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Table of Content

20 May 2014, Volume 41 Issue 5
Effect of Different Concentrations of Scriptaid on the Development and Histone Acetylation Modification of Buffalo Fetal Fibroblasts
RUAN Qiu-yan, SUN Hong-liang, MENG Zhen-miao, LIU Xiao-lin, DU Feng-jiao, SHI De-shun,LU Feng-hua
2014, 41(5):  1-6. 
Abstract ( 249 )  
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The purpose of this study was to investigate the effect of various concentrations of histone deacetylase inhibitor (Scriptaid) on the growth characteristics, cytotoxicity and the level of histone H3K18 acetylation (AcH3K18) of buffalo fetal fibroblasts (BFF), so that to find a novel way to improve the efficiency of buffalo somatic cell nuclear transfer in future. The experiment investigated the effect of various concentrations (0, 250, 500, 750 nmol/L) of Scriptaid on the growth characteristics, cell survival rate through trypan blue staining and the level of histone H3K18 acetylation (AcH3K18) by immunohistochemistry detection. The results showed that there was no effect on the biology characteristics of the BFF treated with Scriptaid, the growth curve was similar with that of the control group, showing “S” distribution, moreover, there was no significant difference on cell survival rate between Scriptaid-treated group and the control group (P>0.05). In addition, immunohistochemistry detection found that the relative fluorescent intensity of AcH3K18 of BFF in the Scriptaid-treated group was significantly higher than that of the non-treated group (P<0.05), and the levels of AcH3K18 of BFF in the 500 and 750 nmol/L groups were significantly improved compared with that of the 250 nmol/L group (P<0.05), but there was no significant difference between 500 and 750 nmol/L treatment groups (P>0.05). These results indicated that an optimum concentration of Scriptaid had little effect on the cell morphology and cell survival rate of BFF, and could significantly improve the levels of histone acetylation.
Cloning and Antigen Epitopes Prediction of Fibronection-binding Protein A Gene A Domain of Bovine Staphylococcus aureus
HE Yan,HAO Yong-qing
2014, 41(5):  7-11. 
Abstract ( 217 )  
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A pair of specific primers was designed on A domain of FnbpA gene sequence of S.aureus published in GeneBank. Then FnbpA-A gene was cloned and sequenced, its protein second structure was analyzed and antigenic epitope was predicted. The sequencing results showed that FnbpA-A gene was 1645 bp in size, encoding 548 amino acids, the homology of coding amino acids between this strain and CIG1835 strain was 100%. Protein secondary structure analysis showed that FnbpA-A protein had alpha helixes 13.69%, β sheet 30.84%, β turn 11.86% and random coil 43.61%. B-cell epitopes were most likely localized in its N-teminal 31 to 46, 54 to 69, 142 to 157, 303 to 318, 324 to 339, 450 to 465, 491 to 506, 533 to 548 sites and T-cell epitopes had the high probability in the regions of 353 to 361 and 245 to 253. To sum up, FnbpA-A could be used as a candidate gene of S.aureus genetic engineering subunit vaccine.
Expression of Truncated E2 Gene of Classical Swine Fever Virus C Strain in E.coli and Preliminary Detection of its Immunogenicity
ZHENG Zhong-hua,LI Dong-sheng,YAO Jun-yong,CHANG Yan,GOU Hong-chao,LIU Wen-jun,ZHAO Ming-qiu,MAO Jing-dan,CHEN Jin-ding
2014, 41(5):  12-16. 
Abstract ( 227 )  
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A truncated gene encoding the A/D antigenic domains (549 bp) of E2 in classical swine fever virus (CSFV) was amplified by RT-PCR from the genomic RNA of CSFV C strain, cloned into pET-32a expression vector to obtain recombinant pET-32a-CSFV-E2, and then sequenced.The result was the right interested gene.The recombinant plasmid was transformed into BL21 and induced with isopropyl-β-D-thiogalactopyranoside (IPTG).After expression,we could see our target band about 39 ku with SDS-PAGE.To purify the His-trap protein in different conditions,and we could get the best one with the most productive target protein.Western blotting indicated that the expressed antigen protein could be recognized by the positive sera of CSFV (the sera from our homemade anti-rabbit one and the clinical swine one).It would benefit the creation of ELISA kit and research on the antibody levels of pigs.
Construction of Bile Acid Receptor TGR5 Eukaryotic Expression Vector and its Expression in Cell
SHI Lin-kai, LIU Qian, ZHANG Zi-ying, LIU Chang, WANG Xu-dong, DING Yue-xia, LI Pei-feng
2014, 41(5):  17-22. 
Abstract ( 255 )  
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The study was aimed to construct a membrane-bound bile acid receptor TGR5 eukaryotic expression vector, which transfected into 293T cell, and expressed in 293T cell. Total RNA was extracted from placenta tissue, and RT-PCR was performed to obtain the TGR5 cDNA, which was inserted into the eukaryotic expression vector pCMV-EGFP. The novel constructed plasmid was confirmed by restriction enzyme digestion and DNA sequencing, then 293T cell was transfected with pCMV-EGFP-TGR5, Real-time PCR and Western blotting were amplified to determine the expression of TGR5. The result showed that the eukaryotic expression vector of pCMV-EGFP-TGR5 was constructed successfully. Green fluorescent was examined under fluorescent microscope, and the expression of TGR5 was increased significantly in 293T cells. The eukaryotic expression vector pCMV-EGFP-TGR5 was constructed successfully and TGR5 was expressed in mRNA and protein levels in 293T cell after transfection.
Prokaryotic Expression and Purification of the Cap Protein of Porcine Toque Teno Virus Type 2
LIU Wei, ZHU Yi-ping, ZHANG Dai-ting, GUAN Min-hui, HE Fei-long, YANG Qian, GUO Xiao-feng
2014, 41(5):  23-28. 
Abstract ( 248 )  
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The study was aimed to highly express Cap protein of porcine torque teno virus type 2 in E.coli, and determine the optimal expression conditions and purify it. ORF1 gene was amplified by PCR and cloned into the prokaryotic expression vector pGEX-4T-1 to construct recombinant plasmid pGEX-4T-1-ORF1, which was identified by restriction and sequence analysis, then transformed into E.coli BL21(DE3) and expressed under the induction of IPTG. The expression product was identified by SDS-PAGE and Western blotting. The induction time, induction temperature and IPTG concentrations were optimized. Both PCR and restriction analysis proved that the recombinant plasmid was constructed correctly. The best induction condition was 37 ℃ for 5 h, whereas IPTG concentration had no significant impact on protein expression. The recombinant fusion protein pGEX-4T-1-ORF1 was approximately 80 ku in a form of inclusion body, which was recognized specially by mouse anti-GST monoclonal antibody. It was the first time to use cutting the gel slice’s method for purification of TTV2 Cap protein, which was cost-effective and convenient. Recombinant plasmid pGEX-4T-1-ORF1 was constructed and the Cap protein was highly expressed in E.coli which provided antigen for indirect ELISA and preparation of monoclonal antibodies.
Research Progress on Major Histocompatibility Complex(MHC)Gene
KANG Xiao-zhen, Erdemtu, JIANG Jian-qiang, CHEN Ze-ming, LIU Tu-ya, Yitegeltu, Sharhu, TU Ya
2014, 41(5):  28-33. 
Abstract ( 400 )  
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Major histocompatibility complex (MHC) is widely found in vertebrate. Because of its high polymorphism, MHC genes have become the focus of attentionhop in relation to genetics and breeding and disease-resistance etc. This article make a retrospective conclusion on the major histocompatibility complex genes polymorphism and the relationship between this gene and disease, and other aspects on the domestic and foreign researchers’ papers in recent decades.Some of these contents as a reference for the next correlation experiments.
Molecular Characterization Analysis of BibA Gene in the Isolate LZQ07006 of Serotype Ⅰa Streptococcus agalactiae from Bovine
CHANG Rui-xiang, WANG Xu-rong, WANG Lei, ZHANG Jing-yan, QIN Zhe, KONG Xiao-jun, WANG Xiao-wu, LI Jian-xi, YANG Zhi-qiang, WANG Xue-zhi
2014, 41(5):  34-38. 
Abstract ( 202 )  
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This study was aimed at analyzing the molecular characteristics of BibA gene and its expression protein, providing theoretical basis for the further study on the expression and immunogenicity of BibA protein. The BibA gene of the isolate LZQ07006 of serotype Ⅰa Streptococcus agalactiae from bovine was amplified using the PCR method, cloned into pMD20-T vector and then sequenced. The molecular characteristics of BibA gene and its expression protein were analyzed using a variety of biological software. The results showed that BibA gene of LZQ07006 was 1185 bp and did not appear the gene deletion, and encoded 395 amino acids. The nucleotide sequence homology of BibA gene derived from LZQ07006 was 45.0% to 99.8% and the amino acid homology was 12.3% to 99.5% among the corresponding sequences from different serotypes Streptococcus agalactiae published in GenBank. The derived amino acid showed that the BibA protein was a stable outer membrane protein, which was secretion protein and hydrophilic highly. The polypeptide contained a signal peptide consisting of 32 amino acids, with cleavage site between 32th and 33th amino acids. In conclusion, it was necessary to study the BibA protein in bovine Streptococcus agalactiae for application in diagnosis or prevention of dairy cow mastitis.
Establishment of TaqMan Real-time PCR for Detection of Porcine Circovirus Type 2
GUO Hui-juan1,2,LI Xiu-li2,ZHANG Guo-wei2,YAN Ming-hua2,LI Hai-hua2,LI Fu-qiang2
2014, 41(5):  39-45. 
Abstract ( 236 )  
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The specific primers and probe were designed according to the nucleotide sequence of porcine circovirus type 2 (PCV2) available in GenBank,CAP gene was amplified by PCR, cloned into the pMD18-T vector and screened positive plasmid standards.By optimization of reaction conditions,we established a TaqMan Real-time PCR method for detection of PCV2. The results indicated that the method was specific, and swine influenza virus (SIV), porcine reproductive and respiratory syndrome virus (PRRSV), pig pseudorabies virus (PRV) and classical swine fever virus detection results were negative. The detection limit of the assay was 4.53×102 copies/μL of plasmid DNA,100 times higher than that of the routine PCR. The standard curve displayed a linear range from 102 to 109 copies/μL and it had a good reproducibility.
Isolation and Identification of Avian Pasteurella Strain Pm-HB
YIN Xiu-feng, WANG Yuan-zhen, DING Mei-juan, XU Xiu-mei, HUANG Xian-ming, ZHANG Hai-tao, MAO Huo-yun,ZHANG Xiao-fei
2014, 41(5):  45-48. 
Abstract ( 268 )  
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A poultry farm suffered an unknown disease suspected as fowl cholera in Hubei province. A bacterium strain was isolated from liver and lung samples. The isolates was identified as Pasteurella multocida by morphological and cultural characteristics, biochemical properties, and sequence alignment based on 16S rRNA, and named as Pasteurella multocida -HB strain. 30 day old SPF chickens were used to test the pathogenicity and the result showed the strain was deadly to chickens at very low dose, 10 CFU per chicken.
Construction of Calves Oral Mucosal Epithelial Cells T7 Phage Display Library
HAN Yan-yan,SONG De-guang
2014, 41(5):  49-52. 
Abstract ( 142 )  
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To screen receptor of vesicular stomatitis virus (VSV), the calves oral mucosal epithelial cells T7 phage display library was constructed. The total RNA of calves oral mucosal epithelial cells was extracted using Trizol reagent. Then, mRNA was separated and purified by mRNA isolation kit and the ds cDNA were synthesized by reverse transcription. ds cDNA ends were ligated EcoR Ⅰ/Hind Ⅲ linkers, then ds cDNA were digested by EcoRⅠ/Hind Ⅲ, so that we got ds cDNA ends containing EcoR Ⅰ/Hind Ⅲ sticky ends. All digested ds cDNA were separated by Mini Column, only ds cDNA fragments more than 400 bp were collected. Then, the collected ds cDNA were ligated into the T7 Select 10-3b vector. After packaging in vitro, the recombinant T7 Select 10-3b vectors were transformed into BLT5403, so we could construct the T7 phage display library. The results indicated that the titer of un-amplified library was 1.3×107 PFU/mL, and the recombination rate was 95.8%. After amplification, titer of library was 2.6×1010 PFU/mL. Randomly picked 100 plaques were identified by PCR, 95% of the inserted cDNA fragments were longer than 400 bp in length. These results indicated that the calves oral mucosal epithelial cells T7 phage display library was successfully constructed.
Cloning and Expression of Sus scrofa Sirtuin 3
WANG Hou-guang,MA Miao-peng,HUANG Kui-ying,LI Hua-zhou, ING Fei-ping,WANG Wei-fang,XIA Feng-geng,LUO Meng-xiao, ANG Jun,CAI Hai-ming,SHI Ju-qing,HUANG Chao-yuan,CHU Pin-pin,DONG Jian-ming,ZHU Hong-xia
2014, 41(5):  53-57. 
Abstract ( 170 )  
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The study was aimed to obtain recombinant Sus scrofa sirtuin 3 (SIRT3) in prokaryotic system and preliminarily evaluate its antigenicity and specificity. The fragment which included the SIRT3 complete coding sequence was obtained by RT-PCR and cloned into pET-28a(+) vector, then the recombinant vector was transformed into E.coil Rosetta (DE3) to induce, express and purify, the antigenicity of recombinant SIRT3 was identified by Western blotting. The double enzyme analysis and sequencing proved that recombinant plasmid pET28a-SIRT3 was constructed correctly. The expressed recombinant protein, with a relative molecular mass of about 39 ku, was recognized by positive serum antibody. The result showed that the SIRT3 was successfully expressed in E.coil, which would provide a useful tool for designing an in-depth investigation of the role of SIRT3.
Establishment of Indirect Hemagglutination Assay for Antibodies Detection of APP
ZHENG Jiao-que,LIU Wan-jing,DENG Shun-zhou,WANG Ping,HE Hou-jun,WU Xiang-dong
2014, 41(5):  57-61. 
Abstract ( 208 )  
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In order to understand the infection status of Actinobacillus pleuropneumoniae in Jiangxi province, we developed indirect hemagglutination assay (IHA) for detection of antibody by using APP isolates of Jiangxi province. IHA consisting of glutaraldehyde-tannic-fixed rabbit red blood cells (RRBC) coated with the supernatant of repetitive freeze-thawed and sonicated APP was established. The positive sera of hog cholera virus, porcine reproductive and respiratory syndrome virus, pseudorabies virus, Escherichia coli, Salmonella, Haemophilus parasuis showed negative results. Different batches of IHA antigen showed good repeatability. The results showed that the specificity and repeatability of IHA were good, which created the foundations for the serological survey.
Cloning and Sequences Analysis of gB and gD Gene of Porcine Pseudorabies Virus Fujian Strain
ZHANG Xin-ping, LIU Jian-kui,WEI Chun-hua,DAI Ai-ling,LI Xiao-hua,YANG Xiao-yan
2014, 41(5):  62-65. 
Abstract ( 237 )  
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The gB and gD antigen epitope genes of pseudorabies virus (PRV) Fujian strain(LY strain) were amplified by PCR, the paraments of gB and gD were cloned into pMD18-T easy and sequenced,getting about 578 and 653 bp DNA fragments, then the gB and gD gene of PRV LY strain were cloned and sequenced. Compared with gB and gD antigen epitope genes in GenBank, the sequence analysis determination show that the gB and gD gene of LY strain shares 74.9% to 98.8% and 97.2% to 98.1% nucleotide homology, 62.9% to 96.4% and 94.5% to 96.8% amino acid sequence homology with other strains. The evolutionary analysis indicated that LY strain had the same evolutionary branch with domestic epidemic strains, and had a close relationship with the Ea strain (Hubei), LA strain (Shandong) isolated from domestic strains other than from abroad.
Development and Application of Pyrosequencing Method for Detection of BLAD
ZHOU Li,JIA Guang-le,LIN Xiang-mei,LIAO Juan-hong
2014, 41(5):  66-70. 
Abstract ( 224 )  
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Bovine leukocyte adhesion deficiency (BLAD) was an autosomal recessive lethal genetic disease, the caused BLAD had a bad effect on the bovine production trait and dairy economic returns. In order to establish a rapid and reliable method for BLAD detection, in this experiment, according to BLAD CD18 gene sequences published in GenBank, PCR primers were designed. After PCR amplification and purification, three standard plasmids including genotypes A/A, A/G and G/G were constructed. Pyrosequencing method were established by using these constructed standard plasmids. Then, the bovine blood samples were detected by the method. The results were compared with Sanger sequencing results to analyze and verify the accuracy of the test results. 300 bovine blood samples of BLAD were detected by pyrosequencing method,the results showed A/A genotype were 294 cases, accounting for 98% of the overall proportion; A/G genotype were 6 cases, accounting for 2%; and the results of two detection methods were consistent. It showed that pyrosequencing method to detect BLAD were specific, sensitive and rapid, and the test results were accurate and reliable. It was suitable for kit development and applied in clinical diagnosis of BLAD.
Development and Preliminary Application of Real-time PCR Assay for Detecting Quantitation of PRRSV in Vaccine
MA Zhi-liang, SUN Ting-ting, YANG Jie, FAN Bin, LIU Xu-chuan, ZHANG Yi-fang
2014, 41(5):  71-76. 
Abstract ( 266 )  
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To establish a rapid, sensitive and specific method for evaluation of porcine reproductive and respiratory syndrome (PRRS) attenuated vaccine, two pairs of primers were designed and synthesized according to porcine reproductive and respiratory syndrome virus (PRRSV) genes in GenBank. PRRSV genes were obtained by RT-PCR and constructed recombinant plasmids pMD19-T-PRRSV. SYBR GreenⅠ method was adopted to improve the Real-time PCR and the method was tested for specificity, repeatability and stability. The results shwed that there was a good linear correlation coefficient (R2=0.9989) as Ct values ranged from 7.43×100 to 7.43×108 copies/μL. Using this method to test PRRS vaccines from different companies, we found that there were significant differences (47.9 times) in the levels of virus among six companies’ vaccines. The Real-time PCR for detecting PRRS vaccine virus can be used for evaluation of vaccine during production process and animals immunization.
Study on Differential Expression of Keratin Associated Protein 8-1 Gene of Different Varieties of Ovis aries in Skin and Hair Follicle
ZHANG Gui-shan,XUE Jing-long,SUN Li-min,ZHAO Jia,XU Jing,JIANG Huai-zhi
2014, 41(5):  76-79. 
Abstract ( 239 )  
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Relative expression of keratin associated protein 8-1 (KAP8-1)gene was determined in different vrieties of Ovis aries skin and hair follicle so as to identify the regulation of KAP8-1 gene on skin and hair follicle development and wool quality difference. Using Real-time PCR SYBR GreenⅠ, Ct of expression of KAP8-1 gene was determined and relative expression of KAP8-1 gene was analyzed by 2-△△Ct. The results showed that there was KAP8-1 gene expression in there different crossbred varieties of Ovis aries (South African mutton Merino sheep(♂)× Northeast Fine wool sheep(♀),South African mutton Merino sheep(♂)× Small-tailed Han sheep(♀),Dorper sheep(♂)× Small-tailed Han sheep(♀)) skin and hair follicle. The ratio of relative expression was 7.56∶2.19∶1. In a word, KAP8-1 gene played a role in regulating skin and hair follicle development and wool quality difference,with difference among different varieties.
Construction and Identification of Outer Membrance Protein H (OmpH) of Coxiella burnetii
QIAO Hai-yan,JIA Guang-le,LIN Xiang-mei, ZHANG Ying,LIAO Juan-hong
2014, 41(5):  80-84. 
Abstract ( 257 )  
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This study was amied to construct and identify the prokaryotic expression plasmids of outer membrance protein H (OmpH) of Coxiella burnetii and analysis its immunogenicity. Amplified OmpH gene from the Nine Miles by PCR, the results showed that the OmpH included 798 bp. Cloned OmpH gene into prokaryotic expression vector pQE-30 after identified by PCR,enzyme digestion and sequence. Constructed the recombinant plasmid pQE-30/OmpH,and then the plasmid was transformed into BL21(DE3) competent cells and induced by IPTG. The results showed that pQE-30/OmpH recombinant proteins were 25 ku by SDS-PAGE, and Western blotting assay indicated that the recombinant protein could express OmpH protein with significant antigencity.
Expression of Nonstructural Protein NS3 and Establishment of ELISA Method of Bovine Viral Diarrhea Virus
FAN Qing, XIE Zhi-xun, XIE Zhi-qin, LIU Jia-bo, PANG Yao-shan, DENG Xian-wen, XIE Li-ji, LUO Si-si
2014, 41(5):  85-89. 
Abstract ( 286 )  
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An indirect enzyme-linked immunosorbent assay (ELISA) method was developed to detect antibody against bovine viral diarrhea virus (BVDV). The nonstructural protein NS3 gene of BVDV was subcloned into prokaryotic vector pET-32a to express. The recombinant protein NS3 of BVDV was used as the coating antigen for the ELISA method, and the reaction conditions were optimized. The specificity, sensitivity and reproducibility all showed good results. Furthermore, four hundred and seventy five bovine serum samples, which had been collected from several cattle farms of Guangxi, were detected by the NS3-ELISA, and the positive rate of BVDV antibody was 24.8%. Compared with commercial kit, the coincidence was 97%. The result suggested that NS3-ELISA was a convenient, rapid method for the diagnosis, antibody level surveillance and epidemiologic investigation of BVDV.
Improvement of Ultrasonic Power on DNA Extraction Method from Milk Bacterial
YAN Xiao-fei,LIN Shan,TONG Ting,WANG Jin-quan,HUANG Ke-xin,DONG Xuan,YUE Mei-ting,YE Dong-dong,YANG Bin
2014, 41(5):  89-93. 
Abstract ( 178 )  
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All dairy cows’ milk were collected from Urumqi Yanerwo cow farm in May 2013.Treated those samples with different ultrasonic power (0,100,120,140,160 W) after mixed,then extracted the milk bacterial DNA with the CTAB and SDS methods,respectively,according the DNA concentration, purity and bacterial universal primers of 16S rDNA V6-V8 region to judging the quality of DNA,finally screen out the best DNA extraction method.The results showed that the DNA which was extracted by the CTAB and SDS methods after treated with ultrasonic power 0 W for 10 min couldn’t amplify the V6-V8 of 16S rDNA.Meanwhile,the V6-V8 of 16S rDNA could been amplified with the other ultrasonic power (100,120,140,160 W),and the result showed multifariously.The highest DNA density was 203.35 μg/mL,which was extracted by the CTAB method and treated with ultrasonic power 140 W.Results indicated that using the CTAB method after treated those samples with ultrasonic power 140 W was the best way to get the significantly higher DNA density,this method could further satisfy the requirements of studying about microbial populations of mastitis from the molecular level.
Cloning and Prokaryotic Expression of Gonadotropin Releasing Hormone 1 Gene in Jinghai Yellow Chicken
ZHANG Tao, WANG Jin-yu ZHANG Gen-xi WANG Wen-hao, WEI Yue, CHEN Xue-sen, TANG Ying, WANG Yong-juan3
2014, 41(5):  94-98. 
Abstract ( 101 )  
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Based on the published mRNA sequence of Gallus gallus gonadotropin releasing hormone 1 (GnRH1) gene, a pair of primers was designed to clone the GnRH1 gene coding sequence of Jinghai Yellow chicken by RT-PCR. GnRH1 gene was cloned in to pET32a vector for expression in E.coil BL21 and bioinformatics analysis. Finally, a 352 bp gene was cloned, which contained CDS region, promoter region and part of the 3′region. It was found that the GnRH1 gene nucleotide sequence shared 93%, 81%, 54%, 58%, 61%, 76%, 76%, 59%, 76% and 66% identities with Meleagris gallopavo, Columba livia, Homo sapiens, Bos taurus, Babirussa, Capra hircus, Ovis aries, Pantholops hodgsonii, Equus caballus and Rattus norvegicus, respectively. Restriction and sequencing analysis confirmed that prokaryotic expression vector pET32a-GnRH1 was successfully constructed. SDS-PAGE analysis showed that the recombinant plasmid was successfully expressed in E.coli BL21 and the molecular weight was approximate 25 to 28 ku. The expression quantity in supernatant was higher than that in the precipitant. Western blotting analysis confirmed that the protein expressed in E.coli BL21 was the interest protein and expressed in soluble form with high activity. The results showed that GnRH1 gene of Jinghai Yellow chicken was cloned and GnRH1 prokaryotic expression system was constructed successfully. The interest protein was expressed successfully, which laid the foundation for further studies.
Establishment and Application of Method for PCR Detection of Beef Ingredient in Meat Products
XIONG Rui,GUO Feng-liu,LIU Xiao-hui,ZHAO Tong-xin,WANG Na,YAN Hong
2014, 41(5):  99-102. 
Abstract ( 209 )  
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This study established a PCR method for detection the beef ingredient in meat products according to the bovine specific mitochondrial DNA fragment,and 67 meat products of cow were detected using PCR.The results showed that a specific amplification fragment of the expected size was detected and demonstrated,buffalo,yak,cow and bovine meat were positive using this method,whereas the 14 kinds of animal meat such as sheep, equine, canis, donkey, rabbit and duck were negative,the detection limit was 53.2 fg/μL DNA.The positive rate of 67 meat products of cow was 100%.The results indicated that the method was quickness,convenience,sensitivity and specificity,and can be used for the identification of bovine source of beef products components.
Truncated Expression and Activity Detection of Porcine Epidemic Diarrhea Virus S Protein,and Preparation of its Polyclonal Antibody
WANG Ling-xiong,ZHANG Jie
2014, 41(5):  103-107. 
Abstract ( 232 )  
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In order to highly express S protein of porcine epidemic diarrhea virus (PEDV) and prepare its specific polyclonal antibody,the main antigen region of S gene was amplified by PCR method,subcloned into pET30a(+) prokaryotic expression vector,transformed into BL21(DE3) expression bacteria,and induced by IPTG.The recombinant S protein was purified by affinity chromatography,its activity was detected by Western blotting,New Zealand White rabbits were immuned using the recombinant S protein to prepare polyclonal antibody,and detection of the antibody titer by indirect ELISA was conducted. After BamHⅠ/HindⅢ double enzyme digestion, we obtained pET30a-S recombinant plasmid,with induction of 1 mmol/L IPTG for 4 h,the recombinant S protein were expressed in inclusion body form,after purification and Western blotting,the protein showed good activity and specificity,antibody titer of polyclonal antibody against S protein was 1∶25600 detected by indirect ELISA. In this study PEDV S protein was successfully truncated expressed and its polyclonal antibody was also prepared,which layed a foundation for further development of rapid immunology detection kit of porcine epidemic diarrhea,and provided a condition for the study of structure and function of S protein and identification of the antigenic epitopes.
Comparative Analysis of Rumen Degradation Kinetics of Soybean Meal and Cottonseed Meal in the Ruminant
CHEN Ao-dong,KONG Ping,ZHAO Wei-li,HE Jia-wen,LIU Chen-li,GAO Wei
2014, 41(5):  108-112. 
Abstract ( 161 )  
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A comparative experiment was conducted to assess the rumen effective degradability of dry matter (DM),organic matter (OM) and crude protein (CP) of soybean meal and cottonseed meal in the ruminant and effective lysine content before and after rumen degradation in this study.Three rumen cannulated Kazakh wethers weighed in average (28.0±2.97)kg body weight were housed in metabolic crate singly.The daily diet was composed of cottonseed hull 200 g,concentrate 500 g,wheat straw 500 g,and the daily refusals were less than 10%.The effective degradability of DM,OM,and CP in soybean meal and cottonseed meal as well as rumen retention time were evaluated by the standard nylon bag technique.The effective lysine content in both feeds were determined by the dye binding method.The results showed that under the conditions of solid phase in the rumen digesta average outflow velocity of 1.84%/h,the rumen degradability and effective degradability of DM,OM and CP in soybean meal were higher than that in cottonseed meal,and rumen retention time of soybean meal was shorter than that of cottonseed meal.Effective lysine content of soybean meal after 16 h rumen incubation increased slightly,while no significant changes for cottonseed meal.Results of this study provide some important parameters for further evaluating intestinal digestion of protein and amino acids for future.
Effect of Different Feeding Conditions on Daily Gain and Serum Biochemical Indexes in Youth Xinjiang Brown Cattle
JULAITI-Abuduwaili,REN Wan-ping,ZHANG Hao,LV Qiang,YU Xiong
2014, 41(5):  112-115. 
Abstract ( 210 )  
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The experiment was aimed to study the effect of different feeding conditions on the changes of daily gain and serum biochemical indexes level in different period of growing youth Xinjiang Brown cattle in growth process.Selected 68 16-months-old Xinjiang Brown cattle,and divided them into scattered group and tethered group.In the experiment,we measured daily gain and tested blood samples contents of albumin (ALB),globulin (GLB),total protein (TP),blood urea nitrogen (BUN),glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) in October,November and December 2012,and January,February 2013.The results showed that the scattered group daily gain was extremely significantly higher than that of tethered group in December 2012,January and February  2013 (P<0.01).The serum content of ALB in scattered group was extremely significantly higher than that of tethered group in January 2013 (P<0.01).The serum content of GLB in scattered group was significantly higher than that of tethered group in October,November and December 2012 (P<0.01, P<0.05).The serum content of TP in scattered group was significantly higher than that of tethered group in December 2012, January and February 2013 (P<0.01, P<0.05).The serum content of BUN in scattered group was extremely significantly higher than that of tethered group in October and November 2012 (P<0.01).The serum content of GOT in scattered group was significantly higher than that of tethered group in November,December 2012 and January 2013 (P<0.01, P<0.05).The serum content of GTP in scattered group was significantly higher than that of tethered group in January and February 2013 (P<0.05). In conclusion,under different feeding mode to fattening youth cattle,compared with tethered feeding mode,ad libitum scattered feeding mode could significantly improve the youth cattle’s daily gain,and also accelerate the digestion and metabolism,and various nutrients that forage provided can effectively be absorbed by the body and the protein synthesis.
Research Progress on Amino Acid Nutritional in Ruminants
HUANG Jian,YANG Fu-he,LI Guang-yu,BAO Kun,WANG Kai-ying
2014, 41(5):  116-120. 
Abstract ( 209 )  
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Protein is the basic nutrition of life,amino acid is the basic composition unit of protein, the main function of the protein is reflected by amino acid.In recent years,the research about amino acids nutrition of ruminant has made great breakthrough and development.In this paper,the source-diversity,limitation and balance of amino acid were reviewed.
Reaserch Progress on Biology Function of Lutein and its Application in Feed
YANG Yu-jiang,ZHANG Hui,CUI Huan-zhong,ZHENG Xin
2014, 41(5):  121-124. 
Abstract ( 204 )  
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Lutein is a carotenoid distributed widely in nature. More and more attentions have been paid to the immunomodulatory function and antioxidant activity of lutein. Numerous researches on lutein were conducted to enhance the color of animal. And lutein was gradually tried to add an application in feed. In this paper, the biology function of lutein and the study of its application in feed were summaried briefly.
Research Progress on Immunoglobulin of Yolk and its Application in Diseases Prevention and Feed
ZHANG Hui,CUI Huan-zhong,YANG Huan,YANG Yu-jiang,CHANG Xiao-bo,YANG Qian,YANG Liang,YIN Jian,ZHENG Xin
2014, 41(5):  124-127. 
Abstract ( 239 )  
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Immunoglobulin of yolk has many significant advantages,such as high stability,easy preparation,high yield and low cost.They have been used widely in animal diseases prevention and cure. With antibiotics disabled,immunoglobulin of yolk will gradually become a new feed additive in feed applications.In this paper,the formation and character of immunoglobulin of yolk and its application in diseases prevention and feed were reviewed briefly.
Comparative Study on the Histological Traits and Physicochemical Properties of Muscles in Small-tail Han Sheep and Crossbred Hanper Sheep
ZHAO Yan-jiao, SUN Hai-yun, CUI Ya-li,CHANG Qing-ling,DUN Wei-tao,SUN Hong-xin,CHEN Xiao-yong,TIAN Shu-jun
2014, 41(5):  128-132. 
Abstract ( 203 )  
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Well-fed Small-tail Han sheep and crossbred Hanper sheep were selected respectively, and the longissimus dorsi, gluteus medius, biceps femoris, triceps brachii muscle samples were collected. We detected the myofiber cross-sectional areas, diameters, crude fat content and the amino acids composition in muscles. The results showed that the diameter and cross-sectional area of longissimus dorsi in Hanper sheep were significant less than that in Small-tail Han sheep (P<0.01). The diameter and cross-sectional area of biceps femoris in Hanper sheep were greater than that in Small-tail Han sheep (P<0.05).Crude fat contents of longissimus dorsi and biceps femoris in Hanper sheep were higher than that in Small-tail Han sheep (P<0.01, P<0.05). There was no difference in crude fat contents of triceps brachii and gluteus medius between the 2 species (P>0.05). Taurine content in Small-tail Han sheep was greater than that in Hanper sheep (P<0.01). The other 17 kinds of amino acids between the 2 species were not difference (P>0.05). Five kinds of flavor amino acids contents in Hanper sheep were greater than that in Small-tail Han sheep. The six kinds of essential amino acids contents except lysine and tryptophan in Hanper sheep were higher than that in Small-tail Han sheep.
Extraction and Identification of Lipopolysaccharide and O-polysaccharide from Chicken S.boydii Type 3
WEI Ya-peng,GUO Hui-juan,ZHANG Mei-ling,CHANG Hong-tao,LIU Hong-ying, WANG Chuan-qing,WANG Xin-wei,YANG Xia,CHEN Lu,ZHAO Jun
2014, 41(5):  133-136. 
Abstract ( 179 )  
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To obtain LPS and its subunit O-PS with high purity from chicken S.boydii type 3, LPS from chicken Shigella was extracted with modified hot phenol-water method, high purified LPS was obtained through sephadex gel chromatography, O-PS was obtained by combination with acid hydrolysis method and sephadex gel chromatography. Biochemical assay showed that the purified LPS and O-PS with high purity, rabbit ideal loop experiment demanstrated that there was high activity in purified LPS.The result indicated that extracted LPS and O-PS could be used in specific pathogenesis research and developed as effective vaccine candidates.
Effect of Oleic Acid and Stearic Acid on the Expression of Fatty Acid Binding Protein 3 and Secretion of Lipid Droplets in Dairy Cow Mammary Epithelial Cells
LIANG Meng-yao, GAO Xue-jun, HOU Xiao-ming, LI Qing-zhang
2014, 41(5):  137-141. 
Abstract ( 193 )  
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To study the effect of oleic acid and stearic acid on the expression of fatty acid binding protein 3 (FABP3) in dairy cow mammary epithelial cells, qRT-PCR was used to detect the expression of genes related with milk fat synthesis. The optimal concentrations and time of oleic acid and stearic acid were 75 μmol/L 12 h and 100 μmol/L 36 h, respectively. Western blotting and immunofluorescence were used to detect the expression of FABP3 and secretion of lipid droplets. These data provided further experimental evidence for studying the effect of FABP3 on the milk fat synthesis signal transduction pathway.
Isolation and Identification of the Cellulose-degradingBacteria Strains from Sheep Rumen
NA-Rentuya,ZHOU Fei-fan,YANG Jin-li,HOU Xian-zhi,WANG Hai-rong
2014, 41(5):  142-146. 
Abstract ( 211 )  
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Three strains of anaerobic bacteria were isolated from the rumen of the Mongolian sheep. The bacterial cellulose degradation detection test showed that all of the bacteria have fiber degradation ability.All of the bacteria were gram-positive bacteria according to the gram stain test.After the physiological and biochemical test, and determination of 16S rDNA gene sequences and the homology analysis,the three strains of bacteria turned out to be Streptococcus bovis,Streptocococcus infantarius and Enterococcus faecium,and were named as E1,E2 and E3,respectively.
Study on Activities and Distribution of Bacterial Colonization in the Intestine of Shaanxi Qingjiaoma Chickens
WANG Jin,HOU Tian-gao,ZHAO Na-na,CHEN Wen-qiang,DENG Bai-wan,HAN Shi-hao2,MU Na-lin4
2014, 41(5):  146-149. 
Abstract ( 205 )  
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In the present study,we determined activities and distribution of bacterial colonization in the intestine of Shaanxi Qingjiaoma chickens.The microorganisms were cultivated and counted by gradient dilution and tissue culture method.The results showed that the middle and downside portions of small intestine contained Bacillus at a concentration of 6.5×105 CFU/mL. Saccharomyces was found in cesium at a concentration of 6.8×104 CFU/mL.Particularly,the downside portion of duodenum contained Lactobacillus at a concentration of 8.6×104 CFU/mL.Clearly,our results indicated that the Bacillus were rich in the intestine of Shaanxi Qingjiaoma chickens.In contrast,Saccharomyces and Lactobacillus as probiotics were less rich in the intestine.
Study on Characteristic of Bartonella infecting Macrophage in vitro
BAI Ya-jie,HONG Jie-hua,YUAN Cong-li
2014, 41(5):  150-154. 
Abstract ( 161 )  
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Bartonella was fastidious gram-negative bacterium, which could cause cat-scratch disease and bacillary angiomatosis in human. Histopathology studies showed that lots of free Bartonella in infected lymph nodes, which indicated that Bartonella was able to escape from host innate immunity or phagocytosis. However, the interaction between Bartonella and macrophage had not been well demonstrated. Here, intracellular survivals of Bartonella tribocorum in J774A,RAW267.4 and C57 mouse peritoneal macrophage were investigated. Gentamycin protection assay demonstrated Bartonella could replicate and survive in 48 hours post infection in J774A, RAW264.7 and C57 mouse peritoneal macrophages, even in LPS induced macrophages.
Study on Pseudolaric Acid B Inhibited Cell Growth through P53, Caspase 8 and Caspase 9 in MCF-7 Cells
TIAN Jie,REN Pei-you,LI Rui,LIU Chun-yu
2014, 41(5):  155-158. 
Abstract ( 169 )  
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The assay was aimed to study the inhibitory mechanism of pseudolaric acid B (PAB) in MCF-7 cell line. MCF-7 cell was human breast cancer cell lacking of caspase 3 function. In this study, it was found that PAB activated caspase 8 and caspase 9 to induce cell apoptosis. Meanwhile PAB up-regulated P53 protein expression. Therefore PAB induced MCF-7 cell apoptosis through increasing P53 protein expression and promoting the activation of caspase 8 and caspase 9.
Molecular Regulation Mechanism of Synthesis of Lactoprotein on Dairy Cattle
LIN Zhong-li,JIANG Ming-feng,REN Hong-hui
2014, 41(5):  158-162. 
Abstract ( 176 )  
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Lactoproteins are main components of the milk which include almost all of the essential amino acids and has very high nutritive value. The synthesis of lactoproteins are regulated by amino acid transporters,Janus tyrosine kinase 2 signal transducer and activator of transcription 5 (Jak2-Stat5) and mammalian target of rapamycin (mTOR) pathway. Amino acid transporters provide materials for the synthesis of milk protein. The Stat family are signal transducer factors to regulate the expression of lactoprotein, especially Stat5. The mTOR pathway can integrate hormone such as prolactin and insulin cytokine such as EIF4E and AKT, to regulate the synthesis of milk protein. It can be deduced the study of mechanism of milk protein synthesis is very important to improve the nutritive value of milk.
Advances on the Studies of Antimicrobial Peptide’s Active Mechanism and its Molecular Design
ZHANG Qing-hua, WANG Qing, SHANG Tian-tian, ZHAO Zhi-yu, XU Yan-zhao, HU Jian-he
2014, 41(5):  163-167. 
Abstract ( 166 )  
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The antimicrobial peptides (AMPs) are a class of defense peptides that widely exist in nature, which are important components of the innate immune system. Compared to antibiotics, AMPs have some good quality, such as small molecular weight, good water-solubility, thermal stability and unique antibiotic mechanism. They can also restrain virus, fungus and even tumour. With the appearance of multidrug-resistant (MDR) bacteria and the increasing of new AMPs, the study focuses on it’s mechanism of biological activity. This paper summarizes the biological activity mechanism and molecular modification of AMPs, so as to provide theory basis for the molecular modification and the application of AMPs.
Histological Observation of Abnormal Laxity on the Nape of the Neck in Sika Deer by Light Microscope
WEI Hai-jun, CHEN Xiu-min, ZHANG Jia-bao
2014, 41(5):  167-170. 
Abstract ( 173 )  
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In this study, elastic fibers and collagen fibers of normal sika deer and abnormal laxity on the nape of neck were observed by Weigert’s-Van Giesson staining and Masson staining for investigating the pathogenesis of cutis laxa in sika deer. The results showed that there were many elastic fibers in the control group arranged regularly dense, wavy, stained uniform in the reticular dermis;However, there were few elastic fibers perpendicular to the epidermis in the papillary dermis. Cuis laxa sika deer showed inelastic fibers in the papillary dermis (No. 0 and 79) or severe hypoplasia in the reticular dermis, appeared accumulation, breakage (No.33 and 79), disorganized, irregular shape (No.0). Collagen fibers of the normal deer were distributed multi-layered regularly in the dermis. Collagen bundles of collagen fibers in cutis laxa was markedly thick, reduced, arranged disorderly and severe sparse compared to the normal collagen fibers. It was exactly similar to the histology of cutis laxa skin in human.
Research Progress of Relationship between P21 Gene and Mammalian Regeneration
GUO Qian-qian, WANG Da-tao, CHU Wen-hui, ZHAO Hai-ping, SUN Hong-mei, LI Chun-yi
2014, 41(5):  171-176. 
Abstract ( 167 )  
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Animals’capability of regenerating multiple tissue types, organs, and appendages after injury are common among invertebrates and urodele amphibians, but notably such regenerative capacity is rare in mammals. Recently, accumulating evidence has proved that mammals may also have regenerative property with the discovery of MRL mouse, and it brings new hope for clinical therapies. The regeneration of appendages depends on cells proliferation, at the same time, cells proliferation depends on cell cycle, so as one of the cell cycle regulatory proteins, we found the deletion of P21 gene could actually result in full MRL epimorphic regeneration phenotype in a new study, providing a firm link between p21 protein and tissue regeneration. This article reviewed the fields of tissue regeneration and p21 protein, and introduced the relationship between tissue regeneration and p21 protein.
Effect of Valproic Acid on in vitro Developmental Competence of Handmade Cloned Pig Reconstructed Embryos
XIE Bing-kun, LV Ling-yan, LU Xing-rong,CUI Kui-qing, SUN Jun-ming, QIN Zhao-xian,SHI De-shun, LIU Qing-you
2014, 41(5):  180-186. 
Abstract ( 168 )  
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To explore the effects of histone deaceylase inhibitors valproic acid (VPA) affecting the development potency of handmade cloned (HMC) embryos, porcine HMC reconstructed embryos were cultured for 24 h with different concentration (0,25,50,75 and 100 nmol/L) of VPA, and the blastocyst development rate, inner cell mass (ICM) cells number and histone acetylation level were compared. The results showed that reconstructed embryos were cultured in PZM-3 containing 50, 75 nmol/L VPA for 24 h, the frequency of the rate of cleavage and blastocyst formation were significantly higher in embryos treated with VPA than 0, 25, 100 nmol/L VPA treatments (P<0.05). In addition, 50 nmol/L VPA treatment significantly increased the number of inner cell mass (ICM) cells compared with the other groups (P<0.05). The AcH3K14 level in embryos treated with VPA was higher than the control group and parthenogenetic activation group. In conclusion, epigenetic modification VPA could enhances the rate of cleavage, blastocyst formation and the ICM cells number of porcine HMC reconstructed embryos, which mean improve the developmental potency of HMC embryos.
Research Progress on Bovine IGF-1 Gene and Association of its SNPs with Milk Production Traits
HUO Qiu-hong,YANG Fang,TANG Yi-bo,JIANG Ming-sheng
2014, 41(5):  186-189. 
Abstract ( 221 )  
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Milk production traits were the important economic traits in the cow production,and were determined by many genes with little effect.Insulin-like growth factor-1 (IGF-1) gene as one of many candidate genes for milk production traits,the mutation of IGF-1 gene might result in the change of IGF-1 structure,and the change in milk production traits.The author reviewed the structure and function of IGF-1 gene and research progress on its single nucleotide polymorphisms and association with milk production traits in bovine,aiming to provide a theoretical reference for relevant studies on milk production traits.
Analysis of SRY Gene Polymorphisms and Patrilineal Phylogenetic Relationships for Bos grunniens and other Bos species
CAI Xin, ZHAO Fang-fang, SUN Lei
2014, 41(5):  190-195. 
Abstract ( 262 )  
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In order to analyze SRY gene polymorphisms and patrilineal phylogenetic relationships for Bos grunniens and other Bos species, SRY gene were amplified and cloned from genomic DNA of five Maiwa yaks, and SRY gene sequences of other Bos species and Bison bonasus were retrieved from GenBank. Polymorphism analyses were performed to examine the variations of SRY gene and the sequences encoding HMG-box among Bos species. The phylogenetic tree and Median-Joining network of Bos species were constructed based on SRY gene sequences, exhibiting the patrilineal phylogenetic relationships for Bos species. The nonsynonymous mutation rate of gene sequences encoding N and C terminals of SRY (flanking HMG-box) (0.57) was significantly lower than that encoding HMG-box (0.69).We deduced that positive selection might have imposed both on gene sequences encoding HMG-box and their flanking regions. Compared with Bubalus bubalis and Bos grunniens, Bison bonasus were more closely related to Bos taurus, which supported the view from a pateilineal aspect that extant cattle species originated from extinct auroch and Bos primigenius. Bos grunnies might have two or more patrilineal origins. Syncerus caffer and Bubalus bubalis showed distinct variations in SRY gene which led to divergent evolution, and phylogenetic analyses supported the sub-species classification of these buffaloes. Therefore, Bos species exhibited specific variation features in SRY gene and the patrilineal phylogenetic analysis supported the viewpoints from previous studies.
Research Progress on Meat Performance and Genetic Parameter of Chinese Sheep
SUN Li-min,ZHAO Jia,ZHAN Wei,JIANG Huai-zhi
2014, 41(5):  196-202. 
Abstract ( 173 )  
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Sheep industry is an important part of animal husbandry in our country. Its main product is lamb which is liked by people with its value of nutrition. Therefore, understanding the meat productive performance and meat quality, genetic parameter of sheep in our country currently is the foundation for idioplasm and selection of sheep. This paper reviewed the current situation and the progress of meat productive performance and meat quality, genetic traits of sheep in our country, hoping provide the theoretical basis for the selection of sheep.
Oocyte Recovery, in vitro Maturation and Embryo Culture Systems Influence the Efficiency of Ovine in vitro Fertilization
MIN Jiang-tao, DU Wei-hua, ZHAO Xue-ming, HAO Hai-sheng,HANG Su-qin, ZHU Hua-bin
2014, 41(5):  202-207. 
Abstract ( 208 )  
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Taking advantage of the ovaries from slaughterhouse, the effects of different kinds of oocyte recovery methods (follicle flushing, slicing, syringe aspiration and pump aspiration), different kinds of gonadotropins (FSH/LH from BIONICHE and Ningbo hormone factory) and serums (estrous sheep serum or fetal bovine serum) during in vitro maturation and different embryo culture systems (mCR and mSOF) on the efficiency of ovine in vitro fertilization were investigated. The results showed that the percentage of grade A and B oocytes recovered using the flushing method was 77.1%, which was significantly higher than using the other methods (P<0.05). In vitro maturation rate of oocytes matured in the medium supplemented with estrous sheep serum (ESS) and FSH/LH from BIONICHE was significantly higher than the other treatments (P<0.05). In mSOF and mCR cultural system, there was no significant difference in the cleavage rate between two groups (P>0.05). However, mSOF provided significantly higher rates of blastocyst and hatched blastocyst than mCR (P<0.05). These results demonstrated that the flushing method introduced in present study was more appropriate for ovine oocyte recovery than other recovery methods;The maturation rate of ovine oocytes could be increased significantly with the supplementation of ESS and BIONICHE FSH/LH into in vitro maturation media;Compared with mCR, mSOF was superior for culturing ovine IVF embryos.
Study on the Effects of Zhike Pingchuan Granules on Expectorant, Antitussive and Antiasthmatic
CHENG Long, XIN Rui-hua, LUO Yong-jiang, WANG Gui-bo, LUO Chao-ying, XIE Jia-sheng, LI Jin-yu, ZHENG Ji-fang
2014, 41(5):  208-210. 
Abstract ( 389 )  
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The assay was aimed to investigate the effects of Zhike Pingchuan granules on expectorant, antitussive and antiasthmatic by animals experiment.Capillary method was used to observe the expectorant effect in rat,the cough model induced by strong ammonia in mice was used to observe the antitussive effect,asthma model induced by spraying method was used to observe the antiasthmatic effect.The results showed that the low dosage group of Zhike Pingchuan granules significantly increased excretion of trachea quantity compared with the blank control group (P<0.05),and there were extremely significant differences between the blank control group and middle,high dosage groups (P<0.01);the three high,middle and low dosage groups of Zhike Pingchuan granules extremely significantly prolonged the latent period of cough and decreased the frequency of cough induced by strong ammonia compared with the blank control group (P<0.01); the low dosage group of Zhike Pingchuan granules had not significantly prolonged the latent period of asthma induced by 4% acetylcholine chloride compared with the blank control group (P>0.05),and middle and high dosage groups had extremely significant difference compared with the blank control group (P<0.01).Zhike Pingchuan granules had the effects of expectorant,antitussive and antiasthmatic.
Isolation and Identification of Haemophilus parasuisand Correlative Research
WANG Xue-min1,2,LU Ming-hua1,3,CHEN Xiao-ling3,LU Ying-ying1,3,ZHANG Pei-jun3,GONG Yu-mei3,WANG Hong-jun3
2014, 41(5):  211-215. 
Abstract ( 222 )  
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In 2013, nineteen small gram-negative bacilli were isolated from pigs with suspected infection of Haemophilus parasuis in Beijing. Isolates were identified as Haemophilus parasuis with the culture characteristics, capsule staining, biochemical method, serotype and PCR identification, and these strains belonged to 4, 5, 7, 12, 13 serotypes. The results of the drug sensitivity test and the pathogenicity test of the nineteen isolates showed that the isolates were multi-drug resistance, and all strains except GS-3 had strong toxicity.
Purifying Effect Analysis of Pullorum between Lines in Erlang Mountain Chickens
WANG Fei, FENG Ze-qing, LIU Yi-ping, ZHU Qing
2014, 41(5):  216-220. 
Abstract ( 133 )  
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In order to explore the breeding process of Erlang Mountain chicken pullorum purifying effect, this study used whole blood glass plate agglutination method to detect lines SD02 and SD03 three generations chicken populations pullorum disease situation, and also for the production of each generation performance data for analysis. The results showed that after three generations of purifying elimination,SD02 and SD03 prevalence among generations were extremely significantly different (P<0.01). SD02 in the rate of prevalence of child health, breeding stage survival rate, egg stage survival rate was significantly negatively correlated(P<0.05). SD03 in the prevalence of brooding survival was significantly negatively correlated(P<0.05). These results suggested that SD02 and SD03 pullorum purifying effect in significantly and improved production performance of flocks, breeding process for the subsequent removal pullorum laid a theoretical foundation.
Survey of Three Kinds of Bovine Viral Diarrhea Pathogens Infection in Some Areas of Qinghai Province
QUAN Ying-cun,LIU Hu-shou
2014, 41(5):  220-223. 
Abstract ( 245 )  
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In order to understand the infection status of 3 kinds of bovine viral diarrhea pathogens,bovine viral diarrhea virus (BVDV),bovine rotavirus (BRV) and bovine coronavirus (BCV) in Qinghai,RT-PCR method was adopted for nucleic acid detection and analysis of BVDV,BRV,BCV on clinical materials 32 with diarrhea and 152 healthy cattle feces samples which collected from 2012 to 2013 in part area of Qinghai province.The results showed that 32 diarrhea disease of cattle feed samples BVDV,BRV,BCV positive rate were 65.63% (21/32),18.75% (6/32),34.38% (11/32),respectively, mixed with 2 or 3 species infection.152 healthy cattle feces samples BVDV,BRV,BCV positive rate were 3.95% (6/152),1.97% (3/152),0(0/152),respectively.These results indicated that BVDV,BRV,BCV widespread infection in cattle in some areas of Qinghai,and mixed infection was serious,which need to further strengthen the comprehensive prevention and control of bovine viral diarrhea pathogens in Qinghai province.
Study on the Anti-inflammatory Effect of Chinese Medicine Compound Gongyanjing Injection
ZHANG Dong-sheng,JIA Shu-hong,LI Xiao-zhen,CHEN-Jie,SUN Yue-bo,LI Bing-qi,GU Xin-li
2014, 41(5):  224-227. 
Abstract ( 252 )  
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To investigate the anti-inflammatory effect of traditional Chinese medicine compound gongyanjing injection on acute inflammation in mice, we induced mouse ear swelling by xylene,induced mouse toe swelling by egg white,induced peritoneal capillary permeability in mice by glacial acetic acid and cotton pellet induced granuloma and then studied the anti-inflammatory effects in four acute inflammation models.The results showed that the traditional Chinese medicine compound gongyanjing injection could significantly reduce the mice auricle swelling ratio and decrease abdominal capillary permeability of mice (P<0.05); and could significantly inhibit the growth of granulation tissue and toe swelling rate (P<0.05); the anti-inflammatory effect of high dose group was the most significant, and had no significant difference with the dexamethasone group (P>0.05). Therefore, the anti-inflammatory effect of traditional Chinese medicine compound gongyanjing injection was significant.
Prevalence of Yak Main Parasitic Diseases in China and Strategies for its Control
YIN Ming-yang,ZHOU Dong-hui,LIU Jian-zhi,CAI Jin-zhong,ZHU Xing-quan
2014, 41(5):  227-230. 
Abstract ( 198 )  
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The yak is a long-haired bovid found mainly in China,the Tibetan Plateau.Yaks can resist cold and are capable of carrying a heavy load long distances,so they have long been an important transportation “tool” on the plateau.The parasitic diseases are the most common diseases for yaks,which severely affect their health.In this paper,the author analyzes the current situation of common parasitic diseases and proposes prevention and control strategies.
Observation and Analysis of Immunization Effects of Porcine Circovirus Disease Inactivated Vaccine
WU Xin, YAO Jing-ming, MENG Fan1, LIU Wen-jun, HAN Yi-chao, WANG Juan-ping, MI Rui-juan, FAN Zhen-hua, CHENG Hai-long
2014, 41(5):  231-235. 
Abstract ( 184 )  
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Porcine circovirus disease (PCVD) was seriously caused by porcine circovirus type 2 (PCV2) in China, it caused huge losses to the pig industry, according to the investigation and literature. In order to effectively control the disease, some new domestic PCVD inactivated vaccine were selected and ELISA method was used to test antibody tracking in immune pigs, and the SPSS software was used to analyze antibody changes. The result demonstrated that it could ensure the fattening pigs against PCV2 infection when pigs were immuned once around 2 weeks of age and strengthened immunity about 10 weeks of age.
Research Progress on Co-infection and Prevention of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome
WANG Yan-feng,ZHANG Ding-hua,CAO Zhi-gao,YIN Ling,GUI Qi-sheng,ZHU Jin-feng
2014, 41(5):  235-239. 
Abstract ( 181 )  
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Highly pathogenic porcine reproductive and respiratory syndrome, a major disease that influenced the pig industry, had spread throughout our country since it came to existence in 2006, causing huge economic losses. Though a lot of measures had been taken to prevent and control the disease, the result was far from satisfaction. The existence of highly pathogenic porcine reproductive and respiratory syndrome co-infection required that comprehensive control measures should be taken. This paper reviewed the research progress in the field of co-infection, vaccines, traditional Chinese medicine prevention and treatment,biological safety and immune suppression relief of highly pathogenic porcine reproductive and respiratory syndrome.
Isolation of Eseheriehia coli Serotype O78 from Sheep and Analysis of 16S rDNA Sequence
WU Yi-chun
2014, 41(5):  240-243. 
Abstract ( 233 )  
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A strain pathogenic bacterium was separated from the tissues of died Wadi sheep,the results of culture characteristics, biochemical identification, serotype test, susceptibility test and animal pathogenic experiment showed that it was the pathogenic Eseheriehia coli,and its serotype was O78.16S rDNA sequence analysis showed that its homology with Eseheriehia coli was up to 100%.
Study on Biological Characteristics of Eustrongyloidesfrom Eel in Hunan Province
CEN Jing,XIA Ning-bo,LUO Xu,LI Kai-wei,GESANG Xi-ruo,CHEN Li,SHENG Xiao-feng,LIU Yi,LIU WeCEN Jing,XIA Ning-bo,LUO Xu,LI Kai-wei,GESANG Xi-ruo,CHEN Li,SHENG Xiao-feng,LIU Yi,LIU Wei
2014, 41(5):  244-247. 
Abstract ( 159 )  
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The eustrongylidiasis caused by Eustrongylides in fish and birds, was a great threat to wildlife conservation and breeding industry. From April 2012 to April 2013, 58839 eel visceras were examined by helminthology post-mortem method. We observed and analyzed Eustrongylides’ mean abundance, the length of the worm and relationship between length and survival time. Meanwhile, we checked eels’ blood pH. The results showed that,Eustrongylides’ mean abundance was 5.45%; the mean abundance in spring and summer (7.85%) was higher than that in autumn and winter (3.73%), and the mean abundance of wild eels (5.77%) was higher than farming eels (1.10%), which indicated Eustrongylides’ mean abundance was affected by the season and farming methods. The averge length of 300 Eustrongylides was 49.69 mm, whose length ranged from 26.80 to 70.50 mm, the length of the worm in spring and summer was 47.10 mm,and in autumn and winter 54.86 mm. It showed that the length of Eustrongylides in autumn and winter was longer than that in spring and summer; the average survival time of Eustrongylides (the length less than 30, 30 to 39.9, 40 to 49.9, 50 to 59.9, more than 60 mm) were 2.5,3.6,4.4,5.4,9.0 d, which indicated that the worm’s survival time was in positive correlation with the length of the worm. Eels blood pH had no apparent seasonal variation. The results of the tests could provide basis for the eustrongylidiasis prevention in Hunan province.
Effect of Lameness on Reproductive Performance in Dairy Cows
LI Xiao-shan,YANG Feng-li
2014, 41(5):  248-251. 
Abstract ( 165 )  
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The cows’ lameness is one of the most common diseases in the modern dairy farms.It is widely believed that the economic losses of cows’ lameness only include reducing milk yield,increasing the cost of treatment and culling rate,and so on.In recent years, some scholars having reported that cows’ lameness has a different degree of negative impact on reproductive performance.In this paper,the latest research on the effects of cows’ lameness on the incidence of ovarian cysts,follicular development,estrus behavior,number of days to first service,the number of days to conception and cull rate were reviewed briefly.
Cloning and Prokaryotic Expression of Outer Membrane Protein 22 Gene in Brucella
DU Zhi-qiang, LIN Tao, LIU Xiao-yan, WU Ya-kun, WANG Jian-ying
2014, 41(5):  252-254. 
Abstract ( 234 )  
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In this experiment, the Brucella outer membrane protein 22 (omp22) gene was taken as the research object. Through the gene sequence cloning, expression vector construction, prokaryotic expression and affinity purification, expression and purification of the protein were studied. The results showed that the nucleotide sequence of omp22 gene contained 639 bp, which encoded 212 amino acids residues with predicted molecular weight of 22 ku. The electrophoresis results showed that molecular weight of recombinant omp22 protein was 47 ku, which was consistent with the theoretical value. All these results provided a good basis for immunity stimulation and immunity protective effect of recombinant omp22 protein in further research.
Effects of Lyophilized Royal Jelly on Antioxidant Abilities in Aged Mice Induced by D-galactose
CHEN Yan-zhen
2014, 41(5):  255-258. 
Abstract ( 164 )  
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To investigate the effects of lyophilized royal jelly (LRZ) on antioxidant abilities in aged mice induced by D-galactose. The subacute aged model mice were made by neck back subcutaneous injection of D-galactose continually. The mice were given three different doses of LRZ, 170, 340, 510 mg/kg,respectively, the effect of LRZ on the content of catalase (CAT), malondialdehyde (MDA), superoxide dismutase (SOD) in serum, liver and brain were observed after 49 d. The CAT, SOD activities in aged model group were significantly decreased (P<0.05), but MDA was significantly increased as compared to normal mice (P<0.05). While in the aged mice treated with LRZ 340 and 510 mg/(kg·bw), the activity of CAT and SOD were significantly increased and the content of MDA significantly decreased as compared to aged mice (P<0.05). LRZ had antioxidant effect in aged mice and showed dose-and-effect relationship.
Generalized Linear Mixed Models Analysis of Mummy Trait in Swine
SHA Ri-nai, JIA Qing, E Guang-xin,WANG Chen
2014, 41(5):  258-261. 
Abstract ( 168 )  
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In order to accurately estimate the fixed and genetic effect of mummy we used generalized linear models analyzed breeds, breeding month, litter month, parities, and estimate the heritability of the mummy. The results showed that impact of the breeds in mummy are not significant difference, P = 0.375(P>0.05).The breeding in the month of August and October appeared high impact than the other months, the value of fixed-effects were 1.592 and 1.658, respectively. The litter month of May and July had produced a greater impact on give birth mummy, the value of the fixed-effects were 1.399 and 0.906. The influence of the first parity on mummy was more previous than first four parities, the effect of the value was -0.507. The fifth parity showed the highest impact on mummy, the fixed value was 0.073. After fifth parity with increase of parity the impact of give birth mummy was decreased. The heritability of mummy was 0.38. The environment had great impact on mummy except disease (P<0.01), next important effect was parity (P<0.05), the breeds didn’t showed significant effect on mummy (P>0.05).
Research Progress on Detection Method for Fluoroquinolones Residues in Edible Animal Products
ZHANG Jia-he,MENG Ting,ZHOU Zuo-hong,GUAN Yuan-hong,HONG Wei-ming,LIU Li,WANG Yong-juan,ZUO Wei-yong
2014, 41(5):  262-266. 
Abstract ( 320 )  
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Animal products for human consumption contain a variety of fluoroquinolones (FQs) residues,that is harmful to production of animal health and brings our animal food safety bad affection.There is a lot of methods to detect these drugs,such as microbiological assay,chemoluminescence method,electrochemiluminescence detection, and so on.This article has reviewed the questions and detection methods of FQs residues,and could provided a reference for future research.
Research Progress on Fibriuretinin
ZHAO Wu,LU Rong-bao,LIU Wei,YIN Hui-hui,QIN Zhen-hua,SUN Jian-hua
2014, 41(5):  267-271. 
Abstract ( 229 )  
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As a plant medicine of pure natural developed by China,fibriuretinin is mainly used to treat gynecological inflammation,surgical infection,bacillary dysentery,enteronitis,respiratory infection,urethra infection,eye conjunctivitis, and so on.To provide references for the further development and utilization of fibriuretinin as a new veterinary drug,the recent studies on the extraction and artificial synthesis,detection methods and pharmacological actions on fibriuretinin were summarized in this paper.
Relations Between Bacterial Count and Absorbance of Streptococcus agalactiae in Bovine Mastitis
XIAO Min,YANG Feng,WANG Xu-rong,LUO Jin-yin,LI Xin-pu,JIA Ning,LI Hong-sheng
2014, 41(5):  271-274. 
Abstract ( 233 )  
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The present study was aimed to establish a method for quickly and accurately calculating the concentration of Streptococcus agalactiae antigen in dairy cow mastitis vaccine. Streptococcus agalactiae were washed from blood plate using physiological saline and diluted into different concentrations of bacteria suspension,which were used to calculate the number of viable bacteria suspension by plate count method.Meanwhile the corresponding absorbance (A450 nm) were determined. Then established the standard curve through the correlation between concentrations and absorbance.The result showed that the viable bacteria counts had a significant linear correlation with the absorbance of Streptococcus agalactiae.Thus,we build a regression equation between Streptococcus agalactiae bacteria suspension concentration and absorbance was y=7.5861x-0.0805,R2=0.9735,through which we could quickly calculate Streptococcus agalactiae concentration by measure the absorbance of its suspension.Compared to traditional plate count method,the absorbance measurement accuracy of the method was verified better by 5 batches of Streptococcus agalactiae bacteria suspension fostered by fermentor,and proved the new method has characteristics of simple,fast and accurate.