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20 July 2014, Volume 41 Issue 7
Cloning, Sequence Analysis and Tissue Expression of Goat CTSL Gene
HUANG Jian-wen,WANG Xiao-ping,WANG Nian-lu,XU Gang-yi
2014, 41(7):  1-9. 
Abstract ( 236 )  
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This study was enforced to acquire and analyze sequence of goat CTSL gene, and to uncover the expression of CTSL mRNA in different tissues of goat. On the basis of other species mRNA, the cDNA of CTSL gene in goat was amplified successfully by using RT-PCR. The structure and function of goat CTSL gene were analyzed by bioinformatics ways. The mRNA abundance of goat CTSL gene was detected in 11 tissues. The results revealed that goat CTSL gene cDNA code area was 1005 bp in length and encoded 334 amino acids. The amino acids homology of goat CTSL were 99.10%, 96.71%, 90.12%, 76.95%, 76.12%, 75.82%, 66.17% and 61.42% comparing to that of the ovis aries, cattle, pig, human, rattus, mouse, medaka and zebra fish, respectively. The CTSL amino acid sequence of goat included a Inhibitor_129 domain and a Pept_C1 domain. The mRNA expression level of goat CTSL gene was tested in all 11 tissues, but it was significantly higher in lung and in kidney than in heart, liver, spleen, pectoral muscle, abdominal muscle, crureus, diaphragm, ocular muscle and soleus(P<0.05).
Expression and Detection of Mycobacterium tuberculosis rv0199 Gene in Mycobacterium smegmatis
CUI Bao-liang, CHEN Li-ping, ZHANG Luan-luan, LI Hua-fang, LIU Si-guo
2014, 41(7):  9-13. 
Abstract ( 246 )  
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To study the role of Mycobacterium tuberculosis(M.tb) rv0199 gene may play in the pathogenicity of the pathogen, rv0199 gene was cloned, the protein was heterologous expressed in M.smegmatis(Ms) and its expression was detected. The E.coli-Mycobacteria shuttle vector pMV261 was improved by introducing the commonly used 6His and Strep Ⅱ recombinant protein tags, the improved vector was named as pMV262. The rv0199 gene was over-expressed in Ms/pMV262 expression system, the recombinant protein was specifically detected by the anti-6His tag antibody and anti-Strep Ⅱ tag antibody. The heterologous expression of M.tb gene in Ms could be easily detected by the shuttle vector pMV262 constructed in this study, without preparing monoclonal or polyclonal antibody against the protein. The recombinant bacteria Ms/262-99 provided material and laid foundation for further study of the potential function of rv0199 gene.
Preparation and Identification of Artificial Antigen for Patulin
WANG Xin-ran, NIE Hong-xia, LI Yan-song, TAN Cai-yun, HU Pan, FENG Xiao-li, REN Hong-lin, GUO Yi-ping2, ZHOU Yu, LIU Zeng-shan, LU Shi-ying
2014, 41(7):  14-20. 
Abstract ( 356 )  
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In order to explore the different antigen synthesis method influence on the preparation of complete antigen and the production of antibodies, the immunogen and coating antigen were synthesized by succinic anhydride and active ester method. Alternative protein carrier of EDA-BSA modifided from BSA was used for coupling the hapten to improve binding rate. Then comparing the difference of mice serum antibody titer immuned by two artificial antigens. Features of the conjugations were identified by agarose gels, native-PAGE, SDS-PAGE and GPC. The study would establish favourable foundation for preparation of antibody against PAT and immunological detecting method.
Cloning and Bioinformatics Analysis of NS1 Gene of Porcine Parvovirus Vaccine Strain
LI Xing-yu, LI Jin-hai, LIN Yi, CAO Ye, LUO Dan-dan, LIAO Dang-jin, WEI Yong, LI Jiang-ling, YU Ji-feng, ZHANG Xian-hui
2014, 41(7):  21-26. 
Abstract ( 259 )  
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In order to study the NS1 protein founction of porcine parvovirus(PPV) further, a pair of primers was designed according to PPV strain NADL-2 NS1 gene. The whole NS1 gene was obtained by PCR, and cloned into pTA2 to sequence. Sequence analysis result indicated that the recombinant plasmid(pTA2-NS1) contained one open reading frame with 1989 bp encoding 662 aa. Compared with PPV strains NADL-2 and Kresse, nucleotide homologies were 99.85% and 99.65%, putative amino acid homologies were 99.70% and 99.70%, respectively. Bioinformatics software(e.g. DNAStar) and net servers were applied to predict and analyze the structures and biochemical properties of NS1 protein. The results showed that the molecular weight of the protein NS1 was predicted as 75.7 ku, the isoelectric point PI was 6.82. It was a soluble protein, mainly located in the nucleus, containing neither signal peptide nor transmembrane region, with T199, Y309, T338, T512, S631, T635 being the potential phosphorylation sites. The construction of pTA2-NS1 and bioinformatics analysis laid the foundation for further functional research.
Research Advance on Angiopoietin-like Protein
LI Pei-pei, WANG Jian-hua, ZHANG Bao-xun,HAO Xiao-jing
2014, 41(7):  26-30. 
Abstract ( 174 )  
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A recently identified new member of the angiopoietin-like proteins(ANGPTLs), angiopoietin-like protein 8(ANGPTL8), this gene encodes a secreted protein with lipid metabolism. Research progress of structure, gene localization, regulation of gene expression, function for ANGPTL8 gene were summarized in this paper, and prospect of treating lipid metabolic diseases for ANGPTL8 were forecasted.
Cloning and Sequence Analysis of TLR5 Gene from Yak
CHEN Ya-bing, LAN Dao-liang,LIN Bao-shan,HUANG Cai,HUANG Yong,LI Jian
2014, 41(7):  30-34. 
Abstract ( 184 )  
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This study aimed to investigate the characteristics of the TLR5 gene from yak. According to the published cattle TLR5 gene sequence in the GenBank, we designed special primers for the TLR5 gene. The results of sequence analysis showed that the length of the cloning TLR5 cDNA was 2582 bp,contained a complete 2577 bp open reading frame(ORF). It encoded a peptide of 858 amino acids whose N-terminal signal peptide contained 21 amino acids, and the protein had a calculated molecular weight of 97.981 ku and a PI of 4.85. The results of protein prediction showed that TLR5 protein perform as the hydrophilic. Sequence homology analysis showed that yak had higher homology with wild yak, cattle, sheep and goat. The results showed that TLR5 gene sequence was conserved in mammals. The successful cloning of TLR5 gene of yak provided important basis for the study of molecular mechanism of innate immune response.
Preparation of Monoclonal Antibody against Swine Influenza Virus HA by Nucleic Acid Immunization
LI Yue, WANG Wei, ZHAO Ying-hui, ZHOU Chang-liang, ZANG Feng-xia, RAN Duo-liang, MENG Qing-wen, CHEN Hong-yan
2014, 41(7):  35-39. 
Abstract ( 199 )  
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For the preparation of monoclonal antibody(McAb) against HA protein of swine influenza virus, DNA plasmid bearing HA gene of A/Swine/Guangdong/2004(H1N1) was used to immunize BALB/c mice followed by generation of a McAb using the hybridoma technique. The generated nine McAbs were identified by ELISA. Functional analysis demonstrated the McAb 4D5 neutralized the virus. Furthermore, the McAb 4D5 didn’t have cross-reactivity with H3N2 or H5N1 influenza viruses, showing highly specific to H1N1. Immunofluorescence assay showed that McAb 4D5 was able to detect infection of cells with H1N1. Over all, McAb 4D5 proved specific and highly sensitive for A/Swine/Guangdong/2004(H1N1) HA and could potentially serve as immunodiagnostic tools for the rapid detection of this specific strain of influenza virus and laid the foundation for the study of single chain antibody fragment.
Cloning and Prokaryotic Expression of Aeromonas veronii Flagellin flaA
ZHAI Xin-xin,WEN Zhen-cai,SHEN Xue-fei,SUN Zhen,JIA Sheng-mei,ZHANG Jun-hui,WANG Wen-dong,YANG Zhen-guo,LU Qiang
2014, 41(7):  40-43. 
Abstract ( 158 )  
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This study was aimed to express Aeromonas veronii flagellin flaA in E.coli BL21(DE3) and preliminarily identify antigenicity of recombinant protein through Western blotting.A pair of primers was designed according to the published sequences in GenBank.Complete ORF sequence of flaA gene was obtained by PCR amplification and cloned into pET-30a vector,and then transformed into E.coli BL21(DE3) to induce,express and purify.SDS-PAGE results showed that the target protein was successfully expressed in E.coli BL21(DE3) and the product was consistent with the theoretical value and seen clearly at approximately 38 ku.Western blotting result showed that the expression of recombinant proteins could produce immune reaction with prepared antibody and His-tag antibody.Therefore,Aeromonas veronii flagellin flaA was successfully expressed, which laid the basis for further study the role of biological function and immune adjuvant of flagellin.
Study on Expression of NK-Lysin Gene in Immune Organs of Sheep Induced by Lipopolysaccharide with Different Concentrations
ZHANG Dian-qing, LI Rui, CHEN Peng-bo, ZHAO Qing-liang, ZHANG Guo-qi, LI You-jian, SHENG Jin-liang
2014, 41(7):  44-49. 
Abstract ( 132 )  
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In order to study the defensive role and action mechanism of antibacterial peptide NK-Lysin during body reaction, when pathogen infect animal body. This study detected the expression of NK-Lysin induced by different concentrations of lipopolysaccharide(LPS) and in different immune organs by Real-time fluorescence quantitative PCR, and explored the generation and response of sheep antimicrobial peptide NK-Lysin in immune organs. The cloning vector of target gene NK-Lysin and reference gene GAPDH were constructed, drew the standard curve and detected the expression of NK-Lysin. The results showed that inducing with the LPS concentrations of 0.01, 0.1, 1 and 10 μg/mL, respectively,the expression of NK-Lysin reached the maximum within 8 h. There was no positive correlation between the expression of NK-Lysin and concentration of LPS. The expression of NK-Lysin mRNA had varying degrees in pre-shoulder lymph, spleen and blood. The expression in spleen was extremely significantly different compared with other organs(P<0.01). Immune organs could express the antimicrobial peptide NK-Lysin in a relatively short time, and NK-Lysin participated in the body’s innate immunes responses.
Cloning and Bioinformatics Analysis of cDNA Expression Libraries in Eimeria acervulina
YAN Xiao-fei, HUANG Bing, HAN Hong-yu, YANG Bin, YUE Cheng, ZHAO Qi-ping, JIANG Lian-lian, DONG Hui
2014, 41(7):  49-53. 
Abstract ( 189 )  
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Analysing the new gene of E. acervulina sporulated oocysts stage by clone and bioinformatics technology. We acquired the full-length cDNA of EST sequence by RACE technique based on the cDNA expression library of E. acervulina. The cDNA of EST sequence coded protein was analysised and predicted by several bioinformatics software, the result showed the EST sequence was a new gene of E. acervulina sporulated oocysts stage, the GenBank ID was EU590120. The EST sequence including a 492 bp open reading frame which coded 163 amino acids residues,the theory of molecular weight was 17.049 ku, isoelectric point was 6.69, 8 acidic amino acids(Asp + Glu), 8 basic amino acids(Arg and Lys),molecular formula was C761H1223N199O219S12, the total average hydrophilic(GRAVY) was 0.731. The signal peptide analysis showed that the ORF encoding protein was an secretory protein containing a signal peptide and four transmembrane structures, its the amino acids residues between 105 to 115, 28 to 32 and 132 to 138 had strong hydrophobicity. At the same time, the protein contains one protein kinase C phosphorylation sites, one N-glycosylation sites, two casein kinase Ⅱ phosphorylation site, four N-end nutmeg acetoxylation site. The bioinformatics analysis showed that the ORF encoding protein possess several biological function sites and potential antigen epitopes.
Mutation Characteristics Analysis of F Genes of Two Strains Genotype Ⅱ Newcastle Disease Virus from Yunnan Province
ZHANG Wen-dong,SONG Jian-ling,ZHAO Huan-yun,LIU Qing-liang,HU Yuan-yuan,ZENG Wei, ZHANG Fu-qiang
2014, 41(7):  54-59. 
Abstract ( 198 )  
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Two field strains of Newcastle disease virus(NDV) from Yunnan province had been isolated and identified by chick embryo inoculation, hemagglutination test(HA), hemagglutination inhibition test(HI) and reverse transcriptase-polymerase chain reaction(RT-PCR). The F genes of two field strains were amplified by specific RT-PCR, then purified and cloned into pMD18-T vector for sequencing. The homologies of F genes of the two field strains showed 99.4% to 99.6%, 99.7% to 99.8%, 89.0% to 89.1% at nucleotide level and 99.3% to 99.5%, 99.5% to 99.6%, 91.8% to 92.0% at amino acid level with La Sota vaccine strain, SP13 and NDV027344, and F48E9 field strains, respectively. Phylogenetic analysis results showed that the two strains were both belonged to genotypeⅡ, and the amino acid sequence of cleavage site was G-R-Q-G-R-L, possessed molecular characteristics of low virulent strains of NDV.
Expression of EGF, IGF-Ⅰand IGF-ⅠR Gene in Mink’s Skin and Hair Follicles of Pelting Period
ZHANG Hai-hua, ZHANG Tie-tao, SONG Xing-chao, CHANG Zhong-juan, ZHAO Jing-bo,ZHOU Ning, LI Guang-yu
2014, 41(7):  59-63. 
Abstract ( 214 )  
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In order to study the mechanism of the development of skin and hair follicle of mink, distribution of epidermal growth factor(EGF), insulin-like growth factor Ⅰ(IGF-Ⅰ) and its receptor(IGF-Ⅰ R) in mink’s skin and hair follicles of pelting period were studied with histological methods and immunohistochemistry. The results showed that there was a strong positive expression of IGF-Ⅰ, IGF-ⅠR and EGF gene in mink’s skin epidermal cells and outer root sheath cells, these three genes were mainly distributed in the nucleus of cells, the positive reaction staining was not observed in vacuole, with hematoxylin counterstained blue, collagenous connective tissue and epidermis fold area under the positive staining was positive result of nonspecific. Widespread expression of IGF-Ⅰ, IGF-ⅠR and EGF gene in the epidermis and hair follicle outer root sheath cells of mink skin, directly involved in the regulation of skin and hair follicle development. So EGF, IGF-Ⅰ and IGF-Ⅰ R gene plays an important role in mink skin and its derivatives development.
Establishment of Loop-mediated Isothermal Amplification for Detection of Swine Influenza Virus
ZHANG Li,YAN Ming-hua,LI Xiu-li,LU Chao,WANG Li-li,HAN Wei,SUN Ying-feng
2014, 41(7):  64-68. 
Abstract ( 235 )  
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The objective of this study was to establish a loop-mediated isothermal amplification(LAMP) method for swine influenza virus (SIV). Six primers were designed according to the SIV NP gene sequence. Then specificity, sensitivity and reproducibility of the test were carried out. In the results, the developed system was specific for the detection of H1N1, H1N2, H3N2, avian-like H1N1 subtype SIV and H1N1 subtype influenza A, there was no cross-reactivity with porcine reproductive and respiratory syndrome virus, porcine parvovirus, classical swine fever virus, pseudorabies virus, porcine circovirus type 2 and Japanese encephalitis virus. The sensitivity of this assay was 100 copies/μL plasmid DNA. These results showed that LAMP method had higher sensitivity and specificity, and could be used for the detection of SIV.
Cloning and Bioinformatics Analysis of F Gene of Genotype Ⅶ Newcastle Disease Virus
WU Yun-pu ZHAO Xiao-tong, XUE Shu-shan SHI Lin, HE Li-kun, ZHANG Xiao-hua, CAO Ming-hui, GU Gui-bo, GU Zhi-da, WEI Shu, CAO Dong
2014, 41(7):  68-73. 
Abstract ( 221 )  
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The assay was aimed to analyze the protein structure and function of fusion(F) gene of genotype Ⅶ Newcastle disease virus(NDV) strain CY1. The total RNA was extracted from NDV CY1 strain, F gene was amplified by RT-PCR and sequenced, then using some related computer software and database, the sequence of F gene of NDV CY1 strain and the deduced amino acid sequence were analyzed, and the secondary structure and hydrophobicity of the protein were determined and predicted. The results showed that F gene of NDV CY1 strain was consisted of 1662 nucleotides encoding a 553-amino acid protein. The protein had two transmembrane domains, and its subcellular localization was in plasma membrane and endoplasmic reticulum. The secondary structure contained several kinds of conformations. Grand average of hydropathicity of F protein of NDV CY1 strain was 0.174. The theoretical pI, estimated half-life, instability index and fatty index of F protein was 8.48, 30 h (mammalian reticulocytes in vitro),31.62 and 108.12, respectively. The maximum and minimum hydropathicities were 3.078 and -2.578,respectively. The protein included a 31-amino acid signal peptide.Motif search showed that F protein had 6 N-glycosylation site, 9 casein kinase Ⅱ phosphorylation site, 7 protein kinase C phosphorylation site, 14 N-myristoylation site,1 amidation site,1 leucine zipper pattern,1 microbodies C-terminal targeting signal, 1 bipartite nuclear localization signal profile. Analysis of phylogenetic tree indicated that the F gene of NDV CY1 strain had the highest genetic relationship with zj1 strain.
Bioinformatics Analysis of Hsp70 Gene of Mycoplasma ovipneumoniae
ZHANG Shuang-xiang,CHENG Zhen-tao,TANG Yu,RAN Long-zhong,LI Tao,ZHOU Bi-jun,WEN Ming,WANG Kai-gong,WANG Hui
2014, 41(7):  74-80. 
Abstract ( 300 )  
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The assay was aimed to establish the ELISA by Mycoplasma ovipneumoniae(Mo) strain Y98 Hsp70 protein which biological characteristics should be analyzed.The physical and chemical parameters,transmembrane,signal peptide,secondary structure,B cell epitopes were predicted and analyzed by DNAStar,Bioedit 7.0,ClustalX 3.0,Mega 4.0 softwares and Protparam,TMpred,IEDB online tools in this study. The results showed that the Hsp70 protein of Mo strain Y98 was stable and soluble which predicted molecular weight was 66.14 ku and pI was 5.08. The protein had no transmembrane and did not secrete signal peptide. It was also proved that the protein was ideal target protein to establish immune method due to the advantage structure of epitope existed. It was indicted that the evolutionary relationships between Mo and Mycoplasma hyopneumoniae was close which was far compared with Mycoplasma mycoides cluster by constructed phylogenetic trees of Hsp70 gene. It would provide theoretical basis to analyze pathogenesis of Mo further.
Development of Real-time Quantitative PCR Method for Rapid Detection of Enterohemorrhagic Escherichia coli O157∶H7
LI Dan-dan,XU Yi-gang,WANG Sui-jia,GAO Shen-yang,LI Yi-jing
2014, 41(7):  81-84. 
Abstract ( 211 )  
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A Real-time quantitative PCR method was developed for the rapid detection of enterohemorrhagic Escherichia coli (EHEC) O157∶H7 based on the primers and TaqMan probes which were designed for the conservative domain of rfbE gene of EHEC O157∶H7.The results showed that the sensitivity of Real-time PCR was 7.3 CFU/mL.20 from 310 samples of meat,egg,milk and its products,animal diarrhea materials and artificial contamination samples were positive detected by Real-time PCR assay,which was in accordance with the testing result according to AOAC standard.The results indicated that Real-time PCR assay was a sensitive,rapid and simple tool.
Application of Armored Virus as Internal Control in the Detection of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus with Real-time RT-PCR
CHENG Bang-zhao,LIU Hua
2014, 41(7):  85-90. 
Abstract ( 213 )  
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To establish a rapid and accurate detection method of highly pathogenic porcine reproductive and respiratory syndrome virus(H-PRRSV), we constructed an expression vector of armored virus for veterinary clinic detection and prepared an internal control for the detection. A pair of specific primers and a TaqMan probe were designed based on Nsp2 gene in the conserved region, the detection method for H-PRRSV was established by optimization of the reaction system and amplification parameter. Mature enzyme protein and capsid protein gene sequence of MS2 phage were inserted into pET-32a vector, and internal control gene of artificial synthesis including primer and probe sequence was cloned into the downstream vector,it was identified by double enzyme digestion and PCR, the recombinant vector was transformed into Escherichia coli BL21, IPTG induced the expression of recombinant vector to prepare armored virus. The result showed that the detection method for H-PRRSV by Real-time RT-PCR was established through the optimization of reaction system and amplification conditions, the proposed method was specific and sensitive, the detection result of 8 known specific viruses was accordant with the fact and the method could detect 1 TCID50/mL H-PRRSV.In the detection of 42 clinical samples,the result was fully consistent with that of the kit designated by ministry of agriculture, the method showed good practicality.
Establishment of Indirect ELISA Detection Method for Toxoplasma gondii Antibody in Dogs
JIA Yan,CHEN Yu-huan,YANG Li-na,LIU Yue-sheng
2014, 41(7):  90-94. 
Abstract ( 221 )  
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The assay was aimed to establish a rapid and specific indirect ELISA method for canine Toxoplasma gondii antibody detection,using expressed protein of recombinant Toxoplasma gondii SAG1 gene transfected into Leishmania tarentolae as the coating antigen. The optimal antigen coating concentration was 6.75 μg/mL, serum optimal dilution was 1∶100, the known positive serum detection was more than 1∶6400, the coefficients of variation for intra-assay and inter-assay were both less than 10%, and the coated ELISA plate could be stored more than 8 months at 4 and -20 ℃. The present indirect ELISA detection method was sensitivity and specificity for detecting Toxoplasma gondii antibody in canine.
Development of Visual Loop-mediate Isothermal Amplification Assay for Bovine and Ovine-derived Materials in Feedstuff
ZHU Zhu,LAN Qing-kuo,ZHAO Xin,CHEN Rui,GUO Yong-ze,WANG Yong
2014, 41(7):  95-98. 
Abstract ( 190 )  
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Two set of primers were designed according to the mitochondrial DNA sequences of the targets,8 meat powder were detected by visual loop-mediate isothermal amplification (vLAMP) for method specificity,and meanwhile the mode of artificially contaminated feed was constructed to evaluate the sensitivity of vLAMP assay.The results showed that the bovine and ovine-derived materials had specific amplification,but the other animal origin submitted negative reactions.Sensitivity of vLAMP assay for bovine and ovine-derived materials was 0.001%.In conclusion,the vLAMP assay developed in the present study was a specific,sensitive,simple and convenient method for the rapid screening of bovine and ovine-derived materials in contaminated feeds.
Effects of Antibacterial Peptide Cecropin on Prouduction Performance,Immune Function and Serum Biological Parameters of Weaner Piglets
LI Bo,HAN Wen-yu
2014, 41(7):  99-103. 
Abstract ( 374 )  
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This experiment was conducted to study the effects of antimicrobial peptide cecropin on production performance,immune function and serum biological parameters of weaner piglets.A total of 240 weaner hybrid piglets (Landrace×Large White×Duroc) with average body weight (8.35±0.12)kg were randomly divided into 4 treatments with 6 replicates of 10 each.The four treatments were control group (fed a basal diet),antibiotics group (fed basal diet+140 mg/kg 50% kitasamycin+100 mg/kg olaquindox +600 mg/kg 10% colistin sulfate),antimicrobial peptide group (fed basal diet+350 mg/kg antimicrobial peptide cecropin),compatibility group (antibiotics + antibacterial peptide group)(fed basal diet+250 mg/kg antimicrobial peptide cecropin+50 mg/kg olaquindox +200 mg/kg 10% colistin sulfate),each group was half male and half female.The pre-test period lasted for 7 days,the trial period lasted for 34 days.The results showed that compared with control group,the average daily gain in compatibility group was significantly increased  (P<0.05),the average daily feed intake in antibiotic group and antimicrobial peptide group was significantly decreased (P<0.05),the feed conversion ratio and diarrhea rate in all three experimental groups were significantly decreased (P<0.05).The antibody blocking rate and serum IgG,IgA contents in antimicrobial peptide group and compatibility group were significantly increased (P<0.05),the serum urea nitrogen and blood sugar contents in antimicrobial peptide group and compatibility group were significantly decreased (P<0.05),but the serum total protein and albumin contents were significantly increased (P<0.05).In conclusion,dietary supplementation appropriate level of antimicrobial peptide cecropin could replace part of the antibiotics,and adding high dose of antimicrobial peptide cecropin could replace antibiotics used in feed completely.
Effect of Different Ratios of Food to Water of Dry Powder Feed on Growth Performance,Nutrient Digestibility and Nitrogen Metabolism of Growing Wusuli Raccoon Dogs
WANG Zhuo,SUN Wei-li,YANG Ya-han,WANG Kai-ying,BAO Kun,LI Dan-li,LI Guang-yu
2014, 41(7):  104-108. 
Abstract ( 239 )  
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This experiment was aimed to explore the effect of different ratios of food to water of dry powder feed on growth performance,nutrient digestibility and nitrogen metabolism of growing Wusuli raccoon dogs.Thirty 11-week-old healthy Wusuli raccoon dogs with average body weight of (3.47±0.20) kg were randomly assigned into 3 groups.The Wusuli raccoon dogs were fed experimental diets with dry powder feed of ratio of food to water of 1∶2.5 (group Ⅰ),1∶3.5(group Ⅱ),1∶4.5 (group Ⅲ),respectively.The pre-test period lasted for 7 days and the trial period lasted for 67 days.The results showed that the average daily feed intake of each week from 11,12 to 13 weeks in groupⅠ was significantly less than that in groups Ⅱ and Ⅲ (P<0.05),the average daily feed intake of each week from 14,15 to 20 weeks was no significant difference among all groups (P>0.05).The total intake in group Ⅰ was significantly less than that in group Ⅱ (P<0.05).The average daily gain of each week from 15 to 17 weeks in group Ⅲ was significantly higher than that in group Ⅰ (P<0.05),while the average daily gain,total weight and the rate of feed intake to body gain in other week-old were no significant differences among all groups (P>0.05).The protein digestibility,fat digestibility and dry matter digestibility were no significant differences among all groups (P>0.05).The amount of urine in group Ⅲ was extremely significantly higher than that in groups Ⅰ and Ⅱ (P<0.01).The nitrogen intake,fecal nitrogen,urine nitrogen,nitrogen deposition,net protein utilization and biological value of protein were no significant differences among all groups (P>0.05).In conclusion,the appropriate ratios of food to water of dry powder feed of growing Wusuli raccoon dogs was 1∶3.5 to 1∶4.5.
Determination of Rumen Degradability and Intestinal Digestibility of Ruminally Protected Lysine by Nolon Bag Technique and Modified in vitro Three-step Procedure
LIU Bao-cang,CHEN Ao-dong,ZHANG Bo-wen,KONG Ping,LIU Chen-li,GAO Wei
2014, 41(7):  108-111. 
Abstract ( 187 )  
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This trial was conducted to evaluate the rumen degradability and intestinal digestibility of the fat dry powder coated lysine by nylon bag technique and standard three-step in vitro procedure.Three and 5 months old 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 400 to 500 g,and the daily refusals were less than 10%.The results showed that the effective degradability of DM and CP of fat dry powder coated lysine in the rumen were all about 20%,the rumen bypass rate achieved 80% or more.After incubation in the pepsin solution and pancreatin solution for 24 h,the in vitro intestinal digestibility of DM and CP were 63.6% and 98.8%,respectively.In summary,This fat dry powder coated lysine could be used as a novel and effective amino acid supplement in the form of ruminant animals.
Research Progress on Processing Technology and Quality Evaluation Methods of Steam Flaking Corn
YUAN Ting-jie,LIU Qiao-xiang,DENG Lu-fang,GUO Liang
2014, 41(7):  112-117. 
Abstract ( 206 )  
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Steam flaking is paid increasing attention for their functions such as enhancing digestibility of corn starch,improving production performance and products quality in cattle,decreasing environmental pollution and cost of feeding. Its application in ruminant diet calls for high processing mechanics and quality standards. The mechanism that steam flaking enhances digestibility of corn,technological process of steam flaking,as well as various techniques and measurements that can be used to appraise the efficacy of flaking are comprehensively reviewed in this article.
Effects of Different Sources and Levels of Selenium on Growth Performance and Tissue Selenium Content of 10~11 months Porcupine
WANG Qian-guang,LIU Qiu,ZHANG Bao-ping,TONG Xue-qin,DONG Li-jun,GAO Hui-lin,REN Guo-ji
2014, 41(7):  118-121. 
Abstract ( 222 )  
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The objective of this experiment was to explore the effects of different sources and levels of selenium (Se) on growth performance and tissue Se content of 10~11 months porcupine.Eighty and 10-month-old porcupine with a similar body weight were assigned to 5 groups with 8 replicates of 2 porcupines each.The porcupine were fed a basal diet without Se complementation (control group) or the basal diet adding 0.30 mg/kg Se in the form of sodium selenite (group Ⅰ),0.20 mg/kg Se in the form of yeast selenium (group Ⅱ),0.30 mg/kg Se in the form of yeast selenium (group Ⅲ) and 0.30 mg/kg mixed Se (0.15 mg/kg sodium selenite and 0.15 mg/kg yeast selenium)(group Ⅳ),respectively.The pre-test period lasted for 7 days and the trial period lasted for 45 days.The results showed that compared with the control group,there was no significant differences in the average daily feed intake (ADFI) in each group (P>0.05),the average daily gain (ADG) in groups Ⅱ and Ⅲ was significantly increased (P<0.05),the feed/gain (F/G) in group Ⅲ was significant decreased (P<0.05).Compared with the control group,the concent of Se in liver and muscle of group Ⅲ was extremely significantly increased (P<0.01),the content of Se in liver and muscle of groups Ⅱ and Ⅳ was also significantly increased (P<0.05 or P<0.01),and there was no significant difference in group Ⅰ(P>0.05).In conclusion,the basal diet supplemented with yeast selenium could improve the growth performance and increase the tissue Se concent in 10~11 months porcupine,and adding 0.30 mg/kg yeast selenium had the best effect.
Effects of Sodium Humate on Survival Rate and Growth Performance of Broiler Chickens
CHEN Guo-shun,LI Chong,YU Rui-shan,FENG Guang-yu,CHEN Sheng
2014, 41(7):  122-125. 
Abstract ( 290 )  
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This study aimed to investigate the effects of adding different levels of sodium humate on survival rate and growth performance of broiler chickens.One thousand Kebao healthy broiler chickens were selected and randomly divided into 5 groups,with 5 replicates for each group and 40 chickens for each replicate.The chickens in control group were fed a basal diet,and the chickens in groups Ⅰ to Ⅳ were fed the basal diets supplemented with flavomycin,1.0% sodium humate,2.0% sodium humate,1.0% sodium humate with flavomycin,respectively.The experiment lasted for 49 days.The results showed that compared with the control group,the survival rate of chickens was increased in experimental group,the average daily feed intake in group Ⅲ was significantly increased (P<0.05),the average daily gain in groups Ⅱ and Ⅳ was significantly increased (P<0.05),the feed/gain in groups Ⅱ,Ⅲ and Ⅳ was significantly decreased (P<0.05).In conclusion,the basal diet supplemented with sodium humate could improve the survival rate and growth performance of broiler chickens,and the optimal supplemental level of sodium humate was 1.0%,and flavomycin was used at the same time had the better effect.
Biological Function and Extraction Process of Glycyrrhizic Acid and its Application in Monogastric Animal
SUN Huan-lin,ZHANG Wen-ju,LIU Yan-feng,TANG Shu-zhen
2014, 41(7):  126-130. 
Abstract ( 278 )  
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Glycyrrhizic acid is the active constituent of glycyrrhiza and the vital indicator of quality assessment.Furthermore,it has several high biological function,such as antioxidant,antitumor and antivirus and so on.In order to provide scientific basis for the further development and utilization of the glycyrrhizic acid in animal husbandry,this article reviews the process of water extraction,ammonia extraction and ammonia ethanol extraction and the application of glycyrrhizic acid in monogastric animals.
Research Progress on Toxicity of Chromium
ZHONG Chuan-de
2014, 41(7):  131-135. 
Abstract ( 275 )  
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Chromium is an essential micronutrient for human and animal.If chromium is ingested too much,it also can directly or indirectly cause severe health problems.This review summarizes the recent research progress on toxicity of chromium,such as toxic mechanism,clinical symptoms,toxicity of performance.
Research Progress on the Effects of Wilms Tumor Gene 1 on the Development of Kidney and Gonads
AN Pei-pei, LI Chun-yi
2014, 41(7):  136-141. 
Abstract ( 173 )  
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The Wilms tumor gene 1(WT1) is first found to be inactivated in Wilms toumor. It encodes zinc finger proteins which act as tumor suppressor and play important roles in the development of the genitourinary system and several other tissues.WT1-null mice do not have kidneys and gonads and have abnormal retinas.In addition, WT1 is also required for normal male sex determination in the context of urogenital development. Hence, WT1 is required for both kidney and gonads development. But the mechanisms underlying these phenotypic abnormalities are largely unknown.
Isolation, Identification and Induced Differentiation of Bovine Skeletal Muscle Satellite Cells
WANG Yi-min, DAI Yang, LIU Xin-feng, LIU Zhong-wei, LI Ji-xia,GUO Hong, DING Xiang-bin
2014, 41(7):  142-147. 
Abstract ( 346 )  
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In this study, bovine skeletal muscle satellite cells were isolated and identified, muscle tissues were digested by collagenase and trypsinase, then the satellite cells were purified and induced differentiate into myotubes. The muscle satellite cells and myotubes were observed, and detected by RT-PCR and immunofluorescent staining technology. The results showed that the muscle satellite cells were in healthy morphology, and more than 93 percent of the cells expressed the marker genes paired box protein 7(Pax7) and myogenic differentiation antigen(MyoD). After myogenesis induction, satellite cells began to convergence, and finally formed multinucleated myotubes expression of myogenin(MyoG) and myosin heavy chain(MHC). Our results indicated that high purity skeletal muscle satellite cells were successfully isolated and identification, which could provide cell source for muscle development and bovine muscle quality improvement study.
Effects of Different Doses of Selenium Protein Powder on Hematological Variables and Blood Biochemistry Indices of Rats
WANG Xin,WANG Yan,QIN Shun-yi,WANG Hong-xiang,WANG Jing-shen
2014, 41(7):  148-152. 
Abstract ( 229 )  
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This experiment was conducted to study the effects of different doses of selenium protein powder on hematological variables and blood biochemistry indices of rats.Eighty healthy SD rats,average body weight was 70 g,were randomly assigned into four groups,each group had two replications,every replication had 10 rats (male and female in half).Rats in control group were fed with basal diet,low-dose group,middle-dose group and high-dose group were fed with the diet which had 3.5,7 and 10.5 mg/kg selenium protein powder (calculated as the contains of selenium) on it,respectively.The experiment lasted for 30 days.The results showed that the hematological variables of low-dose group,middle-dose group and high-dose group were not significantly different compared with the control group (P>0.05),and there were no significant differences in the serum GLU and TP levels among all groups (P>0.05),the serum TG,LDL levels in the low-dose group were significantly lower than that in the control group (P<0.05),the serum A/G,TG,CHO and LDL levels in the middle-dose group were significantly lower than that in the control group (P<0.05),the serum ALT,AST levels in the middle-dose group were significantly higher than that in the control group (P<0.05),the serum ALB,A/G,TG and LDL levels in the high-dose group were significantly lower than that in the control group (P<0.05),while the serum GLB, ALT, AST, BUN, creatinine and HDL levels in the high-dose group were significantly increased (P<0.05).The results indicated that different doses of selenium protein powder had no significant effect on hematological variables in rats,and low-dose selenium protein powder had no significant effect on blood biochemistry indices except for the indices of lipid metabolism,middle-doses and high-doses selenium protein powder had some influence on blood biochemistry indices in rats.
Research Advance and Functional Mechanism on TLR4 in Anomalous Termination of Pregnancy
ZHANG Yi, HE Zi-biao, CHEN Chen, XIAN Hong, ZHANG Ming
2014, 41(7):  153-158. 
Abstract ( 173 )  
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Toll-like receptors(TLRs) as the member of pathogen pattern recognition receptors play important roles in the immune response. TLR4 is the first discovered member of the TLRs, which is the recognition receptors of lipopolysaccharide(LPS), the component from cell wall of Gram-negative bacteria. The binding of LPS and TLR4 in the extracellular induce inflammation by the MyD88-dependent or independent ways. TLR4 plays a pivotal role in the genital tract inflammation, and in clinical treatment,that intracellular signal pathway of TLR4 is blocked by antagonists or drug can achieve therapeutic effects. In the article, molecular structure, tissue expression profiles and intracellular signal transduction were described, and the relationship between its expression and reproductive system disease were stated. After that, we propose some suggestions and outlooks in reproductive clinical medicine by regulating the signal pathway of TLR4.
Study on Two Patterns Comparison of Gold Immunochromatography Test Strip for Rapid Detection of Lincomycin
ZHOU Jie,XU Fei,WU Chao,LIU Wen-li,DING Shuang-yang
2014, 41(7):  159-163. 
Abstract ( 168 )  
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The objective of this study was to develop gold immunochromatography test strips to detect lincomycin in milk. Based on two patterns of direct competitive inhibition and indirect competitive inhibition, we optimized concentration of immunogold labeling BSA-LIN (IgG), pH, concentration of coating IgG (BSA-LIN), pretreatment of milk and so on.The results showed that we got two strips that fulfil practical requirement. The LOD of direct and indirect were 30 and 10 ppb. Comparing two methods, indirect method was better than direct in sensitivity, stability and testing time.
Analysis on the Growth and Development Indexes of Hetian Sheep and Karakul in the Shield
WANG Hai-jun,ZHAO De-liang,LI Shu-wei,XIE Fei,TIE Xin
2014, 41(7):  163-168. 
Abstract ( 232 )  
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This research was analysis the seventeen growth indexes of plain Hetian sheep, mountain Hetian sheep and Karakul sheep in the shield. Samples were collected from 3 months old lamb after weaning. SPSS 17.0 was employed to statistics the differences among them. The results were as following, for the 6 months old lambs, the body high and buttock length had significant differences(P<0.05), while the metathorax wide, tail length, tail wide and tail thickness had higher significant differences between plain Hetian sheep and mountain Hetian sheep(P<0.01). The protothorax wide and metathorax wide had significant differences between mountain Hetain sheep and Karakul sheep(P<0.05). The weight and across chest had significant differences(P<0.05), while the protothorax wide, metathorax wide, tail length, tail wide and tail thickness had higher significant differences between plain Hetian sheep and Karakul sheep(P<0.01). For the average monthly increment of these indexes, the body high had significant difference between plain Hetain sheep and mountain Hetain sheep(P<0.05), while the body high had higher significant difference between mountain Hetian sheep and Karakul sheep(P<0.01). Differences of tail thickness between plain Hetain sheep and Karakul sheep were significant(P<0.05). The results showed that plain Hetain sheep were much better than that of the mountain Hetain sheep and Karakul sheep in those indexes such as weight, body length, body high and across chest.
Relationship between Polymorphisms of Exon 1 of THRSPα Gene and the Carcass and Abdominal Fat Traits in Jinghai Yellow Chicken
XUE Qian,WANG Jin-yu,ZHANG Gen-xi, WANG Ya-nan,TANG Ying,WEI Yue,SHI Hui-qiang
2014, 41(7):  169-174. 
Abstract ( 177 )  
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The objective of this study was to find out the SNPs of THRSPα gene and analysed their effects on the carcass and abdominal fat traits in Jinghai Yellow chicken. This experiment aimed at the exon region of THRSPα gene, using PCR-SSCP combined with sequencing technology to detect the SNPs in 379 Jinghai Yellow chickens at 140 days old. We foud that there was a 9 bp del/ins mutation in the exon 1 area of the gene. For the THRSPα gene, three genotypes(AA, AB and BB) were detected in Jinghai Yellow chicken. The frequencies of A and B alleles were 0.4485 and 0.5515, respectively. The results of the correlation analysis showed that the polymorphsim site had no significant influence(P>0.05) on the 140 day weight, carcass(including carcass weight, head weight, feet weight, wing weight, chest muscle weight, leg muscle weight, eviscerated weight, half eviscerated weight) and abdominal fat traits. Therefore,we concluded that the polymorphsim site could’t be used as a potential marker of carcass and abdominal fat traits in Jinghai Yellow chicken molecular marker-assisted breeding.
Prospects of Generation of Induced Pluripotent Stem Cell Lines from Wuzhishan Miniature Pig
HU Peng-fei,LI Xiang-chen,GUAN Wei-jun,MA Yue-hui
2014, 41(7):  174-177. 
Abstract ( 153 )  
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The predominance of Wuzhishan miniature pig in life science and medical science research were overviewed,current technological development and problems of induced pluripotent stem cell lines from pig were analyzed,advantage of generation of induced pluripotent stem cell lines using recombinant proteins and small molecules were illuminated,prospects of generation of induced pluripotent stem cell lines from Wuzhishan miniature pig were expected.
Genetic Effects of IGF-Ⅰ Gene on the Growth and Reproductive Traits in Jinghai Yellow Chicken
ZHAO Xiu-hua, LI Man-yu, WANG Jin-yu, LIU Guo-jun
2014, 41(7):  178-181. 
Abstract ( 253 )  
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The aim of this study was to analyze the association of SNPs in insulin like factor-Ⅰ(IGF-Ⅰ) gene with chicken growth and reproductive traits. PCR-SSCP approach was applied to assess the single nucleotide polymorphisms(SNPs) and the general linear model was used to analyze genetic effects between genotypes and growth traits of the Jinghai Yellow chicken. For the IGF-Ⅰ gene, three genotypes(AA, AB and BB) were detected in Jinghai Yellow chicken population, the frequency of A and B alleles were 0.613 and 0.387. Sequencing revealed one mutation(A60G) of IGF-Ⅰ gene in BB genotype in comparison to AA genotype. The results showed that BB genotype of the IGF-Ⅰ gene had higher bodyweight at 4 weeks compared to AA and AB genotypes(P<0.05);AA genotype of the IGF-Ⅰ gene had significantly higher bodyweight at 300 day-age-weight compared to BB genotype(P<0.05). Polymorphisms in exon 3 of IGF-Ⅰ gene had certain effect on growth and reproductive traits in Jinghai Yellow chicken. However, whether these polymorphisms were important genetic markers, which related with IGF-Ⅰ gene and reproduction traits in Jinghai Yellow chicken, were needed to be further studied.
Optimization of Miniature Porcine Cloning Embryos in vitro Stage
YANG Xi-xiang, LI Zhi-fang, LONG Chuan, FENG Chong, CHU Ming-xing, PAN Deng-ke, LI Juan
2014, 41(7):  182-186. 
Abstract ( 202 )  
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To improve the in vitro maturation of pig oocytes and to modify the protocols of cloning, the size of cumulus-oocyte and timing of the first zygotic cleavage were mainly studied in the present study. The oocytes were divided into three groups by the number of cumulus cells, subsequently the maturation efficiency was compared, and the matured oocytes were used for the following experiments. According to timing of the first zygotic cleavage, PA and SCNT embryos were divided into three groups to compare the rates of cleavage and blastocysts in the in vitro culturing. Our results showed that a better ability of maturation was observed in the oocytes which had more cumulus cells(39 to 40 h) and few cumulus cells(41 to 42 h) than those oocytes of which the number of cumulus cells(39 to 42 h) didn’t being distinguish. Both of PA and SCNT, the embryos cleaved in 26 h had a better developmental competence compared to the embryos cleaved after 26 h, which was evaluated by the rates of cleavage and blastocysts. Therefore, in the present study, not only the miniature porcine cloning programs was improved, but also that it provided technical reference for organ xenotransplantation.
Study on the SNP of 5′-flanking Region of CISH Gene in Guizhou Black Goat
JIANG Hui-mei, ZHANG Yi-ling, ZHANG Gui-xiang, CAO Juan
2014, 41(7):  186-191. 
Abstract ( 197 )  
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In order to improve the growth performance of Guizhou Black goat, the SNP site in the promoter region of CISH gene in goat was screened and the effect of SNPs on function elements of promoter was analyzed. Guizhou Black goat were selected to construct DNA pools, SNP site was screened by direct sequencing subsequently. Bioinformatics tools were used to predict the core region of the promoter, transfer factor and CpG island. 1 single nucleotide polymorphism(SNP) G-95C was found in the 5′-flanking region of CISH gene. Furthermore, the core region of the promoter and CpG island were obtained. It demonstrated that various novel transcription factors binding sites emerged while a large amount of transcription factors binding sites existed before disappeared based on the SNP found in this study, while it didn’t affect the range of CpG island dramaticlly by using various softwares. There was one SNP existed in the 5′-flanking region of CISH gene that had important effects on promoter elements. This study could lay the experimental foundation for the identification of function of CISH promoter.
Research Progress on Function Genes of Growth Rate in Beef
JIANG Qiu-fei, GU Ya-ling1,LUO Xiao-yu, HONG Long, MA Li-na, FENG Yuan, ZHANG Juan, WU Liang,GONG Xiu-yan,SUN Hao
2014, 41(7):  192-197. 
Abstract ( 187 )  
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This paper will introduce some genes that associate with livestock growth rate by using the candidate gene method and achievement, these gene can be divided into three category which include nucleus-encoded mitochondrial genes and genes related to fat metabolism, some other genes that have influence on beef quality properties were also mentioned. Rely on the results from different kinds of livestock, aims to find advantage genotype of improve beef growth rate, for the development beef cattle breeding scale of Ningxia region and improve the national beef cattle production.
Factors Analysis Influencing Conjugated Estrogens Concentration of Pregnant Yili Horse
GUO Hai-juan,WANG Xu-guang,MENG Jun,LIU Yun-fei,DENG Hai-feng,LUO Jun,YAO Xin-kui
2014, 41(7):  197-201. 
Abstract ( 253 )  
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In this study, effects of horse pregnancy time,age and urine volume on concentrations of conjugated estrogen of Yili horse were analyzed, and the results would provide the basis for animal conjugated estrogen production. Total 35 pregnant Yili horses with same rearing conditions were divided by horse age into 3 groups. Within the 28 to 38 weeks of horse pregnancy, urine samples from Yili horse were collected dividedly and urine volumes of 24 h were recorded weekly. Concentrations of conjugated estrogens in the samples were determined by HPLC. Effects of the pregnancy time and horse age on concentrations of conjugated estrogens were statistic analyzed by least squares analysis process with SAS 8.1. Correlation between urine volume and concentrations of conjugated estrogens was determined by correlation analysis process.The results showed that concentration of conjugated estrogens was affected extremely significantly by pregnancy time (P<0.01); concentrations of sodium estrone sulfate and sodium equilin sulfate were affected extremely significantly by horse age (P<0.01), but concentration of sodium 17α-dihydroequilin sulfate was not affected by horse age (P>0.05). The correlation analysis showed that there were extremely significant negative correlations between urine volume and 17α-dihydroequilin sulfate, sodium equilin sulfate (P<0.01). There was no significantly negative correlation between urine volume and sodium estrone sulfate (P<0.05). Thus the pregnancy time was the key factor to affect concentration of conjugated estrogens. Concentrations of conjugated estrogens could be improved by collecting urine from the pregnant horse under 13 ages and extremely significantly negative correlation with urine volume.
Research Progress on Radio Frequency Identification in Pork Safety Traceability System
ZHU Shi-ping,DONG Wen-hua,LIU Ying,DAI Xiu-zhen,WANG Yong,HUA Jin-di,BAO Wen-bin
2014, 41(7):  202-207. 
Abstract ( 155 )  
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So far,since the quality and safety traceability system is the most effective food safety management,radio frequency identification (RFID) technology has become the main technical means for pork safety traceability system in China.The author summarized the concept of traceability system and RFID technology,and focused on the application of RFID in pig breeding,slaughtering,transportation,storage,marketing,and so on.Finally,the article proposed corresponding countermeasures for the problems of RFID’s application,so as to provide a reference for establishing a pork safety traceability system based on RFID technology in China.
Identification and Genetic Evolution Analysis of Salmonella Isolated from Pigeon in South China
LI Xue-yan,FU Tao,JI Gui-yun,ZHANG Jin-jin,JIANG Yi-rong,LIN Xiao-shuang
2014, 41(7):  207-211. 
Abstract ( 219 )  
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Salmonella was isolated using BS selective medium from liver and intestinal contents of diseased pigeon raising in South China and made the purified culture, biochemical test, serotype experiment, sequencing analysis of 16S rRNA and regression test. A single colony of Salmonella was isolated, identified by biochemical experiments. Serotype experiment result combined with the biochemical testing results were showed that the strain was the result of A polyvalent F. After aligning the sequence of 16S rRNA fragments with the sequences in GenBank, the results of genetic evolution analysis showed that the similarity of pigeon Salmonella and Salmonella paratyphi was 100%.In the animal regression test,it showed that the Salmonella isolated in pigeon was pathopoiesis.
Effect of Kidney-tonifying and Blood Circulation-promoting Recipes on Matrix Metalloproteinase-1 and Prostaglandin E2 in Osteoarthritis SD Rats
LIN Yong-kai, HUANG Yu-xin, LIANG Gui-hong, SUN He, LIN Huan-teng, LIANG Zu-jian
2014, 41(7):  212-215. 
Abstract ( 198 )  
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The study was aimed to observe the influence of Kidney-tonifying and Blood Circulation-promoting Recipes on the level of prostaglandin E2(PGE2) in joint fluid and matrix metalloproteinase-1(MMP-1) in articular cartilage of osteoarthritis model SD rats, so as to investigate the protection mechanism of articular cartilage of osteoarthritis by Kidney-tonifying and Blood Circulation-promoting Recipes. Twenty-four SD rats were randomly divided into three groups, sham operation group, traditional Chinese medicine(TCM) group and western medicine group, the sham operation group was treated with normal saline, the western medicine group was treated with celebrex, the Chinese medicine group was treated with Kidney-tonifying and Blood Circulation-promoting Recipes in 4 weeks after operation. In 8 weeks after operation, rats were sacrificed, with the OARSI to evaluate the lesions of articular cartilage, observed the growth of chondrocytes by light microscopy, using immunohistochemical method for determining the content of MMP-1 in articular cartilage in 4 weeks after operation, measuring PGE2 in the fluid in 4 and 8 weeks after operation, compared the datas of the three groups. The cartilage degeneration of sham operation group, western medicine group and TCM group reduced sequentially, observed under light microscope, there were only a small amount of cartilage cells in the sham operation group and western medicine group, and a lot of cartilage cells were seen in the TCM group. In the western medicine group, almost all layers of chondrocytes could see a lot of positive expression of MMP-1, expression of MMP-1 in TCM group and sham operation group were extremely significantly lower than that of western medicine group(P<0.01); PGE2 of synovial fluid in TCM group and sham operation group were significantly and extremely significantly lower than that of western medicine group(P<0.05, P<0.01), respectively; compared with the ones which in the 4 weeks after operation the PGE2 of synovial fluid in TCM group and western medicine group were extremely significantly increased(P<0.01). Kidney-tonifying and Blood Circulation-promoting Recipes could significantly inhibit the expression of MMP-1 in osteoarthritis articular cartilage and reduce the level of PGE2 in the joint fluid, promote the growth of chondrocytes, and to slow the progression of osteoarthritis, which had the potential for the treatment of osteoarthritis.
Isolation and Identification of Duck Salmonella and Screening of Sensitive Antimicrobials in Shandong Province
QI Jing, WANG Shou-rong, LI Lan-bo LUO Yan-bo, LI Lu-lu, HU Ming, WANG Jin-wen, LI Hong-da, JIA Ji-mei, ZHAO Min, YANG Sheng-fu, LI Yu-bao, SHAN Hu, LIU Yu-qing
2014, 41(7):  216-220. 
Abstract ( 245 )  
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317 tissue samples of sick and dead ducks from various regions of Shandong province were collected for bacterial isolation, staining microscopy and biochemical identification. Totally, 105 strains of Salmonella were identified and the separation rate was 33.1%. Antimicrobial susceptibility testing of all isolates to 17 kinds of antibiotics was carried out by setting up different concentration gradient (1/4 L, 1/2 L, L, 2 L and 4 L) of the clinical recommended dose. The sensitive antimicrobials to the isolates were screened according to the inhibition ratio greater than 80% at the concentration of L gradient. The result showed that Salmonella was one of the main pathogenic bacteria in duck breeding of Shandong province, and with severe multiple drug resistance. Several sensitive antimicrobials including ampicillin, norfloxacin, kanamycin, florfenicol and pefloxacin could be used as clinical recommended drugs for the prevention and control of this disease.
 Research Progress on the Diagnostics of Capripoxvirus
WANG Man, LU Chang, ZHAO Zhi-xun, WU Guo-hua, YAN Xin-min, ZHU Hai-xia, LI Jian, ZHANG Qiang
2014, 41(7):  221-225. 
Abstract ( 212 )  
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Capripoxvirus, which belongs to Chordopoxvirinae genus, are important animal poxvirus. There are three members in capripoxvirus, sheep poxvirus, goat poxvirus and lumpy skin disease virus. And capripoxvirus seriously affect the development of wool-growingindustry and international trade. The pathogenic features and clinical symptoms of capripoxvirus were introduced in the present, as well as the research progress on the classification of capripoxvirus. Besides, the development of new detection technology was prospected.
Surveillance of Antibiotic Resistance of E.coli Isolated from Chickens and Pigs at Slaughter
RAO Li-li, LUO Dong-mei, CHEN Pei-ling, CHEN Xiao-jie, HE Dan-dan, YUAN Qi-ming, ZENG Li, LIU Jian-hua
2014, 41(7):  225-229. 
Abstract ( 219 )  
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The study was aimed to determine the antimicrobial susceptibility of E.coli strains to common antimicrobials, so as to evaluate the potential food safety.E.coli strains were isolated from chicken and swine fecal samples which were randomly collected from pig and poultry markets in Guangzhou. Antimicrobial susceptibility of 15 antimicrobial agents was determined by agar dilution method. A total of 606 and 125 E.coli strains were isolated from 658 swine samples and 133 chicken samples, respectively. 731 E.coli stains all displayed a different degree of resistance to 15 antimicrobials and showed multiple-antibiotic resistant phenotype. The resistance rates of sulfamethoxazole and tetracycline were over 90.0%. There was a relative low frequency of resistance to cefoxitin, colistin and amikacin among these isolates, and resistance rates were less than 3%. Chicken isolates showed significantly higher frequency of resistance to cefotaxime, ceftriaxone, neomycin, amikacin, nalidixic acid and ciprofloxacin than pig isolates(P<0.05). About 97.60% chicken isolates and 94.72% pig isolates showed resistant to more than 3 kinds of antimicrobials. Antibiotic resistance of E.coil recovered from pigs and chickens at slaughter was serious, most of them showed multiple resistance phenotypes. It indicated that these resistant bacteria carried by food animals at slaughter might pose great threat to food safety and human healthy.
Effect of Vaccination with Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Attenuated Live Vaccine at Gestation on Immunity in their Piglets
ZHANG Xue-mei, ZHANG Yan-yan, JIANG Ying, HAO Mao-xian, ZOU Min-fa, WANG Qin-fu
2014, 41(7):  230-234. 
Abstract ( 250 )  
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To investigate the impact of vaccination with highly pathogenic porcine reproductive and respiratory syndrome(HP-PRRS) attenuated vaccine(TJM-F92 strain) at gestation of 60 days on immune competency for their piglets. The field test was performed using sows vaccinated with HP-PRRS attenuated vaccine (TJM-F92 strain) as the immunized group, and the unimmunized sows were injected with saline under the same condition as control. The serum and anti-coagulated blood was taken randomly from their piglets at days 10, 20 and 30, for detection of maternal antibodies against PRRSV, CSFV, PRV by ELISA, and T, B lymphocyte function by MTT assay, NO level by Griess’s method and lysozyme activity by agar plate method. The results showed that the vaccination with HP-PRRS attenuated vaccine (TJM-F92 strain) on the sows at gestation of 60 days enhanced the immunologic defense ability for their piglets against PRRSV, the maternal antibody against PRRSV was significantly higher than that of control group at day 10(P<0.05), and it was benefit for the ability of anti-CSFV and anti-PRV abilities, the blocking antibody against CSFV at day 10 was extremely significantly higher and the infectious antibody against PRV(gE) at day 20 was extremely significantly lower compared with the control group(P<0.01). The results also showed no significant immunosuppression on T, B lymphocyte transformation assay, NO level and lysozyme content in serum from the piglets between two groups(P>0.05). The results indicated that the vaccination with HP-PRRS attenuated live vaccine (TJM-F92 strain) at gestation increased maternal antibodies against PRRSV and CSFV, whereas no suppression on the non-specific immunity and T, B cell immunity in their piglets.
Optimization of Extraction Process of Yellow Oral Liquid
ZHANG Hui,YIN Wen-shuai,QIN Zhi-hua,ZOU Ming,SHAN Hu,ZHANG Qi-di
2014, 41(7):  234-237. 
Abstract ( 196 )  
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The assay was aimed to obtain the best extraction conditions of the yellow oral liquid. The article determinated the content of berberine hydrochloride in the yellow oral liquid in different extraction conditions by UV method, selected the highest content extraction conditions as the best ones. A good linear correlation of berberine hydrochloride was observed within the range of 1.5625 to 25 μg/mL. The regression equation was y=0.1493x+0.0583, R2=0.9996. The best technology of water extraction was soaking for 45 min, adding 10 times water, decocting 3 times and 45 min each. Considering the practical production and cost factors, the water extracting technology of Chinese herbal compound was soaking 30 min, adding 10 times water, decocting 3 times and 30 min each time which all closed to the maximum result in the orthogonal design.
Research Progress of Shigella Antibiotic Resistance Mechanism against β-lactam
ZHU Zhen, WANG Jing, LIU Cui-cui, GUO Xiao, ZHANG Ji-yu, WEI Xiao-juan,ZHOU Xu-zheng
2014, 41(7):  238-242. 
Abstract ( 170 )  
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Shigella is the main pathogen of infectious diarrhea. In recent years, with the abuse of beta-lactam antibiotics, Shigella drug resistance has increased year by year. In addition, it shows the multi-drug resistance and cross-resistance. The resistance mechanisms is complex since it produces inactivated enzyme, changes action target and membrane permeability, and so on. Resistance genes are located in the plasmid, chromosome or transposable element, so it is easy to get spread. The study of resistance mechanism has a guiding significance for controlling the diseases caused by Shigella and new drug development.
Research Review of T-2 Toxin
DU Ji-hong,TU Menbayaer, LI Fu-yuan, LU Feng-qi, HOU Xiao-jiao, WANG Xiu-min
2014, 41(7):  243-247. 
Abstract ( 266 )  
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T-2 toxin is one of the most poisonous materials of trichothecene mycotoxins. It has strong hazard to the health of animal and human being. It causes the toxic effects on multiple system organs. The review focused on the widely toxicity. On the basis of recent study, the future research direction was discussed.
Research Progress on Natural Immunomodulatory Medicine for Tumor Adjuvant Therapy
GONG Lei, ZHOU Zheng-hong, ZHOU Jun-ping, YAN Ting-liang
2014, 41(7):  247-251. 
Abstract ( 183 )  
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Seeking effective tumor therapy medicine or tumor adjuvant therapy drugs has always been a hotspot for doctors and medical scientists. But in a long time, surgery and radiotherapy and chemotherapy are still the main forms of treatment in tumor therapy. Since great immunosuppression come with these treatments, curative effects could be depressed. With low toxicity and slight side effects, natural medicine could enhance the body’s immunization to defeat the side effects. Therefore natural medicine becomes an ideal adjuvant for tumor treatment. To provide that the references for natural medicine could be proper adjuvant in tumor therapy, the mechanism and research progresses of natural immunomodulatory drugs such as traditional Chinese immunomodulatory prescriptions and the active ingredients of some Chinese medicine were reviewed in present paper.