Table of Content

31 May 2009, Volume 29 Issue 5

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Whole Brain Slices Culture and Anoxia/hypoglycaemia Model Building

YANG Li-jun1，CUI Hong1，YANG Ai-jun1，HUANG De-zhuang2，CUI Yan1，WEI Tian-li1

2009, 29(5):  283-285. 

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Objective To investigate how to build up a single and convenient anoxia/hypoglycaemia model using neonatal rat brain slice culture. Methods On base of brain slice culture, nitrogen injection was used to acquire the anoxia/hypoglycaemia model. After frozen section on the brain slices, HE stain was introduced to detect the effect of anoxia/hypoglycaemia. Results Whole brain slices grew well. Karyopyknosis and nuclear fragmentation were observed in anoxia/hypoglycaemia group compared to control. Conclusions The anoxia/hypoglycaemia model was established by using brain slices in vitro.

Establishment of Hypoxia-ischemia Brain Damage Model and Pathologic Change of Brain in Rats

SHI Zhi-yong1，NIU Ting-xian1，Luo Jian-jun2，WAN Zhong-jun1

2009, 29(5):  286-288. 

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Objective To establish hypoxia-ischemia(HI) brain damage(HIBD) model in rats and investigate the pathologic change in brain tissue after establishing model at the different time point. It would provide the evidence of diagnosis, therapy and repair in clinic. Methods Sixty Wistar rats were randomly divided into control and experimental group. Then the experimental group was also divided into 5 sub-groups as the different time point (4 h，8 h,12 h, 24 h and 72 h ) after the model established. The HIBD model was established by occluding both common carotid arteries and enforcing hypoxia. The pathologic change of brain tissue was observed. Results The obvious pathologic change of brain tissue in experimental group was observed, but it had diversity in the different sub-groups. It was not evident difference among 4 h，8 h and 72 h sub-groups. But the pathologic change in 12 h and 24 h sub-groups became more serious than that in the other sub-groups. Conclusions HIBD model was established successfully in rats. The pathologic change in brain tissue was diversity after building model at the different time point.

Polymorphism Analysis of PPAR-γ Gene Exon 1 in Three Strains of Chinese Native Minipig

ZHAO Yu-qiong1，YAO Chun-tao1，ZHAO Quan2，CHEN Hua3

2009, 29(5):  289-292. 

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Objective To analysis single nucleotide polymorphism (SNPs) of peroxisome proliferative activated receptor-T gene exon 1 in three strains of Chinese minipigs, Bama minipigs, Wuzhishan minipigs, Nongda minipigs, to provide basic data for their uses on research of diabetes mellitus and metabolic disease. Methods Genomic DNA was extracted from the blood samples of the minipigs, and served as templates for target gene segment and amplified and selected by polymerase chain reaction (PCR). The PCR products were sequenced and analyzed for documenting single- nucleotide polymorphisms (SNPs). Results One SNP was found in the target region, and the variation of ATG to GTG cause a encoding change of Met59Val. The sequencing results showed that the variations were found in 4 of 6 Bama minipig with 2 homozygous mutations and 2 heterozygous mutations, in 6 of 6 Wuzhishan minipig with homozygous mutations, in 4 of 10 Nongda minipig with heterozygous mutations. Conclusion There are variations exist among the strains and individuals in native minipigs on the SNP in PPAR-Y gene exon 1.

HBsAg Immune Response on Mouse Models Enhanced by Recombinant Murine Interluekin-12

XIA Shu-qi，WANG Zeng-song，WANG Cui-ling，JIE Yan，XIE Fei-qun，ZENG Zheng-fei，DUAN Xiang，ZHANG Yi-jun

2009, 29(5):  293-297. 

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Objective To investigate the enhanced immunity of hepatitis B surface antigen (HBsAg) vaccine in response to interleukin-12 (IL-12) stimulation. Method C57BL/6J mice were injected with rmIL-12 plus HBsAg combination，after which the serum IFN-T level as well as the response of murine spleen lymphocytes to HBsAg was detected. Results Serum IFN-T level in high dose combination of HBsAg and rmIL-12 group after two injections was significant higher than that in HBsAg or rmIL-12 alone (尸<0.05). The ability of murine spleen lymphocytes producing IFN-T in response to HBsAg was significantly improved (P<0.05), and IFN-T mainly originated from CD4+ & CD8+ T cells detected by flow cytometric analysis. Conclusion The immunity response of C57BL/6J mice to HBsAg can be rapidly and specifically enhanced by rmIL-12, which may be a reference of IL-12 plus HBsAg clinical application on CHB infection.

Observation of Anesthetic Effect of Katamine and Sodium Pentobarbital on Tree Shrew

SUN Xiao-mei，GAO Jia-hong，YE You-song，HAN Yuan-yuan，DAI Jie-jie

2009, 29(5):  307-309. 

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Objective To observe and compare the anesthetic effect, side effect of ketamine and sodium pentobarbital on tree shrew for establishing the safe, reliable anesthesia methods. Methods One hundred and ten adult tree shrews were divided into two groups: 60 were anesthetized with i.p of ketamine (40 mg/kg), another 50 with i.p of Sodium pentobarbital (50 mg/kg). Results In ketamine group，the induction period of anesthesia was 2.78±1.28 min, maintain duration of anesthesia was 10.12士2.13 min, and the animal with heavy salivation. The induction period of anesthesia by sodium pentobarbital was 4.84±2.15 min, maintain duration was 51.95±12.94min，all animals keep a stable state but body temperature dropped notable in this group. Conclusion Ketamine and sodium pentobarbital are safe, suitable for tree shrew in these dose. The induction period and maintain of ketamine anesthetic were shorter than those of sodium pentobarbital. More stable anesthesia was obtained by using sodium pentobarbital.

Analysis on Blood Gas and Electrolytes of WHBE Rabbits and Compared with Japanese White Rabbits and New Zealand White Rabbits

CHEN Min-li，L（U） Jian-min，XU Xiao-ping，PAN Yong-ming，XU Jian-qin，WANG De-jun

2009, 29(5):  331-333. 

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Objective To investigate the difference of blood gas and electrolytes among WHBE rabbits, Japanese white rabbits, New Zealand white rabbits. Methods WHBE rabbits, Japanese White rabbits and New Zealand White rabbits were selected and the blood sample were taken from center artery in ear, the blood gas and electrolytes indexes were detected by NOVA blood gas and electrolytes analyzers. Results Compared with Japanese White rabbits，pH, PCO₂，PO₂，SO₂，BE-ECF, BE-B, SBC, HCO3, TCO₂，a/A and PO₂/FI indexes were significantly lower in WHBE rabbits than those of Japanese White rabbits(P＜0.05，P＜0.01)，while Osm, Na+ and Mg⁺⁺ indexes were higher in WHBE rabbits than those of Japanese white rabbits(P＜0.01). Compared with New Zealand White rabbits, pH index was significantly lower in WHBE rabbits than that of New Zealand White rabbits(P＜0.01)，while Mg++ index was higher in WHBE rabbits than that of New Zealand White rabbits(P＜0.01). Conclusion The results above could provide a reliable data for the standardization of WHBE rabbits in China, meanwhile, it could provide an experimental basis for the research application of WHBE in respiration system and cardiovascular disease.

Genetic Analysis of Outbred and Inbred F5 of WHBE Rabbit by Microsatellite Loci

ZHU Liang，YING Hua-zhong，CHEN Min-li，CAI Yue-qin，YU Jia，XIAO Hui，TAO Tao，PAN Yong-ming

2009, 29(5):  334-340. 

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Objective To assess and verify the identity of outbred and mbred F₅ of WHBE rabbit on molecular level, polymorphism of microsatellite loci of WHBE rabbit genome were analysed. Methods Twenty-one microsatellite loci of rabbit were used to analyze and compare the genetic polymorphism in two rabbit population. Results Eleven microsatellite loci which were successfully amplified with high polymorphism were selected in 21 microsatellite loci. For the outbred WHBE rabbit, the number of alleles ranged from 3 to 8; the average number of effective alleles was 3.0344 and the average observed heterozygosity was 0.4956. The average value of polymorphism information content or the power of cumulative discrimination for the 11 polymorphic loci was 0.6130 and 1.0，respectively. The average exclusion probability for the 11 polymorphic loci in the absence or in the presence of genetic information on the first parent was 0.9683 and 0.9980，respectively. For the mbred F₅ generation of WHBE rabbit, the number of alleles ranged from 3 to 9; the average number of effective alleles was 1.8132 and the average observed heterozygosity was 0.3808. The average value of polymorphism information content or the power of cumulative discrimination for the 11 polymorphic loci was 0.5241 and 1.0，respectively. The average exclusion probability for the 11 polymorphic loci in the absence or in the presence of genetic information on the first parent was 0.9264 and 0.9933，respectively. Eight alleles in 7 micro satellite loci were specific in both outbred and F₅ of WHBE rabbit. The average number of effective alleles and the average observed heterozygosity of F₅ were lower than those of outbred. It showed that the purity of gene in F₅ was higher than that in outbred of WHBE rabbit. Conclusion 11 microsatellite loci we selected were suitable for monitoring cultivation of inbred strain of WHBE rabbit.

The Optimization of RAPD Reaction System and Primers Screening in Experimental Rabbit

YU Jia，CAI Yue-qin，XIAO Hui，CHEN Min-li

2009, 29(5):  341-346. 

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Objective The optimal reaction system and primers of RAPD-PCR in experimental Rabbits were established. Methods One hundred and fifty samples were collected from WHBE rabbits, JW rabbits and NZW rabbits，and their DNA was extracted by standard procedure of phenol/chloroform. The optimization of its RAPD-PCR reaction system, the most appropriate concentration of Mg²⁺, dNTP, Taq polymerase, random primers and DNA template, was studied. Sixty primers were used to amplify DNA of experimental rabbits with RAPD-PCR methods. Results The optimal PCR system for RAPD analysis was carried out in 20 ul reaction volumes containing the following: 10×loading buffer 2ul, Mg2+ 1.5 mmol/L, dNTP 0.25 mmol/L, Taq polymerase 1.25 Units, random primer 5 jumol/L, DNA template 40 ng and sterile distilled water to total volume. On the basis of preliminary experiments, twenty-five primers with high polymorphism were selected in our study. Conclusion The selected 25 primers and optimalizing PCR reaction system can applied to RAPD reaction in experimental rabbit.

Progress on Transgenic Animal Techonology

GU Rui-huan，CHEN Xue-jin，GUO yi

2009, 29(5):  347-351. 

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Transgenic animal technology should be traced back to the late 1970s .Alone with the rapid development of biological science, transgenic animal technology has made a great progress. Transgenic animals plays an important role in biomedical, agriculture and so on. The paper reviews transgenic animal technology on its development, methods, application and problems.

The in vivo Research Methods of Endocrine Disruptors' Biological Effects in Rats

DU Yuan-sheng，JIANG Xue-wu

2009, 29(5):  352-352. 

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Due to the complexity of in vivo study for endocrine disruptors (EDs)，biological effects, the related research results are usually incongruity. In the recent years, a detection system with more authority is being gradually set up. Rat model was especially emphasized in this system. This article reviewed the significance of rat model in the study of EDs’ biological effects, and also, the detail methods, mechanism, and evaluation of rat model in these studies.