Table of Content

28 February 2013, Volume 30 Issue 01

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Preparation and Preliminary Application of Sendai Virus Antigen

LIU Xian-Ju, TONG Wei, XIANG Zhi-Guang, ZHANG Li-Fang, LI Yu-Han, WANG Yan-Rong, WEI Qiang

2013, 30(01):  1-4. 

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Abstract: Objective To prepare and purify the murine Sendai virus ( SeV) antigen so as to establish the indirect ELISA and Western blot test for detecting the antibody against SeV． Methods SeV was inoculate into the allantoic cavity of SPFchick embryo，and then the allantoic fluid was added to the BHK-21 cell culture to proliferate SeV． Sendai virus antigen was prepared and purified by the differential centrifugation and ultrasonication，and its purity was identified by SDS-PAGE． The best coating antigen concentration for the indirect SeV-ELISA was selected and confirmed． SeV antibody in murine serum was screened by the indirect immunofluorescence assay ( IFA) ，and the SeV-ELISA specificity was verified by Western-blot test． Results The HA titer of Sendai Virus from BHK-21 cell culture was 1∶ 128，and SeV antigen electrophoretic purity reached to 80 percent． While the best coating antigen concentration for the indirect SeV-ELISA was assigned to 2. 5 μg /mL，the detection rate of positive Sev antibody in murine serum by the SeV-ELISA was absolutely compatible with that by IFA． Conclusions The SeV antigen prepared by differential centrifugation and sonication broken method can be used for detection of Sendai Virus antibody in mice，and the antigen-specific and activity of Sendai Virus have been verified by the Western-blot test．

Transfection Efficiency of Lentiviral Vector-based SMGT for Dog Testicular Germ Cells in vivo

WANG Hong-Jun, MA Xiao, ZHAO Ming, DU Xiao-Yan, LI Yi-Chen, CHEN Zhen-Wen

2013, 30(01):  6-10. 

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Abstract: Objective To observe the transfection efficiency of lentiviral vector for dog testicular germ cells so as to provide the foundation for generating the transgenic animal by lentiviral vector-based sperm-mediated gene transfer． Method LV-GFP with the titers 5 × 108 ，1 × 108 ，2 × 107 TU / mL was injected respectively into the bilateral testis of Beagle dogs in high，medium and low-dose group at a dose of 2 ml / dog． Sperm-collecting was set about from the second week after the testicular injection． The integration and expression of GFP gene were assessed by PCR tests of semen DNA and sperm detection with fluorescence microscope． Results 1) In high-dose group the integration and expression of GFP gene firstly appeared at the 4th week and maintained till the 17th week，and it appeared at the 5th week in the medium and low-dose group and lasted till to the 14th and 12th week respectively; 2) The peak level of GFP expression occurred in 7 to 10 weeks after the testicular injection; 3) GFP expressed in the entire sperm，but expressed highly in the acrosomal back zone and the neck section of sperm tail; 4) There was ascent trend of dog sperm deformity rate in high and low-dose group during 2 weeks to 6 weeks after the injection; 5) During the peak period of GFP expression，the percentage of GFP sperm in high，medium and low-dose group was respectively 43. 33% ，35. 53% ，4. 55% ，with a significant difference． Conclusion The lentiviral vectorbased SMGT can successfully generate transgenic dog sperm，and the transgenic rate，the intensity and the duration of transgenic expression were related to the titer of lentiviral vector．

Effects of Mutation Gene on the Structure and Function of Immune Organs in Hairless Mice

SHI Chong-Min, DU Chun-Yan, WANG Jun-Min, ZHU Kui-Cheng

2013, 30(01):  11-14. 

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Abstract: Objective To probe into the effects of hairless mutant gene on the structure and function of immune organs of hairless and haired mice． Method The observation of histological changes in immune organs and determinations of lymphocyte subset，proliferation of lymphocyte，and sIL-2R level in serum in hairless and haired mice，and comparison of the these indexes between hairless mice and haired mice were carried out． Result There were certain differences in structure and function of immune organs between hairless mice and haired mice． Conclusion It is suggested that the hairless mutant gene affects not only the hair structure but structure and function of immune organs，and that the gene in heterozygosity also exerts certain effect，indicating that it has certain codominance．

Pathological Investigation and Analysis of Health Status of Laboratory Rodents of Beijing Area in Autumn 2012

WANG Ji-Hong, YAO Hao, SONG Zhi-Qi, YUAN Zhen, LIU Chun-Fa, XUE Zhi-Xin, FU Yong-Yao, 吕Yue , ZHU Ting, ZHOU Yang, YANG Li-Feng, YIN Xiao-Min, ZHAO De-Ming, ZHOU Xiang-Mei

2013, 30(01):  15-18. 

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Abstract: Laboratory animal become more and more important in various fields with the development of science and technology． In October 2012，our lab took laboratory rodents from Beijing area to investigate the health status of laboratory rodents． Their heart，liver，spleen，lung，kidney，large intestine and small intestine were sampled，and then prepared into two different sections for optical microscopy including all paraffin specimens stained with HE and the frozen sections stained with Oil Red O and PAS． The results showed that the vast majority of laboratory mice were pathologically diagnosed as healthy animals． Liver fatty degeneration and slight lung lesion were main lesions， which might affect the health of laboratory animals． Protein cast of kidney，fibrin exudation occured in the Bowman's capsule and parasitic infections in the large intestine could be seen in several individuals． The result might be related to breeding，feeding environment and transport stress．

Research on Effects of Fenofibrate Tablets on Chronic Nonalcoholic Fatty Liver Disease in Rats

XU Li-Ying, XIAO Xiao-Peng, ZHENG Guo-An, DING Qi, LI Liang, XIAO Xiao-Hua

2013, 30(01):  19-21. 

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Abstract: Objective To investigate the effects of fenofibrate tablets on nonalcoholic fatty liver disease ( NAFLD) in rats． Method NAFLD Wistar rat model was established by high fat and sugar diet，and the effects of fenofibrate tablets on the serum and hepatic lipids were observed． Result Fenofibrate tablets obviously increased liver coefficient，reduced TG in serum and hepatic tissue，increased TCH and LDL，reduced serum HDL，and enhanced liver cell injury． Conclusion Fenofibrate tablets reduced serum and hepatic tissue lipids in NAFLD rat mode1，but increased liver injury．

Observation on Infectiousness of Echinococcus multilocularis by Different Inoculation Methods in Cricetulus migratorius

LIAO Li-Fu, LIU Bin, XU Yi-Mei, YAN Shun-Sheng, SHI Shen

2013, 30(01):  22-25. 

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Abstract: Objective To observe infectiousness of Echinococcus multilocularis by different inoculation methods in Cricetulus migratorius so as to provide the foundation for developing hydatid disease animal model in variety ofcavity，heart or liver． The development status and parasitic parts of the cysts were surveyed by dissection，and cysts coefficient ( cyst weight / body weight × 100% ) and infection rates were computed． Result The infection rate in 29 animals was 100% with a cyst coefficient of 7. 1% at 100 days after intraperitoneal inoculation． The cyst parasitism in the abdominal cavity was found in all 7 sacrificed animals，and liver cyst infiltration was found in 3 animals，spleen cyst infiltration in 2 animals and kidney cyst infiltration in 2 animals with total cyst coefficient of 51. 7% at 300 days after intraperitoneal inoculation． At 116 days after armpit inoculation，the cyst parasitism in armpit was revealed in all 19 sacrificed animals with a cyst coefficient of 3. 27% ，and at 290 days after armpit inoculation the cyst parasitism in armpit was revealed in all 5 sacrificed animals with a cyst coefficient of 22. 95% ． At 100 days after liver inoculation in 17 animals，the cyst parasitism in the abdominal cavity revealed in 8 animals， the cyst parasitism in liver in 5 animals，the cyst parasitism in heart in 1 animal，and the cyst parasitism in lung in 3 animals with a cyst coefficient of 7. 1% ． At 115 days after heart inoculation in 26 animals，the cyst parasitism in heart revealed in 6 animals，the cyst parasitism in lung in 9 animals，and the cyst parasitism in thorax in 1 animal with a cyst coefficient of 2. 91% ． Conclusion The celiac environment of Cricetulus migratorius may be hospitable for the growth and development of Em，which is appropriate for developing of the short-period animal model of hydatid disease． Em grows and develops slowly in the forelimb armpit of Cricetulus migratorius，so that the armpit inoculation is suitable for bread conservation of Em． Because of the lower in fecting rate for inoculating heart and liver in the animal models，the inoculation technology and methods should be improve，so as to increase the infection．

A Study on Acute Toxicity of Crude Polysaccharide Extracted from Juglans Mandshurica Maxim to Mice

WANG Guang-1, WU Yi-Yan, LEI Tao

2013, 30(01):  26-27. 

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Abstract: Objective To estimate the acute toxicity of crude polysaccharide extracted from juglans mandshurica maxim to mice． Method Crude polysaccharide extracted from juglans mandshurica maxim ( concentration of 5. 4 mg /mL) was orally administrated to the mice at a dose of 40 mL / kg，three times per day，and was injected intraperitoneally to the mice at a dose of 20 mL / kg，two times per day． After administrated for 1 days，continuous observated 7 days． Result In the course of the test，the activity and diet of mice were normal，and there were no death of the mice． Autopsy revealed nothing remarkable． The maximum dosage of oral administration and the maximum dosage of intraperitoneal injection were 6. 48mg /10g and 2. 16 mg /10g respectively． Conclusion Toxic effect of the rude polysaccharide extracted from juglans mandshurica maxim on the mice in the two administration routes was low．  crude polysaccharide extracted from juglans mandshurica maxim; mice; intragastric administration; intraperitoneal injection

Application of Ivermectin in Expelling and Killing Internal and External Parasites of Laboratory Animal

QU Ping, JING Hai-Jiang, CHOU Zhi-Hua

2013, 30(01):  28-30. 

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Abstract: Objective To find safe and effective drugs for expelling and killing internal and external parasites of laboratory animal． Method The male mice carrying Myocoptes musculinus were randomly divided into 3 groups， and female mice were randomly divided into 3 groups; the male mice carrying Syphacia obvelata egg were randomly divided into 3 groups，and female mouse were randomly divided into 3 groups． The expelling and killing effects were observed through drinking 0. 1% ，0. 5% and 1. 0% ivermectin for three cycles． Result Three concentrations of ivermectin could exert expelling and killing effects for Myocoptes musculinus and Syphacia obvelata of laboratory animal． Conclusion Ivermectin can be used as a safe and effective drug for expelling and killing internal and external parasites of laboratory animal．

Investigation and Analysis of Animal Experiment Ethics Cognition in Cardiovascular Disease Researchers

CUI Yong-Chun, TIAN Zhong, TANG Yue, SUN Jia-Kang, WU Ai-Li, LIU Xiao-Peng, LUO Fu-Liang

2013, 30(01):  31-34. 

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Abstract: Objective To find out animal experiment ethics cognition in the cardiovascular disease researchers to provide new strategies in training for animal experiment ethics． Method Self-designed questionnaire about animal experiment ethics was completed in 197 cardiovascular disease researchers performed animal experiments in animal experimental center，Fuwai Hospital from January 1，2009 to April 30，2012． Result All of the subjects believed that animal experiment ethics was necessary for animal s' health and happiness． Good score of animal welfare and ethical knowledge was found in 54. 3% of subjects，qualified score in 35. 5% ，and unqualified score in 10. 2% ． All the unqualified subjects were the graduate students of 20 ～ 30 years old and without animal experiment research experience． Conclusion It is necessary to strengthen the training on experimental animal ethics，especially for the graduate students who would take part in animal experiments．

A Comparative Study on Determining Methods for Crude Fat in Extruded Feed

WANG Man-Hong, ZHANG Hai-Feng, LIU Jian-Guo, HUANG Wei-Dong

2013, 30(01):  35-37. 

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Abstract: Objective To investigate proper determination method for crude fat in extruded feed． Method Crude fat contents in experiment feeds including non-extruded dog feed，extruded dog feed，non-extruded primate feed， extruded primate feed，non-pelleted rodent feed，pelleted rodent feed，non-pelleted rabbit feed and pelleted rabbit feed were determined by acid hydrolysis method and Soxhlet extraction method， respectively． Result The determined level of crude fat in extruded primate feed by acid hydrolysis method was very significantly higher than that by Soxhlet extraction method ( P ＜ 0. 001) ． The determined level of crude fat in extruded dog feed by acid hydrolysis method was significantly higher than that by Soxhlet extraction method ( P ＜ 0. 05 ) ． However，the determined levels of crude fat in non-extruded primate feed and non-extruded dog feed had no differences between two the methods( P ＞ 0. 05) ，and The determined levels of crude fat in pelleted feeds and non-pelleted feeds also had no differences between the two methods( P ＞ 0. 05) ． The loss of crude fat determined by acid hydrolysis method was less than that by Soxhlet extraction method during feed processing． Conclusion The acid hydrolysis method is suitable for determining crude fat in extruded feed．

The Exploration of Construction of Data Quality in Laboratory Animal Resources Database

WANG Xiao-Ming, LIU Wan-Ce, CHEN Mei-Li, LI Hui-Ping, HUANG Ren

2013, 30(01):  38-41. 

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Abstract: The data quality is the heart of shared data resourse and the necessary guarantee of the value of data resources． In the construction of laboratory animal resources database，the key point of data quality guarantee is the establishment of data measurement and collection and storage pross standardization． The article discusses the data quality conrol measures on laboratory animal resources database，focusing on the design of data acquisition system and data storage audit process，and also raising some helpful thoughts on improving the data quality during the late period of database running，which will provide some reference for data information construction．

The Current Situation and Countermeasures in Laboratory Animal Industry of Sichuan Province

ZOU Yi-Xing, JIANG Jia, WANG Jing-Dong

2013, 30(01):  42-45. 

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Abstract: Laboratory Animal Science is the basic disciplines of life sciences，also the important support conditions in bio-medicine，pharmacy，life sciences and areas’development． ，On the basis of systematic analysis in industry structure ，production and usage situation，the existing problems in industry development of Sichuan Province，the article put forward corresponding countermeasures and suggestions to promote healthy and sustainable development of the province's laboratory animal industry．

The IACUC’s Built and Operation in Hospital

XU Hu-Feng, LIU Yun-Bo, LI Xue-Yong, ZHANG Zhong-Min, XIE Miao-Rong, YOU Hong, GE Ru-Jun, ZHANG Bing

2013, 30(01):  46-49. 

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Abstract: Laboratory animal welfare and ethics has got more and more attention in China． The paper is based on the feature of laboratory animal in hospital and the five“M”concept( Manager /Men /Market /Mechanism /Money) ，to discuss how to built and operate the Institutional Animal Care and Use Committee in hospital．

Progress in Researches on Application of Tree Shrews to the Relevant Field of Hepatitis C Virus

KUANG De-Xuan, JIANG Qin-Fang, SUN Xiao-Mei, TONG Pin-Fen, GAO Jia-Hong, LU Cai-Xia, DAI Jie-Jie

2013, 30(01):  55-60. 

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Abstract: Persistent infection with hepatitis C virus ( HCV) has become a global public health problem． The lack of proper study models has brought difficulties in the study of the mechanism of viral infection，life cycle and pathogenic mechanism of HCV． Tree shrews are classified into the primate animal and many biological characteristics are close to human． In recent years，Tree shrews has aroused the common interest in the biomedical researchers and the relevant regulators． The domestic and foreign scholars have made many positive attempts in order to determine whether or not tree shrews is used as the model of HCV infection． This article will introduce the newly progress of application of tree shrew in HCV experimental model research．