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    28 August 2021, Volume 38 Issue 4
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    Research Progress and Application of Genetic Quality Detection Technology in Laboratory Animal
    DU Xiaoyan, HUO Xueyun, CHEN Zhenwen
    2021, 38(4):  1. 
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    Abstract:Genetic quality monitoring is one of the most important composition of laboratory animal quality control.It is essential method to find the genetic variation in laboratory animal and to confirm standardized animal quality. With the development of science and technology, especially with biological science progress, the technique of laboratory animal quality monitoring boosts from morphological genetic markers to DNA molecular level, and become more accurate, convenient and speedy. We sort out the history about methods in laboratory animal quality control, compare and analyze the advantages and limits of these methods, discuss the bright prospect of their application as well.
    The Roles and Significance of Pathology in the Evaluation of Animal Models of Human Diseases
    YANG Lifeng, ZHAO Deming
    2021, 38(4):  6. 
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    Abstract:Diseases have always coexisted with humans, threatened human health, and brought new challenges to the development and progress of human medicine. Animal models are “ essential tools” and “ important carriers” for revealing the pathogenic mechanism of human diseases and developing prevention and treatment technologies and products. The success of the development of animal models can directly affect the scientificity, accuracy and reliability in medical research result. Therefore, objective assessment for the animal models is particularly  important. As the “ gold standard” for disease diagnosis, pathology not only provides the most direct evidence for the evaluation of experimental animal modeling, and then assists in animal modeling design, optimization of modeling conditions, and optimization of modeling animals, etc. , but also helps to evaluate the characteristic lesions of animal models and directly reflect location, type, stage, degree, etc. , and at the same time, discover the lesions that may be associated with modeling, and exclude non-characteristic or background lesions. It is helpful to analyze the correlation between the modeling method and the disease, and screen out the most suitable animal model. Therefore, pathology plays an indispensable role in the assessment of animal models.
    Histopathological Effects of 137Cs-γ External Irradiation on Laboratory Red Crucian Carp
    TAO Hongting , LIAO Xiaoli , PENG Zanping, LI Dengke, CHEN Kejie, WU Duansheng
    2021, 38(4):  11. 
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    Abstract: Objective To investigate the histopathological effects of 137Cs-γ external irradiation on laboratory red crucian carps. Method The laboratory red crucian carps were subjected to 137 Cs-γ one-time external irradiation in HXFS-IA biological irradiator, with the irradiation doses 1. 94 Gy, 3. 88 Gy, 7. 76 Gy and 15. 53 Gy. On the 14th day after irradiation, these carps were dissected and the tissues of heart, liver, kidney and spleen were taken out. Paraffin sections of the heart, liver, kidney and spleen were stained by HE using routine method. Result The red crucian carps showed some histopathological changes after irradiation from 1. 94 to 15. 53 Gy 137Cs-γ. The arrangement of myocardial fibers is disordered, and the structure of myocardial fibers is uncompleted. Some hepatocytes appeared edema, and then necrosis. In the spleen, lymphocytes gradually decreased and reticular cells and fibrous tissue proliferated. The epithelial cells of the renal tubules were edema, then appared cell necrosis. The heart, liver, spleen and kidneys were damaged with the increase of nuclear dose. Conclusion In the irradiation doses from 1. 94 Gy to 15. 53 Gy, the laboratory red crucian carps were exposed to 137 Cs-γ for one-time external irradiation, and the heart, liver, kidney and spleen got histopathological damaged. The degree of tissue damages with the increase of radiation dose was exacerbation.
    Establishment of an Inducible Cardiomyocyte-specific Ppara-deficient Mice#br#
    WANG Jingjing, CHI Yafei, QIAO Xin, LU Jing
    2021, 38(4):  16. 
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    Abstract: Objective To establish a stable and effective inducible cardiomyocyte-specific Ppara ( gene of peroxisome proliferator - activated receptor α, PPARα ) -deficient mice model, the effects of different tamoxifen administration on PPARα knockout efficiency and cardiac function in these inducible cardiomyocyte-specific Pparadeficient mice. Method Pparafl / fl, myh6-ERT2Cre mice and the littermate control ( Pparafl / fl ) mice were obtained by mating and backcross between Pparafl / fl mice and Myh6-ERT2Cre mice. Group A ( mice were intraperitoneally injected with a single dose of 2 mg tamoxifen) , group B ( mice were intraperitoneally injected with 20 mg / kg· d tamoxifen for 5 days) and group C ( mice were intraperitoneally injected with 40 mg / kg· d tamoxifen for 5 days) . Two weeks after administration of tamoxifen, the cardiac function was monitored by echocardiography, myocardial fibrosis was observed with Masson staining and the PPARα expression was detected by Real-time qPCR and Western blot. Result Compared with the vehicle group, the cardiac function of mice in group A was not affected, as Real-time qPCR showed that the knockdown efficiency of Ppara in the myocardium was insufficient. While, in group B and group C, results of Real-time qPCR and Western blot confirmed the loss of PPARα in cardiomyocytes, and the echocardiography and histology indicated that the cardiac function of mice was normal in group B, but injured in group C. Conclusion These data suggested that intraperitoneal injection of 20 mg / kg tamoxifen for 5 consecutive days on Pparafl / fl, myh6-ERT2Cre mice can induce Ppara knockdown in cardiomyocytes sufficiently, leading to a successful establishment of inducible cardiac-specific Ppara deficient mice model.
    Similarities and Differences between the Self-established Tg. C57-ras V1. 0 Transgenic Mice Model and Japanese Tg. rasH2 Mice Model
    LIU Susu, CAO Yuan, YANG Yanwei, WANG Cenfei, WANG Sanlon, FAN Changfa
    2021, 38(4):  22. 
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    Abstract: Objective Compared the genetics of the independently established Tg. C57-ras mouse and Japanese Tg. rasH2 mouse model, and the biological characteristics of the first generation of hybrid mice between Tg. C57-ras transgenic mice and BALB / c mice ( CB6F1-NIFDC) and Japanese Tg. rasH2 transgenic mouse model ( CB6F1-CIEA) , which provided theoretical support for applying self-established mouse model in new drug preclinical carcinogenicity evaluation. Method The genetic characteristics, biological characteristics and sensitivity to positive carcinogens of the 2 models and F1 models were analyzed. Result The 2 models was used to construct by the genetically modified method. Analysis of genetics features showed that c-Ha-ras gene fragment has no actual difference, and only 1 copy difference in the copy number. mRNA expression level shows that genes in different organs of the two models are similar. But although the lung tissue protein levels in the Japanese model is slightly higher than the independent model, it reached no statistical difference. According to literature review and analysis, the growth curves of the 2 models were different, but there was little difference in body weight during the carcinogenic experiment cycle. There are some differences in blood physiological and biochemical indexes, and these differences were estimated to be related to genetic background. Conclusion Due to the comparing the characteristic data from 2 aspects, it is suggested that Tg. C57-ras transgenic mice established by ourselves may be a promising candidate animal model for development of a rapid carcinogenicity testing system for preclinical safety evaluation of drugs.
    Determination of Blood Physiological and Serum Biochemical Values of R26-hSCARB2 Mice
    ZHOU Jiaqi, WEI Jie, WANG Hong, LIU Susu, GUANG Jiaona, XIONG Rui, WANG Chenfei, FAN Changfa, YUE Bingfei
    2021, 38(4):  28. 
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    Abstract: Objective To examine blood physiological and serum biochemical values of R26-hSCARB2 transgenic mice and analyze the influence of genders and ages on these values. Method Four weeks and 8 weeks R26-hSCARB2 mice were taken respectively, with a gender ratio of 1 ∶ 1 10 mice in each group. After venous blood was collected, 22 blood physiological values and 12 serum biochemical values were determined, and statistical method were used to analyze the genders and weeks differences of each value. Result There were significant differences (P<0. 05) in LYM, NEUT% and BUN, and the extremely significant differences (P<0. 01) in WBC, CHO and TG between male and female of 4 weeks R26-hSCARB2 mice. There were significant differences ( P < 0. 05) in EOS%, NEUT, PDW, PLT, ALB, ALP, GLU and TG, and the extremely significant differences ( P< 0. 01) in BAS%, LYM, MON%, WBC, CHO, P and TP between male and female of 8 weeks R26-hSCARB2 mice. There were significant differences ( P < 0. 05 ) in MPV, NEUT, NEUT% and PCT, and the extremely significant differences (P<0. 01) in LYM, LYM%, MON, MON%, PLT, RDW, WBC, ALB, ALP, ALT, AST, BUN, Ca, CHO, CREA, GLU, P, TP and TG between different ages of R26-hSCARB2 mice. Conclusion Both genders and ages can affect the blood physiological and serum biochemical values of R26-hSCARB2 mice, and the effect of ages is greater than that of genders. This study provides data support for the application of R26-hSCARB2 mice.
    Vascular Endothelial Growth Factor Combined with Degradable Chitosan for the Treatment of Myocardial Infarction in Rats
    LI Yanmin, FENG Yan, WEI Yanyun, LIU Jing, WANG Xianzhong, LIU Junfa, ZHOU Wei
    2021, 38(4):  33. 
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    Abstract: Objective To study the therapeutic effect of vascular endothelial growth factor ( VEGF) combined with degradable chitosan ( DC) on myocardial infarction MI rats. Method Thirty two SD rats were randomly divided into the sham operation group, the model group,the VEGF group and the VEGF +DC group, with 8 rats in each group. These rats were modeled by ligating the left anterior descending coronary artery, and the sham operation group was only threaded but not ligated. After modeling, 100 μL of normal saline was injected into the myocardium of rats in the control group, rats in VEGF group were injected with the same volume of VEGF solution at the same location, and rats in VEGF+DC group were injected with VEGF+DC solution. After 4 weeks of modeling, all rats were subjected to echocardiographic function testing to investigate the left ventricular systolic and diastolic anterior wall thickness ( LVAW; s, LVAW; d) , left ventricular systolic and diastolic diameter ( LVID; s, LVID) ; d) , left ventricular systolic and diastolic volume ( LV Vol; s, LV Vol; d ) , left ventricular ejection fraction rate ( EF%) and short axis shortening rate ( FS%) , hemodynamic study left ventricular end diastole pressure ( EDP ) and the maximum rate of increase of left ventricular pressure during isovolumic systole and diastole ( + dp / dt, -dp / dt) , left ventricular index was calculated and the myocardium was stained with HE to observe the pathological changes of the myocardium in each group of laboratory animals. Result The myocardial infarction of rats in the VEGF group and the VEGF + DC group was effectively improved, and the improvement effect of the VEGF + DC group was more significant. The VEGF combined with DC can significantly improve various cardiac function indexes in rats with myocardial infarction. Conclusion In this study, it can be known that VEGF combined with DC can improve the cardiac function of rats with myocardial infarction and have certain therapeutic effects on myocardial infarction.
    The Function of MAOA in Inducing the Neuroendocrine Differentiation of Prostate Cancer
    SHUI Xue, XU Rong, XIE Qinghua, ZHANG Caiqin, TAN Dengxu, SHI Changhong, ZHAO Jumei
    2021, 38(4):  40. 
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    Abstract: Objective The cell and animal model of neuroendocrine prostate cancer ( NEPC ) were established respectively to explore the function and mechanism of monoamine Oxidase A ( MAOA ) in the neuroendocrine transdifferentiation ( NED ) of prostate cancer ( PCa ) . Method Human prostate cancer cell lines C4-2 were selected to establish the cell model of NEPC by long-term induction of Enzalutamide ( ENZ) treatment. The change of MAOA expression was detected by the enzyme activity experiment, and the marker of neuroendocrine differentiation were analyzed by Real-time fluorescent quantitative PCR and Western blot technology. The PCa cells 22RV1 were implanted subcutaneously into nude mice to establish xenograft model and to observe the effect of MAOA inhibitor clorgyline ( CLG) on neuroendocrine markers, to explore its underlying mechanism. Result ENZ could be used to induce the NEPC cell model. During the process of NED, the expression and activity of MAOA was increased. When CLG inhibit the activity of MAOA in the NEPC animal model the process of NED was delayed. The reason may be MAOA suppression of hypoxia relative signal. Conclusion MAOA is a key target to promote NED and maintain the characteristics of neuroendocrine cells. MAOA inhibitor CLG could delay the
    occurrence of NEPC and may be a new potential therapeutic drug for prostate cancer patients.
    Genetic Modified Mouse Models of Alzheimer’ s Disease
    LI Zhiping, YANG Lifeng, ZHOU Xiangmei, ZHAO Deming
    2021, 38(4):  47. 
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    Abstract:Alzheimer’ s disease ( AD ) is a neurodegenerative disease that affects more than 40 million peopleworldwide and it is predicted to exponentially increase in the coming decades. The typical pathological feature of AD is the abnormal accumulation of neurotoxic amyloid beta ( Aβ) peptides in brain, the clinical manifestations including memory loss, cognitive regression and abnormal behavior. While AD pathophysiology has fundamentally been identified, the underlying mechanisms which leading to initial disease onset and disease progression still remain open to speculation. Because no curative treatment and lack of systematic and completed pathophysiological studies, animal models is the most valuable scientific tool for research. This article summarizes the details of some
    kinds of genetic modified mouse models of AD, such as APP, PS, Apo E and ob genetic modified mice, and discusses the phenotypic characteristics, pathological changes and the application of them in the AD related research in a bid to provide references for the further studies on AD as well.
    Operation and Management Practice of Laboratory Animal Center Based on Internet of Things and Big Data Technology
    LI Zifa, GENG Xiwen, XU Kaiyong, HU Minghui, WANG Kezhou, SU Xixi, WEI Sheng
    2021, 38(4):  52. 
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    Abstract:In this study, by introducing the Internet of things into the management of the laboratory animal center, the direct collection of relevant data inside and outside the barrier environment is realized. Furthermore, the system can automatically trigger the relevant internet of things hardware to control according to the preset conditions. At the same time, big data analysis is introduced to graphically display the existing data in the laboratory according to management and business requirements, and the relevant data can be viewed at a glance through mobile phones, computers and large screens. The introduction of the internet of things and big data technology makes the management of experimental animals more intelligent and Real-time, and adds new management means by
    introducing related technologies, which provides real and Real-time data as support for scientific decision-making.
    Comparison of the Effects of Three Recombinant Hepatitis B Vaccines on Animals for Abnormal Toxicity Test
    CAO Chunran, LIU Hongyan, WU Wei, LI Xuemei, ZHU Chunyue, WANG Jiashuai
    2021, 38(4):  56. 
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    Abstract: Objective To observe and compare the effects of 3 recombinant hepatitis B vaccines on the weight gain and post-injection status of animals and to explore the effects of different pharmaceutical excipients on animals used for abnormal toxicity testing. Method The animal weight data and the observation status of the animals in the abnormal toxicity test of recombinant hepatitis B vaccine (Hansenula ploymorpha) , recombinant hepatitis B vaccine (Saccharomyces cerevisiae) and recombinant hepatitis B vaccine ( CHO Cell) , were collected from March 2016 to February 2018. The weight data statistically analyzed using IBM SPSS Statistics 23 analysis software to summarize the state of animals after vaccination. Result All of the vaccine passed the inspection. After the vaccination, the animals showed no abnormal state. Compared the weight gain of mice in the blank control group and the 3 hepatitis B vaccine administration groups, the differences were not statistically significant ( P > 0. 05) . The average weight gain of mice in the recombinant hepatitis B vaccine (S. cerevisiae) administration group was greater than that of the recombinant hepatitis B vaccine ( CHO Cell) and the recombinant hepatitis B vaccine ( H. ploymorpha) , the difference is significant (P<0. 05) . To compare the weight gain of guinea pigs in the blank control group and the 3 hepatitis B vaccine administration groups, the difference between guinea pigs in the recombinant hepatitis B vaccine ( CHO Cell) group and the blank control group was statistically significant (P<0. 05) , and the differences between the other 2 groups of animals and the blank control group were not statistically significant (P>0. 05) . To compare the administration of the 3 vaccines, there was no significant difference in the weight gain of guinea pigs in the medicine group (P>0. 05) . Conclusion There was no obvious difference in the general state of the animals, and none of the peritoneal irritation symptoms in mice caused by aluminum adjuvants recorded in the literature were observed; the effect of three recombinant hepatitis B vaccines on the weight gain of mice within 7 days after administration. In contrast, there was no significant effect on the weight gain of guinea pigs within 7 days after administration.
    Improved Method for Establishment of Porcine Superior Mesenteric Artery Thrombosis Model
    CONG Mingchi, JING Yuchen, XIN Shijie
    2021, 38(4):  62. 
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    Abstract: Objective An animal model of acute superior mesenteric artery embolism was established to evaluate the effect of self-developed thrombectomy and thrombectomy. Method Animal autologous blood clots were prepared in vitro. Interventional method were used to inject autologous blood clots into the superior mesenteric artery, resulting in arterial embolism. Embolization was performed using a thrombectomy device. Result Laboratory pigs successfully established acute intestinal ischemia-induced intestinal ischemia models, which are closer to the actual clinical situation than traditional method. Conclusion Using this experimental method, an animal model of acute superior mesenteric arterial embolism suitable for follow-up thrombectomy can be successfully established, which can better simulate the embolism of clinically-onset patients.
    Study on the Genotoxic Toxicity of Isobutyl Nitr
    ZHAO Shaohao, ZHANG Yuelong, JIANG Qingqing, TAN Yao, LIU Yanni, CHEN Xuejun
    2021, 38(4):  65. 
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    Abstract:To explore the mutagenic effects of isobutyl nitrite, according to the “ Technical Specifications for the Identification of Chemical Toxicity” , Salmonella typhimurium reverse mutation test ( Ames test) and mouse bone marrow polychromatic erythrocyte micronucleus test are used for the genotoxicity of isobutyl nitrite. The strains used in the Ames experiment are the histidine-deficient strains of S. typhimurium TA97a, TA98, TA100 and TA102. There were 10 dose groups in the experiment, and the dose range is from 250 to 5 000 μg / dish, setting up negative and positive control groups at the same time. Adopting plate incorporation method to observe the number of colonies per dish. The micronucleus test has three dose groups of 11 mg / kg, 41 mg / kg and 103 mg / kg. A negative control group and a positive control group are also set up to detect micronucleus rate of bone marrow polychromatic erythrocytes. The result indicate that isobutyl nitrite did not show mutagenic effects in the Ames experiment, and in the micronucleus experiment, there was mutagenic effect on mice at 103 mg / kg.
    Bowel Preparation before Colonoscopy on Macaques
    TANG Chuanjun, GONG Hui , HUANG Zhiyin , FENG Zhe , GAO Jinhang, TAN Qinghua , ZHAO Xudong , LV Longbao , LIU Chao , LV Weimin , WANG Yun, HU Bing, TANG Chenwei, ZHANG Qiongying
    2021, 38(4):  69. 
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    Abstract:Macaques are very important and precious laboratory animals. The aim of this study was to examine the entire colon on alive macaques repeatedly. In order that,it is particularly important to establish a method about colon preparation before colonoscopy. But until now there are no reports on specific preparation method for colonoscopy on macaques. The aim of this study was to develop a method of bowel preparation before colonoscopy on macaques. Thirty macaques were fed with glauber’ s salt through nasogastric tubes several times before colonoscopy. Intestinal cleansing was observed by colonoscopy. Three macaques were prepared for grade Ⅲ and the rest of the macaques were grade Ⅰ and grade Ⅱ . All macaques did not have serious complications such as dehydration, shock, and death during the process of washing the intestines. The colonoscopy was successfully completed on intestinal preparations of grade Ⅰ and grade Ⅱ macaques.
    Establishment and Research Progress of Animal Model of Chronic Wound in Elderly
    XU Feng, FAN Weijing, ZHANG Yinfeng, YAN Shimeng, LIU Guobin
    2021, 38(4):  73. 
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    Abstract:In the global environment of aging, the prevalence of chronic wounds in the elderly population is increasing. In order to better explore the pathogenesis of common chronic wounds in the elderly and more effective means of diagnosis and treatment, appropriate animals need to be established. This article summarizes the three animal disease models with high compressive ulcer wounds, venous ulcer wounds, and diabetic foot wounds, as well as their existing problems and future development.
    Advances in Animal Models of Chronic Bronchitis
    CUI Shuang, ZHANG Mingqian, LIANG Wulin, ZHANG Shuofeng
    2021, 38(4):  78. 
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    Abstract:To study the chronic bronchitis animal model of experimental research progress and pathogenesis. The influencing factors of chronic bronchitis are complex and changeable, which are related to long-term stimulation of infectious or non-infectious factors. Its pathogenesis is not only related to harmful gases and dust, bacteria, viruses, but also related to age, climate, organism immunity and other factors. This paper summarizes the method of building animal models of chronic bronchitis, such as fumigation, chemical drug and compound stimulation, in order to provide research progress and theoretical support for other scientists in animal models of chronic bronchitis.
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