Table of Content

15 October 2011, Volume 31 Issue 5

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Measurement of Some Blood Physiological and Biochemical Parameters in Cloned Bama Minipig

CHEN Hui-lan, ZHANG De-fu, WU Cai-feng, DAI Jian-jun, ZHANG Ting-yu, ZHANG Shu-san, GU Xiao-long, WU Bin

2011, 31(5):  330-332.  DOI: 10.3969/j.issn.1674-5817.2011.05.005

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Objective To investigate the basic data of blood physiological and biochemical characteristics in cloned Bama mini-pig. Methods To get a number of blood physiological and biochemical indexes of cloned Bama minipig, the blood samples of minipigs were measured by the automatic blood counting instrument and the automatic biochemical analyzer. Result The physiological and biochemical indexes were not significant differently in cloned Bama mini-pig and common Bama mini-pig, and quite close to those of human beings. Conclusion Some physiological and biochemical characteristics of cloned Bama mini-pig are similar to thoes in human beings.

Studies on Construction and Transfer of Cloned Minipig Embryos

DAI Jian-jun, LI Yao, GU Xiao-long, ZHANG Ting-yu, WU Cai-feng, ZHANG Shu-shan, WU Bin, CHEN Hui-lan, CHEN Xue-jin, ZHANG De-fu

2011, 31(5):  333-337.  DOI: 10.3969/j.issn.1674-5817.2011.05.006

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Objective To establish the somatic cell nuclear transfer technology of minipig and provide technical support for studies of transgenic animal models. Methods Using somatic cell nuclear transfer techniques, nuclear transfer was finished with fetal fibroblast cells and ear fibroblast cells (donor cells) from minipig and Fengjing pig respectively. Embryo transfer was carried out using the two kinds of reconstructed embryos. Result There were no significant differences between the two kinds of donor cells in vitro development (P>0.05). Chemical activation after electric fusion could partially improve the blastocyst rates, but the difference was not significant (P>0.05). Four healthy cloned piglets were borned after embryos transfer by the mixture of the two kinds reconstructed embryos. Microsatellite identification of cloned piglets showed whole same with their donor cells in heredity, and had nothing to do with their surrogate sows. Conclusion The method of our study group could be used to produce clone pigs.



Effects of Different Donor Cells Treatment with Aphidicolin and Donor Cell Passages on the Preimplantation Development of Somatic Cell Nuclear Transfer Embryos in Minipig

ZHANG Ting-Yu, DAI Jian-Jun, ZHANG Shu-Shan, WU Cai-Feng, ZHANG De-Fu

2011, 31(5):  338-344.  DOI: 10.3969/j.issn.1674-5817.2011.05.007

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Objective To optimize somatic cell nuclear transfer (SCNT) procedures in mini-pigs, the present study was designed to examine the effects of donor cell types, cell passage number and aphidicolin (APC) treatment on in vitro development of reconstructed embryos. Methods The Oviduct epithelial cells (OEC), ear fibroblast cells (EFC) and cumulus cells (CC) derived from mini-pigs were treated with serum starvation only or serum starvation followed by treatment of 0.1 ?g/ml APC. The reconstructed embryos were cultured for 7 days to evaluate their developmental competency. Results Cleavage and blastocyst formation rates of reconstructed embryos derived from the OEC by APC treatment were significantly higher than the serum starvation (61.82% vs 56.25%, 24.55% vs 17.86%; P<0.05). The cleavage rate from the EFC was significantly increased by APC treatment compared to serum starvation only (63.36% vs 57.01%; P<0.05). In the ooctyes with the CC, the reconstructed embryos could yield high blastocysts formation rate by APC treatment (29.63%; P<0.05). In the presence of APC, CC gave rise to highest cleavage and blastocyst formation rates among the three cell types. All of the three donors can be used as donor karyoplasts for nuclear transfer, and the efficiency is not influenced by culture passages. Conclusion The treatment of CC with serum starvation plus APC prior to nuclear transfer is more suitable in SCNT of mini-pigs.

Selection and Breeding of Experimental Wuzhishan Minipigs

WANG Xi-long, MIN Fan-gui, PAN Jin-chun, YUAN Wen, CHEN Rui-ai, HUANG Bao-song, WANG Feng-guo

2011, 31(5):  345-348.  DOI: 10.3969/j.issn.1674-5817.2011.05.008

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Objective To establish the closed colony and inbred strains of Wuzhishan Minipigs (WZSP). Methods For the purposes of establishing closed colony with genetic diversity and inbred strains with high homozygous genes, traditional breeding techniques and modern molecular genetic technology were employed for the seed selection and standardization of experimental WZSP. Results Under the premise of maintaining small size, one closed colony and two inbred strains of WZSP were fostered from those introduced outbred WZSP with rich genetic diversity and inbred WZSP with high genetic homogeneity. At the same time, the databases of biological properties of the established WZSP strains were created. Conclusion The newly established outbred and inbred WZSP strains, which maintained small size, steady physiological, biochemical, genetic parameters, and high productive performance, were proved to be ideal laboratory animals with a wide range of environmental adaptability. The WZSP strains would meet the needs of a variety of research areas and have potential application value and broad prospects.

Determination of Reproductive Performance of Outbred Wuzhishan Minipigs

MIN Fan-gui, PAN Jin-chun, YUAN Wen, CHEN Rui-ai, WANG Feng-guo, WANG Xi-long

2011, 31(5):  349-351.  DOI: 10.3969/j.issn.1674-5817.2011.05.009

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Objective To measure the reproductive performance of outbred Wuzhishan minipigs (WZSP). Methods All reproductive performances were determined according to national and industry standards. The statistical analyses were performed by the SAS soft. Results The reproductive behavior parameters of WZSP were stable and the females showed early sexual maturity. There were some differences between birth frequence and born and survival number of the generations, but most of the differences were not obvious. Correlation analysis showed that there were negative correlations between birth survival number & body weight of outbred WZSP, which were enhanced with the increase of birth frequencies. Conclusion WZSP exhibited higher reproduce performance and dressing percentage. And as experimental animals, WZSP will show a better value. Some differences in coefficient of variation showed us further more work on seed selection and purify.

Determination of Growth and Development of Outbred Wuzhishan Minipigs

WANG Xi-long, MIN Fan-gui, PAN Jin-chun, YUAN Wen, CHEN Rui-ai, WANG Feng-guo

2011, 31(5):  352-355.  DOI: 10.3969/j.issn.1674-5817.2011.05.010

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Objective To determine growth and development parameters of outbred Wuzhishan Minipigs (WZSP) from birth to 12 months old. Methods All growth and development parameters were determined according to national and industry standards and statistically analyzed by the SAS soft. Results The growth and development parameters were body weight, body height, body length, chest circumference, abdominal circumference, crown-rump length, ankle circumference, chest depth, chest width, hip width, head length, forehead width, jaw distance, tail length and tail circumference. Results showed that the above parameters of six months old WZSP were 12.11±2.99 kg, 50.93±5.02 cm, 32.64±3.81 cm, 52.13±5.43 cm, 50.64±5.77 cm, 58.30±5.88 cm,18.77±2.29 cm, 8.51±0.74 cm, 16.80±2.09 cm, 13.95±1.61 cm, 1.95±1.31 cm, 17.75±1.47 cm, 7.93±0.54 cm, 9.74±0.90 cm, 14.90±2.40 cm and 5.10±0.68cm respectively. Conclusion The growth and development parameters of WZSP were stable. The WZSP had a greater adaptability on climatic conditions of Guangdong and would be suitable for breeding in Guangzhou as well as southern China region.

Intravenous Glucose Tolerance Test and Analysis of Blood Fat in Wuzhishan Minipigs

PAN Jin-chun, MIN Fan-gui, YUAN Wen, WANG Xi-long, Chen Hua, Chen Rui-ai, WANG Feng-guo, LUO Shu-ming, YE Jian-cong

2011, 31(5):  356-359.  DOI: 10.3969/j.issn.1674-5817.2011.05.011

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Objective To screening the level of intravenous glucose tolerance test (IVGTT) and determine blood fat in Wuzhishan Minipigs (WZSP). Methods The WZSPs without castrationwere injected i.v. 50% glucose injection by 1.2ml/kg after 12 hours with water but fasting. The blood-glucoses were tested by fast blood glucose meter at the time of fasting, and 10, 30, 60, 90, 120 minutes after glucose injection, respectively. Meanwhile, the fasting serum samples were taken for testing biochemical parameters such as GLU, TG, CHOL, HDL-CH, and LDL-CH. Results The level of blood-glucoses was significantly fluctuant after intravenous glucose in WZSP. The level of blood-glucoses reached peak value at 10 minutes after glucose injection, then dropped gradually, and recovered at the time of 90 minutes after injection . There are 7 minipigs whose partial blood-glucoses exceed the upper bound of 95% degree of confidence in the IVGTT. And the blood serum biochemical parameters of 12 minipigs exceed the upper bound. It was found that GLU is significantly relevant to TG, CHOL, HDL-CH and LDL-CH by correlation analysis. Multiple linear regression analysis shows that GLU is significantly correlative to TG and HDL-CH. Conclusion The minpigs were screened out, which were abnormal in glycometabolism, lipid metabolism, and impaired glucose tolerance. And the minpigs could be bred which are susceptive to type 2 diabetes.

The Comparison of Genetic Polymorphism of SLA-DQA Gene Between Diannan Small-Ear Pig and Yunnan Wild Boar

LIU Rong-hui, LU Li-yi, TAO Xuan, YU Hui, LI Hua

2011, 31(5):  360-364.  DOI: 10.3969/j.issn.1674-5817.2011.05.012

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Objective Analyzed genotype and allelic polymorphism of SLA-DQA gene in Diannan small-ear pig and Yunnan wild boar (YWB, S. s. silvianus) to detect whether the genotype frequency of restriction enzyme sites was fit with Hardy-Weinberg equilibrium. Methods PCR-RFLP was used to analyze the partial sequences of intron 1 and intron 2 and the complete sequence of extron 2 of SLA-DQA in Diannan small-ear pig(DN) and Yunnan wild boar. Result Both EcoRⅠand AluⅠdigested sites had three kinds of genotypes, There were nine composite genotypes in Banna pig(BN) and eight in Mingguang pig(MG), but only four composite genotypes in Yunnan wild boar. The χ² test value of genotypes at both EcoRⅠand AluⅠsites in Diannan small-ear pig wasn’t fit with Hardy-Weinberg equilibrium (P<0.05), versus in Mingguang pig and Yunnan wild boar was fit with Hardy-Weinberg equilibrium, respectively (P>0.05). Conclusion Genetic polymorphism of SLA-DQA at both EcoR˘ńand AluⅠsites in Diannan small-ear pig is moderate, at EcoRⅠsite in Yunnan wild boar is moderate versus at AluⅠsite in Yunnan wild boar is low; the disequilibrium of Hardy-Weinberg in Diannan small-ear pig is majorly resulted from Banna pig.



Dynamic Observation On Growth of Subcutaneous and Orthotopic Gastric Tumor by in vivo Fluorescence Imager

PENG Xiu-Hua, SHEN Yan, XU Chun-Hua, YANG Yu-Qin, ZHOU Wen-Jiang

2011, 31(5):  371-375.  DOI: 10.3969/j.issn.1674-5817.2011.05.015

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Objective To observe the growth profile of MKN-45-Luc gastric tumor in nude mice by fluorescence imaging system of small animals in vivo , after the tumor transplantation into the nude mice through subcutaneous or orthotopic transplantation. Methods To inoculate subcutaneously the MKN-45 gastric cancer cells which were transfected Luc into nude mice, when the tumor formed, it were harvested and inoculated subcutaneously or directly transplanted into the stomach of nude mice. AT the same time, we used the fluorescence imaging system of small animals in vivo to monitor the dynamic changes of MKN-45-Luc cells in mice. Results From the first week to the fifth week, both the luciferase expressed area of the subcutaneous and orthotopic tumor and the fluorescent intensity were gradually increased with the time increase of tumor transplantation . A positive correlation existed among the tumor volume, fluorescence area and fluorescent intensity (r=0.882, P<0.001), which disappeared after 5-6 weeks following the inoculation. Conclusions The fluorescence imaging systems of small animals in vivo were untilized to fully observe the growth and metastasis profile of gastric cancer, which provided the basis for investigating the biological characteristics of the MKN-45 gastric cancer.



Comparative Observation on Pulmonary Fibrosis Models between Two Strains of Mice

LIU Rong, ZHAO Jin, LI Hong-tao, TAN Ya-Xia, CHEN Guo-qin

2011, 31(5):  376-380.  DOI: 10.3969/j.issn.1674-5817.2011.05.016

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Objective To provide a better animal model of pulmonary fibrosis by comparing the model establishment in different strains of mice. Methods BALB/c and C57BL/6 mice were divided into model and control group respectively, which received intratracheal injection of bleomycin (BLM) and normal saline , respectively. The mice were sacrificed 28 days after treatment. The masson mass and the degree of fibrosis in the left pulmonary lobe were observed by light microscope. Hydroxyproline contents in right pulmonary lobe were measured. Results The pulmonary consolidation and collagen deposition appeared in all bleomycin groups especially in the C57BL/6 group. Hydroxyproline contents in pulmonary tissue in C57BL/6 mice was the highest and normal saline group was the lowest one. Conclusion The pulmonary fibrosis model can be established in BALB/c and C57BL/6 mice by intratracheal injection of bleomycin, while the C57BL/6 mice was more suitable.



The Application of Reverse Transcriptase Assay in Golden Hamster

FU Rui, GONG Wei, YUE Bing-fei, HE Zheng-ming

2011, 31(5):  381-383.  DOI: 10.3969/j.issn.1674-5817.2011.05.017

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Objective To detect the reverse transcriptase (RTase) activity in the serum of golden hamster, rabbit, guinea pig, rat, mouse, simian and in Vero, BHK21, PERV cell on the basis of RTase assay. Method A pair of primer is designed according to the BMV RNA sequence. As reverse transcriptase, the serum and cells are tested by reverse transcript PCR (RT-PCR), and the specific band is observed by electrophoresis. Results The RTase activity is positive in rabbit, guinea pig, rat and golden hamster serum. Conclusion This assay has fine specificity and sensitivity, and it can be used in detection for RTase activity in animal serum.



Studies on Black Line Hamster (Phodopus sungorus) Used for Research and Their Biological Characteristics

HUANG Zhen-zhen, ZHAO Yuan, FAN Hai-ting, TANG Jia-ming

2011, 31(5):  384-388.  DOI: 10.3969/j.issn.1674-5817.2011.05.018

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Objective To raise the pet black line hamsters(Phodopus sungorus) for research use according to the standard breeding practise of laboratory animals, thus to expand the experimental hamster species resources, and fulfill the needs of biomedical researches. Methods The species was identified according to the rodents classification and identification methods. The animal was bred according to laboratory animal standardization requirements. The growth, development, reproduction, anatomical features were observed. The main viscera weight, hematology and blood biochemistry normal values were determined. Results The species is in accordance with the black line hamster (Phodopus sungorus)characteristic of hamsters subfamily by examination of the appearance, skull determination and chromosome check. The black line hamsters grow well in conventional environment with full nutrition granule feeds, and can be long-term male-female paired and bred. Hematology and blood biochemistry detection showed no significant differences between the sexes except a few indexes. Conclusion After breeding and standardizations, the black line hamster can be used for experiment. The observation indexes of the growth and breeding, and the determination of main viscera weighing, hematology and blood biochemistry normal values provide the reference for its biomedical research use.



Primary Emerging and Prevalent Infectious Agents in Rodents

LIU Ji-hong, SHEN Zhi, WANG Jian-fei

2011, 31(5):  392-398.  DOI: 10.3969/j.issn.1674-5817.2011.05.020

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With the rapid development of Bio-pharmaceutical industry and the broader and deeper international intercommunion in laboratory animal, there is a new feature expressed in disease prevalence in lab animals, especially in rats and mice. One hand, some emerging infectious agents are becoming more prevalent in lab animal colonies. On the other hand, a few traditional infectious agents which are required to be monitored routinely from national standard still keep high infection rates in rodents. Many of these infections may result in significant changes in immune function, the physiology and biochemistry of animals, and animal reproduction, even presence of clinical and pathological signs in animals. These alterations are serous variables that can have a devastating effect on animal colonies and research results. This article provides an overview of the prevalence, discovery, diagnosis, and research impact of the primary emerging and prevalent infectious agents in other countries recently in laboratory rodents. Hopefully, this article could be useful information to laboratory animal veterinarian or laboratory animal facility management in China.



Progress on Study of Rotavirus Mouse Model

YANG Meng-meng, ZHANG Yan, LIU Xin-you, LUO Jia-bo

2011, 31(5):  399-403.  DOI: 10.3969/j.issn.1674-5817.2011.05.021

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To summarize the study progress of rotavirus infected mouse model by analyzing the difference of virus replication, clinical performance, pathology change and pathophysiological mechanism between mice infected with homologous and heterogeneous virus.