Table of Content

28 December 2007, Volume 24 Issue 06

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Article

cDNA Cloning and Sequence Analysis of Partial Sequence of Activin-β_A and-β_B Gene from Guangxi Bama Mini-pig

LAN Gan-qiu,HUANG Xiong-jun,GUO Ya-fen,LI Fang-fang,WANG Ai-de,JIANG He-sheng

2007, 24(06):  1-5. 

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In this study,total mRNA isolated from the ovary tissue of Guangxi Bama mini-pig were used as templates to synthesize cDNA by reverse transcription polymerase AMV.The partial sequence of activin β_A and β_B cDNA with length of 964 bp and 791 bp respectively,was obtained by PCR.The homology analysis showed that they are highly conservative.The homology rate was 99.79% and 99.62% respectively,compared with the activin β_A and β_B gene of other pig breeds reported in GenBank.Two nucleotides mutation were found in activin β_A gene,of which one mutation occurred in β_A mature peptide and the other in none coding part. Three nucleotides mutations occurred in the signal peptide of β_B,of which one is silent mutation.

Cloning and Sequence Analysis of Growth Hormone Release Hormone(GHRH) Mature Peptide from Guangxi Bama Minipig

GUOYa-fen,LI Fang-fang,LAN Gan-qiu,HUANG Hiong-jun,WANG Ai-de,QIN Xiao-e

2007, 24(06):  6-10. 

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Objective To clone and analysis the sequence of GHRH mature peptide gene of Guangxi Bama minipig.Methods The sequence of GHRH exon3 was amplified by a specific pair of primers using Guangxi Bama minipig genome as template.After sequencing the GHRH exon3,the GHRH mature peptide was cloned by over-bang extension PCR protocol and then ligated into sub-clone vector for sequencing and restriction analysis.Results It was showed that the cloned GHRH exon3 and mature peptide gene from Guangxi Bama minipig contained 132 and 105 bp,respectively.Conclusion Sequencing confirmed that the sequence of GHRH exon3 and mature peptide of Guangxi Bama minipig were successfully cloned in the study.

Cloning and Sequence Analysis of Ghrelin cDNA from Guangxi Bama Minipig

GUO Ya-fen,LUO Qin,WANG Shi-kai,LAN Gan-qiu,WANG Ai-de

2007, 24(06):  11-14. 

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Objective To clone and analyze the sequence of Ghrelin cDNA of Guangxi Bama minipig.Methods The sequence of Ghrelin cDNA was amplified by a specific pair of primers using Guangxi Bama minipig belly total RNA as template.The Ghrelin cDNA was cloned by RT-PCR and then ligated into pMD18-T vector after purification.The recombined pMD18-T vector was transformed into the coliform DH5α for sequencing.Results The cloned Ghrelin cDNA gene from Guangxi Bama minipig contained 357 bp.The homology analysis showed that compared with the sequence of other pig breeds reported in GenBank,the homology rate of the Ghrelin cDNA sequence were 99.7%.Conclusion Sequencing confirmed that the sequence of Ghrelin cDNA of Guangxi Bama minipig was successfully cloned in the study.

cDNA Cloning and Sequence Analysis of Myostatin Gene from Guangxi Bama Minipig

WANG Shi-kai,LAN Gan-qiu~*,LUO Qin,GUO Ya-fen,WANG ai-de

2007, 24(06):  15-19. 

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The total RNA was purified from Guangxi Bama minipig muscle tissue.The complementary DNA(cDNA) encoding Myostatin protein was amplified by the reverse transcription polymerase chain reaction(RT-PCR) method.The amplified cDNA fragment was ligated into pMD18-T vector after purification.The recombinant was verified by the method of PCR,restrictive cleavage and sequencing,respectively.The result showed that the Bama minipig Myostatin gene cDNA was successfully cloned in this study.The homologous analysis indicated that the homologous rate was 99.7% compared with the porcine Myostatin gene reported in GenBank and this gene has high conservativeness among mammalian animals(>92.3%).Two synonymous mutations and one missense mutation were detected in Myostatin gene cDNA sequence of Bama minipig.

Cloning and Sequence Analysis of Growth Hormone Gene from Guangxi Bama Minipig

YANG Xiu-rong,LAN Gan-qiu~*,GUO Ya-fen,WANG Ai-de

2007, 24(06):  20-25. 

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In this study,Bama minipig growth hormone gene was amplified by PCR.The target segment was sub-cloned into pMD18-T vector.One positive recombinant clone was sequenced and analyzed.The result showed that Guangxi Bama minipig growth hormone gene with a size of 2 007 bp was successfully cloned in this study.The homology of nucleotides sequence of Bama minipig growth hormone gene to that of Wuzhishang pig,Taihu pig and Sus scrofa was 99.4%,98.3% and 93.9%,respectively.

Cloning and Prokaryotic Expression of Follistatin Gene from the Guangxi Bama Minipig

GUO Ya-fen~1,LAN Gan-qiu~1,MO Yi~1,LIANG Fang-fang~2,YANG Xiu-rong~1,JIANG Qin-yang~1,JIANG He-sheng~1

2007, 24(06):  26-29. 

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In this study,the total RNA was isolated from Guangxi Bama mini-pig ovary tissue with RNA_(watson) extraction kit.The complementary DNA(cDNA) encoding Follistatin protein was ampilified by the reverse transcription polymerase chain reaction(RT-PCR) method and a pair of differential primers.The amplified cDNA fragment was inserted into pMD18-T vector.The recombined vectors were verified through the sequence survey.The results showed that the cloned Follistatin cDNA was highly homologous with the reported nucleotide encoding swine Follistatin in GenBank(99.4%).Meanwhile,the identified Follistatin cDNA was inserted in the multiple cloning sites of plasmid pET 32a+ to construct prokaryotic expression vector,the latter was transformed into the E.coli.BL_(21) induced by IPTG.Then a fusion protein was obtained(about 58.4 ku) in SDS-PAGE and Western blotting.These results showed that the Follistatin cDNA was expressed in prokaryotic cells.

Analysis of Prepuberty and Semen Characteristics of Male Guangxi Bama Minipig

LU Sheng-sheng,LUO Long-xing,LAN Zong-bao,LI Zong-qiang,GUO Ya-fen,LAN Gan-qiu,WANG Ai-de

2007, 24(06):  30-32. 

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The objectives of this study were to examine the prepuberty and semen characteristics of male Guangxi Bama minipig.Testes of 15,20,25,26,27, 28,29,30,45,55,62,75 days old Bama minipig were formalin-fixed and paraffin-embedded and stained with hematoxylin and eosin,and were examined if sperms exist.Semen of Bama minipig were collected and sent to Guangxi Center for analysis and test research to analyze its physical characteristics and chemical compositions.The results showed that:(1) the prepubertal age of male Bama minipig is preliminarily determined to be 45 days old approximately;(2)the physical characteristics(ejaculate volume 107.5 mL±39.0 mL,pH 7.6±0.3,osmolarity 316±3 mOsm,sperm viability 0.754±0.079,motility 0.376±0.168,abnormality 0.256±0.159,concentration 73.2×10~6/mL) and chemical compositions(DNA 127.4 μg±8.0 μg/mL,protein 6.0 g±0.7 g/100 mL,phospholipids 8.7 mg±0.2 mg/100 mL,fructose 117.4 mg±81.0 mg/100 mL,prostaglandin 3.6 pg±0.8 pg/ mL,citric acid 45.6 mg±77.6 mg/100 mL,Na~+ 1.56 mg±0.13 mg/mL,K~+ 0.78 mg±0.06 mg/mL,Cl~-1.62 mg±0.18 mg/mL)of Bama minipig semen were analyzed.

Analysis of Reproductive Hormone Concentrations of Guangxi Bama Minipig at Different Age

LU Sheng-sheng,LUO Long-xing,GUO Ya-fen,LAN Gan-qiu,WANG Ai-de

2007, 24(06):  33-35. 

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Objective The reproductive hormone concentrations of Guangxi Bama minipig at different age were investigated.The results showed that:(1) in male Bama minipig from two to six months serum testosterone level increases and reaches 12.2 ng±8.9 ng/mL at six months old,after which it decreases;from two to six months old serum FSH level increased and reached 64.6 ng±6.1 ng/mL at six months,after which it decreased;from 2 to four months serum LH level increased and reached 46.8 ng±12.5 ng/mL at four months,after which it decreased;(2)in female BaMa mini-pig,from two to four months old serum levels of FSH,LH,E2 increased significantly,and reached 51.9 ng±54.0 ng/mL,23.2 ng±31.6 ng/mL and 79.3 pg±93.0 pg/mL,respectively,after which the levels of FSH,LH,E2 decreased.

Determination of Protein and Energy Concentration in Diets for Grower of Bama Minipig

LAN Gan-qiu,GUO Yan-fen,WANG Ai-de

2007, 24(06):  36-38. 

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Objective Three trials were carried out to evaluate the crud protein(CP) and digestive energy(DE) concentration in diets for growing Bama minipig.Methods Forty Bama minipigs weaned at age of 40 d with similar body weight were assigned randomly into four groups of 10 pigs each.In trial Ⅰ,the pigs in group 1,2,3 and 4 were fed with diets containing 13%,14%,16% and 18% CP from age of 45 to 80 d,respectively,and the digestive energy(DE) level in the four diets was set as 12.95 MJ/kg.In trial Ⅱ,the animals in group 1,2,3 and 4 of trial Ⅰ were fed with diets contained 11%,12%,13% and 14% CP from 81～117 d of age,respectively,and all diets had the same DE concentration(12.95 MJ/kg).In trial Ⅲ,another 40 pigs with similar body weight weaned at 40 d of age were assigned randomly in 4 groups with 10 pigs each.Four dietary treatments were used:(1)15% CP and DE 13.37 MJ/kg;(2)15% CP and DE 12.95 MJ /kg;(3)13% CP and DE 13.37 MJ/kg and(4)13% CP and DE 12.95 MJ/kg.Results The results showed that there was no difference in body weight gains between the fours groups of animals in trial Ⅰ(P>0.05).In trials Ⅱ,pigs fed diet with 11% CP had significant low body weight gain compared to the animals in the other three groups(P<0.05).However,no significant difference in body weigh gain was observed between the animals from the other thee groups(P>0.05).In trial Ⅲ,no significant difference in body weight gain was detected in the four dietary treatments(P>0.05).Conclusion The appropriate concentrations of CP and DE for growing Bama minipig were proposed as 12%～13% CP and 12.95 MJ/kg based on the results in this study.

The Diet-Induced Atherosclerosis In Wuzhishan Minipigs

CHEN Hua,XIE Zhong-chen,HUANG Guang-yong,LIU Ya-qian,LI Chun-hai,GUO Xiong-ming,XIN Ning-bo

2007, 24(06):  39-43. 

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Objective To induce atherosclerosis in Wuzhishan minipigs,and establish the formula of experimental atherosclerosis in the minipigs.Methods Thirty Wuzhishan minipigs were randomized to 3 groups,one group of 6 animals was set as control,the other 2 groups of 12 animals each were set as test groups fed with 1.5% cholesterol,high-fat diet(group T1) and 3.0% cholesterol,high-fat diet(group T2),respectively.The serum TG,TC,LDL-C and HDL-C were analysed monthly.At months 4,6,8 and 12 after high-cholesterol,high-fat diet feeding,three swine in each test group were subjected to a full gross necropsy to exam the atherosclerosis by macroscopic and microscopic evaluation.Three animals from the control group were necropsied at months 6 and 12.Results After feeding with the high-cholesterol,high-fat diet,the values of cholesterol were elevated significantly and the atherosclerosis lesions were observed in swines.The mean values of LDL-C,TC and HDL-C were increased significantly after 1 month high cholesterol feeding,and were approximate 13～15 folds,8～10 folds and 3～4 folds respevtively comparing to the control group at months 3 and 4.Most of the atherosclerosis lesions were found in abdominal aorta,iliac artery,an coronary artery,and were seldomly distributed in other examined arteries.after 4～8 months high-cholesterol feeding.The atherosclerosis lessions were more extensive and severe after 12 months administration.The abdominal aorta,iliac and coronary arteries were the region most severely involved by fibrosis and intimal thickening.The carotid,mesenteric and kidney arteries were slightly involved area and free of atherosclerosis at early period.Group T2 has higher serum LDL-C,TC and HDL-C values,and more severe atherosclerosis than group T1,but the differences were slightly.Conclusion 1.5%～2.0% cholesterol in high-fat feed is reasonable for inducing atherosclerosis in minipigs.To study primary atherosclerosis need 4～8 months high-cholesterol,high-fat feeding,and 12 months should cost for research secondary atherosclerosis in minipigs.

The Pathological Observation on Diet-Induced Atherosclerosis In Wuzhishan Minipigs

CHEN Hua,LI Chun-Hai,XIE Zhong-chen,LIU Ya-qian,GUO Xiong-ming

2007, 24(06):  44-48. 

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Objective To observe the morphological features of atherosclerotic lesions and its distribution and progression of induced atherosclerosis in Wuzhishan minipigs.Methods 30 Wuzhishan minipigs were divided into 3 groups,one group of 6 animals was set as control,the other two groups of 12 animals each were set as test groups with 1.5% cholesterol,high-fat diet(group T1) and 3.0% cholesterol,high-fat diet(group T2) respectively.At 4,6,8 and 12 month after high-cholesterol,high-fat diet feeding,each test group had 3 swine undergoing a complete necropsy to characterize the atherosclerosis both by gross and microscopic examination.The control swine were necropsied at 6 and 12 month with 3 animals each time.Results The high-cholesterol,high-fat feeding led to the development of an accelerated atherosclerosis.The atherosclerotic lesions had developed at thoracic aorta,abdominal aorta,iliac,and coronary arteries after feeding inducing diet 4 months,and the atherosclerotic lesions developed further during 6 and 8 months.At 12 months after inducing diet feeding,atherosclerotic lesions were observed in entire arterial tree.The abdominal aorta,iliac,and coronary arteries were the region most severely involved by fibrosis and intimal thickening.The carotid,mesenteric and kidney arteries were slightly involved area and free of atherosclerosis at early period.The atherosclerotic lesions can be divided into 3 kinds primarily,which was fatty streaks was representative by thoracic aorta,fibrous or complicated plaque was representative by abdominal aorta,and fibrofatty plaque was representative by coronary artery respectively.The morphological features of initiation and progression of atherosclerotic lesions varied in different arteries and different regions.The period of atherosclerosis initiation and progression also varied among the testing animals.Conclusion The pathological features of induced atherosclerosis in minipigs were closely similar to that of patients,and some specialties existed in minipigs,the inducing period and arteries needed to be selected according to the research purposes when making use of inducing atherosclerosis in minipigs.

Comparison of Three Strains of Minipigs with Induced Type Ⅱ Diabetes Mellitus

CHEN Hua,LIU Ya-qian,LI Chun-hai,GUO Xiong-ming,HUANG Li-jie

2007, 24(06):  49-55. 

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Objective To establish and compare the typeⅡdiabetes mellitus models in three strains of native minipigs for promoting their use on type Ⅱdiabetes mellitus research.Methods Six Bama minipigs,six Wuzhishan minipigs and six Nongda minipigs were fed with a high-sucrose high-fat diet to induce type Ⅱ diabetes mellitus.Four Nongda minipigs were assigned as controls fed with a normal diet.Body weight,fasting serum glucose and insulin concentrations were measured monthly,and serum TG,CHO,HDL-C,LDL-C and intravenous glucose tolerance test(IVGTT) were tested at 2 month intervals during the 8-month inducing period.Results Bama minipigs and Wuzhishan minipigs were susceptible to the high-sucrose high-fat diet.Obesity,hyperinsulinemia and impaired insulin tolerance were noted at month 5 and 6.One of six Bama minipigs and one of six Wuzhishan minipigs developed diabetes mellitus and three Bama minipigs and two Wuzhishan minipigs reached the pre-diabetes stage at month 8.Nongda minipigs were relatively resistant to the high-sucrose high-fat diet.Fasting serum glucose,insulin concentration,insulin sensitivity index and IVGTT remained normal during the 8 months high-sucrose high-fat feeding.Only slightly obesity and despaired insulin tolerance were observed at month 7 and 8.All the parameters related to blood lipid metabolism(TG,CHO,HDL-C,LDL-C) were increased significantly after feeding high-sucrose and high-fat in the three strains of minipigs with Bama minipigs increased more significantly.Conclusion Bama minipigs and Wuzhishan minipigs were susceptible to the high-sucrose and high-fat diet inducing typeⅡdiabetes millitus.Susceptibility differences existed among the animals in the same strains.Nongda minipigs were less susceptible to the dietary inducing.The results provided foundamental data for further research on typeⅡdiabetes mellitus susceptibility relative genes in the minipigs and for selective breeding of family stocks with high typeⅡdiabetes mellitus susceptibility.

Production of Transgenic Cloned Embryos by Nuclear Transfer Using FetalFibroblast and Marrow Stromal Cells of Laboratory Miniature Pig

PAN Deng-ke~1,PAN Qiu-zhen~2,MU Yu-Lian~1,WAN Rong~2,LIU Ji~1,ZHENG Mao-en~

2007, 24(06):  56-59. 

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Objective The efficiency of production transgenic cloned embryos using the different types of somatic cells was examined.Methods Fetal fibroblast cells(FF) and marrow stromal cells(MSCs) of Wuzhishan pig(WZSP) were transfected with pEGFP-N1 plasmid DNA by lipofectin-mediated way.Clones were obtained by G418 selection.MSCs,FF,and GFP-MSCs,GFP-FF were used as nuclear donors for somatic cell nuclear transfer(SCNT).Results There were no significant differences in the development rates to the blastocyst stage of SCNT embryos from nontransgenic and transgenic cells between MSCs and FF(7.1% vs.8.3%,P>0.05;9.9% vs.9.6%,P>0.05).Blastocyst rate of reconstructed embryos with GFP transfected MSCs or FF in comparison to those derived from non-transgenic somatic cells did not decreased(P>0.05).Conclusion It was suggested that porcine transgenic blastocysts could be efficiently produced by SCNT using transgenic MSCs as well as FF.

Cloning and Eukaryotic Expression Vector Construction of Growth Hormone Receptor Gene from Chinese Inbreeding Wuzhishan Pig(WZSP)

LI Ji-tao,MU Yu-lian,ZHANG Li,YANG Shu-lin,LI Kui,FENG Shu-tang

2007, 24(06):  60-63. 

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Objective To disclose the molecular mechanism of dwarfism for Chinese inbreeding Wuzhishan pig(WZSP).Methods Growth hormone receptor gene form WZSP were cloned to pGEM-T vector through RT-PCR,which based on the total RNA obtained from WZSP's liver tissues.The segment containing WZSP GHR gene was linked with the segment of pEGFP-C1 after digestion by SalⅠ and XbaⅠ enzyme and purification.Results The results of sequencing showed that the coding sequence of GHR gene for WZSP contained 639 amino acids including the signal peptide of 18 amino acids.The eukaryotic expression vector named WZSPGHR-pEGFP-C1 was constructed successfully after enzyme digestion and sequencing.Conclusion Five substitutions of amino acids were found in the cytoplasmic domain of GHR,which possibly influenced GHR signal transduction.The study laid a foundation for the research of function and signal transduction of GHR.

Study on the Monitoring of Pig's Chimera with Embryonic Stem Cell(EG)

MU Yu-lian,FENG Shuang,LIU Li-xin,DONG Xiao,CHEN Hong-ping,KUANG Ling,ZHANG Qing-feng,ZHANG Yong

2007, 24(06):  64-65. 

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Microsatellite was used to check the various tissues of chimeric pig generated by injecting EG cells into blastocysts.The results indicated that WZSP EG cells contributed to these tissues: skin,brain,pancreas,thyroid gland and blood.Contributions of the WZSP EG cells to development are not detectable in these tissues: heart,lung,liver,spleen,kidney,jejunum and ovary.

Trying to Obtain Cleaning WZS Minipigs by Spontaneous Labour and Hand-feeding

SHAN Tong-ling~1,JIN Er-hui~1,YANG Shu-lin~1,CUI Li~2,TIAN Ke-gong ~3,FENG Shu-tang~

2007, 24(06):  66-68. 

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Objective To study whether or not obtaining cleaning WZS minipigs by spontaneous labour and hand-feeding.Methods Two hours after Spontaneous Labour,the handling young pigs were transmitted into isolator,and was raised with milk powder,rice flour and forage.In the hand-feeding phase,intestinal bacteria was given to the pigs seven times,and iron and selenite were added two times.After 42 days,pathogens were detected.Results The survival rate of pig is 75%.Many pathogens were undetectable.Conclusion Cleaning WZS minipigs were obtained by spontaneous labour and hand-feeding.

Determination of Clinical Normal Physiological Parameters in Chinese Wuzhishan Pig

JIN Hong-tao~1,FAN Chun-rong~1,LI Hui~1,LI Jin~1,LI Ji-tao~2,WANG Xue-feng~2,FENG Shu-tang~2,WANG Ai-ping~1

2007, 24(06):  69-73. 

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Objective To obtain the reference normal physiological parameters of Wuzhishan pig(WZSP).Methods The normal physiological parameters of 24 WZSPs,male and female in half,were detemined and compared with different sexes,including body weight,body temperature,20 normal values of hematology,21 normal values of biochemistry,4 normal values of cardiograph,urine,organ weight and organ relative weight.Part of parameters were detemined continuously for 4 months.Results The results showed that there were no significant between male and female except some hematological parameters such as WBC,LYM%,GRAN%,PCT and biochemistry such as ALP,P,Na,Cl.Some values of biochemistry such as GGT were in conformity to those of human.The four-month continuous determination showed most physiological parameters were stable.Conclusion The normal ranges of physiological parameters of WZSP were therefore established and these data may give a useful reference for further studies and use of WZSP in medical field.

Establish Meat of Hepatic Fibrosis Model of Guizhou Minipig by Multiplex Factor

ZHAO Ju-hua~1,WU Shu-guang~2,WU Quan-sheng~2,GUO Ke-nan~2,DONG Xiao-ju~2,WEI Jian-lin~3,QIAN Ning~

2007, 24(06):  74-77. 

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Objective To establish the hepatic fibrotic model of Guizhou minipig by multiplex factor.Methods Sixteen Guizhou minipig(8 females and 8 males) were divided into two groups including one control group and one experiment group at random.Control group was injected into peritoneum with olive oil and 0.9% NS,0.5 mL/kg twice a week,fibrosis model of experiment group was duplicated by peritoneal injection of 40% carbon tetrachloride and rabbit serum 0.5 mL/kg twice a week,for nine weeks.They were sacrificed at the end of the ninth weeks,then serum marker levels of liver function of ALT,AST,TP,ALB,A/G were determined.The liver were taken out and paraffin sections were stained by hematoxylin-eosin,Masson,Van Gieson,and were examined under light microscope(LM).The liver from experimental group were observed under electron microscopy(EM).Results Compared with the control group,serum ALT,AST activity of experiment group elevated significantly(P<0.01),ALB,A/G content decreased markedly(P<0.01);the pathological changes of liver of experiment group were observed under LM and EM.Conclusion\ Hepatic fibrosis model of Guizhou mini-pig was duplicated by peritoneal injection of 40% carbon tetrachloride and rabbit serum.

Ultrastructure Observation of Cardiac Muscle Cells of Guizhou Miniature Pig

QIAN Ning~1,WU Shu-guang~1,DENG Hong~2,ZHANG Yu-qin~2

2007, 24(06):  78-80. 

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Objective To observe the ultrastructure of cardiac muscle in Guizhou mini-pig.Methods The observation was obtained by transmission electron microscope. Results Cardiac muscle cells of Guizhou miniature pig exhibited regularly,a cross-striated banding pattern distinguishing Z line,I band,A band,H band and M line.It was obvious that numerous mitochondria around myofilaments and the sarcoplasmic reticulum were pretty well developed and wanders irregularly through the myofilaments,with little transverse tubules existing and possessing only one T tubule at the Z line.At the level of the Z lines between adjacent cardiac muscle cells,the intercalated disk composed of 3 main junctional specializations complexes.Conclusion Though possessing its different characteristic,cardiac muscle cells of Guizhou miniature pig exhibit the same ultrastructures of human being.

Preventive Effects of "Hu-Shen No.1"on Cisplatin-Induced Nephrotoxicity in Guizhou Miniature Pigs

DONG Xiao-jun~1,GUO Ke-nan~1,WU Shu-Guang~1,LI Shu-chai~1,ZHANG Yu-qin~2,DENG Hong~2,QIAN Ning~

2007, 24(06):  81-83. 

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Objective To study the preventive effects of "Hu-Shen No.1" on acute nephrotoxicity of cisplatin in Guizhou miniature pigs. Methods Guizhou miniature pigs were randomly divided into three groups,namely the control group,the model group and the prevention group.The prevention group received "Hu-Shen No.1" by gavage for five days.After one hour of the 5th administration of "Hu-Shen No.1",animals were given cisplatin(4 mg/kg) by a single peritoneal injection.Urine samples were collected by a bladder fistula.Urinary output,urine level of creatinine and urea nitrogen were dynamically observed on the 1st hour before injection and the 1st,3rd,6th,24th,48th,96th hours after cisplatin treatment.The excretion of creatinine and urea nitrogen were calculated on the basis of the above determination.Results Urinary output,excretion of creatinine and urea nitrogen of the model group were significantly lower than that of the control group.The results of the prevention group were markedly improved. Conclusion "Hu-Shen No.1" may protect kidney against cisplatin-induced nephrotoxicity.

Established Alveolar Cleft Model on Guizhou Miniature Pigs

TIAN Kun~1,SHE Xiao-ming~2,QIAN Ning~3,WU Shu-guang~3,ZHANG Qian~2,JIN Xin~2

2007, 24(06):  84-85. 

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To set up a animal model of cleft alveolus by surgical approach. Methods The alveolus model was made surgically in 20 minipig.Two "L" mucoperiosteal flaps were designed at both labial and palatal sides.A segmental bone,about 8 mm×9 mm,from the first lateral medial tooth to the medial of the canine was excised.The raw bony surface was covered by the mucoperiosteal flaps.The samples were examined by gross radiation,histological analysis.Results In all experimental minipig,hard tissue was not closed up and soft tissue was healed up.Conclusion The animal alveolar cleft model by surgical technique on Guizhou minipig was set up.

The Investgation on The Carrying Status of Main Pathogenic Organism in Guizhou Miniature Pig

TIAN Wei-yi,QIAN Ning,WANG Ping,CAI Kun,WANG Wen-jia,WU Shu-guang

2007, 24(06):  86-88. 

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Objective To test the carrying situation of pathogenic organism of Guizhou miniature pig. Methods Collecting the samples of hair,skin,secretion and blood by using morphology,biochemistry and serology test,36 microorganisms and parasites were checked on the different position of Guizhou miniature pigs. Results There were 25 pathogenic organisms indentified in Guizhou miniature pigs.Conclusion The presence of athogenic organism in Guizhou miniature pig indicates the needs for the monitoring and controlling microorganism and parasite.

Prokaryotic Expression of ORF2 Gene and Its Four Different Fragments of Porcine Circovirus Type 2

QU Su-jie,HUANG Wei-jian~*,FENG Jian-yuan,ZHU Yun,CHEN Qiong,XIE Li-hua

2007, 24(06):  89-92. 

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Based on the porcine circovirus type 2(PCV2)LC strain,5 pair of primers were designed according to the Luchuan sequence,the ORF2,ORF2a,ORF2b,ORF2c and ORF2d gene were amplified from the PCV2-LC plasmids by PCR,and inserted into the prokaryotic expression vector PET-32a.The constructed recombinant expression plasmids were transformed into the receptive cells of E.coli BL21 and induced by IPTG with a finial concentration 1 mmol/L,then the recombinant protein was expressed in the form of inclusion bodies.The protein expression could react with PCV2 positive serum by Western-blot,sharing a good antigenicity.This study will provide a good fundament for development of PCV2 diagnostic ELISA diagnosis assay kit.

Sequences Comparison and Phylogenetic Tree Analysis of 288 Strains of Porcine Circovirus

YIN Ye-shi,HUANG Wei-jian~*,CHEN Qiong,LIU Hai-peng,XIE Li-hua,FENG Jian-yuan,WANG Ai-de

2007, 24(06):  93-99. 

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In order to know the heredity disciplinarian of porcine circovirus(pcv) and pave the basement for preventing and controlling the disease.In this research,nucleotide sequences of 26 strains of PCV1 and 262 strains of PCV2 were analyzed,also,the ORF1's and ORF2's nucleotide sequences and deduced amino acid sequences were analyzed.At the nucleotide level,the results show that PCV1sequences and PCV2 sequences were divided into two different branches,and among the PCV1 sequences there were no obvious regularity,but among PCV2 sequences,there were divided into two different sub-gene types.The sub-gene typeⅠwas mainly consisted of sequences from American,Canada and Australia,and the sub-gene typeⅡwas mainly consisted of sequences from Franch and Newzealand.At the amino acid level,the phylogenetic tree was very complexity,and there were no obvious regularity.It was found statistically that,although nearly ten years pasted, the sequences homology of PCV had no large changes,the results showed that PCV was very conserved.After analyzed the phylogenetic tree,we discovered that the distributing of PCV had no geographic and time restriction.Also,we analyzed the mutant dots of the PCV2 nucleotide sequences and deduced amino acid sequences,we discovered that,PCV2 nucleotide sequences were easy to has base transversion.

The Application of BLS Cell Fusion Instrument on Porcine Parthenogenetic Activation

LIU Xiao~

2007, 24(06):  123-125. 

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Objective To establish and optimize the electric parameters of BLS cell fusion on porcine parthenogenetic embryo,being for the cloned animals.Methods MⅡ oocytes matured in vitro were activated by electrically stimulation with different pulses and different strength and different field duration to induce parthenogenetic activation and further development.Results\ Parameter (1.2 kV/cm,100 μs,2 pulses) supported higher percentages of cleavage and blastocyst(70% and 30% or so);BLS fusion instrument was efficient and stable for production of porcine embryos in vitro.

The Research of Methods of Superovulation and Embryo Recovery on Minipigs

DAI Qi~

2007, 24(06):  126-128. 

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\ Miniture pigs were used for superovulation and the recovery of early embryo.The results of this paper were: the rate of synchronization of PG+PMSG+HCG on weaning pig was 100%,while the rate of non-weaning pig was only 72.73,but there appearanced obviously distance on average ovulation number(P<0.05);the recovery of early embryo of slaughter and surgery were 84.29% and 76.92%(P<0.05),but no obviously distance on the number of recovery embryo(P>0.05).