Table of Content

31 May 2008, Volume 28 Issue 5

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A Study on Keratonosus Pathogens in B6-C0 Mice

SHAO Yi-xiang1, YIN Xiao-mm2，WANG Chun-zhong1，JIANG Ymg-mei1，GUAN Huai-jin2

2008, 28(5):  277-281. 

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Objective Keratonosus pathogen and nosogenesis of B6-C0 mouse was studied to infer the cause of human keratonosus pathogeny and to control the disease. Methods Get cornea secretion or cornea scraping from the eyes of normal B6 and B6-C0 mouse at different ages, and introduce them to blood plates and nutrient broth respectively, cultivate them for 24 hours at constant temperature. Pick the bacterium colonies and the suspension in the broth to prepare films, stain the films and examine them under microscope, finally identify the purified bacteria. Results There was difference in cornea bacteriology between normal B6 and B6-C0 mouse. Conclusions The result showed that there was a relationship between the cloudiness of cornea and microbe infection.

Nonlinear Equation Construction between Emotion and Micro-index Using Female Macaque

ZHANG Hui-yun, WEI Sheng, QIAO Mmg-qi

2008, 28(5):  282-288. 

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Objective Construct nonlinear equation between emotion and micro-index using female experimental macaque to provide subject supporting for revealing the famous TCM theory of “liver controlling dispersion”. Methods Adopt the project described by Carol A.Shively to select different status macaque enter experiment,use Emotional Evaluation Scale about Female Experiment Macaque to score macaques’ emotion，collect blood and urine to test transmitter and hormone content in vivo, and construct nonlinear equation by using mathematics modeling method. Results The liver-qi invasion macaques had a atypia negative correlation between 5-HT content and indignation emotion,while with the increasing of P content，the score of indignation factor descended rapidly in protophase and slowly in anaphase applying Scale method to investigate the relationship of expression&behaviour and transmitter&hormone level in vivo on macaques model. Conclusion There is a very complicated relationship between emotion and micro-index in vivo, and the emotion fluctuation maybe related to P、 PRL、5-HT content in vivo closely.

Analysis and Clone of Apolipoprotein E Extron 4 in Mongolian Gerbil

LIU Yue-huan, SHI Qiao-juan，GUO Hong-gang, MA Wei-feng

2008, 28(5):  289-298. 

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Objective To clone the apolipoprotein E gene Extron 4 of the Mongolian gerbil. Methods The primers was designed according to the published sequences in GenBank. A RT-PCR method was applied to clone the apolipoprotein E Extron 4 in liver of the Mongolian gerbil. The PCR products was sequenced and blast with related sequences. Results The results of PCR and sequence presented that the sequence of the apolipoprotein E gene Extron 4 in the Mongolian gerbil was a 987 bp sequences. The result of blast showed the identification between Mongolian gerbil and rat or mouse is 73%~74%, between Mongolian gerbil and human is 62%, between Mongolian gerbil and pig or bovine is above 60%, between Mongolian gerbil and no-human primates is 57%. The coding protein is 263 amino acid residuals. Conclusions The RT-PCR products was the sequence of the apolipoprotein E4 gene of Mongolian gerbil.

Study on Sequence Variation of Mitochondrial D-loop Gene and Polymorphism among four populations of Reed Vole (Microtus fords)

Xie Jian-yun1’2，Feng Jie1，Bai Xiong3, Hu Jian-hua1，Gao Cheng1

2008, 28(5):  299-303. 

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Objectives To discuss the genetic polymorphisms of the mitochondrial DNA (mtDNA) D-loop among four populations of Reed voles (Microtus fortis) in different regions of China. Methods A total of 34 specimens representing 4 populations were sequenced for the entire D-loop gene (922bp). All sequences were aligned by Clustal W software, and The neighbor-joining (NJ) phylogenetic tree was formed by the MEGA Version 3.1 software. The polymorphic site, haplotype diversity and nucleotide diversity were analyzed by Dnasp 4.0 software. Result 19 haplotypes were determined among 34 individuals, the haplotype ratio was 55.88%. The haplotype diversity and nucleotide diversity was 0.934士0.024 and 0.04334士0.00285 respectively. In reference to mtDNA D-loop sequences of Microtus kikuchii (GenBank AF348082) as a control, there were 172 genetic mutation sites, accounting for 18.66% of 922 bp sequence.The phylogenetic trees showed that 34 vole individuals clustered into two distinct branches: voles from Heilongjiang (smaller) were one branch, the other three vole populations comprised as the other branch, and in this branch, there were two sub-branches, all vole from Ningxia and some voles from Heilongjiang (lager) buildup one sub-branch, voles from Hunan and the rest voles from Heilongjiang (lager) belonged to the other sub-branch. Conclusion These findings suggested that the mtDNA D-loop in Reed voles from China existed informative variations within the four populations, there were some genetic diverse among intra- and inter-populations of Reed voles, but voles from Heilongjiang (smaller) showed more diversity from the other three populations.

The Comparative Studies on in vitro Fertilization and Development of Embryos in Mice

JIA Qmg1’2，GAO Juan1’2，YANG Bo2’3，LIU Li-jun2, YU Li-li2，XU Pmg2, RUI Rong1

2008, 28(5):  304-308. 

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Objective To compare the fertilization in vitro in various mouse strains and the effects of different culture medium for development of embryos in C57BL/6J mice in vitro. Methods Females from 7 strains of mice were superovulated and operated by IVF, and the average number of superovulation and the fertility rate were observed. HTF、CZB、KSOM、Ml6、 HECM were used as the culture medium to culture the2-cell of C57BL/6J mice in the same condition and the development rate from 4-cell to blastcyst were observed respectively. Results there is a significant difference (P＜0.01) in the average number of superovulation markedly among 13.0—38.7, and the rate of fertility of individual mouse with a range of 70.2% to92.8% (P＜0.05). The development rate of 4-cell, 8-cell, morula and blastcyst in the 5 culture mediums with a range of 60.3%?95.8%、52.8?91.1%、40.6—87.5% and 5.1%?75.4% . Conclusion the genetic background is a primary factor for the effect of superovulation and fertilization in vitro in mice. Glucose can result in 2-cell interdiction of the embryo development; it is not primary energy substance before 8-cell. Phosphate can result in retardarce of the embryo development. Amino and EDTA are beneficial for overcoming the block of the embryo development.

Comparison on Physilogical and Blood Biochemical Index of German Shepherd Dog before and after Exercise

MIAO Qin，ZHANG Hm-dong，WEN Hai, GU Jin-qiao

2008, 28(5):  309-312. 

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Objective To investigate the physilogical and biochemical response of German Shepherd Dogs to exercise. Method Five German Shepherd Dogs were choosed randomly to run a distance of 1200 meters in 3 minutes. Arterial blood samples were collected before and immediately after exercise, and at 30 minutes of recuperation. Meantime，rectal temperature, heart rate and respiration rate were tested. Result The mean rectal temperature,heart rate and respiration rate increased after exercise and recovered to the pre-exercise values by 30 minutes after exercise measurement. Serum biochemical changes reflected some fluid movement from the vascular compartment, there being significant increases of lactate dehydrogenase and blood urea nitrogen. And serum creatine kinase increased slingtly. Other changes included significant decreases of serum glucose, serum creatinine and calcium, serum. 30 minutes after exercise, serum glucose, lactate dehydrogenase,blood urea nitrogen.,creatinine and calcium returned towards pre-exercise values. Conclusion The rule of the physilogical and biochemical response of German Shepherd Dogs to exercise provides a theoratical basis for the training.

Study on Irritable Bowel Syndrome Model of Spleen Deficiency Type in WHBE Rabbits

PAN Yong-ming, ZHANG Li-zhong, CHEN Mm-li, XU Jian-qin,CHEN Liang, PEN Dmg-guo，WANG De-jun, YING Hua-zhong

2008, 28(5):  313-317. 

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Objective To establish irritable bowel syndrome animal model of spleen deficiency type in WHBE rabbits for providing a certain tool for medicine development and basic research on irritable bowel syndrome of deficiency type. Methods 18 WHBE rabbits and 18 Japanese white rabbits were randomly divided into two groups respectively, with 6 rabbits in normal control groups and 12 rabbits in model groups. Using the methods of moist heat，stress and stimulation adding taking senna decoction in model groups of rabbits, the rabbits model of irritable bowel syndrome of spleen deficiency type was made, and 6 rabbits in each model group recovered naturally for 10 days after the successful establishment of model. To observe the change of clinical syndrome, weight, abdominal circumference indexes, water content of tools，thymus indexes，spleen indexes，visceral sensitivity and small intestine movement rate in rabbits. Results Compared with normal control group, weight of the model rabbits, thymus indexes and spleen indexes were significantly decreased (P＜0.05，P＜0.01) in model group, abdominal circumference indexes, water content of tools, visceral sensitivity and small intestine movement rate were significantly increased (P＜0.05，P＜0.01) in model group, and its recovered slowly. Conclusion The irritable bowel syndrome of spleen deficiency type rabbit model accords with the clinical characteristics of IBS, and so WHBE rabbits are ideal experiment animal more suitable for the study of irritable bowel syndrome of spleen deficiency type.

Construction of Eukaryotic Expression Vector of pSecTag-EGFP and Research of Chicken Embryonic Fibroblast Cells

ZHU Jmg1，DOU Ru-hai1，JIN Dong-qmg1, YANG Na-na2

2008, 28(5):  318-320. 

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Objective To study the transfection efficiency of pSecTag in chicken embryonic fibroblast cells, obtained the optimum condition of transfection, and established the foundation for the target gene expression in embryonic fibroblast cells of chicken. Method EGFP gene was cloned by PCR and constructed secretive eukaryotic expression vector by digestion, ligation and transformation, the relative content of the green fluorescent protein were detected by experimental design optimization. Results The optimum condition of transfection was with 2μl lipofectamine and 1μg expression vector in medium. Conclusion This method can effectively transfect the chicken embryo fibroblast.

Effect of Allantoxanic Acid on Serum Uric Acid Contents in Rats

ZHANG Liang, SHI Le, YUAN Dong-pmg, XU Li

2008, 28(5):  321-323. 

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Objective To observe the fluctuation of uric acid in rat serum with hypoxanthine, uricase inhibitor, and to establish a new method to replicate rat hyperuricemia model. Method Adopted the combination of hypoxanthine and uricase inhibitor to establish rat hyperuricemia model, and compared the uric acid contents in serum when using hypoxanthine singly with that of using the combination of hypoxanthine and allantoxanic acid. Simultaneously those factors of research methods have been evaluated, such as those proposed model-leading drug’s dose-effect and time-effect relationship. Results The combination of hypoxanthine and uricase inhibitor could increase the content of uric acid in rat serum by means of appropriate dosage and method of administration (1000 )amol/L). It could create the rat hyperuricemia model successfully. Conclusion The combination of hypoxanthine and allantoxanic acid could increase uric acid contents in rats rather than using hypoxanthine singly. Meanwhile, by repeatedly dosing regimen，uric acid contents in serum of rats could increase during rather longer time (P<0.01). So it has made some progress in replicating steady hyperuricemia model at present experiment.