Table of Content

28 February 2021, Volume 38 Issue 1

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The Key Points of Genotype Identification in Genetically Modified Mice Model

TAN Dengxu, MA Shuaijun, WU Pengpeng, ZHANG Caiqin, ZHAO Yong, BAI Bing, QIN Jing, SHI Changhong

2021, 38(1):  1. 

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Abstract: Objective To study the therapeutic effect on prostate cancer and possible application in living imaging of synthetic compounds DZ1-ABi and 783-ABi derived from fluorescent dye heptamethine carbocyanine ( nearinfrared fluorescence, NIRF ) and abiraterone acetate respectively. Method DZ1-ABi and 783-ABi were chemically synthesized; Human prostate cancer cells PC-3, LNcaP and C4-2 were cultured to the logarithmic growth phase, then DZ1-ABi, 783-ABi and ABi were added to observe the inhibitory effects on tumor cell

proliferation; Human-derived prostate cancer PDX tumor ( B45354) was subcutaneously transplanted into nude mice, randomly grouped and treated with DZ1-ABi, 783-ABi and ABi to monitor tumor growth. Tumor-bearing mice were injected with DZ1-ABi and 783-ABi ( 0. 1 nmol / mouse) for monitoring metabolism and tumor-targeting ability of these compounds by in vivo imaging. Result Compared with ABi and 783-ABi, better inhibitory effect of DZ1-ABi was observed on the proliferation of human prostate tumor cells; Moreover, DZ1-ABi exhibited better therapeutic effects on the treatment of tumor-bearing mice; In addition, in vivo imaging disclosed that DZ1-Abi has an obvious advantage over 783-ABi in terms of tumor-targeting ability or fluorescence signal intensity. Conclusion Compared with 783-ABi and the clinical drug ABi, DZ1-ABi has obvious advantages in tumor inhibition and in vivo imaging, which potentiates it as a new dual-targeted drug for tumor imaging and treatment.

Repairment of Cartilage Defects through BMSCs Overexpression BMP-4 with Allogeneic Bone

ZHANG Fei, WEN Fuli, DANG Yuan, XUE Laien, ZHENG Heping

2021, 38(1):  7. 

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Abstract: Objective In our research, we used BMSCs modified by BMP-4 through adenovirus and combined with allogeneic bone scaffold in order to have an effective regeneration of articular cartilage. Method Firstly, manufacturing the allogeneic bone cartilage column grafts, then repairing the articular cartilage defects with allogeneic bone cartilage column grafts which are modified by BMSCs overexpressing BMP-4. After that, taking a general observation and analysis of knee joint specimens. Lastly, taking a quantitative analysis of the regeneration of articular cartilage through the HE, saffron O, Masson and immunohistochemistry. Result The result of the HE, safranine O, Masson and immunohistochemistry indicated that the experimental group could significantly increase the content of cartilage fibers in the regeneration of articular cartilage. Conclusion The composite of allogeneic bone scaffolds with BMSCs overexpressing BMP-4 can significantly promote the regeneration of cartilage cartilage and the repairment of cartilage damage.

Research of Skin Transplantation and Biochemical Marker Genes in Spontaneous Marker Genes in Spontaneous Congenital Cararact Rats

YOU Jinwei, DONG Min, WU Zhenglin, LIANG Lei, TIAN Xiaoyun

2021, 38(1):  13. 

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Abstract: Objective To investigate the anti-rejection effect of xenogenic skin transplantation and biochemical marker genes in congenital cataract rats ( TSC) . Method Ten male TSC rats (170-200 g) were chosen for tergal skin transplantion of the autologous and allogeneic. One week later, bandage was removed and skin groeth was observed. We chose 6 TSC rats (200-220 g) from the same nest of basic group F29 generation, half male and half female, aged from 6 - 8 weeks. Tests were measured according to Chinese national standards GB / 14923—2010( laboratory animal-genetic quality control of mammalian laboratory animals) and GB / T 14927. 1—2008 ( laboratory

animals-method for biochemical markers of inbred mice and rats) . Result The skin graft survival with conclusious observation of 100 days from TSC rats. The marker loci were Akp1 a, Cs1 a, Es1a, Es3 a, Es4 b, Es6 b, Es8 b, Es9 c, Es10 a, Hbb b and Alp1 b, respectively. Referring to the biochemical genetic profile of the inbred rat strains commonly used at present, we did not find exactly the same biochemical markers with our TSC rats. Our TSC rats have unique genetic profiles. Conclusion There was no rejection after skin transplantation, which indicated that the histocompatibility gene of inbred TSC rats was consistent. The result of 11 biochemical markers

such as esterase-1 were homozygous, which met the quality standards of inbred lines.

Effect of Nutritional Supplements during Pregnancy on Spontaneous Motor Activity and Explorative Behavior of Mother and Filial Mice

ZHANG Changyong, BAI Jingcui, BAO Guo, HE Jialing, LI Nan, WANG Shangming, ZHANG Bin, ZHANG Shucheng, SUN Deming, WANG Ning, WANG Tianqi

2021, 38(1):  17. 

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Abstract: Objective To observe the effect of adding multiple vitamins and mineral nutritional supplements during pregnancy on the autonomy and exploration behavior of mother and filial mice rats, as well as, on the development of filial mice. Method SPF Kunming mice were randomly divided into low-dose group, medium-dose group, high-dose group, and vehicle control group after mating and seeing the suppository. The nutritional supplements added during pregnancy were converted into mice according to the recommended daily intake of 2 g / d. Comparing to the solvents as the control group, the experimental group divided into 3 groups, whose effective dose were 150 mg / ( kg· d) , 300 mg / ( kg· d) and 600 mg / ( kg· d) , respectively. Observing the birth weight, weaning weight and

weaning weight index of 1 week after weaning, at the same time, we conduct the open field experiment on the mother and filial mice 1 week after weaning to observe their behavioral differences. Result The birth weight of the 3 dose groups increased compared with the control group, and the difference between the high dose group and the control group was statistically significant ( P<0. 05) . The body weight of the filial mice on the day of weaning was significantly higher than that of the control group ( P<0. 05,P<0. 01) . One week after weaning, the weight of the 3 dose groups increased significantly compared with the control group ( P<0. 01) . There was no significant changes in the 2 analysis indexes of horizontal and vertical movements of postpartum mother rats ( P> 0. 05 ) , and the 2 analysis indexes of the staying time in the central area and the numbers of crossing the central area showed an increasing trend. One week after weaning, there was no significant difference between the horizontal and vertical movement indexes of female filial mice ( P > 0. 05) , but the horizontal movement indexes showed an increase of 5. 16% and 14. 79% in the medium and high dose groups; in the medium and high dose groups, the residence time of the central district increased by 4. 59% and 72. 95% respectively, and the high-dose group increased significantly ( P<0. 01) ; the number of crossings the central area increased by 8. 58% and 56. 89%, which the high-dose group increased significantly ( P<0. 05) . In male filial mice, 3 experimental groups compared with the control group had an increase in the number of vertical exercise times, horizontal exercise time and horizontal exercise number of times. Among them, the difference in the number of vertical exercises between the high-dose group and the control group was statistically significant ( P< 0. 05 ) . Conclusion The addition of nutritional supplements to pregnant mothers during pregnancy can increase the body weight of the filial mice, which is benefit to the growth and development of the filial mice, and can increase their exploratory behavior without increasing the autonomous activity of the experimental animals.

Effect of miRNA-125a Targeted Regulation of Bcl-2 on Neuronal Apoptosis in Epileptic Rats

PEI Jing, CHEN Ming, GAO Hua, BAI Yang, WANG Ping

2021, 38(1):  23. 

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Abstract: Objective To investigate the effect of miRNA-125a targeting Bcl-2 on neuronal apoptosis in epileptic rats. Method Thirty two male SD rats were randomly divided into normal control group ( group A) , epilepsy group ( group B) , miRNA-125a antigomir control group ( group C) and miRNA-125a antigomir group ( group D) . The expression level of miRNA-125a was detected by qRT-PCR. HE was used to detect the pathomorphological changes of CA-1 region in hippocampus in each group. TUNEL method was used to detect the apoptosis number of neurons in CA-1 region in hippocampal. Western blot and qRT-PCR were used to detect the expression of Bcl-2 in brain tissue of rats in each group. The targeting relationship between miRNA-125a and Bcl-2 was detected by luciferase activity. Result Compared with group A, the expression of miRNA-125a in brain tissue of group B and group C increased significantly ( P<0. 05). In group A, hippocampal cells were arranged orderly, cell morphology and structure were clear and complete, nucleolus was obvious, interstitial tissue had no edema; in group B, necrosis was seen, cell arrangement was disordered, nuclear pyknosis and nucleolus disappeared, cell gap widened and edema appeared. The result of TUNEL showed that the number of apoptotic cells in hippocampus of group B, C and D was significantly higher than that of group A, while the expression of Bcl-2 mRNA and protein was decreased. The expression of miRNA-125a in group D was lower than that in group C, while the expression of Bcl-2 in group D was higher than that in group C ( P<0. 05) . The result of luciferase reporter gene experiment showed that miRNA-125a and Bcl-2 could target to combine. Conclusion The expression of miRNA-125a in brain tissue of epileptic rats can be further regulated by Bcl-2, which can further lead to brain horse neuron injury and a large number of neuron apoptosis; inhibition of miRNA-125a expression level can improve the neuron injury of epileptic rats.

Study on the Application of COMT Gene SNP Loci in the Identification of the Timid Temperament of Labrador and Golden Retriever

BAI Jing, MA Xuena, CHEN Shuting, LUO Dong, ZHOU Zijuan, WANG Jingyu, WANG Fujin

2021, 38(1):  29. 

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Abstract: Objective To detect the polymorphism of the single nucleotide polymorphisms ( SNP) loci of catecholO-methyltransferase ( COMT ) gene in Labrador and Golden Retriever for statistically analyzing the association between the genotypes and haplotypes of the SNP loci and the timid temperament, in order to provide effective genetic biomarkers for pre-selection of guide dogs. Method One hundred and eighteen dogs (91 Labrador and 27 Golden Retriever) were selected. The dog courage assessment ( DCA) was used to assess the courage behavior of dogs. High resolution melting ( HRM) was used to detect the genotype of the 4 SNPs of COMT gene: -1 666 bp C>G, c. 39 A>G, c. 216 G>A and c. 482 G>A. Finally, the SPSS 19. 0 and PHASE 2. 1 software were used to analyze the correlations between the genotypes and haplotypes of the 4 SNPs of COMT gene and courage behavior. Result No significant correlation between the breed, sex, or age and courage behavior of Labrador and Golden Retriever ( P >0. 05) was detected. However, a significant collection between the genotypes of the 4 SNPs of COMT gene and courage behavior was found ( P< 0. 01) . In addition, the haplotypes of the 4 SNPs loci of COMT gene were also found to be significantly associated with the timidity of Labrador and Golden Retriever ( P<0. 01) , and the A alleles at the c. 216 G>A and c. 482 G>A sites played a dominant role. Conclusion There were significant correlations between genotypes and haplotypes of the 4 SNPs of the COMT gene and courage behavior of Labrador and Golden Retriever. This suggests that the SNP of COMT gene may significantly affect the dog’s timid temperament and can be used as an effective genetic molecular marker to assist early identification of timid dogs.

Study on Rabbit Model of Hepatitis E Virus Infection by Blood Transfusion

LIANG Chunnan, LIU Zhiguo, HUANG Weijin, ZHAO Chenyan, XING Jin, FU Rui , WANG Youchun, SHE Ruiping

2021, 38(1):  35. 

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Abstract: Objective To establish a rabbit model of hepatitis E virus ( HEV) infection via blood transfusion in order to investigate the possibility of HEV transmission through whole blood transfusion and observe the dynamics of HEV markers during infection. Method Anticoagulant-treated blood samples collected via posterior vena cava between the hepatic veins and diaphragm from four rHEV-infected specific pathogen free ( SPF ) rabbits, were transfused to five naive SPF rabbits ( three of the blood samples were transfused to 3 recipient rabbits and the other sample to the 2 remaining recipient rabbits, respectively) through ear edge vein, with a dose of about 10 mL / kg. HEV RNA, antigen and anti-HEV antibody of the recipient rabbits were monitored weekly for 13 consecutive weeks. Result After intravenous transfusion, fecal HEV shedding occurred in all five rabbits; three of them had viremia and antigenemia. Anti-HEV antibody was negative or weakly positive; the other two rabbits were positive for anti-HEV antibody, but negative for antigen and viremia. The total success rate of the experimental infection was 5 / 5 (100%) . Among them, 2 / 5 ( 40%) rabbits had acute HEV infection, and 3 / 5 ( 60%) rabbits showed chronic HEV infection as the infection persisted more than three months. Conclusion The rabbit model of HEV

transfusion infection was successfully established. Acute and chronic HEV infection could occur in the infected rabbits.

Analysis and Enlightenment of Policy System and Standard System of Laboratory Animal Management under Different Systems at Home and Abroad

GONG Wei, LU Shengming, CHEN Hongyan, CHEN Zhenwen, FAN Chun, ZHAO Deming, LI Genping, HE Zhengming

2021, 38(1):  43. 

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Abstract: Objective To expound the advantages of the management system of experimental animals in China, find out the existing shortcomings, and put forward countermeasures and suggestions to improve and strengthen China's policy system and standard system for the management of laboratory animal. Method To analyze the policy system and standard system of laboratory animal management at home and abroad. Result There is no law on the management of experimental animals in China, which is far behind the international general requirements. Compared with the international advanced standards, the content of the standards of experimental animals in China is relatively backward, and there is a lack of systematization and coordination between standards. Conclusion Strengthening the legal management of experimental animals, promoting the revision of the regulations, promoting the formulation of local standards, and lay the foundation for further improving the service capacity of laboratory animals for the development of science and technology in China.



Dissemination Mode Research on Science Popularization of Laboratory Animals in the New Media Environment

2021, 38(1):  49. 

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Abstract:The study analyzes the advantages and characteristics of the new media, and expounds the importance of

science popularization and dissemination of laboratory animals, and takes the dissemination activities in the form of

new media created by the author􀆳s unit as an example, such as the laboratory animal network platform, the

laboratory animal WeChat public account platform, the digital science popularization exhibition platform,

“ interesting experimental” “ scientist live broadcast” , online and offline science popularization lectures and video

broadcasting, etc. This study researches and analyzes science popularization dissemination mode of “ Internet + new

media + laboratory animals” , with a view to providing experience and reference for other units in the industry to

conduct extensive laboratory animal science popularization activities in the new media environment.

Technological Process of Waste Gas Treatment Equipment for Laboratory Animal Facilities

DIAO Ruiguo, ZHANG Wenming, LI Na

2021, 38(1):  53. 

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Abstract: Objective The impact of technological process of waste gas treatment equipment on environment. Method The optimal plan of waste gas treatment for laboratory animal facilities has been found out by analyzing the experimental result of water spray absorption method, photooxidation, low-temperature plasma decomposition method and water spray dosing method. And the most effective deodorization method has been selected by comparing the concentration of gas such as ammonia, hydrogen sulfide, and odor in the inlet and outlet of equipment. Result The result showed that the concentration of ammonia, hydrogen sulfide and odor in the exhaust gas is better than that in the emission standard of odor pollutants ( GB 14554—93) by adding sodium hypochlorite water or using photooxidation plasma to deodorize. Conclusion The application of sodium hypochlorite in spray water or the combination of photooxidation plasma deodorization and water spray deodorization can meet the need of deodorization in animal facilities, which are feasible from economic and technical perspective. Thus, these method can reduce the odor content in the outlet of equipment of laboratory animal facilities and effectively solve the problem of odor around the facility, which have the advantages of high efficiency and energy saving, and can be promoted and applied in laboratory animal facilities.




A Modified Method for the Transgastric Administration of Rabbit

LI Yanjin, CAI Xianhua , FENG Ruibing, CHEN Fang

2021, 38(1):  56. 

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Abstract: Objective Introducing a modified drug delivery method for the transgastric administration of rabbit.Method On the basis of previous experimental experience, the author introduced a modified drug delivery methodfor the transgastric administration of rabbit and compared with the prior method. Result Compared with the prior method, the modified method showed simple operation,short learning curve, limited damage, high efficiency and can be done by one person. Conclusion The modified drug delivery method conforms to the principle of “3R” for experimental animals and is beneficial to take large sample study.




Analysis of Carrying Status in Common Bacteria Identified from the Upper Respiratory Tract of SPF Mice

WEI Tianwen, HU Qinbao, XUAN Junwen, LI Chenjiang, ZHOU Xiaojiang, LV Ming, ZHANG Xinchuang

2021, 38(1):  59. 

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Abstract: Objective To analysis the carrying status in common bacteria identified from the upper respiratory tract of SPF mice. Method The bacteria were isolated and cultured, stained, examined under microscope, biochemical reaction, and identified by Vitek 2 compact automatic bacterial identification. Result There were 8 species of common bacteria in the respiratory tract of SPF mice, which were divided into 4 classes according to carrying rate: the first category was 98. 6% carrying rate of Granulicatella adiacens; the second category were 45% - 65% carrying rate of Staphylococcus sciuri and Sta.xylosus; The third category were Enterobacter cloacae, Escherichia coli, Proteus singularis, which carrying rate were 25% -45%; the fourth category were less than 10% carrying rate of Acinetobacter, Klebsiella pneumoniae. Conclusion The species and carrying rate of common bacteria from the upper respiratory tract from SPF mice were obtained by Vitek 2 compact. This result would provide support for the evaluation of the microbial quality of SPF mice and the selection of SPF mice for scientific research projects.




Research Progress on Modeling Methods in Gestational Diabetes Rats

JIANG Xinyong, GE Li, PANG Shuqin, XIE Yong, LAI Yuting, HUANG Pingping, WANG Guandong, CHEN Fan

2021, 38(1):  63. 

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Abstract:The animal model of gestational diabetes mellitus ( GDM ) is the experimental basis for studying the development, treatment and nursing mechanism of GDM. Whether the GDM rat modeling method is consistent with the pathogenesis of human GDM and its modeling rate and mortality is essential. This article reviews the GDM rat modeling method based on domestic and foreign kinds of literature. At present, the commonly used modeling method in GDM rats mainly include drug induction, diet induction, and drug combination diet induction, but they're still much difference among them. There was a large difference in dose, lack of uniform GDM rat modeling standards and comparison between different modeling method and neglected the influence of activity factors on the GDM rat model.




Review on Rapid Detecting Approaches of Avian Salmonella

FANG Huanxin, LI Zhili, HUANG Shujian, HE Cheng, YUAN Sheng

2021, 38(1):  69. 

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Abstract:Salmonellosis is an important zoonotic disease, threating to animal industry and food security. Current eradication of avian Salmonellosis and Leucosis are implementing across breeder chickens in China. However, both efficacy and procedure are hampered due to lack of rapid, highly specific and high sensitive test assay. The conventional methods are not satisfied for highly demand in light of rapid detection, animal health and food safety due to time-cost duration, differentiation of serotype and pathogenicity. Confronting with technical dilemma, the review will engage in comprehensive analysis of bacterial isolation, serodiagnosis, PCR method and microarray approaches and sheds light on new technologies of detecting avian Salmonellosis in future.

The Key Points of Genotype Identification in Genetically Modified Mice Model

LIU Susu, WANG Chenfei, YUE Bingfei, LI Guanming, FAN Changfa

2021, 38(1):  74. 

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Abstract:Recently, with the development of biomedical research and gene editing technology, there are more and more animal models have been established. Due to the different production methods, the genotype identification technology is also different, resulting genotype identification disordered phenomenon. This paper is based on the method of the genetically modified mice model,introduced the genotype identification key points of the random transgenic mice, conditional knockin mice, and knockout mice. Through this paper, we want to achieve standardize the genotype identification method and ensure the accuracy of animal experimental materials, so as to promote the application and development of animal models in basic and applied science and technology fields.