Laboratory Animal Science

Preparation and Preliminary Application of Monoclonal Antibodies Against JEV

WANG Ji,WEI Li,GONG Wei,HE Zheng-ming,Xing Rui-chang

2006, 23(03): 1-4.

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Objective\ To prepare Monoclonal Antibodies against JEV and apply preliminary the antibodies to the detection.Methods\ BALB/c mice were immunized with purified Japanese Encephalitis Virus(JEV).The immunized spleen cells of these mice were fusing with SP2/0 myeloma cells.Four hybridoma secreting monoclonal antibodies(McAbs) against JEV were prepared after screening by ELISA and cloning three times by limiting dilution.Results\ Four hybridomas were obtained,named A5,Gl0,F4,and C10 respectively,which were able to secrete McAbs stably.The immunoglobulin subclass of G10 and F4 were IgG1 and A5 was IgM,the subclass of C10 was not identified;the ELISA titers of C10 and F4 ascites were above 10~5,while the ELISA and IFA titers of C10 and A5 ascites were above 10~2.Conclusion\ Monoclonal Antibodies against JEV were successfully prepared,the method of IFA and ELISA based on McAb was developed for the detection of JEV in the biological products from the swine for human use.

Establishment and Application of Assay for the Detection of LCMV Nested Polymerase Chain Reaction

XING Jin,WEI Li,GONG Wei,HE Zheng-ming,XING Rui-chang

2006, 23(03): 5-8.

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Objective\ To establish PCR test method that monitor Lymphocytic Choriomeningitis Virus in Laboratory.Methods Design two series primers of Nested PCR according to S segment(GPC and NP genes) of LCMV cDNA.To clone and measure sequence for our LCMV strain though PCR.Use Nested PCR to test 15 mice brains.Results That is 99% homology between the target segment and LCMV cDNA.The test result of mice brains PCR is 10 positive and 5 negative.Conclusion The test result is correct on LCMV infected mice brains,therefore,it indicates the Nested_PCR method can be used for testing LCMV in Laboratory.

Effect of Genetic Contamination of BALB/c Mouse on the Preparation of Monoclonal Antibodies

LIU Shuang-huan,MA Li-ying,WANG Xiu-qing,WANG Hong,HE Zheng-ming,XING Rui-chang

2006, 23(03): 9-11.

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Objective\ To monitor the influence of the genetic contamination of BALB/c Mouse on the preparation of monoclonal antibodies.To strengthen the management and breeding of the inbred strains mouse and improve the detection on the genetic quality.Methods\ The method of biochemical label was used to detect the 13 alleles(Akp-1,etc)of two groups of BALB/c mouse.Results \ After the injection of hybridomas cells,no ascites was produced in these of the BALB/c mouse.Four heterozygosis at the points of Ce-2 etc were found in the first group of mouse,and 8 heterozygosis at the points of Ce-2 etc were found in the second group of mouse.Conclusion\ The results proved that there were shortomings in management and breeding of the laboratory animals,and it is important to enhance the management about laboratory animals and strengthen the detection of genetic quality.

Effects of Cilostazol on Blood Lipoids and Haemorheological Indexes in Rats With Diabetes Induced by Streptozotocin

CHEN Wang~1,LI Hong~1,GU0 Jian~1,CAI Xiao-pin~2

2006, 23(03): 12-14.

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Objective\ To investigate the effects of cilostazol on blood lipoids haemorheological indexes in rats with diabetes induced by streptozotocin.Methods\ 24 eight_week_old male SD rats were randomly divided into normal control group,diabetes model group,and diabetes group treated by cilostazol,and there were 8 rats in each group.Experimental diabetes was induced by a sigle dose(45mg/kg) of injection of streptozotocin via the tail vein.After the onset of diabetes,cilostazol was orally administered into each rat in the treated group at 50mg/kg/day.Haemorheological indexes,plasm toatal cholesterol and triglyceride were determined after being treated by cilostazol for 24 weeks.Results\ As compared with normal control group,plasm triglyceride in diabetes model group was increased by 85%,and the treatment by cilostazol could produce a significant increase in the washed erythrocyte viscosity at low shear(an index of erythrocyte deformability) and a significant decrease in plasm triglyceride.Conclusion\ It was suggested that cilostazol could produce a significant increase in the washed erythrocyte viscosity at low shear and a significant decrease in plasm triglyceride in the diabetic rats.In addition,the inhibitory effect of cilostazol on platelet aggregation may provide a new perspective in the field of prophylaxis for chronic complications of diabetes.

Effects of Shenzhu Capsules on Cancer Cellular Differentiation in the Mouse of Gastric Carcinoma Metastasis with Deficiency of the Spleen

YE Bing~1,PENG Cheng~2

2006, 23(03): 15-18.

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Objective\ To survey effects of Shenzhu capsules on cancer cellular differentiation in the mouse of gastric carcinoma metastasis with deficiency of the spleen,and delve into.Methods\ The animal model of gastric carcinoma metastasis with deficiency of the spleen was developed in the nude mice,and effects of Shenzhu capsules on cancer cellular differentiation in the mouse of gastric carcinoma metastasis with deficiency of the spleen were observed with HE staining.Result\ Shenzhu capsules could exert the regulating effect on cancer cellular differentiation of gastric carcinoma.Conclusion\ Shenzhu capsules could improve the heterotype of the cancer cell and induce cancer cellular redifferentiation,which may be one of the mechanisms governing treatment of gastric carcinoma metastasis by Shenzhu capsules.

A Comparative Experiment on Infections of Exorchis Dongting huensis to Red Crucian Carp and Goldfish

WU Duan-sheng~1,SHI Meng-zhi~2,JIANG Qian-qiu~1,HU Dai-yan~2,TAN Xiang-wen~1

2006, 23(03): 19-21.

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Objective\ To develop the infectious models by Exorchis Dongting huensis. Methods\ Red crucian carp(Carassius auratus red variety) and goldfish(Carassius auratus van) were respectively put into the chlorine_free tap water with the oncomelanias so as to be infected naturally with Exorchis Dongting huensis cercariae,and the anatomic observations were carried out at different times.Results \ All the fishes were infected with Exorchis Dongting huensis cercariae(the infectious rate being 100%) and the cercariae were developed into the mature metacercariae in the fishes.During the experiment,three goldfishes were dead and no red crucian carp died.Conclusion\ The red crucian carp and goldfishes could be the host animals in developing the infectious model with Exorchis Dongting huensis, and the former may be superior to the latter.

The Effect of Rabbit Sex and Body Temperature on Pyrogen Test

XU Wen-yu,REN Jun,DU Ying,HE Zheng-ming,XING Rui-chang

2006, 23(03): 22-25.

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Objective\ To analyze the effects of rabbit sex and body temperature changes on pyrogen test.Methods According to the 《Chinese Requirements for biologics》 edited in 2000.Results The statistic showing the rabbit body temperature are difference when the sex are different and the difference are significant(P<0.05).Conclusion The female has been able to bring the sham result more then the male,the inverse proportion about 6∶1.So,the stabilization of temperature is better of the male then female which the rabbit on pyrogen test.

Determination of Blood Physiological and Biochemical Indexes in Growing Beagle Dogs

SUI Li-hua,WU Na,WANG Dong-ping,LI Gui-jun, WU Xiao-yan,FAN Wei,LI Wen-long,HONG Bao-qing, ZHAN Da-wei,SHANG Shi-chen,CHEN Zhen-wen

2006, 23(03): 26-30.

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Objective\ To obtain the basic data of hematological indexes of closed colony beagle dogs in our center,and compare the difierences in these indexes.Methods The blood physiological and biochemical indexes were determined in 180 female and male beagle dogs with different ages.Results There were significant difierences in part of hematological indexes between female and male with same age;there were no significant differences in most of hematological indexes between 5_month and 6_month old dogs;and when comparing biochemical indexes of 4_moth old dogs with 5_month and 6_month old dogs,orderliness of the data were poor.Conclusion There were differences in a few hematological indexes between male female dogs,and there were no difierences in most of hematological indexes between the dogs with different ages.These showed that the hematological parameters of the closed colony beagle dogs have been in a stable status.

Evaluations of Developing Acute Pulmonary Respiratory Distress Syndrome by Detergent in Piglets

ZHAO Xiao-wei~1,LIU You-ning~1,WANG Xiao-guang~1,ZHANG Jian-peng~2,XIE Jing-hong~2,MA Hong-jie~2

2006, 23(03): 31-33.

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Objective\ To evaluate feasibility of developing acute pulmonary respiratory distress syndrome by detergent in piglets.Methods\ Detergent was injected into tracheae of 16 healthy piglets,and the change of oxygen partial pressure from artery blood,chest CT,pathological change of the lungs,and heart rate were observed.Results\ After injecting detergent into trachea,oxygen partial pressure from artery blood was markedly decreased,oxygenation index was less than 100mmHg,and sudation of inflammatory cells in the trachea and alveolus,hemorrhage and edema of alveolus,and collapse of alveolus were found.Vital indexes were stable.Conclusion\ Injection of detergent into trachea was able to induce acute pulmonary respiratory distress syndrome in the piglet.

Study on the Effect of Dietary Levels of Calcium and Calcium Phosphorous Ratio on Bone Mineralization by Growing Dogs

LIU Yuan,ZHAO Shuang,QIU Zheng-liang,WANG Zhi-hong,AN Xing-lan,FU Qiang,SUN Ai-ping

2006, 23(03): 34-36.

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Objective\ To study the effect of dietary levels of Calcium and calcium phosphorous ratio on bone mineralization by growing laboratory beagal dogs.Methods Feeding test,digestive test,slaughter test.Results 0.6% dietary levels of calcium gain less bone mineralization(P0.05).High dietary calcium level has no influence on phosphorous content and high dietary phosphorous level has no influence on calcium content in bone.But high dietary calcium affect magnesium content in bone.Conclusions 1.2% dietary calcium level and 1.2∶1 dietary calcium phosphorous ratio are optimal for bone mineralization.

Preparation and Standardization of Diagnostic Serum for Detecting Pathogenic Bacteria in Laboratory Animals

GAO Zheng-qin,WANG Chun-ling,XING Jin,HE Zheng-ming,XING Rui-chang

2006, 23(03): 54-56.

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Objective To prepare and standardize diagnostic serum which can be used to detect pathogenic bacteria in laboratory animals.Methods Thirty-two SPF rabbits were immunized with sixteen reference strains through intravenous and subcutaneous injections.All rabbits were immunized for five times with 5.0×10~(-8)～10×10~8/mL suspension made of inactivate reference strains after 18～24h culture in 37℃.The titers of antiserum were tested with bacteria agglutination.Results The antisera passessed a high efficiency of agglutination titers.The antisera had no cross-reaction with other reference strains and with a good specificity.Conclusion The antisera produced in this study would provide the useful diagnostic reagents for detecting pathogenic bacteria in laboratory animals.

Research on Control of Technological Index in the Construction of Laboratory Animal Barrier Facility

XUE Jing-li~1,XING Jin-shan~1,WANG Chun-yao~1,DU Chun-yan~1,JIN Shu-xing~1,SHI Chong-min~1,YIN Zhi-gang~1,LI Hong-tao~1,SHI Guo-chun~2

2006, 23(03): 57-60.

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In the light of the practice in the construction and operation of barrier facility for laboratory animal and on the basis of incorporating the experience of the biological cleaning buildings in other trades(such as medical institution and pharmaceutical factory),several technical measure of ensuring an implement of the national standard(GB/T14925-2001) in construction of barrier facility for laboratory animal were approached from the angle of engineering construction(especially air conditioning and purifying,control of temperature and humidity,air circulation, gradient pressure difference,saving energy,and high efficiency.At last,the matter of construction standardization of laboratory animal facility was proposed.

Growth Curve of Cynomolgus

LI Bing

2006, 23(03): 61-62.

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Objective To describe the growth curve of cynomolgus from ablactation to three-year old.Methods\ The data in our center were collected in the past few years.Result\ It showed the growth rules of cynomolgus in our center before three-year old.Conclusion\ The results are similar to the foreign research.It will afford a useful reference to breeding and animal experiments.

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