Table of Content

25 September 2005, Volume 25 Issue 3

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Establishment of a Hygromycin B Resistant NIH3T3 Cell Line as Feeder Layer

ZHANG Mei-Ying, LI Hua, DONG Wan-Wei, YANG Wei, QIN Ying, WANG Ying, ZHOU Sheng-Lai, YU Yang, WANG Wei, WANG Tai-Yi, WANG Lu-Zeng

2005, 25(3):  131-135. 

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Objective In order to obtain a feeder layer of ES cells transfected by pTRE-Ins-human gene by means of the hygromycin-resistant NIH3T3 cell line established.Methods The plasmid pHyg containing hygromycin phosphofransferose gene was purified and transfected into NIH3T3 cells with Lipofectin.The transfected cells would be survived in further culture medium containing hygromycin B antibiotic as the hygromycin phosphofransferose gene could express a hygromycin B resistant products.The identification of the hygromycin B resistant NIH3T3 cell line was conducted by polymerase chain reaction with the specific primers of hygromycin phosphofransferose gene. Results Under the condition of hygromycin B（300μg/ml ），hygromycin B resistant NIH3T3 cell clones have been successfully selected.The hygromycin B resistant NIHNIH3T3 cells were no differences from normal NIH3T3 cells in morphology and propagation rate.The hyg phosphofransferose gene DNA fragment could be amplified by PCR with specific primers in the genomic DNA of hygromycin B resistant NIH3T3 cells；Southern blot analysis showed that hygromycin phosphofransferose fragment was integrated into NIHNIH3T3 cells. Conclusion The hygromycin B resistant NIH3T3 cells were established successfully by Lipfectin,which may be useful of selecting ES cells transfected by pTRE-Ins-human gene.

Antagonistic Effect of Selenium on the Change of Total Anti-oxidative Capacity of Liver and Gill Tissues in Swordtail Fish Initiated by Mercury

FANG Zhan-Qiang, WANG Chun-Feng

2005, 25(3):  136-139. 

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Objective The effects of mercury （Hg） and selenium （Se） on the total anti-oxidative capacity （T-AOC） of liver and gill tissues and the antagonistic effect of Se on the change of T-AOC of liver and gill in swordtail （Xiphophorus helleri Heckel） initiated by Hg were investigated in order to test if the liver or gill T-AOC change in fish can be considered as a valid biomarker for environmental risk assessment （ERA）. Methods Swordtails exposed to Hg with 0,0.21 and 0.42 mg/L, Hg plus Se with （0.21+0.0664） and （0.42+0.0664） mg/L, Se with 0.0664 mg/L using the addictive index method, the changes of T-AOC of liver and gill tissues in 5 days were determined quantitativelly. Results The T-AOC of liver and gill tissues were decreased in different level when swordtails were exposed to Hg during the experimental period；high Hg concentrations could remarkably （P＜0.01） decrease the T-AOC of gill and liver tissues to 39% and 30% of the control group at fifth day, however, the T-AOC of gill and liver tissues were significantly increased （P＜0.05） when swordtails were exposed to Se first day after exposure. Compared to the single Hg treated, the mixed exposure （low Hg concentration plus Se） could restrain the fall of the T-AOC in gill tissues all through the exposure period； however, the T-AOC in liver tissues was significantly decreased （P＜0.05） at fifth day. High Hg concentration plus Se the mixed exposure could only restrain the fall of T-AOC in gill and liver tissues at first day, and then the T-AOC of gill and liver tissues were significantly decreased （P＜0.05 or P＜0.01） as the time increased. Conclusion The Se could prevent the decrease of T-AOC in liver and gill tissues from the poisoning of Hg in fish at a certain extent.In addition, the T-AOC of gill and liver tissues in swordtails c6uld be considered a valid biomarker for environmental risk assessment （ERA）.

Establishment of Allergic Asthma Model in Mice and Expression of OX40 in the Lung

HUANG Li-1, HU Yu-Min-2, CHEN Xu-Yan-1, SHI Qin-2, JI Wei-1

2005, 25(3):  140-142. 

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Objective To establish allergic asthma model in BALB/c mice for investigating the expression of 0X40 in asthma. Methods Mice were sensitized by i.p. injection of OVA ,two weeks late mice were challenged with aerosolized OVA,saline was administered to the mice of the control group in a similar manner. Total number of bronchoalveolar lavage cells and the percents of eosinophils、lympho-cytes were counted； Part of the lung tissues were stained by HE and PAS to show if the murine model can reflect the pathological changes of asthma, and the another part of lung tisuuses were stained by immunohistology to evaluate the expression of OX40. Results Total number of cells and the percents of eosinophils lymphocytes in the BALF were increased in the model group than that of the control group，the difference was statistically significant；the expression of OX40 in model group were more manifest than that of the control group. Conclusion The B ALB/c mice were ideal animals to establish the model of allergic asthma, the expression of OX40 was increased in the lung of mice in asthma.

L1 Gene of Local Strain HPV16 Induces Immuno-humoral Response in Mice

GAO Hui-1, 2 , SUN Qiang-2, HUANG Zheng-Fang-2, WU Bao-Jin-1, 2 , LI Hou-Da-2

2005, 25(3):  143-146. 

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Objective To test the expression of recombinant plasmid pcDNA-L1 and evaluate its efficiency to induce specific humoral immune responses. Methods The local HPV16-L1 gene was generated by polymerase chain reaction, inserted into pGEM-T Easy cloning vector, and then subcloned into the eukaryotic expression vector pcDNA3.1（-）. The expression of recombinant plasmid pcDNA-L1，transferred in Cos-7 cells, was identified by immunofluorescence. The 6-week-old BALB/c mice were immunized via muscular 3 times with pcDNA-L1, thereafter the sera antibody of were measured by immunofluorescence and western-blot. Results With the immunofluorescence technique, L1 protein was found to express transiently in the transfected cells of the pcDNA-L1. Specific anti-HPV16-L1 antibodies could be detected by IFA at the end of 30,60 and 180 days after the first immunization, No specific antibodies could be detected in the control group of pcDNA3.1（-）. Conclusions The recombinant plasmid pcDNA-Ll DNA constructed by us could induce specific immuno-humoral response in mice.

Model of Diffuse Pulmonary Hemorrhage in Immature Rats

WU Chun-Ying, LU Jian-Sheng, SHU Tao, HUANG Jing-Ning, ZHOU Quan

2005, 25(3):  147-150. 

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Objective Imitating clinical etiology of congestion of pulmonary vessels, the model of diffuse pulmonary hemorrhage was established in immature rats for research on pulmonary hemorrhage in the newborn. Methods Twenty four rats were divided randomly into experimental group and control group （n=12 each ）.The experimental group received 0.1% epinephrine by subcutaneous injection b.i.d. The dosage of epinephrihe was 0.2 mg/kg /dose at first day，0.3 mg/kg/dose at the second day，and was increased gradually to 0.8 mg/kg/dose at the seventh day，The control group received normal saline at the same volume. After rats died or were killed，the lungs were removed and the degree of pulmonary hemorrhage was divided into five grades according to macroscopy：Grade Ⅰ （normal lung），Grade Ⅱ （pulmonary edema ），Grade Ⅲ （ spotty pulmonary hemorrhage ），Grade Ⅳ （local pulmonary hemorrhage ）and Grade Ⅴ （diffuse pulmonary hemorrhage ）. The lung samples were also observed by optical and electron microscope. Results The average days injected epinephrine are 5.25±1.82 day.Macroscopic examination showed Grade V in 10 rats，Grade IV in 1 rat and Grade Ⅲ in 1 rat in the experimental group , and all the control group was Grade I . By the RIDIT analysis，a significant （X²=18.00,P＜0.01）. Optical microscopy exam showed ex-tensive accumulation of red blood cell in the alveolar spaces and serious dilatation and congestion in the small vessels and capillaries for Grade V. Electron microscopy exam showed local disintegration of epithelial layer, thinning and discontinuity of blood-gas barrier for Grade V. Conclusions The model of diffuse pulmonary hemorrhage in immature rats can be established with epinephrine by subcutaneous injection from serious congestion of pulmonary vessels .

Disorders of Glucose and Lipid Metabolism and Pathological Changes of Pancreas, Liver and Kidney in New Zealand White Rabbits Induced by High Sucrose,High Fat and High Cholesterol Diet

LIU Yi-1, ZHANG Chi-1, XIAO Jun-Xia-2, XI Shou-Min-1, YIN Wei-Dong-1, WANG Zong-Bao-1

2005, 25(3):  151-156. 

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Objective To investigate the levels of blood glucose, triglyceride and total cholesterol, and pathological changes of pancreas,liver and kidney in New Zealand white rabbits fed with high sucrose, high fat and high cholesterol diet. Methods Male New Zealand White rabbits were divided into two groups： control group （n= 10,with normal fodder） and high sucrose, high fat and high cholesterol diet （HD） group （n=10, with 37% sucrose, 10% lard,2% cholesterol and 51% normal fodder）, and observed for 16 weeks. Once 4 weeks after an overnight fasting, the blood samples of the rabbits were tested for the levels of serum glucose, triglyceride and total cholesterol. At the end of 16 weeks all the rabbits were sacrificed,and a part of pancreas, liver and kidney tissue were sampled for histopathological examination. Results The rabbits in the HD group emerged hyperglycemia, hypercholesterolemia and hypertriglyceridemia subsequently, and no obvious changes were observed in control group. The pancreatic islets from the HD group became atrophied, with loss in number of the islets and cells within them. The nucleus of islet cells were small and deep-dyed. There was an abundance of large vacuolations in cytoplasm. Hepatic steatosis and fibrosis, as well as glomerular hypertrophy and glomerulosclerosis, could be observed in the rabbits of HD group.It is suggested that high sugar, high fat and high cholesterol diet can increase the blood glucose, triglyceride and total cholesterol, and induce pathological changes of pancreas, liver and kidney in New Zealand White rabbits.

The Original Study on the Complex Rat Model with Chronic Obstructive Pulmonary Disease and Kidney Weak Syndrome

ZHANG Wei, BI Xiao-Li

2005, 25(3):  157-161. 

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Objective To set up the method of the complex rat model with chronic obstructive pulmonary disease and kidney weak syndrome. Method Smoking with SO₂gas and feeding with adenine to establish the complex rat model. Histomorphyological changes were observed. The micrometer of computerized image analysis system was used for measure the diameter of the bronchioles. Testosterone and estradiol were detected. One-way AVON A was used for checkout. Result The lung tissues of the rats in model group showed the typical change of chronic obstructive pulmonary disease （COPD）. Compared to the normal group, they also showed that the outer diameter of the airway was increased, and the inner and the odds of the inner and outer were decreased, which was the basic pathologic Changes of COPD. Meanwhile, content of the testosterone and estradiol were all decreased, which was the experiment character of kidney weak syndrome. Conclusion The rat rnodel with chronic obstructive pulmonary disease and kidney weak syndrome could be established by smoking with SO₂gas and feeding with adenine.

Analysis on Sexual Dimorphism of the Scapula and Humerus Variables of Macaca mulatta in Taihang mountains

ZHAO Xiao-Jin, LEI Meng-Yun, ZHAO Yu-Ting, HU Chun-Hong, WU Su-Hui

2005, 25(3):  162-164. 

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Objective To discuss the sexual dimorphism of the related variables of scapulas and humeri. Method Fifteen variables of scapulas and humeri of the twenty-six adult Macaca mulatta （female 16， male 10） in Taihang Mountains were measured and carried out SPSS 11.0 One-Way ANOVA, Bivariate correlations and allometry analysis. Result （1） there are nine variables which had statistically significance in sexual dimorphism； （2） the results of bivariate correlations and allometry analysis showed that the variables of sexual difference had different correlation with weight； It was found that the variables that were non-significantly or negatively correlated with weight corresponded to negative allometry（b＜l.00）,and that were significantly or highly significantly correlated with weight corresponded to positive allometry（b>l.00）. Conclusion The sex dimorphism of the adult Macaca mulatta in Taihang Mountains are related to the times of sexual maturation, growing rates, living manners and movement behaviors.

Establishment of SPF Golden Hamsters Breeding Colony by Cesarean

XIA Fang, ZHANG Hua-Qiong, HUANG Lin, ZHANG Shui-Tian, LUO Su-Lan, HE Fan

2005, 25(3):  165-167. 

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Objective To establish a breeding colony of SPF golden hamsters for improving quality of Japanese Encephalitis vaccine. Methods The golden hamsters were cleansed by cesarean operation and the baby golden hamsters were fostered in isolators by SPF golden hamsters from the Harlan in Germany. Results In the end of last year, there were more than 200 pairs of SPF golden hamsters in the isolators. The examination of the pathogens showed, the colony was upgraded to the standard of SPF golden hamsters. Conclusion The SPF golden hamsters breeding colony is established successfully by cesarean operation and have been mass-produced in the barrier.

Establishing Great Cardiac Vein Occlusion Model in Pig

LIU Fan-1, CUI Wei-1, HAO Yu-Ming-1, DU Guo-Ying-1, ZHANG Tao-1, ZHANG Qing-1, MIAO Hua-Wei-1, LI Bao-Hua-1, WU Jin-Feng-1, XIE Rui-Qin-1, LIU Kun-Shen-2, DU Jun-1

2005, 25(3):  168-169. 

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Objective To establish the model of the Great cardiac vein （GCV） occlusion by COOK DMBOLIZATION coils in pigs. Methods By using percutaneous transluminal technology, Coils were delivered in GCV transcatheter in 20 pigs. Results The coils were deliveried into GCV at all of animals. Operation successful rate reached to 100%. One pig died 40 minute after the operation. The GCV occlusion completely in 18 of 19 pigs by fluoroscope 1 hour after. One week later, all pigs，GCV were occlusion totally. Conclusion Using percutaneous transluminal technc5logy, COOK EMBOLIZATION coils were deliveried in GCV can established an animal model of the GCV occlusion.

Recent Studying Situation of Several Experimental Fishes

LONG Hua-1, 2 , WANG Deng-Qiang-2, CHEN Jian-Wu-2, ZHANG Yan-2, LIU Jiang-Dong-1, ZHAO Gang-1, CHEN Ji-1, LIU Wei-1, YU Qi-Xing-1

2005, 25(3):  175-181. 

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The recent studying situation on hematology, genetics, environment science, developmental biology and molecular biology of Danio rerio, Oryzias latipes, Gobiocypris rarus Ye et Fu, Fugu Obscurus Abe, Carassius auratus and Xiphophorus helleri Heckel was summarized on the point of view of experimental animals. The biologiccal characteristics of Liobagrus marginatus Gunther, which has the developmental perspective as a experimental model fish, was evaluated.

Microsatellite Technology and their Applicationin

CHEN Xi-Wen, GUAN Min-Qiang

2005, 25(3):  182-185. 

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Microsatellite marker has many advantages such as a great quantity, extensive dispersion around the genome, high polyomorphism, Mendelian codominant inheritance, rapid and convenient detection. In this paper，it were briefly introduced principle and structure, the application of microsatellite technique was reviewed in the study of animal, for instance, construction of genetic linkage map, molecular marker assisted breeding, evaluation of genetic diversity, genetic monitoring and so on were summarized.