Acta Veterinaria et Zootechnica Sinica

The Research Progress in the Physiological Structure and Hereditary Factors Influencing the Reproduction Performance of Meishan Pig

XIE Xiaoxian;WANG Qishan;HE Kan;CHEN Zhenliang;MA Yufang;PAN Yuchun

2012, 43(3): 329-335. doi:

Abstract ( 405 )

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Meishan pig, one of the Chinese Taihu pig breeds, is well known all over the world for the high prolificacy. The recent research progress in the physiological structure and hereditary factors affecting the high litter size of Meishan pig was summarized in the article. The role of the uterus and placenta on the high litter size, and QTL fine mapping affecting the high farrowing performance, the association of polymorphisms of the major genes with high litter size, and the effect of early embryonic development function on high prolificacy were described. Furthermore, how to investigate the causes of high litter size at the genome level was discussed, so as to understand the molecular genetic mechanism of high prolificacy in Meishan pig.

The Relationship between Oncogene and Oncogenesis Mechanism of Avian Leukosis/Sarcoma Viruses

CHEN Hao;WANG Yixin;ZHAO Peng;LI Jianliang;LI Deqing;CUI Zhizhong

2012, 43(3): 336-342. doi:

Abstract ( 303 )

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Avian leukosis/sarcoma viruses were divided into two categories according to the difference of the activated oncogenes. Nonacute transforming virus can cause tumors by insertional activation of cellular protooncogene with a long latency, while acute transforming virus can induce a fast tumorigenesis through direct transcription of viral oncogene. This paper will review the relationship between the oncogenesis mechanism and oncogenes.

Association of Polymorphisms in the 5′ Regulator Region of the VIPR1 Gene with Chicken Broodiness

ZHOU Min;HE Jun;XIE Xiujuan;SHEN Xu;XU Haiping;ZHANG Xiquan

2012, 43(3): 343-352. doi:

Abstract ( 325 )

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: The objective of this study was to investigate the effects of polymorphisms or haplotypes in the 5′ regulatory region (-496~-1 bp) of the vasoactive intestinal peptide type I receptor (VIPR1) gene on broodiness in Qingyuan Partridge chicken, and then to look for the genetic marker inhibiting broodiness. The polymorphisms in the 5′regulatory region of the VIPR1 gene were detected by directly sequencing in 498 Qingyuan Partridge chicken under cage condition. The association between mutation sites and broodiness was analyzed by the least square mean analysis. The PHASE software was used for haplotype association analysis. A total of 12 SNPs were found in -496-1 bp region of the VIPR1 gene. Significant associations (P<0.05) were found between G359T, G266T, A134G, A94G, C72G and duration of broodiness from 18 to 43weekold. A94G was significantly associated with broody frequency (%) from 18 to 54weekold (P<0.05). Haplotypes based on 12 polymorphism sites in -496-1 bp of VIPR1 gene significantly associated with duration of broodiness from 18 to 54weekold (P<0.01). The least square analysis showed that the birds with CCTGGGAAGCAG/TTGGGGAAGCAC had significantly longer duration of broodiness than those with other haplotypes (P<0.01). Markertrait association analysis indicated that genetic marker for chicken broodiness in the 5′ regulatory region of VIPR1 gene may be exist. The individuals with CCTGGGAAGCAG/TTGGGGAAGCAC had a great genetic effect on broodiness in Qingyuan Partridge chicken, which can be a candidate marker for molecular markerassisted selection.

Expression of Porcine Globular Adiponectin Gene in Lactococcus lactis

LIU Aisha;LI Yan;HU Wenfeng;WU Tongshan;LI Jiaqi;CHEN Zhenwei;JIANG Guanyao;LI Li

2012, 43(3): 353-357. doi:

Abstract ( 270 )

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This experiment was conducted to clone, express porcine globular Adiponectin (gAd) gene in recombinant Lactococcus lactis NZ9000 and analyze the biological activity of expression product. The total RNA was extracted from porcine adipose tissue, and the Adiponectin gene was cloned into T plasmid for gene sequencing. Primers containing the corresponding restriction digestion sites and Histag sequence were designed, and then gAd gene was subcloned into expression vector pNZ8048 and expressed in recombinant L. lactis NZ9000. After purification, the immunological acitivity of recombinant protein was determined by Western blotting. To evaluate the biological activity of recombinant gAd, purified gAd was injected into the mice fed highglucose diet. The results showed that the gAd gene was successfully obtained and expressed in recombinant L. lactis NZ9000. The molecular weight of gAd protein is 17 ku. The recombinant gAd significantly decreased blood glucose of mice. This work established a good foundation for further study on the regulation of porcine lipometabolism by recombinant globular adiponectin.

Polymorphism in 3′UTR of DLX3 Gene and Its Association with Wool Quality Traits in Chinese Merino Sheep

RONG Enguang;WANG Zhipeng;ZHANG Zhiwei;YANG Hua;LI Hui;WANG Ning

2012, 43(3): 358-367. doi:

Abstract ( 342 )

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This experiment was designed to study the effects of polymorphism of DLX3 gene on wool quality and growth traits in sheep. Five strains of Chinese Merino (Xinjiang Junken Type) sheep were used in the present study. The polymorphism in 3′ untranslated region (UTR) of DLX3 gene was detected and identified using DNA sequencing, and SNP genotyping was performed by PCRRFLP analysis. Allele frequencies were analyzed by Chisquare test. Haplotypes were constructed by linkage disequilibrium analysis. The association between the SNPs and haplotypes and wool quality and growth traits were assessed by JMP4.0 software, respectively. The result showed that the allele frequencies for these four SNPs were significantly different among the five Chinese sheep strains (P<0.01). The genotype distribution in super fine wool strain were different from that in the other four strains, and the genotype distribution in two prolificacy strains were significantly different from that in other three noprolificacy strains (P<0.01); Association analysis displayed that the four SNPs and their haplotypes had consistently significant effect on wool curvature (P<0.05), but no significant effect on other traits (P>0.05). The result indicate that polymorphisms of 3′UTR of DLX3 gene are significantly correlated with wool curvature in Chinese Merino (Xinjiang Junken Type) sheep, and these SNPs can be used in molecular MAS for wool quality traits.

Comparison of Three Methods to Predict Carcass Traits in Bovine

ZHANG Limin;ZHANG Meng;ZHOU Zhengkui;LIU Xidong;CHEN Cui;CHEN Xiaojie;LI Jiao;YUAN Zhengrong;GAO Xue;GAO Huijiang;XU Shangzhong;LI Junya

2012, 43(3): 368-375. doi:

Abstract ( 300 )

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To search for a method to predict accurately carcass traits in bovine, in this study, DPS and SAS software were used to compare the methods of partial least squares regression, GM(1,N) gray system and BP neural network, in order to observe their accuracy in predicting carcass traits in bovine. Seven preslaughter growth traits including body height, body length, chest circumference, abdominal circumference, cannon bone circumference, live weight and average daily gain were used to predict the carcass weight and meat weight. The results showed that the partial least squares regression gave the highest accuracy, while the average relative errors of GM(1,N) gray system and BP neural network were lower. In this study, the three predicted results were combined and their mean value were calculated as the predictive values, which would greatly improve the accuracy of prediction. The results would provide some scientific references to beef production.

Detection of CN and DUMPS Carriers in Holstein Bulls and Pedigree Analysis

XIE Yan;FAN Xuehua;WU Xiaoping;ZHANG Yi;LIU Lin;GONG Weijia;CHEN Shaohu;SUN Dongxiao;ZHANG Shengli;ZHANG Yuan

2012, 43(3): 376-381. doi:

Abstract ( 319 )

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The aim of the present study was to detect the proportion of Holstein bull carrying two recessive genetic defects including citrullinemia(CN) and deficiency of uridine monophosphate synthase(DYMPS) and carry out the pedigreed analysis, a more easier diagnostic method was developed as well. The genetic defects of CN and DYMPS was detected by PCRRFLP in 591 Holstein bulls, and the ASPCR technology was constructed for detecting the CN recessive deleterious gene. 2 CN carrier bulls and 1 DUMPS carrier bull were found, and the proportion of carrier bulls was 0.34% and 0.17%, respectively. Through pedigree analysis, the two CN carrier bulls were the descendants of Linmack Kriss King, and the DUMPS carrier bull was Skokie sensation Ned’s offsprings. The results indicate that it is necessary to establish early monitor system on genetic defects in all Holstein young, proven bulls and suspect cows, and put such information in their pedigree records. Through young bull preselection and selection and assortative mating system, carrier bulls could be avoided to enter into the national genetic improvement program, frequencies of the undesired recessive alleles could be greatly decreased as well.

he Development of Dorsal Hair Follicle and Expression of Lef1 in Postnatal Murine

WANG Haidong;YANG Lei;WANG Yi;YU Xiuju;DONG Yanjun;HE Xiaoyan;MA Shuhui;DONG Changsheng

2012, 43(3): 382-387. doi:

Abstract ( 346 )

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To study the histological characteristics of murine hair follicle and the function of Lef1 at different hair follicle growth stages, the expression and localization of Lef1 were investigated. HE staining was used to observe the morphology of the murine hair follicle at each stage, Western blotting and immunohistochemistry were used to determine the expression level and localization of Lef1 protein at different hair follicle cycle stages. The result suggested that the gowth of murine hair follicle showed cyclic variation. For each stage, murine hair follicle had stagespecific characteristics. The result of Western blotting showed that Lef1 was expressed in murine skin at different growth stages, and the expression level in hair follicle early growth period was higher than that in hair follicle middle and late growth periods. The result of immunohistochemistry showed that Lef1 was expressed in sebaceous gland, root sheath and dermal papilla of hair follicle at different growth stages in different degree. The results indicate that cyclic variation of hair follicle depend on the expression level of Lef1.

Effects of Propylene Glycol on Energy Balance, Blood Parameter, Concentration of Glucose and Adipose in the Liver during Peripartum Period in Dairy Cows

LIU Qiang;WANG Cong;ZHANG Yanli;ZHANG Shuanlin;PEI Caixia;LI Xin;SHI Caiye;BAI Yuansheng;SHI Zhouge

2012, 43(3): 388-396. doi:

Abstract ( 363 )

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The objective of this study was to observe the effects of propylene glycol on energy balance, blood parameter, concentration of glucose and adipose in the liver during peripartum period in dairy cows. Multiparous Holstein dairy cows with similar body weight ((683.1±14.9)kg), lactation number (2.5±0.2), corrected milk yield in last lactation ((7 339±18)kg) and expected calving date ((24.5±0.2)d ) were allocated into four treatments at random. Graded levels (0, 150, 300 and 450 mL·d-1) of propylene glycol were supplemented in the four treatments from day 19 prepartum. The results showed that body condition score at 7, 21 and 35 d postpartum increased, body weight loss from 7 to 35 d postpartum reduced, energy balance at 14 d prepartum and 7, 21 and 35 d postpartum was improved significantly by the 300 and 450 mL·d-1 treatment compared with 0 mL·d-1 treatment (P<0.05). Cows receiving 300 and 450 mL·d-1 propylene glycol had a higher plasma glucose and insulin at 14 d prepartum and 7, 21 and 35 d postpartum, lower plasma NEFA, βhydroxybutyrate and triacylglycerol at 7, 21 and 35 d postpartum than that of 0 mL·d-1 treatment (P<0.05). 300 and 450 mL·d-1 propylene glycol supplementation decreased the contents of triacylglycerol in the liver and increased the contents of glycogen in the liver compared to 0 mL·d-1 propylene glycol treatment (P<0.05). The results indicated that propylene glycol supplementation was beneficial to peripartum cows in reducing the lipolysis, enhancing the gluconeogenesis and improving energy balance status, the optimum dose was about 300 mL·d-1.

Study on the Major Microorganism Changes during the Silage Processing of Two Kinds of Corn Silage

YANG Yungui;ZHANG Yueli;DU Xin;LIU Guiyao;CAO Shehui

2012, 43(3): 397-403. doi:

Abstract ( 269 )

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The aim of this study was to observe the number of major microorganisms and change trends during the corn silage processing. The corn silage in bottles was made by crosscutting of corn and rubbing stalk, the raw materials of corn silage were got and the silage samples at 0.5, 1, 2, 3, 4, 5, 6, 7, 9, 11, 13, 15, 20, 25, 30, 35, 40, 45 and 50 days during silage processing were collected, the samples with aseptic water were soaked, and then the main microorganisms(lactic acid bacteria, yeast and mold) were cultured with selective culture medium. The results showed that the moisture content changed very little during the 60 days after the corn silage sealed. pH reduced to <4 at the 2nd day and then stayed at about 3.5. pH of corn silage was lower than that of rubbing stalk silage. The number of lactic acid bacteria increased rapidly and reached to the peak of 109 orders of magnitude at the 6th or 7th day, and later declined slowly and stabled at 107 orders of magnitude at 15th20th day. The number of lactic acid bacteria of corn silage with a peak at 6th day was higher than that of the rubbing stalk silage with a peak at 7th day. The number of yeast had fluctuations initially and reached the peak to 107 orders of magnitude, then reduced with the treating time prolonged. The number of yeast of rubbing stalk silage reduced rapidly and disappeared after the 40th day. But the number of yeast of corn silage increased within 12 hours at the beginning of silage and then reduced slowly, finally disappeared after the 50th day. The number of mold reduced sharply because of the shortage of oxygen after sealed, and it was not found after 11th day. The number of mold of rubbing stalk silage reduced sharply and no mold was found after 4th day, while the number of mold of corn silage reduced slowly and no mold was found after 11th day. In the process of corn silage, the number of major microorganisms decreased gradually with the treating time prolonged; rubbing was beneficial to make the corn silage.

The Effects of Boiling Time on the Determination of NDF and ADF in Corn Silage with the Filter Bag Technique

AN Guilong;;CHENG Cheng;CAO Chunmei;DIAO Qiyu

2012, 43(3): 404-409. doi:

Abstract ( 293 )

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To study the effect of boiling time on the content of NDF and ADF in corn silage by the filter bag technique, the result got from traditional Van Soest method was used as a standard, the different boiling time (30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130 min) were screened using the filter bag technique. The results showed that the optimum boiling time were 110 and 120 min for determination of NDF content in wholeplant corn silage and corn silage without ear, and 60 and 70 min were the optimum boiling time for determination of ADF content in wholeplant corn silage and corn silage without ear. Therefore, the optimum boiling time for determination of NDF content was very different from that for determination of ADF content in corn silage by the filter bag technique.

Expression of Main Epitope Domains of CD151 from Marc145 Cells and Relationship with Infection of Porcine Reproductive and Respiratory Syndrome Virus

XIAO Yihong;WANG Weiwei;ZHANG Lu;GAO Jiming;MA Xiaochun;ZHOU Enmin

2012, 43(3): 410-415. doi:

Abstract ( 291 )

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To study the role of CD151 in porcine reproductive and respiratory syndrome virus (PRRSV) infection, primers specific to CD151 were designed according to the similar gene sequences submitted in GenBank. Total RNA was extracted from Marc145 cell and a 294 bp of CD151 was amplified by RTPCR. The target gene was cloned into pGEX4T3 vector, then was transformed into E. coli and expressed with IPTG inducer. SDSPAGE and Western blot were employed to identify the expressed protein. The protein was purified and used to immunize mice. The antiserum titres were determined by the ELISA and IFA, and were used to block PRRSV infection of Marc145 cells. The results showed that CD151 gene was amplified and cloned into pGEX4T3CD151 vector. The recombinant CD151 protein with MW of 37 kDa was produced. The antiserum reacted specifically with the recombinant protein and blocked PRRSV infection of Marc145 cells. These results provide useful information for further understanding the role of CD151 in PRRSV infection.

Isolation and Identification of a Pig Bovine Viral Diarrhea Virus

DENG Yu;;SUN Chunqing;ZHANG Hongbiao;LIN Tao;;ZHANG Rong;LONG Jinxue;HUANG Lv;CAO Sanjie;YUAN Shishan;WEN Xintian;ZHENG Hao

2012, 43(3): 416-423. doi:

Abstract ( 340 )

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The aim of this study was to isolate virus from bovine viral diarrhea virus (BVDV) positive sample of the piglet and to study BVDV originating from pigs. BVDV positive samples from the piglet were inoculated in MDBK cells and a pig BVDV strain (SD0803) was obtained. In the study, cell culture, direct immunofluorescence assay, PCR amplification of 5′UTR，electron microscopy and TCID50 were performed, moreover，bioinformatics software were used to analyze the characteristics of molecular evolution. The results showed that the virus strain hadn’t caused cytopathic effect (CPE) until the 13th passage on MDBK cell. SD0803 infected cells showed strong fluorescent signal. DNA fragment of 307 bp and 428 bp were amplified by PCR from 5′UTR and Npro of SD0803. Under electron microscopy, virus particles were near round and enveloped with size of 50 nm in diameter. Virus titer of SD0803 was 10-6.5 TCID50·0.2 mL-1. The homology comparison and phylogenetic analysis of 5′UTR and Npro between SD0803 and representative BVDV isolates showed that SD0803 belongs to BVDV1. However, SD0803 had an obvious distance from the subtypes of BVDV1 and formed a separate group according to the phylogenetic relationship and this showed that the isolate might be a new subtype of BVDV1. So a pig originated BVDV SD0803 was successfully isolated and identified, and belongs to NCP BVDV1 and maybe a probably new subtype of BVDV1.

The Prevalence of Torque Teno Sus Virus and Porcine Circovirus 2 Coinfection in Pigs

XIA Yingju;ZI Zhanchao;CAI Lin;HAN Xue;ZHAI Xinyan;TIAN Kegong;NI Jianqiang

2012, 43(3): 424-430. doi:

Abstract ( 336 )

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The aim of this study was to investigate the prevalence of porcine circovirus 2 (PCV2) and Torque teno sus virus (TTSuV) coinfection in China, 1898 swine serum samples were collected from 29 provinces in China in 2009 and detected for TTSuV1, TTSuV2 and PCV2 by PCR. 1 103 samples were TTSuV positive (58%), 435 samples were PCV2 positive (23%). Among the positive samples, 275 (14%) samples were both positive for PCV2 and TTSuV, including 249 (13%) infected with both TTSuV1 and PCV2, 200 (10%) infected with both TTSuV2 and PCV2, and 174 were triple infection. These results suggested that coinfection of TTSuV and PCV2 was highly prevalent in China. Distribution analyses of positive pig herds demonstrated that there were significant difference in different regions, and the breed pattern might not be a factor of aggravation in the coinfection.

Expression of P46 Protein of Mycoplasma hyopneumoniae in Escherichia coli and Used for Indirect ELISA

SHEN Qingchun;WANG Fang;HAN Mingyuan;QIN Qingsong;FAN Xuezheng;YANG Hanchun;NING Yibao

2012, 43(3): 431-437. doi:

Abstract ( 329 )

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Mycoplasma hyopneumoniae is the agent of Enzootic pneumonia of swine. Part of the hydrophilic region of the surface protein P46 of M. hyopneumoniae was cloned, and three TGA coding Trp in the ORF of P46 gene were point mutated to TGG. The modified DNA fragement was coloned to vector pET28a(+), and highefficiency expressed in E. coli BL21 (DE3). SDSPAGE showed that the target DNA was expressed in inclusion body, the molecular weight of the recombinant protein was 31 kDa, owed 35% of the total proteins. The recombinant protein was proved to be well reacted with the high immune serum against M. hyopneumoniae from rabbit specially by western blotting. After wash and purification with Ni2+ column, the recombinant protein was used as coating antigen of indirect ELISA for detection the antibody against M. hyopneumoniae in swine serum. The ELISA detection kid was developed by optimizing the relational parameters and reagents. The further compare detection test indicated the recombinant P46 protein based ELISA was satisfactory and high complicated with the marketable products.

Sequence Analysis and Homology Modelling on Peptidoglycanassociated Lipoprotein A of Haemophilus parasuis

ZHOU Lunjiang;CHE Yongliang;CHEN Rujing;JIANG Bin;WANG Longbai;WEI Hong;WU Xuemin;ZHUANG Xiangsheng

2012, 43(3): 438-445. doi:

Abstract ( 351 )

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To lucubrate bionomics of PalA in Haemophilus parasuis (Hps), the complete palA gene was amplified by PCR with a pair of specific primers designed and synthesized, and was cloned and sequenced. The Hps PalA was analyzed and predicted by the tools of bioinformatics in the following aspects: the composition and alignment of nucleic acid sequences and amino acid sequences, molecular structure, physical and chemical characters, secondary and tertiary structure of protein and so on. The results revealed that 462 bp fragment of the palA gene was amplified by PCR; There were less similarity in nucleic acid sequences than amino acid sequences among five Hps strains; The palA gene preferred mostly to GCA and AAA; The PalA(PI=6.28) showed weak acidity; and main component of all secondary structures included α spiral, random coil and extended strand; The homology of the amino acid of Hps PalA and Haemophilus influenza Pal is 71.97%; The 3D models of Hps PalA were successfully constructed based on the template of Pal of H. influenza; Like Pal, the tertiary structure of Hps PalA also contained a Binding pocket region which included Y80, F39 and L84. The homology modelling successfully of Hps PalA provided a refer to lucubrate the PalA.

Construction of a Recombinant Adenovirus Expressing NcSRS2 Protein of Bovine Neospora Caninum and Immunogenicity Analysis of the Protein

JIA Lijun;YU Longzheng;ZHANG Shoufa

2012, 43(3): 446-452. doi:

Abstract ( 318 )

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In order to construct a recombinant adenovirus expressing NcSRS2 protein of bovine Neospora Caninum, NcSRS2 gene of bovine Neospora Caninum was amplified by PCR, pMD18TNcSRS2, pCR259NcSRS2 and TransposeAdNcSRS2 were constructed in this research. Coated with liposome, TransposeAdNcSRS2 was transfected into QBIHEK293 cells to package recombinant adenovirus Ad5NcSRS2. Recombinant adenovirus NcSRS2 gene was detected by PCR. The expression of NcSRS2 gene in QBIHEK293 cells was detected by IFAT and Western blotting. After the virus titer was determined, the virus fluid was collected to inoculate BALB/c mice and IgG antibody levels in the sera were measured by indirect ELISA. The size of NcSRS2 gene was 1 227 bp. The nucleotide sequence of the gene shared 99% homology with that in GenBank (AF061249). Recombinant adenovirus Ad5NcSRS2 was successfully packaged in 293 cells. The protein expressed by Ad5NcSRS2 was 43 kD and had good reactogenicity. The titer of recombinant adenovirus Ad5NcSRS2 was 109TCID50·mL-1. Three weeks post the second immunization, IgG antibody titers in the sera of BALB/c mice were up to 1:2 048, measured by indirect ELISA. A recombinant adenovirus Ad5NcSRS2 which have good immunogenicity was successfully constructed. This settles a solid foundation for the development of recombinant adenovirus vaccine against Neospora Caninum based on NcSRS2 gene.

Cloning and Sequence Analysis of the ITS and 5.8 S rDNA of Larval Eustrongylides spp. Isolates from Monopterus albus (zuiew)

DUAN Liuchun;LIU Wei;XU Pingyuan;HE Xin;ZHOU Xiaojun;LIU Zhilian;ZHENG Yanqiong;LIU Yi

2012, 43(3): 453-459. doi:

Abstract ( 315 )

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The objectives of the present study were to examine sequence variation in the internal transcribed spacer (ITS) and 5.8S rDNA of Eustrongylides spp. isolates from Monopterus albus, and to reconstruct their phylogenetic relationship using ITS sequences. The ITS sequences were amplified from each Eustrongylides spp. samples and the amplicons were cloned into pGEMT Easy vector, respectively. The inserts were successfully sequenced, and the length of ITS in 32 Eustrongylides spp. isolates were different（922927 bp）. Sequence analysis revealed that the ITS1, 5.8S and ITS2 rDNA of these samples were 350351 bp, 102 bp and 340344 bp in length, respectively. The results indicate that the ITS sequences provide useful genetic marker for molecular identification of Eustrongylides spp. It was the first time that the complete sequence of ITS and 5.8 S rDNA of Eustrongylides spp. were reported. The results of this study lay down the foundation for further study on molecular epidemiology and diagnostics of Eustrongylides spp.

The Role of HDAC1 in the Signal Pathway of TGFβ1 / Smad7 in AAN Model

YANG Yang;ZHANG ZhongwenYAO Hua;GAO Lei;LI Peifeng;WU Guojuan

2012, 43(3): 459-468. doi:

Abstract ( 234 )

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The aim of this study was to study the changes in the expression of transforming growth factor β1 (TGFβ1), the TGFβ1 receptor (TβRI), Smad7, and histone deacetylase 1 (HDAC1) in mice with aristolochic acid nephropathy (AAN), and analyze the relationship between HDAC1 and Smad7. RTPCR was applied to detect the mRNA levels of TGFβ1, Smad7 and HDAC1 in kidneys. The TGFβ concentrations were determined by ELISA. HADC1 and Smad7 proteins were localized and detected by immunohistochemistry method. The tissue section technique was used to observe the histopathologic changes, such as kidney damage and renal tubules interstitial fibrosis degree. Results were as follows: In mice with induced AAN, the mRNA levels for TGFβ1and HDAC1 were increased with treatment time. The mRNA levels for TGFβ1 were higher than that of their control (normal mice without treatment) (P<0.05). Starting from 28 day, mRNA of HDAC1 became significantly higher than the level of the control (P<0.01). The levels of Smad7 protein and its mRNA were decreased with time, and lower than that of control. HDAC1 protein expression was increased with time post treatment. Compared with the control group, HDAC1 positive stains were mainly distributed in the epithelial and stromal cell nucleus. Histopathologic changes of kidney showed acute tubular interstitial damage, such as turbidity, swollen, degeneration and fall off of renal tubular. With extension of treatment time, renal tubular damage was aggravated progressively. During the development of AAN in mice, Smad7 can cause renal tubular damage, impare renal tubular renewable repair ability, restrain Smad7 mRNA expression. The weak expression of Smad7 in renal tissues can promote TGFβ1 signal transduction, help the fibre formation and maintain the proliferation condition. One of the mechanism happened is about HDACl, which mRNA and protein high expressions plays an important role in the development aristolochinc acid nephropathy, may be a new potential target.

Study on the Tissue Structure and Mucosal Immunityassociated Cells in Abomasum of the Yak

LU Yangang;GAO Shijie;FANG Mei;XIANG Meng;JIA Ning

2012, 43(3): 469-475. doi:

Abstract ( 314 )

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The aim of this study was to reveal the tissue structure and the distribution and number changes of abomasum mucosal immunityassociated cells in the yak. The tissue structure and intraepithelial lymphocytes, plasma cells and mast cells of abomasum in the yak were investigated with histochemical method, the scion image software analysis techniques method and the electron microscope technique. The results showed that there were mucousmembrane, submucosa, tunica muscularis and serosa in the abomasum of yak. The gastric pit was deepest in the pyloric gland region of abomasum, and this glandular region has also the longest gland and the thickest muscular layer. The thickness of muscular and glands length in each glandular region of abomasum are significantly different (P<0.01). The gastric pit of the pylorus gland region is significantly different (P<0.01) with the fundus gland region, and it has also significantly difference compared with the cardiac gland region, but the gastric pit of the fundus gland region is quite similar (P>0.05) to the cardiac gland region. The number of intraepithelial lymphocyte and plasma cell in each glandular region of abomasum are not significantly different（P>0.05). The number of the mast cells between the fundus gland region and the pyloric gland region are significantly different (P<0.01), the fundus gland region has most mast cells, and the pyloric gland region has the lest mast cells, but both of them are similar to the cardia gland region (P>0.05). There are a lot of diffuse lymphatic cells and solitary lymphoid nodules in the lamina propria of abomasum. Under the electron microscope, gastric pit columnar epithelial cells are aligned tightly. There are lots of mucous cells in the lamina propria of pylorus gland region, these cells are tall columnar or cone shape, their nucleus are located basally part of the cell, there are the high electrons density particles are gathered in the top of these cells. There are a large number of parietal cells and chief cells in the fundus gland region and cardiac gland region. The structure of abomasum in the yak is similar to other ruminant animals, but each layer has its characteristics. There are quite a lot of diffuse lymphatic cells and solitary lymphoid nodules in the lamina propria of abomasum in the yak, it make the abomasum mucosal immunity function of the yak stronger than other ruminant animals.

Effect of Sex Hormone on the Cells Secretion Activity of TNFα and TGFβ in Goat Endometrial Cells

QU Yanyan;QI Xuefeng;NAN Zhichun;WANG Aihua;ZHAO Xianjun;JIN Yaping

2012, 43(3): 476-481. doi:

Abstract ( 297 )

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The aim of this study was to study the effect of sex gland hormones on the secretory activity to endometrial cells of goats. Based on the hanging cell coculture insert system, immortalization of goats endometrial epithelial cells (EEC) and stromal cells (ESC) has trained coculture in vitro studies model, the different levels and combinations of progesterone (P4) and estradiol (E2) were added in medium, without or with ESC together, TNFα and TGFβ secretory activity of EEC were determined by ELISA. The results showed that compared with no hormone control group, the secretion of TNFα on EEC apical was enhanced markedly by P4 (P＜0.05) when added alone or together E2 and P4, but not significant on basolateral; TGFβ secretory activity of both EEC apical and basolateral were reduced markedly by different concentrations and combinations of E2 and/or P4 (P＜0.05), but TGFβ on EEC apical was restrained significantly by E2 alone (P＜0.05). About coculture with ESC together, both apical and basolateral of TNFα secretion were repressed significantly by sex hormones E2/P4 (P＜0.05); TGFβ on EEC apical was restrained significantly by E2/P4 (P＜0.05), but not significant on basolateral. Our results indicated that the sex hormones in coculture mode alone or together with ESC, all can regulate the secretion of TNFα and TGFβ with different levels, suggested that ESC play an important regulatory role to the secrete direction and secretion levels of goat EEC when coculture with sex hormones.

Research on the Role of Myostatin in Dexamethasoneinduced Myofibril Damage

QIN Jian;DU Rong;YANG Yaqun;ZHOU Ying;ZHANG Hongqiang;LIU Wenhua

2012, 43(3): 482-487. doi:

Abstract ( 274 )

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In order to clarify the roles of myostatin in myofibril damage and investigate the measures to improve the production and quality of muscle mass in livestock or control the muscle atrophy in human, the serious stress models of mice were made with highdose dexamethasone, and the effect of physiologicaldose insulin on the dexamethasoneinduced myofibril damage and the relationship between the effect and myostatin expression were researched. In addition, the intervention effect of myostatin gene immunization on the dexamethasoneinduced myofibril damage was studied. The results showed that: dexamethasone upregulated the expression of myostatin gene and caused the serious damage of myofibril and swelling of mitochondria, while insulin attenuated the effects of dexamethasone; myostatin gene immunization reduced the myofibril and mitochondria damages induced by dexamethasone. The results suggest that myostatin is one of key factors for myofibril degradation and the role of myostatin may be attributed to the stimulation of mitochondrial function by myostatin.

Construction of the Recombinant Adenovirus of Malonyl/Acetyl Transferase of Xinong Saanen Goat

ZHU Jiangjiang;LUO Jun;WANG Ziqian;XU Huifen;SUN Yuting;SHI Hengbo;HAO Juan;ZHAO Liyuan

2012, 43(3): 489-495. doi:

Abstract ( 326 )

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The aim of this study was to observe the role of the Malonyl/Acetyl transferase (MAT) region of goat fatty acid synthase (FASN) in fatty acids metabolism of goat mammary, and to construct the recombinant adenovirus vector with goat MAT for observing gene overexpression in primary goat mammary epithelial cells. The primers were designed according to the MAT sequence in GenBank, and the gene was cloned and sequenced by RTPCR. The goat gene fragments containing both MAT region and HIS sequence were inserted into shuttle vector pAdTrack/CMV to construct recommbinant shuttle vector pAdTrack/CMVMATHIS. After identifying by digestion and sequencing, pAdTrack/CMVMATHIS plasmid was linearized by PmeⅠ, and then it was transformed into E.coli Bj5183 competent cells containing backbone vector pAdEasyⅠto obtain recombinant vector pAdEasyMATHIS by homologous recombination. pAdEasyMATHIS plasmid was linearized by PacⅠ, and transfected into HEK 293 cells for virus packing, amplification and titer testing by LaSRT. The MAT sequence cloned had a GA mutation at 601 bp by comparing with the corresponding sequence in GenBank, which caused an AlaThr mutation at the 201st amino acid residue. The results of PCR detection and Western blot showed that recombinant overexpression vector containing MAT was successfully constructed. The titer of virus obtained was 2×109 PFU·mL-1. The recombinant adenoviral vector pAdEasyMATHIS was constructed successfully in this experiment.

Analysis of Outer Membrane Proteins Phenotype of Haemophilus parasuis

ZHU Bifeng;YANG Xufu;PENG Ling;WEI Zhaoyu

2012, 43(3): 496-502. doi:

Abstract ( 241 )

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The aim of this study was to explose relatioship between outer menbrane protein and virulence of Haemophilus parasuis. The outer membrane proteins (OMP) of 82 isolates of Haemophilus parasuis were determined by SDSPAGE. We compared the diversity of the OMP phenotype of isolates from diffent clinical background to explore the associativity between OMP and virulence pathogen, and analyzed relationship between the OMP and virulen isolates by cluster analysis based on Rf of electrophoresis and contents of OMP. The results of analysis of OMP phenotye showed that isolates were divided into two major PAGE type groups as differece of Rf. PAGE I was characterized from 36 to 40 kD OMP and PAGEⅡwas characterized 4245 kD OMP, and included about 34% isolates recovered from disease pigs and 8.7% isolates recovered from healthy pigs belong to PAGE I; PAGE II was characterized by absence of the 3640 kD proteins. The result of cluster as Rf indicated that isolates were divided into three types: PAGEⅠ, PAGEⅡand PAGEⅢ. Analysis of cluster as contents of OMP obtained five classsifications of isolates. It is prompted that the 3640 kD OMP was correlated with virulence.

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