo, FENG Yufang, WANG Shujing, GAO Qiang, WANG Xile, GONG Wei, HE Zhengming, LI Genping, YUE Bingfei
    2021, 38(5):  19. 
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    Abstract: Objective To provide reference to production, management and detection of laboratory animals by comparative analyzing of the recently 3 years’ test result from 2017 to 2019 on microbes and genetics of laboratory animals in Beijing area. Method According to the national and local standards, relevant law and regulation, the laboratory animal production units in Beijing area were randomly inspected and the tested. According to the data, 8 species of the laboratory animals including mice, rats, guineas, shrews, rabbits, dogs, monkeys and minipigs were classified according to their breeds, grades, units, etc. , and their quality problems were analyzed. The change in trend analysis of the quality of the laboratory then was analyzed and compared with the same period from 2014 to 2016. Result From 2017 to 2019, there were 144, 131 and 135 batches of random testing annually, while 30, 15 and 17 unqualified batches, respectively. The main reasons for disqualification were the immunity level dissatisfaction (20 batches in rabbits, dogs and minipigs) and 20 batches were infected by pathogens. No genetic variation was found. Compared with the relevant results from 2014 to 2016, no genetic problems have been detected. The status of substandard immunity has improved and pathogenic infections have increased. Conclusion This result can provide support to the healthy development of the laboratory animal industry.
    Effects of High-fat Diet on Body Weight and Bone Mass of Growth Period Rats
    YAN Guoqiang, LIU Lili, LIANG Zhen, REN Juan, SHU Xiong, ZHAO Danhui
    2021, 38(5):  28. 
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    Abstract: Objective To study the effect of high-fat diet on body weight and bone mass of rats during growing period. Method Twenty-four SD rats in 21-day-old were randomly divided into 2 groups: the general diet group and the high fat group (12 in each group, half male and half female) . After 6 weeks of feeding with the general diet or the high fat diet, the body weight was measured and the right femur was scanned by Micro-CT. The changes of body weight and bone mass were compared between these 2 groups. Result After 6 weeks of feeding, their body weight in the high fat group was significantly increased compared with that in the general diet group, and the growth rate of the body weight in the male group was significantly higher than that in the female group( P<0. 05) ; BMD was reduced, but there was statistical difference in the female group ( P < 0. 05 ) ; BV / TV and BS / TV decreased, but Tb. Sp increased ( P < 0. 05) , while BS / BV and Tb. Th did not change significantly in both rats ( P>0. 05) . Conclusion The obese rats fed with high-fat diet were more suitable to be male obese rats model, but the female obese rats were more likely to have osteoporosis and the whole loss of bone mass.
    The 240 SNP Panel Genetic Monitoring Analysis of CD ( SD) Rats from 2017 to 2019
    LONG Yanmeng, ZHENG Tianwei, ZHANG Qiaozhi, MIAO Xiangnan, TIAN Jun, YIN Lianghong, HAN Xue
    2021, 38(5):  32. 
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    Abstract: Objective From 2017 to 2019, Beijing Vital River has consecutively joined the 240 SNP panel genetic monitoring program for Charles River CD ( SD) rats to assess the levels of inbreeding and genetic divergence among colonies from barriers of various locations. Method DNA was extracted from CRL: CD ( SD) rat’ s ear specimens with QIAGEN spin columns. SNP loci were genotyped by PCR assays in a microarray. The pair of alleles at each locus were identified and labeled with 2 different fluorescent dyes, VIC and FAM, respectively. Signals were measured and reactions were analyzed. SNP genotypes were analyzed using GenA1Ex 6. 5 population genetics analysis software to evaluate the percentage of polymorphic loci, heterozygosity and inbreeding coefficients. Result From 2017 to 2019, polymorphism of Beijing’ s CD rat colonies were 73. 6%, 76. 9% and 74. 2% respectively, which staying relatively high and stable. The inbreeding coefficient indicated that the mating was random within colonies with data of -0. 02, -0. 06 and -0. 07. Fixation index for subpopulations and total of 3 years fluctuated around 0. 05. Conclusion CD ( SD) rat colonies of Beijing Vital River has kept high and stable polymorphism and the mating was random. In addition, they performed minimal to moderate genetic differentiation with the foundation colony of Wilmington, US. Regular introduction has been acting as an essential role in polymorphism maintenance and stability.
    The Role of Long-chain Non-coding RNA H19 in Bortezomib-resisted Multiple Myeloma
    XIAO Ping, CAI Guicheng, CHEN Wenting
    2021, 38(5):  37. 
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    Abstract: Objective To explore the role of long-chain non-coding RNA H19 ( lncRNA H19) in bortezomibresisted multiple myeloma ( MM) . Method Twenty six newly diagnosed MM patients, 25 chemotherapy relieves MM patients, and 22 bortezomib-resisted MM patients were selected. The human multiple myeloma OPM2 cells were transfected by siRNA-H19 and lentivirus. lncRNA H19 was detected by Real-time PCR analysis. The sensitivity of OPM2 cells to bortezomib was detected by CCK-8 cell viability analysis. Result The 2-△ △ Ct values of lncRNA H19 in the control group, the newly diagnosed MM group, the remission MM group and the drug-resistant MM group were (0. 107±0. 032) , (0. 324±0. 067) , (0. 218±0. 042) , and (0. 486±0. 095) , respectively. There were significant differences among these groups ( P < 0. 01) . Which was significantly higher in MM patients than those in the control group (P<0. 01) , in the newly diagnosed MM patients than those in the remission MM patients ( P<0. 01) , and in the drug-resistant MM patients than those in the newly diagnosed MM patients (P<0. 01) . The IC50 values of bortezomib to the control group, the OPM2 / si-H19 group, the OPM2 / NC group, the OPM2 / H19 group and the OPM2 / Blank group were ( 17. 86 ± 1. 64) nmol / L, ( 6. 83 ± 1. 05) nmol / L,( 18. 04 ± 1. 72) nmol / L, (31. 14±3. 57) nmol / L and (17. 75±1. 68) nmol / L, respectively. There were no significant differences among the control group, the OPM2 / NC group, and the OPM2 / Blank group ( P> 0. 05) . The IC50 values were lower in the OPM2 / si-H19 group than that in the control group, and in the control group than that in the OPM2 / H19 group (P<0. 01) . Conclusion High expression of lncRNA H19 may be involved in bortezomib resistance.
    Transcriptome High-throughput Sequencing of Ginsenoside Rb1 on DSS-induced Colitis in Mice
    DONG Jianyi, Li Changyi, CHEN Dapeng, WANG Liang, WANG Fujing, WANG Jingyu, ZHOU Zijuan
    2021, 38(5):  42. 
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    Abstract:In China, the incidence of Inflammatory Bowel Disease ( IBD) has increased significantly for the past few years. This disease is not easy to cure and comes with high expenditure. In order to find a cheap and efficient therapeutic method Illumina high-throughput sequencing techniques on mice with DSS-induced colitis. In this study, 15 C57BL / 6 mice were divided into the normal group, the model group and the Rb1 group. The treatment of model group and Rb1 group was 4 % ( m / v) DSS ( added in water) the first day, and 40 mg / kg ginsenoside Rb1 was fed to group Rb1 on the 2nd day. All mice were killed on the 9th day for samples of colon. The highthroughput sequencing platform Illumina was used to sequence the transcriptome of group normal, model, and Rb1. The Rb1 genes were compared with the model gene sets, and then the screened differential gene sets were analyzed by GO and KEGG pathway. The result showed that all indicators have improved after using Rb1. A total of 10,450 differentially expressed genes were GO annotated by Gene Ontology, containing 13. 5% molecular function, 68. 73% biological process, 17. 77% cellular component. Among the first 20 KEGG pathways that were most significantly enriched, the Neuroactive ligand-receptor interaction, the Calcium signaling pathway, and the cAMP signaling pathway were closely related to Rb1. Based on the result, we came to the conclusion that ginsenoside RB1 can alleviate the symptoms of DSS-induced colitis in mice. This work constructed the transcriptome sequence database of Rb1 on DSS-induced colitis in mice, which provides the sequence basis for future research on the gene mining function and metabolic pathways of ginsenoside Rb1.
    Application of Combined Anesthesia with Zolitel, Sumianxin Ⅱ and Lidocaine for Craniotomy in Beagle Dogs
    JIAO Kun, LU Jing
    2021, 38(5):  49. 
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    Abstract: Objective Zolitel and sumianxin Ⅱ were used for general anesthesia combined with lidocaine for local infiltration anesthesia. To study the effects of anesthesia for craniotomy in Beagle dogs. Method Atropine (0. 1 mg / kg ) was injected subcutaneously firstly. Fifteen minutes later, zolitel ( 2 mg / kg ) was injected intravenously, and sumianxin Ⅱ ( 0. 02 mL / kg ) was injected intramuscularly. The last, 0. 5% lidocaine (0. 1 mL / kg) was injected subcutaneously at the surgical site. Result The analgesic effect of anesthesia was
    evaluated, and the changes in animal heart rate, respiration and body temperature before and after anesthesia was detected. Conclusion This combined anesthesia method has high safety and good analgesic effect, which can meet the requirements of craniotomy in Beagle dogs. This method of anesthesia has a greater impact on the heart rate, so experiments with higher heart rate requirements should be carefully selected.
    Application of Curcumae Longae Rhizoma in Labeling Certain Laboratory Animals
    JIAO Jian, ZHANG Tingting, LU Lin, LIU Jinpeng
    2021, 38(5):  54. 
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    Abstract: Objective To examine the skin toxic reaction of extracts from Curcumae longae rhizoma by alcohol, thereby investigating the effect of the pigmentation on some rodents. Method Fine granules or powder of Curcumae longae rhizoma was soaked in appropriate medicinal alcohol overnight and the upper liquid was reserved. The dermal irritation test, skin phototoxicity test and skin sensitisation test were employed on guinea pigs with the upper liquid. Result Skin reaction score showed that Curcumae longae rhizoma had no obvious skin irritation, phototoxicity and other abnormal reaction. The color was visible until day 21 after administered. Conclusion The edible pigment extracted from traditional Chinese medicine Curcumae longae rhizoma is suitable for marking laboratory animals such as rodents.
    Feeding Management and Analysis of Laboratory Ferrets
    HE Zilin, YAO Minhua, WEI Tao, ZHAO Bohou, QI Yixin, JI Qingchao, CHANG Wenwen, LIU Xiaodong, SUI Hongshu
    2021, 38(5):  59. 
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    Abstract:Not only feeding conditions can affect the growth and development of laboratory animals, physiological functions and metabolism of the body, but also the standardization and feasibility of the operation will affect the collection of experimental data, which will affect the authenticity, accuracy and scientificity of the experimental results. The respiratory system of ferret is similar to human′s, which is an ideal animal model for studying influenza virus and common lung diseases. There are no detailed reports on the feeding and management of ferret in China. By improving ferret living environment, feeding method and experiment operation, the experiment result can be more real and accurate. In this paper, the feeding management and operation of labortary ferrets were reviewed in the combination with practical operation and relevant literature. It is expected to provide researchers engaged in ferret experiments with a set of standardized ferret feeding management and experimental operation program.
    Discussion on the Function and Ethical Review Process of IACUC in Drug GLP Organization
    SUN Changhua, ZHANG Yanmei, LI Shuixian, FAN Zhiyun, LI Zhikun, ZHU Qingfen
    2021, 38(5):  63. 
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    Abstract:In the certification system of Good Laboratory Practice ( GLP ) and the licensing management of laboratory animals, the Institutional Animal Care and Use Committee ( IACUC ) , as the internal supervision organization of the unit, plays an important role in the management of laboratory animals, the implementation of animal welfare and the occupational safety. In combination with the operation situation of IACUC in the center of drug safety evaluation of Shandong institute for food and drug control, our study aims to discuss the function and review process of IACUC, in order to provide information for relevant works.
    Exploration of the Intelligent Electrical Stimulation System Applied in the Mouse Electrical Irritation Experiment to Improve the Modelling Rate
    WANG Qiujing, LU Jinfeng, YU Xiaoya, LI Tianshu, SONG Xiangfu, AN Gang
    2021, 38(5):  67. 
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    Abstract: Objective In order to improve the success rate of electric stimulation model in mice, an intelligent electrical stimulation system was developed to meet the requirements of electric irritation experiment. Method The electrical stimulation system inputs the user's control parameters through the computer window interface. Through the RS232 communication between the computer system and STM32F103 single chip microcomputer, the control parameters input by the user are transmitted to the STM32F103 single chip microcomputer, and the high voltage output circuit is controlled to output the high voltage pulse required by the user to the cage electrode device, so as to irritate mice and apply it in animal experiments. Result The result showed that the success rate of the model reached 90%, which was in line with the 3R principle of animal welfare, and could meet the observation of the effect of chlorpromazine on the anti electric irritation of mice. Conclusion The developed electrical stimulation system can realize the intelligent control of stimulation parameters by computer, and its technical performance can meet the requirements of experimental teaching of electric irritation in mice.
    Research Progression of Animal Model in Schizophrenia
    WANG Fengjuan, YU Xiaoya, DING Yu, ZHANG Jinyu, YUAN Bao, HU Jinping
    2021, 38(5):  70. 
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    Abstract:Schizophrenia is a complex and persistent psychosis. Patients often can’ t control their emotions, which leading to appear delusional, world-weary and even suicidal behavior. The establishment of animal models in schizophrenia plays an important role in studying the pathogenesis and treatment of schizophrenia. In order to promote the improvement of animal models in schizophrenia and provide theoretical basis of studying the pathogenesis and the development of new medicines in schizophrenia, this article introduces schizophrenia, the ideal animal models in schizophrenia, the existing animal models in schizophrenia and so on.
    Overview of the Generation of Mice Carrying Point Mutation Based on CRISPR / Cas9 Technology
    LIU Suli, ZHU Yun, WU Huijuan
    2021, 38(5):  75. 
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    Abstract: CRISPR ( Clustered Regularly Interspaced Short Palindromic Repeats ) / Cas ( CRISPR-associated ) system has been widely used in biomedicine since it was first demonstrated to be able to perform genome editing in human cells and other eukaryotic cells in 2013. In this paper, the applications of CRISPR / Cas9 in the generation of mice carrying point mutation will be reviewed, providing theoretical guidance for the design and in vitro transcription of single guide RNA ( sgRNA ) and single strand oligonucleotide ( ssODN ) , for microinjection of zygotes and for genotyping of founder and progeny mice.
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