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25 October 2016, Volume 36 Issue 5
    

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  • Establishment and Evaluation of Spontaneous Abortion Mouse Models of Progesterone Insufficiency Induced by Mifepristone
    YANG Qian, YU Lin, GU Yan, SHI Yan, HE Ya-ping, WANG Jian
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 327-333. https://doi.org/10.3969/j.issn.1674-5817.2016.05.001
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    Objective To establish spontaneous abortion mouse models of progesterone insufficiency induced by mifepristone (RU486) to provide an ideal animal model for the mechanism of spontaneous abortion and clinical practice of supplying progesterone in pregnancy period. Methods Adult female mice were randomly divided into a RU486 high-dose group (50 μg/100 μL propylene glycol/only), middle-dose group (25 μg/100 μL propylene glycol/only), low-dose group (10 μg/100 μL propylene glycol/only) and solvent control group (100 μL propylene glycol/only). A subcutaneous injection of RU486 or solvent was given on pregnancy day 8, and the phenotype of pregnant mice and neonatal mice were observed on pregnancy day 10(d10) and after their delivery. The peripheral blood and uterus tissues of the mice in each group were collected on d10 to detect the levels of their serum estradiol (E2) and progesterone(P4). The uterine tissue differentially expressed genes between the experimental and control groups were screened and analyzed by Gene Chip Technology, and three of them were validated by Western blot. Results The abortion proportions were 100%, 46.0% and 13.9% respectively in the high-dose group, middle-dose group and low-dose group, and some embryos stopped developing and were absorbed in the uterus were observed in the middle-dose group, whereas the newborn mouse birth weights in the low-dose group were significantly lower than those in the control group (P<0.05). Compared with those of the control group, the E2 levels of the pregnant mice serum of the middle-dose group were increased (P<0.05) and P4 levels were decreased (P<0.001), while the E2 levels of the pregnant mice serum of the low-dose group were decreased (P<0.05), and P4 levels were increased (P<0.05). The Gene Chip and Western blot analysis and verification discovered that the expression of CDK4 in the uterine tissues was significantly reduced in the middle-dose group (P<0.05) compared with the control group, the expression of Bax was also significantly decreased (P<0.05), whereas the expression of CCND2 significantly increased and was statistically significant (P<0.05). Conclusion The appropriate dose of RU486 able to induce the pregnancy loss of pregnant mice after embryo implantation or neonatal weight loss. It might become a new animal model to be used for study of pathological mechanisms of the abortion or intrauterine growth restriction induced by progesterone deficiency in women pregnancy period.
  • Expression, Purification and Antigenicity of P Domain and Its Variant of Monkey GII.17 Norovirus Genome
    LIU Bo, LI Chao, TAO Yu-fen, LI Xin-tong, LIU Jian-sheng, HE Zhan-long, LIU Hong-qi
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 334-339. https://doi.org/10.3969/j.issn.1674-5817.2016.05.002
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    Objective To express P domain and its variant of monkey GII.17 norovirus genome, purify protein and analyze antigenicity. Methods The gene of P domain was synthesized, following designation based on the nucleotide sequence that was obtained from monkey GII.17 norovirus genome and bias of E. coli. PCR with these specific primers was performed to amplify P domain and make its variant, followed by cloning into prokaryotic expression vector via in-Fusion ligation. Protein expression was induced by IPTG, purified through GST affinity column and analyzed by Coomassie staining, immunoblot and ELISA assay. Results Sequencing results showed that P domain and its variant were successfully cloned into the expression vector. Analysis of SDS-PAGE revealed that P domain was observed in both the supernatant and pellet, and its variant was mostly expressed in the supernatant. The expressed and purified proteins were further confirmed by immunoblot analysis via the anti-GST antibody. The results of immunoblot analysis and ELISA assay indicated the good antigenicity of the proteins. The ELISA titers of antibodies induced by these proteins were 1:32000. Conclusion The P domain and its variant of monkey GII.17 norovirus genome were successfully expressed. The antibodies were obtained after immunization of mice with these two proteins. The findings of this study pave the way for the development of viral detection kit, viral identification and assay of virus-receptor binding.
  • Establishment of Over Active Bladder Model Associated with Chronic Cystitis in Rat
    GAO Feng, ZHANG Shi-geng, ZHANG Nan, ZHANG Bu-yi, DAI Yu-liang, LI Shao-jiang
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 340-344. https://doi.org/10.3969/j.issn.1674-5817.2016.05.003
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    Objective To build the over active bladder (OAB) related to chronic cystitis model which is similar to clinical process including pathogenesis and urodynamic performance. Methods Forty female SD rats were randomly divided into model group and saline control group. Model group: the bladders were instilled with Escherichia coli ATCC35218 (1×108~1.2×108CFU/mL) 100 μL, and a urine test were done after 3 days to confirm cystitis. The rats were given by gavage with Gentamicin (0.5 mL, 2 million units) 4 times a day × 7 days and urine were retested 10 days a cycle. The rats were done urodynamics and HE staining for bladder tissue after 4 cycles. Saline control group: were given by gavage with the same amount of saline solution instead of Escherichia coli, the remaining process was the same as the model group. The animals were raised under the same conditions. Results A small amount of epithelial was found degeneration, apoptosis, loss, and scattered lymphocytes in the lamina propria in model group. Thirteen rats were successfully modeled. Conclusion The OAB animal model can be established after cystitis by repeated E.coli infusion into bladder of female SD rats’. It offers ideas for pathogenesis, treatment and research about OAB associated with chronic cystitis.
  • Histopathological Observation on a Cross-territory Flap Choke Area of Ear in Mouse
    WEN Fu-li, LIANG Cheng, ZHUANG Yue-hong, WU Jian-wei, ZHENG He-ping
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 345-349. https://doi.org/10.3969/j.issn.1674-5817.2016.05.004
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    Objective To observe the histological and morphological changes of mouse’s cross-territory ear flap model of vascular. Methods The right-ear of mice as experimental group, the caudal of the mouse ears were cut for 2/3 with eye scissors from caudal to cephalic direction, ear’s flap based on a vascular pedicle but crossed three vascular territories was created.The left-ear as the control group, just clearing the hair. The ear of mice were surgically cut in 0 d, 1 d, 3 d, 5 d, 7 d respectively post-operation, and fixed in 10% formaldehyde for histological observation of the changes of choke I area and choke II area in the ear’s flap. Results At different time points, the average value of choke I area of blood vessels in the experimental group was larger than that of the other groups (P<0.01). The amount of venous blood vessels in choke I and choke II area respectively reached peak value at 3 d and 5 d, and then decreased gradually, and this trend may be related to the establishment of the venous reflux in the basal part of the ear flap.In choke I and choke II area, the mean values of arterial and venous blood vessel filling degree were 38.85%, 40.64% and 57.02%, 61.48%, respectively. Conclusions ①After establishment of extended flap, the dilation of veins seemed to passive , whereas the dilation of arteries seemed to active; ② The number of the choke vessels between the dynamic and potential territories that are involved in dilation and extent of the dilation are much smaller than that of the choke vessels between the anatomic and dynamic territories; ③ The change of vascular diameter has positive effect on the survival of skin flap.
  • Pathological Observations on Canine Mammary Tumors
    LI Yan-shu, CHU Fang, YANG Li-feng, ZHAO De-ming, ZHOU Xiang-mei
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 350-356. https://doi.org/10.3969/j.issn.1674-5817.2016.05.005
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    Objective To provide a evidence for the diagnosis and treatment of mammary tumors by pathological classification and description of the collected canine mammary tumors. Method The canine mammary tumors were obtained from China Agricultural University Animal hospital by surgical treatment, which were classified described according to Pathological observations. Results The benign and malignant mammary tumors were pathological described one by one, which were including the all common and uncommon canine mammary tumors. Conclusion There was greatly different pathology in different types of canine mammary tumor, so the treatment was greatly different.
  • Mechanism Analysis of Qiji Shenkang Total Granule Flavonoids on Regulating Glomerular Extracellular Matrix Dynamic Equilibrium through Inhibition of Inflammatory Response
    YANG Guan-qi, ZHANG Jun
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 357-360. https://doi.org/10.3969/j.issn.1674-5817.2016.05.006
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    Objective Through in vitro mesangial cell culture experiments, to explore if the total flavonoids of Qiji Shenkang Granule is the key effective component in the Fufang, and to further explore its mechanism. Method Used the macroporous resin adsorption method to extract total flavonoids in Fufang, and then use vascular tension angiotensin II to induce apoptosis in cultured mesangial cell membrane, which increased the extracellular matrix, after the intervention of total flavonoids of Qiji Shenkang Granule, analyzed the effect of interleukin 6 (IL-6) and monocyte chemotaxis protein-1 (MCP-1), fibronectin (FN) and type IV collagen (ColIV) protein expression. Results the Qiji Shenkang Granule flavonoids in high, dose group significantly reduced IL-6 and MCP-1 protein expression, FN and collagen content were also reduced, compared with Ang II group, P<0.01. Conclusion By regulating the expression of pro-inflammatory factors, the total flavonoids in the Qi Ji Shen Kang Granule can inhibit the excessive accumulation of extracellular matrix.
  • High-fat Diet Induced Type 2 Diabetes Mellitus and Obesity in C57BL/6 Mice
    LIANG Yong-jun, ZHANG Peng, WANG Yue-qian, GAO Li-li, CAO Ting, QIAO Zheng-dong
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 361-364. https://doi.org/10.3969/j.issn.1674-5817.2016.05.007
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    Objective To investigate the effect of high fat diet on Type 2 dialbetes mellitus (T2DM) and obesity, the model of T2DM was established by using prolonged high-fat diet (HFD) feeding on C57BL/6 mice. Methods C57BL/6 mice were fed with 60F10S diet for 12 weeks, the fasting blood-glucose (FBG) and animal body weight were monitored at six different time points. Moreover, oral glucose tolerance test (OGTT) was carried out to detect the blood glucose regulation ability and reflect the β cells dysfunction. Results Compare to the control group, the FBG had significantly increased from fourth week (P<0.001), while body weight grew steadily until the 12th week to formation of obesity (P<0.001). Correlation analysis reveals that the changes of FBG and body weight were correlated (P<0.001). After twelve weeks HFD feeding, the area under the curve of the OGTT (in mmol·h-1·L-1) was extraordinarily enlarged which means that the mice had poorer ability of blood glucose regulation (P<0.001). Conclusion Obese models of T2DM can be successfully induced by 12 weeks’ HFD feeding in C57BL/6 mice. As the simulation of high-fat diet in modern life, this model can reproduces the T2DM caused by diet disorders, and provides a powerful tool for the study of diabetes.
  • Detection of Main Immune Indexes in SCID. Beige and NOD.CB17-Prkdcscid Mice
    YUAN Shui-juan, SHI Guo-jun, TANG Hui-qing, XU Wen-jing, FU Jie, SHI Mei-lian
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 365-368. https://doi.org/10.3969/j.issn.1674-5817.2016.05.008
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    Objective To measure the immunoglobulin, complement and main immune indexes in SCID. Beige and NOD.SCID mice and analyze their difference. Methods Thirty SCID.Beige and thiry NOD.SCID mice, half male and female, at the age of 4 weeks, 8 weeks and 16 weeks were selected for detecting the main immunoglobulin and complement indexes. Another fourteen SCID.Beige and fourteen NOD.SCID mice, half male and female, at the age of 8 weeks were also selected for detecting the T, B lymphocyte function and NK cell activity. Results The IgG of SCID.Beige at all the age stages were significantly lower than those of NOD.SCID, but the IgA were significantly higher than those of NOD. SCID.The serum C4 of SCID.Beige mice at the age of 4 weeks and 16 weeks were significantly higher than those of NOD.SCID. The IgG of SCID.Beige and NOD.SCID mice at the age of 8 weeks were significantly lower than the other two age stages. In the T cell subsets, the CD4+CD8+ percentages of SCID.Beige mice at the age of 8 weeks were significantly lower than those of NOD.SCID. Among the same sex and age group of NOD.SCID and SCID.BG, the immune indexes show no obvious differences. Conclusions The antibodies, complement and immune cell levels of SCID.Beige and NOD.SCID are significantly lower than normal mice. They all belong to severe immunodeficiency animals. Among the same age and sex group of the two strains, the immune indexes were different. So the animals should be selected according to the research purpose in the experiments.
  • Comparison on Effects of Single Caged and Social Group on Physiological Indexes of Female Macaca Fascicularis
    LI Wei, JIN Yi, BAI Xiong, WANG Xiao-dong, XING Zheng-hong, WANG Xiao-quan
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 369-373. https://doi.org/10.3969/j.issn.1674-5817.2016.05.009
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    Objective To understand the differences on physiological indexes of single caged and social group female macaca fascicularis. Methods Nine female cynomolgus monkeys at the age of 4-6 years old were singly raised in cage at the first 5 months,and group raised in next 5 months.The body weight, body temperature, blood routine, urine routine, blood biochemistry were measured in single caged rearing and group breeding, repeated measures analysis of variance was used to compare whether there were differences between the two stages. Results Body weight in single caged stage was significantly lower than that in group phase (P<0.05). In blood routine, white blood cell count (WBC), neutrophil count (NEUT), neutrophil percentage (NEUT%) in single caged stage were significantly higher than that in group stage (P<0.01); monocyte count (MONO), the percentage of mononuclear cells (MONO%) in single caged stage were significantly lower than that in group stage (P<0.01); lymphocyte percentage(LYMPH%) in single caged stage was significantly lower than that in group stage (P<0.05). In blood biochemistry, total bilirubin(TBIL) in single caged stage was significantly lower than that in group stage (P<0.01); aspartic acid amino transferase (AST), triglyceride (TG) in single caged stage were significantly higher than that in group stage (P<0.05). The positive rate of urine protein in single caged stage is higher than that in the group stage(P<0.05). Other indexes between the groups had no statistical difference. Conclusion The macaca fascicularis was more suitable for the breeding of group, if the long-term single caged feeding, it could have a greater impact on body weight, hematology and urine routine of some indicators.
  • Effects of X-ray Irradiation with Different Dosages on Immune System in Mice
    YU Chun-miao, ZHAO Li-song, GUO Xu, MENG Dan, YU Dong-hua
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 374-377. https://doi.org/10.3969/j.issn.1674-5817.2016.05.010
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    Objective To explore the effects of the different dosages irradiation of X-ray on immune system in mice. Method A total of 192 mice, which were randomly divided into 6 groups: the normal control group, single irradiated experimental groups with the radiation dosages of 2.0 Gy, 3.0 Gy, 4.0 Gy, 5.0 Gy, 6.0 Gy respectively, irradiated by linear accelerator. 24 h, 48 h, 96 h, 192 h after irradiation, blood samples were taken from 8 mice of each group for detection of red blood cells (RBC), white blood cells (WBC), platelets (PLT) and hemoglobins (HGB). Results Compared with the control group, the content of RBC decreased in the different sampling time point. The content of WBC had significant decreased, during the same detecting time, the radiation dosage and the reduction extent of the content of WBC show a positive correlation. The content of PLT decreased, with a positive correlation between the radiation dosage and reduction extent. During different measuring time of each experimental group, the content of HGB decreased with the extension of time. Conclusion There are different changing rules of contents among different peripheral blood cells after radiation, radiation had great influence on the content of WBC, while had no significant influence on other peripheral blood cells.
  • An Introduction of Application of a Simple “X” Shaped Immobilization for Rats
    GAO Yuan, LIU Ding-long, XU Ping, HU Li, ZHANG Wei-bo, WEN Pei-tong
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 378-381. https://doi.org/10.3969/j.issn.1674-5817.2016.05.011
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    Objective To introduce a simple and convenient method for relative long time immobilization for rats. This method can meet the requirements of acupuncture or electroacupuncture experiments on the rats. Methods Imitating the external immobilization for clavicular fracture in orthopedics, the rats were immobilized with two strips of cotton threads about 110~120 cm long, one roll of scotch tape about 6 cm wide and one strip of silk ribbon about 20 cm long and 1cm wide by “X” shaped immobilization. Result The rats are immobilized effectively within a short time and the efficiency of the experiment is improved greatly by means of “X” shaped immobilization. Conclusion The simple, economical, effective and relatively safe method for immobilization the rats, “X” shaped immobilization is helpful to retain needle with the longer time and worth of popularizing.
  • Research Progress on Premature Ovarian Failure Animal Model induced by Vinylcyclohexene Diepoxide
    GENG Li-hua, TAN Yong
    Laboratory Animal and Comparative Medicine. 2016, 36(5): 404-408. https://doi.org/10.3969/j.issn.1674-5817.2016.05.018
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    Premature ovarian failure(POF) is a reproductive system disease as the main characteristics of ovarian function decline in advance.Although the high morbidity, there are few patients found and diagnosed early,which limits the study of the disease. Accordingly establishing the POF animal model which is similar to anthropic reproductive endocrinology and histological changes is of great value to POF pathogenesis reseach and further treatment.In experiments, POF model can be prepared by many methods, but current animal models of POF fail to adequately replicate this particular stage in female life.The article states the research progress of POF model induced by 4-vinylcyclohexene diepoxide (VCD) mainly on the diverse injection concentration, different injection time, disparate day age, distinct species and a large amount of regulatory factors which closely participated in follicular recruitment, growth and apoptosis.
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