Table of Content

29 June 2014, Volume 31 Issue 03

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Preliminary Study on the Interaction between S1 Gene of a New Type Reovirus and Its Cell Receptor

2014, 31(03):  1-4. 

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Abstract:Objective To study the key fragment which was importent for the interaction between S1 gene of a new type reovirus and its cell receptor, and provide the basis for receptor screening. Method Three fragments,which coding full-length S1 and N and C terminal of S1 gene were cloned to pIRSE2-EGFP vector to get recombinant plasmids pGS, pGSN and pGSC. L929 and Hela cells were transfected with three recombinant plasmids, respectively, and the fragment which was involved for interaction of S1 gene and receptor was indicated by analysis the expression of reporter gene EGFP. Result When transfecting with equal amount of three recombinant plasmids, pGSN-treatment group showed the highest level of EGFP expression and pGSC-treatment group produced the lowest EGFP expression. Moreover, when co-transfecting with pGS and pGSN at different ratio, the higher ratio of pGSN the admixture comtained, the higher level of EGFP expression could generate. Conclusion N terminal of S1 gene (62-406 bp) was essential for interaction between S1 gene and its cell receptor.

Establishment of the Genetic Monitoring System in the Mouse and Rat by Using Selected Microsatellite Markers

2014, 31(03):  5-9. 

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Abstract: Objectives To discriminate frequently-used 5 inbred mouse strains and 3 inbred rat strains respectively for genetic monitoring with easiness and credibility, we screened the combinations of the microsatellite loci with polymorphisms in those mouse and rat strains. Method According to the detected strains, by querying the database as well as reported literatures with definite records, the microsatellite data base composed of microsatellite loci panel ( 37 loci for mouse, 24 loci for rat) were compared and screened. By using PCR amplification and 5% agarose gel electrophoresis, microsatellite loci combination could be identified and optimized for distinguishing one strain from another. Result For mouse, 11 loci was optimized to be used for identification of 5 inbred strains, and a microsatellite panel containing 6 loci was selected for genetic monitor those strains. For rat, 6 microsatellite loci were optimized and were suitable for identification of 3 inbred strains. Conclusion The microsatellite monitoring system were successfully established for identification of 5mouse inbred strain and 3 rat inbred strains.

The Observation of the Success Rate in the Model of Diabetic Retinopathy on db /db Mice

2014, 31(03):  10-14. 

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Abstract: Objective To observe the model of diabetic retinopathy(DR) on db /db mice． Method During the seven months，the mice were weighted and the caudal vein was taken to test fast blood glucose once a month． Seven months later，F-ERG was examined on ten db /db mice and six db /m mice ( normal control group). Result The body weight of db /db mice rised obviously，while the fast blood glucose rised unremarkably． 97% db /db mice were under the standard of the fast blood glucose of the model of DR mice( 11. 1 mmol /l). The statistical result of F-ERG showed that the latencies of a-wave and b-wave were delayed significantly in the experimental group( P <0. 01), and the amplitude of b-wave was significant lower in the experimental group( P < 0. 01), compared with normal control group. There were significant difference( P < 0. 01) in the total amplitude of Ops and O2 between normal control group and experimental group． Conclusion Although most of their fast blood glucose are under the standard in this experiment，the retina of db /db mice have changed as the early lesion of DR． So the standard of the model should consider the change of the retina，instead of just depend on fast blood glucose． The standard of model by db /db mice should be modified，through more researches are needed．

The Effect of Caffeine on Mice Behavior in Chronic Unpredictable Mild Stress Model

2014, 31(03):  15-20. 

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Abstract: Objective To investigate the effect of caffeine on behavior of mice，which suffered from chronic unpredictable mild stress ( CUMS) ． Method 48 C57BL /6J mice were divided into NC + NS group，NC + caffeine 30 mg / kg group，CUMS + NS group，CUMS + caffeine group ( 5，15，30 mg / kg) six groups． By the CUMS animal model，six groups were given physiological saline as a control or the intervention of different doses of caffeine for continuous 22 days of modeling． The body weight gain，in open field test and fluid consumption changes in mice were observed． Result During 22 days of modeling，compared with the normal + NS group，the body weight，fluid consumption，behavior indicators in model + NS group were not statistically significant ( P ＞ 0. 05) ． Compared with model + NS group，in 5mg / kg 和 30mg / kg caffeine intervention groups the 1% sucrose preference and sucrose consumption showed significant difference( P ＜ 0. 05) ． Conclusion It is difficult to make successfully CUMS model of depression on C57BL /6J mice． The partially protective effect of caffeine on CUMS mice is dose related，and is not caused by excitement role of caffeine on cerebral cortex．

Morphological Observation of Willis Ｒing Structure of the Kunming Mouse by Using a Perfusion Technique

2014, 31(03):  21-22. 

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Abstract: To establish a fine morphological structure of Kunming mice arterial ring ( Willis ring) ． Select Kunming mice by perfusion technology． Briefly infusing plasticizer about 2 ml from the left ventricle to cerebral vessel was performed，then skull bones were removed，and the structure of Willis ring was observed． This method made a fine cerebral vessel very clear and Willis ring clearly was revealed in the three-dimensional． We successfully adopted the perfusion technology to show the morphological structure of mice cerebral vessel which would play an important role for manufacturing the model of mice cerebral vessel．

Study for Effect of the Palygorskite Corncob Bedding on the Safety of KM Mice

2014, 31(03):  23-25. 

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Abstract: Objective To investigate the effects of bedding materials consisted of wood shavings，corn and the palygorskite corncob on the body weight，the blood biochemical parameters，pathological changes of KM mice． Method The 80 KM mice were divided into four groups randomly，20 in every group，with half male and half female，using the above-mentioned bedding materials and no bedding materials，respectively． The body weight were Weighed weekly，the blood biochemical parameters，including ALT，TP，ALP，LDH，T-SOD，ADH，were measured． Four weeks later，the pathological tissue organs were observed． Ｒesult Comparing with animals used the palygorskite corncob bedding，the weight of the male and the female KM mice in poplar bedding group was reduced from the third week，P ＜ 0. 05． Compared with the KM mice in the palygorskite corncob bedding group，LDH of KM mice in poplar bedding group was raised，significantly，P ＜ 0. 05; Compared with the KM mice in the palygorskite corncob bedding group，ADH of KM mice in poplar bedding group was reduced，significantly，P ＜ 0. 05; Compared with the KM mice in the palygorskite corncob bedding group，ALT of KM mice in poplar bedding and corn bedding groups were raised，significantly，P ＜ 0. 05; Significant pathological changes were not observed in the examined organs． Conclusion The palygorskite corncob bedding material has little effect on body weight，the blood biochemical parameters in KM mice，and the palygorskite corncob bedding material can be safely used in KM mice breeding．

The Effect of Dahuang Mudan Decoction on Exocrine Function of Pancreatic Acinar Cells in Acute Pancreatitis Model Rats

2014, 31(03):  26-29. 

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Abstract: Objective To study the effect of Dahuang Mudan Decoction ( ＲPDP) on exocrine function and intracellular Ca2 + concentration( FI) of pancreatic acinar cells in acute pancreatitis ( AP) model rats． Method Dahuang Mudan Decoction was administrated to SD rats and the serum containing Dahuang Mudan Decoction ( ＲPDP-S) wasprepared． SD rats randomly divided into three groups: the normal control group，the sham operation ( SO) group and the AP group． Separate pancreatic acinar cells were isolated and co-incubated with ＲPDP-S， amylase secretion of pancreatic acinar cells and the change of FI were analyzed． Ｒesult Compared with the SO group，amylase secretion of pancreatic acinar cells was significantly increase in AP group( P ＜ 0. 05) ; and amylase secretion was significantly decrease in the pancreatic acinar cells treated with ＲPDP-S( P ＜ 0. 05) ，and the FI was raise up along with the prolonged disease ( P ＜ 0. 05 ) ． Conclusion ＲPDP can decrease intracellular Ca2 + concentration，and inhibit the overload of FIand the exocrine function of pancreatic acinar cells．

Comparison of HI Antibody Titers of Ducks with Different MHC Haplotype Vaccinated to AIV Ｒecombinant Vaccine

2014, 31(03):  30-32. 

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Abstract: Objective Different species of ducks respond differently to AIV vaccination． To investigate whether the different response to AIV vaccination is related with different species of ducks with MHC genetic background． Method In this study，four haplotypes determined by TAP gene alleotypes located in MHC loci of SPF ducks were used to compare HI titers changes post vaccination with recombinant live attenuated H5 avian influenza vaccine． Ｒesult The result showed that B3 haplotype SPF ducks produced significantly higher HI antibody titers than others during the 9 wks experiment． Conclusion These results hints that duck MHC genetics may affect the immune responses to AIV vaccination．

Establishment of Experimental Model for Neurotropic Ｒegeneration Ｒesearch of the Multi-branch Facial Nerve Defect

2014, 31(03):  33-36. 

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Abstract: To create experimental model for neurotropic regeneration research of the multi-branch facial nerve defect． Method Twelve rabbits were used in this study． The rabbit’s facial nerve was explored and cut near the crotch，leaving a 5 mm gap． Then，the gap was repaired by multi-branch chitosan chamber． 1，2，3，8，16，24 weeks later，ethology of the rabbit’s face was evaluated． Ｒesult Eleven rabbits were still alive，24 weeks after the operation． Swell or rejection was not found． Conclusion The experimental model is stable and reproducible． Tthe use of multi-branch chitosan conduits for nerve regeneration study is feasible．

Investigation of Coat Color Gene MC1R ( R306ter) and TYＲP1 ( Q331ter) SNP Loci in Labrador and Golden Retriever

2014, 31(03):  37-41. 

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Abstract: Objective To establish the method for detecting SNP loci of coat color gene MC1R R306ter ( c． C916T) and TYRP1 Q331ter ( c． C991T) in canine． Method In order to establish the SNP detecting method，the SNP of coat color gene MC1R c． C916T and TYRP1 c． C991T in canine were detected by using PCR / sequencing method． And then，using the established method，the genotypes of coat color genes in 19 Labrador and 15 Golden Retriever dogs were investigated． Result The established PCR / sequencing method could detect the SNP loci for TYRP1 c． C991T clearly and effectively． However，there was a specific peak in the PCR / sequencing results for MC1R gene c． C916T SNP loci． Using cloning / sequencing method，we confirmed that the specific peak in the PCR / sequencing results of MC1R gene was heterozygous SNP loci． The genotype analysis for color genes showed that Golden Retriever dogs were all homozygous eeBB ( n = 15) genotype，no other genotypes were detected． For Labrador， black dogs showed EeBB ( n = 6) and EeBb ( n = 2) genotypes，yellow dogs showed eeBB ( n = 9) and eeBb ( n = 2) genotypes． No others genotypes such as EEBB and EEBb ( black ) or eebb ( yellow ) were detected． Furthermore，because there was no chocolate Labrador dog，so no corresponding Eebb or EEbb genotypes were detected． Conclusion The PCR / sequencing method for detecting the SNP loci of coat color gene MC1R R306ter ( c． C916T) and TYRP1 Q331ter ( c． C991T) in canine is successfully established． These works provide the basis for investigating the correlation between coat color genotypes and training success rates of guide dogs．

Polymorphism of Exon2 Sequence in ApoE Gene of Guangxi Bama Mini Pig

2014, 31(03):  42-45. 

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Abstract: Objective Cloning and analysis of ApoE gene sequence in Guangxi Bama mini-pig were carried out to establish the basic genetic data for Atherosclerosis model in Guangxi Bama mini-pig． Method A 237bp sequence of the ApoE gene was cloned，polymorphism analysis was performed using PCＲ-SSCP． Ｒesult The PCＲ-SSCP analysis found that in the 229 loci existed polymorphisms of ApoE gene fragments，characterized by AA，the AT and TT type three genotypes． And the cloned sequencing was 100% homology compared with Sus Scrofa． Conclusion ApoE gene sequence of Guangxi miniature pig was successfully cloned． And the gene fragment showed AA ，AT and TT three genotypes，and the genotype frequences were 0. 302，0. 489 and 0. 209．

The Influence of Negative-pressure Process on PCR-DGGE Results

2014, 31(03):  46-49. 

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Abstract: Objective To compare gel solidification time of DGGE before and after the negative-pressure process and to explore if the change of the negative-pressure treatment of the experiment can affect the PCR-DGGE results． Method Using amplified PCR products from two different primers ，run the denaturing gradient gel electrophoresis of the two products before and after different negative-pressure treatment，respectively． Result Compared with the results without the negative-pressure treatment process，the experimental results with negative-pressure treatment were significantly different，but the denaturing gel solidification time is not significantly affected after the vacuum treatment． Conclusion Negative-pressure process has significant influence on PCR-DGGE results，and it is an indispensable part of the experiment．

Nursing and Preoperational Preparing for the Establishment of A Hemi-Parkinsonism Model in Monkey

2014, 31(03):  50-52. 

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Abstract: Objective To study the nursing technique experience and preoperational preparing for the establishment of a hemi-Parkinsonism model in rhesus． Method 10 monkeys were injected with MPTP in right side of internal carotid artery ，the experimental skill was important，and careful nursing and preoperational preparing were carried out，include routine care before and after surgery，prevention of complications and husbandry． Result All monkeys were successfully injected，and the post-operative survival rate was 90%． Conclusion Although sophisticated surgical method plays an important role in the modeling，delicate nursing care and pre-operation strategy is the key to safety and rapid postoperative recovery of monkeys．

Experimental Animal Welfare and the Advantage Reflected of IVC System

2014, 31(03):  53-55. 

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Abstract: With the further advance of the work of life science and medical research，the status of experimental animals in the scientific research has been paid more and more attention to． The purpose of this paper is to investigate the welfare of the experimental animal and the present state，both the domestic and foreign; compare the normal laminar flow rack， shelter system and the IVC ( individual ventilated cages ) system． This paper demonstrates that the IVC system has its specific characteristics on the increasing of experimental animal welfare during the laboratory construction and feeding．

Advances in Research on the Structure and Function of TAP Using Antigen Processing

2014, 31(03):  56-59. 

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Abstract: The transporter associated with antigen processing ( TAP) is a heterodimeric protein，and the TAP1 and TAP2 subunits are encoded by closely linked genes in the MHC． TAP is essential for peptide delivery from the cytosol into the lumen of the endoplasmic reticulum ( ER ) ， where these peptides are loaded on major histocompatibility complex ( MHC) I molecules to form a complex． The lack and mutation of TAP protein may infect the presentation of antigen peptide and further harm the function of immune response．

Advances of Microsatellite and Single Nucleotide Polymorphism Analysis for Genetic Monitoring in Mouse and Rat

2014, 31(03):  60-65. 

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Abstract: With the rapidly advances in molecular biology technology，microsatellite and SNP has become an important means of analysis for genetic monitoring of laboratory animals． Microsatellite and SNPS，as a kind of new molecular markers，has the accurate and fast advantages，compared with the biochemical and immune markers． In this paper，we will have a detailed development of microsatellite and SNP analysis for genetic monitoring of mouse and rat has been reviewed．