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25 November 2011, Volume 42 Issue 11
综述
Molecules of Disturbing Host Immune Response Encoded by Orthopoxviruses and Their Action Pathways
JING Zhizhong;JIA Huaijie;ZHOU Tao;HE Xiaobing
2011, 42(11):  1503-1512.  doi:
Abstract ( 407 )   PDF (1506KB) ( 848 )  
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Members of Orthopoxviridae (OPXV) include cowpox virus (CPXV), vaccinia virus(VACV) and monkeypox virus (MPXV), they are more important pathogens of animalborne diseases to human. This review focuses on the immunomodulatory molecules encoded by OPXV and their action pathways,more detailly introduced the different strategies with blockading of host interferon responses, inhibiting of host TNFinduced responses and inactivating of key immune components of the host defense responses. This will improve our acquaintance about the molecular mechanism of evading the host immune responses, host specificity of infection and pathogenesis for poxviruses.
遗传繁育
Genetic Polymorphisms of the Porcine Developmental Pluripotency Associated 5 Gene and Its Relationship with Reproductive Traits
LI Wen;LI Yong;ZHANG Longchao;WANG Ligang;YAN Hua;WANG Lixian
2011, 42(11):  1513-1518.  doi:
Abstract ( 472 )   PDF (447KB) ( 500 )  
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The purpose of this study was to detect the genetic variation of the porcine developmental pluripotency associated 5(Dppa5) gene and its relationship with reproductive traits in swine. The nucleotide sequence was determined by pooled genomic DNA sequencing approach to identify polymorphic sites. These sites were genotyping by PCRRFLP, and then their genetic effects on reproductive traits in Large White pigs and Beijing Black pigs were analyzed. The results were follows: (1) Two singlenucleotide polymorphisms sites (g.363 T>C and g.844 G>T) in exons were detected. (2) The 2 sites displayed polymorphism in 2 pig populations. (3) The association analysis showed that both the g.363 T>C site and g.844 G>T site had significant effects on litter weight at birth in later parities, but the difference of total number born and number born alive among different genotypes was not significant. These results indicate that Dppa5 gene could be regarded as a candidate gene affecting reproductive traits in sows.
The Expression of Follicle Stimulating Hormone and Its Receptor Genes in Certain Nonreproductive Tissues of Chicken
CUI Huanxian;ZHAO Guiping;LIU Ranran;ZHENG Maiqing;CHEN Jilan;WEN Jie
2011, 42(11):  1519-1525.  doi:
Abstract ( 433 )   PDF (2068KB) ( 591 )  
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The mRNA and protein expressions of FSH and FSHR were detected in some nonreproductive tissues of Beijingyou chickens by PCR and immunohistochemistry in this study. Subsequently, their correlation analysis also was performed. The results revealed that all of the FSHR mRNA, FSHR and FSH protein levels in five detected tissues (liver, heart, kidney, lung and duodenum) had the same distribution trend, and their expression level from high to low were the liver, heart, kidney, lung and duodenum. But FSHR mRNA, FSHR and FSH protein were not expressed in the spleen, stomach, breast muscle and thigh muscle. And FSH mRNA was not expressed in nine tissues. There was the significant positive correlationship (P<0.05 or P<0.01) between the tissue weight and FSH level in liver, heart, kidney, lung and duodenum, and there were also the significant positive correlations (P<0.05 or P<0.01) between the level of FSHR mRNA or protein and the FSH level in different tissues. In conclusion, it had been preliminarily revealed that FSH, coming from blood circulation, might induce the expression of the FSHR gene to promote the tissues development of the heart, liver, lung, kidney and duodenum in chicken.
Light Microscope Immunocytochemical Studies of the Distribution of Caspase-9 in the Reproductive Organs of Meat Sheep
ZHANG Taojie;CUI Yan;YU Sijiu;FAN Jiangfeng;LIU Ben;ZHENG Kai
2011, 42(11):  1526-1531.  doi:
Abstract ( 447 )   PDF (4337KB) ( 598 )  
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The expression of the apoptotic gene caspase9 in the reproductive organs of meat sheep was observed and its physiological significance was determined by using immunohistochemistric technology. Caspase9 was only found in the cytoplasm. The majority of positive cells in the ovary were observed in primordial and primary follicles.In the later luteal phase, Caspase9 was positive in granular lutein cells but negative in theca lutein cells;during the luteal phase, Caspase9 was positive in endometrial epithelium and shallow glands of the oviduct, the lymphatic nodule of the cercix uteri, the ciliated cell and secretory cell of the mucous epithelium in ampulla tubae uterinae ,and the epithelium of serous gland of the tubal isthmus; during the follicular phase, no obvious positive expression was found. All results mentioned above indicate that Caspase9 participate in the regulation of apoptosis and immunological tissue of the major reproductive organs of meat sheep. It is important for the stability of reproductive organs in meat sheep.

Cloning of CS-1 in Ovine and Tissue Expression in Liangshan Semifine Sheep
SU Qiangqiang;WU Dengjun
2011, 42(11):  1532-1536.  doi:
Abstract ( 375 )   PDF (571KB) ( 440 )  
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The aim of this study was to investigate CS1 gene sequences, protein structure, function and its expression in different tissues, so as to predict its association with meat quality traits. Calsarcins (Calcineurinassociated sarcomeric protein,CS)is a family of calciumbinding proteins,CS1 gene is closely related with meat quality in livestock and poultry. Based on the bovine, Mus musculus and Homo sapiens CS1 mRNA sequences, the cDNA of CS1 gene in ovine was amplified successfully by using comparative genomics technology. The results showed that CS1 of ovine was 951 bp in length with an open reading frame (ORF)793 bp in length and encoded 264 amino acids, and there were three conserved domains in amino acids sequences. Bioinformation analysis indicated that its secondary structures mainly were random coil and helical regions, and contained rich hydrophobic regions, certain phosphorylation sites and protein kinase C (PKC) sites. Meanwhile, tissue expression of the CS1 gene in the Semifine wool sheep were analyzed, the results showed that it was expressed in heart and muscle. When the Semifine Wool sheep was at fifteen days old, the expression was the highest in heart, and there was a significant difference between heart and other tissues (P<0.01). Furthermore, the expression of CS1 decreased with the rise of the age and became the highest when the Semifine Wool sheep was at sixty days old in heart. This result could provide scientific basis for improve meat quality traits.

遗传繁育
Genetic Diversity and Origin Based on Y-SNPs in Chinese Cattle
CHANG Zhenhua;WEI Lixuan;ZHANG Runfeng;GUO Benling;HE Cheng;LAN Xianyong;CHEN Hong;LEI Chuzhao
2011, 42(11):  1537-1542.  doi:
Abstract ( 464 )   PDF (686KB) ( 439 )  
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To determine the genetic diversity of Ychromosomal SNPs (single nucleotide polymorphisms) and paternal origin of Chinese cattle,284 bull samples from 16 Chinese native cattle breeds and 4 bulls from Burmese cattle were analyzed using PCRSSCP and sequencing methods based on four YSNPs markers of DDX3Y7,UTY19,ZFY9 and ZFY10.The result showed that 16 Chinese native cattle breeds contained Y2 haplotype (Bos taurus) and Y3 (Bos indicus) haplotype,which corresponded to two patrilines Y2 and Y3.However, no Y1 (Bos taurus) haplotype was detected in Chinese cattle.Y3 (Bos indicus) haplotype was detected in 4 Burmese cattle.Among 16 Chinese cattle breeds,the frequencies of Y2 haplotype and Y3 haplotype were 57.0% and 43.0%,respectively.Y2 haplotype dominated in the northern Chinese cattle group (98.3%),while Y3 haplotype dominated in the southern Chinese cattle group (76.1%),Central Chinese cattle group had the higher frequency of Y2 haplotype with 63.8% and the lower frequency of Y3 haplotype with 36.2%.The result indicates that Chinese cattle breeds have two distinct paternal origins of Y2 and Y3 haplotypes. The Y2 haplotype frequency of Chinese cattle decreases from North to South,and Y3 haplotype frequency increases from North to South,so the Central China become a hybrid zone of zebu(Y3) and taurine(Y2) cattle.
Unfolding of Genetic Characterization of Binglangjiang Buffalo by Microsatellite DNA Markers
LIU Wei;MIAO Yongwang;LI Dalin;TANG He;ZHANG Huifang;ZHANG Chunxi
2011, 42(11):  1543-1549.  doi:
Abstract ( 521 )   PDF (653KB) ( 509 )  
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Binglangjiang buffalo is the first indigenous river type buffalo which has been found in western Yunnan province of China in recent years. In order to elucidate genetic background information on its genetic diversity, genetic composition, genetic relationship with typical river and swamp buffalo, and the gene introgression from swamp buffalo, 141 random samples from Binglangjiang buffalo population and 24 samples from control river buffalo population (Murrah) and 41 samples from control swamp buffalo population (Diandongnan buffalo) were genotyped by employing 30 microsatellite DNA markers. The result showed that a total of 253 alleles were identified in Binglangjiang buffalo (including 54 alleles shared among the three populations assayed, 58 alleles only shared between Binglangjiang and Murrah buffalo, 73 alleles only shared between Binglangjiang and Diandongnan buffalo, and 68 alleles unique to Binglangjiang buffalo). The genetic diversity parameters in Binglangjiang buffalo were much higher than that in the control groups, its mean number of alleles per locus, mean observed heterozygosity, mean expected heterozygosity and polymorphism information content(PIC) were 8.433 3, 0.640 5, 0.600 9 and 0.594 7, respectively. Inbreeding coefficient (FIS) in Binglangjiang buffalo was 0.061 9. Genetic distance(DA) between Binglangjiang and Murrah buffalo was the lowest(0.114 4), while genetic distance(DA) between Diandongnan and Binglangjian buffalo(0.382 9) or between Diandongnan and Murrah buffalo(0.555 3) was much larger than that between the two formers, which make Binglangjiang buffalo and Murrah buffalo, grouped together and Diandongnan independently clustered in the NJ tree. Structure analysis revealed that Binglangjiang buffalo displayed a genetic mixed pattern by river buffalo and swamp buffalo, most of the individuals having some proportion of swamp buffalo component (membership coefficients =0.067 2). The results indicate that Binglangjiang buffalo is of high degree of genetic diversity and is unique genetic resource, but this buffalo is under the conditions of heterozygote deficit and genetic mixture with swamp buffalo.
动物营养
Dietary N-carbamylglutamate Supplementation on the Reproductive Performance of Sows during Late Pregnancy
LIU Xingda;WU Xin;YIN Yulong;LIU Yaqian;YANG Huansheng;LI Tiejun;HUANG Ruilin
2011, 42(11):  1550-1555.  doi:
Abstract ( 409 )   PDF (369KB) ( 787 )  
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This study was conducted to investigate the effects and potential mechanisms of dietary Ncarbamylglutamate (NCG) supplementation during late gestation on the reproductive performance of sows. Thirtytwo Landrace × Large White sows on d 80 of gestation were assigned randomly to 4 groups (control group and experimental group 1, 2, 3), 8 replicates in each group. Sows in the control group were fed with basic diet, and those in the experimental groups were fed with basic diet supplemented with 0.04%, 0.08% and 0.12% NCG. Blood samples were obtained on d 110 of gestation to examine the plasma concentration of free amino acids, NO, NOS, hormones and biochemical indices. The results showed that supplementation with 0.08% NCG increased the number of piglets born alive by 11.75% (10.75 vs. 9.62, 0.05<P<0.1) and live litter birth weight by 13.23% (16.52 kg vs. 14.59 kg,P<0.05), decreased piglets born dead per litter by 57.14% (0.75 vs. 1.75,P<0.05) and plasma urea nitrogen concentration compared with the control group, and increased plasma concentration of arginine, NO, NOS, growth hormone and Zn+ too (P<0.05). 0.04% and 0.12% NCG enhanced the reproductive performance of sows. The result suggest that dietary NCG supplementation could enhance reproductive performance of sows, and the potential mechanisms is that the supplemented NCG increases endogenous arginine synthesis in sows, so as to improve intrauterine environment and fetal nutrition, and guarantee fetus survival and growth.
Cow Injecting Lipopolysaccharide into External Pudic Artery
LI Changhao;YUAN Tingjie;WANG Jiaqi;YANG Yongxin;BU Dengpan;LI Shanshan;ZHANG Yangdong
2011, 42(11):  1556-1561.  doi:
Abstract ( 418 )   PDF (804KB) ( 478 )  
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The aims of this study were to investigate the effects of injecting low dose bacterial lipopolysaccharide (LPS) into external pudic artery on plasma proteins in lactating dairy cow, and explore the mechanism of host response to LPS challenge. All 6 cows were injected LPS(0.01 μg·kg-1 body weight). Twodimensional electrophoresis gel (2DE) coupled with MALDITOFTOF spectrometry was used. Plasma proteins at 0, 6, 12 and 24 h after LPS injecting were separated by twodimensional electrophoresis. After visualization proteins with Coomassie Brilliant Blue G250, the different protein spots were detected by PDQuest 8.0.1 software and differential proteins were identified by MALDITOFTOF spectrometry. The result showed that milk SCS started to increase after injecting LPS and peaked at 6 h, and then gradually decreased to the level before injecting LPS at 24 h. Eight protein spots were identified to be four proteins including vitamin Dbinding protein precursor, serpin A36, alpha1 antitrypsin and serpin A31 precursor. These protein expression levels were upregulated at 6, 12 and 24 h in cows after injecting LPS, but there was not significantly different among 6, 12 and 24 h. Vitamin Dbinding protein precursor, serpin A36, alpha1 antitrypsin and serpin A31 precursor play important roles in immune response, the identification of this proteins may be helpful to elucidate the molecular mechanism of host response to LPS challenge.
预防兽医
Construction of Genetically Engineered Vaccine Candidate of Type O FootandMouthDisease Virus of Pig and Analysis of Its Immunogenicity
LI Pinghua;BAI Xingwen;SUN Pu;LI Dong;LU Zengjun;BAO Huifang;CAO Yimei;FU Yuanfang;CHEN Yingli;XIE Baoxia;YIN Hong;LIU Zaixin
2011, 42(11):  1562-1569.  doi:
Abstract ( 490 )   PDF (535KB) ( 523 )  
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The outbreaks of type O footandmouth disease (FMD) severely affect development of Chinese hog industry, its causative agent is type O footandmouth disease virus (FMDV), which has been evolved several lineages. The available FMD vaccines in China cannot afford sufficient protection against viruses of multilineages, which exert important obstacle to disease control. In order to develop vaccine candidate with characteristics of good immunogenicity and broad spectrotype, we constructed a fulllength cDNA clone with genetically modified in capsid protein VP3 (58 amino acid) and VP1 (43, 48, 137, 139, 140, 141, 142 amino acid) based on the infectious cDNA clone of O/HN/93 vaccine strain. Upon cotransfection of BHK21 cells with linearized recombinant plasmids and plasmids expressing T7 RNA polymerase, the virus was rescued. The results of RTPCR and sulk mice pathogenicity showed that the rescued virus could be stably maintained during serial passages and has similar pathogenicity with parental virus for sulk mice. To test if the genetically modified virus can be used as a vaccine candidate, the pigs inoculated with inactivated FMD vaccine which made from rVOSyNa and O/HN/93 viruses were challenged with swineisolated strains of Chathay, PanAsia and Mya98 topotypes, respectively. As a result, the rVOSyNa vaccinated pigs were fully protected against three viruses challenge after 28 days postvaccination, in contrast, 16/16 O/HN/93 vaccinated animals fully protected against viruses of PanAsia and Mya98 topotypes challenge, but only 12 /16 O/HN/93 vaccinated animals got protection from virus of Chathay challenge. Our results demonstrated that genetically engineered FMDV is a good vaccine candidate with improving protection against virus of Chathay topotype challenge and expanding spectrotype.
Establishment and Preliminary Application of Classical Swine Fever Virus Pseudovirions Microneutralization Assay
ZHOU Bin;LIU Ke;WEI Jianchao;CHEN Puyan
2011, 42(11):  1570-1576.  doi:
Abstract ( 382 )   PDF (908KB) ( 593 )  
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Three plasmids, namely pcDNAE0/2/012, pHIT60 (including the structural genes of MuLV) and pHIT111 (including the retroviral genome, containing LacZ as a reporter) were cotransfected into HEK293T cells for the production of pseudotyped virions with E0/2/012 glycoproteins of classical swine fever virus (CSFV) Shimen strain. The retroviral supernatants were harvested at 48 hours posttransfection and used in western blot and infection assays. Westernblotting revealed only E012 could be expressed on the virions, indicated that the glycoprotein E012 was incorporated onto the retroviral virions. Infection test were performed on SK6, PK15, ST, BHK21, Vero, COS7, HEK293T and CEF cells. The results showed that SK6, PK15 and ST infected were LacZ positive, indicating viral entry, and revealed that the pseudtype virions of MuLVE012 were infectious. To assess whether the CSFV pseudotyped virus entry is pHdependent, PK15 cells were treated with wellcharacterized lysosomotropic agent NH4Cl. Treatment with 30 mmol·L-1 NH4Cl caused >90% inhibition of infection by MuLVE012 or MuLVVSV G. These data indicated that the MuLVE012’s entry may be pHdependent. The pseudotyped MuLVE012 particles were used to develop an in vitro microneutralization assay that was both sensitive and specific for CSFV neutralizing antibody. Neutralization titers measured by this assay were highly parallel with those measured by the assay using live CSFV high virulence strain. Because the pseudotype assay does not require handling live CSFV virus, it is a useful tool to determine serum neutralizing titers during natural infection and the preclinical evaluation of candidate vaccines.
Influence of the Mutations of VP2 on the Replication Efficiency of Infectious Bursal Disease Virus in vitro
QI Xiaole;GAO Li;WU Guan;DENG Xiaoyun;;YU Fei;ZHANG Lizhou;QIN Liting;GAO Yulong;WANG Yongqiang;GAO Honglei;LIU Di;HUA Yuping;WANG Xiaomei
2011, 42(11):  1577-1583.  doi:
Abstract ( 493 )   PDF (1016KB) ( 462 )  
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To study the molecular basis for the replication efficiency and virulence of infectious bursal disease virus (IBDV), two amino acid mutations (A222P, D279N) was introduced in to VP2 of IBDV, respectively, based on the information of sequence analysis. Using the reverse genetic research system, two responding mutated strains of IBDV were rescued and their biological characteristics were studied. The replication kinetic curves showed that the rescued virus including A222P replicated over 6 times higher than its parent strain did, while D279N couldn’t. Animal experiment of SPF chickens showed that residue 222 and 279 of VP2 did not contribute to virulence of IBDV. The relations between the two residues of VP2 (222, 279) and the replication and virulence of IBDV were reported firstly. The findings provide necessary information for further understanding the gene function of IBDV and the design of new tailored IBDV vaccines.
Comparison of Antigenicity and Immunogenicity of H9N2 Avian Influenza Virus Strains Isolated from different Years
WANG Xinwei;YANG Daguang;WANG Chuanqing;WANG Zelin
2011, 42(11):  1584-1590.  doi:
Abstract ( 432 )   PDF (374KB) ( 436 )  
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Antigenicity and immunogenicity of 25 H9N2 subtype avian influenza viruses isolated from China during 19982008 were compared and analyzed by crossinghemagglutinin inhibition test(HI), cells neutralization test and challenging test. HI test results revealed that the 25 H9N2 AIVs were divided into three classes, the first class including 12#, 17#, 18#, whose HI titer was from 4.0 log2 to 5.0 log2, the second including 2#, 5#, 11#, 15#, 16#, 21#, 22# and 25#, whose titer was from 6.0 log2 to 7.0 log2, the rest strains formed the third whose titer was from 7.0 log2 to 8.0 log2. It was indicated that the antigenicity of different isolation strains of H9N2 AIV displayed variant, and Maximum difference of them in HI titer reached to 3.15 log2. Neutralization test demonstrated that the antigen correlation(R) among 8 representative H9N2 AIV strains selected were from 0.42 to 0.84, the antigenicity between 3# and 14#, 11# and 17# showed diversity (the R was 0.46 and 0.42), but that of all the rest strains showed inconspicuous difference. Cross challenging protection test among 6 representative H9N2 AIV strains selected made clear that the antibody induced by the vaccine from the viruses could protect the chicken against infection. The birds vaccinated with Hp vaccine could be protected against the challenge from both three H9N2 AIV strains (10#, 12#, 17#) displaying different antigenicity, and jointchallenge with different viruses (9# and15#,16# and 17#). It was suggested that the radical change in immunogenicity of the prevalent H9N2 AIVs did not happen up to now.
Development of Blocking ELISA for Mycoplasma suis with Peroxidaselabelled Monoclonal Antibody Against MSG1 Protein
ZHANG Changying;LI Yufeng;JIANG Ping
2011, 42(11):  1591-1597.  doi:
Abstract ( 392 )   PDF (443KB) ( 438 )  
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To provide a serological method for detecting antibodies against Mycoplasma suis (M. suis), a blocking ELISA was developed based on a monoclonal antibody (MAb) to M. suisMSG1 protein. The conditions for each step were optimized. The optimal concentration of the coating antigen was 0.5 μg·mL-1; sera samples diluted 1 fold and incubated 1.5 h at 37 ℃; MAbHRP dilution is 10 000 fold and incubated 1 h at 37 ℃. The blocking results of one hundred M. suis negative sera samples were statistically analyzed, and the cutoff of blocking ELISA was determined that the samples presenting a percentage inhibition of ≥ 23.71% were considered positive; samples with a calculated percentage inhibition of ≤ 36.35% were rated negative and those presenting a blocking effect between 23.71% and 36.35% were considered inconclusive. The Blocking ELISA proved to be specific, sensitive and it showed high reproducibility and low variability. This method will be useful in clinical detection and epidemiological study on M. suis.
基础兽医
Transcription Change of Chicken NLRC5 Receptor and IFN Genes Initiated by Bacterial Endotoxins
CHANG Guobin;CHEN Rong;LUAN Deqin;ZHANG Ying;LIU Xiangping;MA Teng;DAI Aiqin;ZHOU Wei;CHEN Guohong
2011, 42(11):  1598-1604.  doi:
Abstract ( 420 )   PDF (837KB) ( 548 )  
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This study was conducted to investigate the transcription change of chicken NLRC5 receptor and IFN genes initiated by different bacterial endotoxin, aiming to provide a theoretical basis on poultry NLRC5 receptor function and its regulation mechanism on innate immune. Different concentrations (0, 0.1, 1.0, 10.0 μg·mL-1) of endotoxin from Escherichia coli(E.coli,EC) and Salmonella enteritidis(S. enteriditis,SE) were used to stimulate the chicken macrophage cell line (HD11) for 2,4 and 6 hours, respectively. Total RNA was isolated from cells at different time points, then the transcription difference of NLRC5, IFNA and IFNB genes between different treatments were determined by quantitative realtime RTPCR, using SYBR Green RTPCR, and immune response for HD11 cells to different bacterial endotoxin were finally analyzed. In comparison with the control group, 1.0 and 10.0 μg·mL-1 of the ECLPS, SELPS could induce immune responses in HD11 cells, whereas 0.1 μg·mL-1 group failed to induce responses successfully. The immune response time induced by different bacterial LPS was different. NLRC5 and IFNB gene transcription induced by ECLPS, SELPS showed upregulation and reached the peak at 6 hours poststimulation, which presented significant difference with that of 2,4 hours poststimulation, but IFNA gene transcription induced by ECLPS reached the peak at 2,6 hours, while was down regulated at 4 hours poststimulation, showing as "V"type transcription trend. The transcription of all three genes induced by SELPS reached the peak at 6 hours poststimulation and presented significant difference with that of 2, 4 hours and control group, showing as one up trend line with time going on. The results showed that the IFNA, IFNB and NLRC5 receptors participated jointly the immune response to bacterial endotoxins, and NLRC5 genes may have regulate function on innate immune in chicken.
Effect of Monochromatic Light on the Distribution and Expression of Gonadotropinreleasing Hormone (GnRH) in Broiler Brain
WANG Yao;SONG Zhiqi;CHEN Yaoxing;CAO Jing;WANG Zixu;YANG Guang
2011, 42(11):  1605-1609.  doi:
Abstract ( 381 )   PDF (689KB) ( 471 )  
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24 0 dayold male broilers were reared under four light treatments (n=6), white (400760 nm), red (660 nm), green (560 nm) and blue (480 nm) by using LED (lightemitting diodes) as light sources. Light intensity was 15 lx at the height of birds’ heads and scheduled for 23 h of light and 1 h of dark during the entire experiment. Using the method of immunohistochemistry, the distribution and expression of GnRH were shown in different light groups at 49 day of age. The results showed GnRH neurons and fibers were distributed in hypothalamus, thalamus and mesencephalon, and GnRH expression in nucleus preopticus periventricularis (POP), nucleus preopticus medialis (POM), nucleus periventricularis hypothalami (PHN), nucleus paraventricularis magnocellularis (PVN), nucleus ovoidalis (OV) and nucleus rotundus (ROT) in blue light group was higher than those of in red and green light groups. The results suggested that color lights could affect the expression and secretion of GnRH in hypothalamus and thalamus of avian.
PCNA Expression in the Bursa of Fabricius (BF) in Chicken Early Embryo Development
YUAN Yuan;LIU Shuqiang
2011, 42(11):  1610-1614.  doi:
Abstract ( 379 )   PDF (3544KB) ( 457 )  
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The bursa of Fabricius (BF) is a special organ in birds, being the site of mature B lymphocytes. This study investigated proliferating cell nuclear antigen (PCNA) expression in the BF in chicken early embryo development (5 to 18 days). The results showed that the BF of chicken embryos comes into being at the deepest point of the chicken cloaca at embryos of 6.5 days (E6.5), commencing from small vacuoles that develop into one large capsule at 8 days (E8), which continues growing from 8 to 10 days. The BF begins to exhibit small ruga in the capsule at E11, the ruga then become increasingly evident from 11 to 18 days, come through the whole BF, and the BF forms. When the BF profile formed at E9, with the strongest PCNA expression in the mucous epithelium, there is also broad PCNA expression in the lamina propria as the early embryo develops (9 to 18 days), the relative percentages of PCNAimmunopositive cell accounting for the total mucous epithelium were 100%, 100%, 100%, 99%, 99% and 99% at E9, E10, E11, E12, E14 and E18, respectively; the relative percentages of PCNAimmunopositive cell accounting for the total lamina propria cells were 97%, 93%, 71%, 55%, 56% and 20% at E9, E10, E11, E12, E14 and E18, respectively, which suggesting that lamina propria cells proliferation of BF decrease gradually in chicken early embryo development.
临床兽医
Analysis on the Dynamic Changes of Lymphocyte Subsets and Genetic Transcription of Cytokine IL-2, IL-4 in Peripheral Blood of Dairy Cows during Perpartum
CUI Qunwei;GUO Ning;ZHOU Xuan;WANG Zhenyun;WANG Yachun;WANG Genlin
2011, 42(11):  1615-1619.  doi:
Abstract ( 395 )   PDF (472KB) ( 526 )  
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To reveal the mechanism of impaired immune function in perpartum period, the dynamic changes of CD3+, CD4+ and CD8+ in lymphocyte subsets as well as gene transcription level of cytokine IL2 and IL4 in peripheral blood of dairy cows during 21 days before calving to 21 days after it were investigated in the present study. The dynamic changes of Lymphocyte cells CD3+, CD4+, CD8+ and the mRNA expression of cytokine IL2, IL4 in peripheral blood were determined by flow cytometry and RTPCR respectively. The results showed that significant difference was found in the proportion of CD3+ in peripheral blood 21 days before calving (50.8±4.61)% and 14 days after caving (39.21±4.98)%, and also was found in the proportion of CD4+ in blood before (21.38±0.65)% and after (6.89±0.32)% calving. The proportion of CD8+ increased to the highest level (16.9±2.02)% during calving (P<0.05). The ratio of CD4+/CD8+ was >1 before calving, and <1 after calving. The mRNA expression of IL2 was the lowest (P<0.05) during 014 days after calving, then gradually increased. The mRNA expression of IL4 increased to the peak during calving, and declined to the lowest level on the 7th day (P<0.01). It was suggested that the immune function of dairy cows was impaired within 21 days before and after calving, especially during the 014 days after caving. This might be caused by the quantity and ratio changes of lymphocyte subsets (CD3+, CD4+and CD8+) and the downregulation of the cytokine IL2, IL4 during this critical period.
研究简报
Characteristics of the Spatial and Temporal Expression of SPMI Gene in the Genital Tracts of Boars and Gilts
SONG Chengyi;GAO Bo;WANG Xiaoyan;WU Han;ZHOU Huiyun;LI Bichun;CHEN Guohong;MAO Jiude
2011, 42(11):  1620-1624.  doi:
Abstract ( 373 )   PDF (739KB) ( 436 )  
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To investigate the spatial and temporal expression profiles of seminal plasma motility inhibitor (SPMI) gene in the genital tracts of boars and gilts, the qualitative RTPCR was used to detect tissue expression pattern along reproductive tracts and semiquantitative RTPCR was applied to determine the developmental expression of SPMI in seminal vesicle and bulbourethral gland in the current study. The qualitative RTPCR revealed that the expression of SPMI mRNA was highest in seminal vesicle, moderate in bulbourethral gland and uterine horn, weak in prostate, cervix and ovary, no signal was found in the other reproductive tissues. The semiquantitative RTPCR revealed that the similar developmental expression pattern of SPMI in seminal vesicle and bulbourethral gland was found, and the expression of SPMI both in seminal vesicle and bulbourethral gland initiated on day 1 and improved until sexual maturity (150 d) with age increase. The expression of SPMI in seminal vesicle increased significantly on days 60, 90 and 150 d (P<0.05); while the expression of SPMI in bulbourethral gland increased significantly on days 30, 90 and 150 d(P<0.05), the expression increased on day 60 d compared with that on day 30 d, but the difference was not significant (P>0.05). These results indicate that the SPMI express extensively along the genital tracts of boars and gilts, and in general, the temporal expression of SPMI during boar development is age dependent and the expression level is improved significantly from puberty to sexual maturity.
The Distribution of LH Receptor on the Ovaries among Three Different Sheep Breeds by Different Efficiency of Superovulation
NI Hemin;ZHANG Hongbo;LIU Yunhai;DENG Guixin;LU Tiangang;GUO Yong
2011, 42(11):  1625-1631.  doi:
Abstract ( 374 )   PDF (1979KB) ( 532 )  
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In this study, it was aimed to investigate the distribution of luteinizing hormone receptor (LHR) on the ovaries among three different sheep breeds by different efficiency of superovulation. Three local sheep breeds (Smalltailed Han sheep, White Tan sheep and Ujumqin sheep) were selected depending on their different sensitivity to superovulation by LH as following: Smalltailed Han sheep, White Tan sheep, and Ujumqin sheep. At 8, 16, 24 hour after the last injection with LH, the sheep ovaries were collected and treated by immunohistochemical staining to observe the distribution of LHR on their different ovarian follicles. The results showed that: ① LHR were expressed in both granulosa and theca cells of antral follicles,but did not express on the preantral ones. ②At 8, 16, 24 hour after the last injection with LH, the LHR levels on the prematured follicles of the three local sheep breeds were significantly higher than those on the corresponding secondary ones respectively(P<0.05). ③The expression change of LHR in the prematured follicles had significant difference among the three sheep breeds, the order as following: Smalltailed Han sheep>White Tan sheep>Ujumqin sheep(P<0.05). The result indicate that: ①The induction of LH for the development of the primary ovarian follicles may not be through the specific LH receptor signal way, but through some other nonspecific ones. ②The expression change of LHR in the prematured follicles is consitent with the effect of superovulation, which indicate that the effect of superovulation is linked with the distribution of LH receptor on ovaryes in sheep.
Analysis on Polymorphism and Expression in Different Coat Color Skin of Syntaxin17(STX17) Gene in Mongolian Horse
LI Bei;HE Xiaolong;MANG Lai
2011, 42(11):  1632-1637.  doi:
Abstract ( 389 )   PDF (1227KB) ( 405 )  
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In order to research the relationship between syntaxin17(STX17) gene and Mongolian horse coat color and mechanism, the polymorphism in intron 6 and the relative expression quantity in different colors skin of Mongolian horse were analyzed by seminested PCR and realtime quantitative PCR. The results showed that 4.6 kb duplication segments existed in intron 6 of STX17 gene in gray Mongolian horse, but not existed in nongray Mongolian horse by seminested PCR. The expression level of STX17 gene was highest in gray skin and lowest in bay skin by realtime quantitative PCR. The results indicate that the STX17 gene has typical relationship with gray Mongolian horse.
Prokaryotic Expression of Reindeer β-defensin-1(reBD-1) Gene and Bioactivity Identification of the Recombinant Protein
SU Lina;YANG Yinfeng;JING Lan;CAO Guifang
2011, 42(11):  1638-1642.  doi:
Abstract ( 412 )   PDF (940KB) ( 425 )  
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The aim of this experiment was to construct prokaryotic expression vector of reindeer βdefensin1(reBD1), induce the expression of reBD1 fusion protein in E.coli, and evaluate the bioactivity of the expression products. The prepropeptide of reBD1 was amplified by RTPCR. The mature peptide of encoding reBD1 was amplified from recombinant cloning vector PMD19T/reBD1, and then cloned into pET32a(+), in which reBD1 fusion protein expression was induced by IPTG in E.coli BL21(DE3). The expressed product was further cultured and purified for experiment of bacteriostasis in vitro. The results showed that the amplified products of prepropeptide and mature peptide were 215 and 138 bp, respectively, and the homology of the sequences of the targeted gene and reBD1 mRNA was up to 100%. The molecular weight of fusion proteins of prepropeptide and mature peptide were 28 and 24 ku.The agar diffusion method has demonstrated that 0.08 mg·mL-1 purified mature peptide protein has obvious antimicrobial activity against S. aureus and E.coli. From the results we can conclude that the prepropeptide and mature peptides have high expression level in E.coli, and the mature peptide has resistance to both Gram negative and Gram positive bacteria.
New Pathogenicity Characters of Natural Coinfection ALVJ and REV in Layer Breeding Chickens
WANG Yue;JIANG Yanping;YU Linlin;WANG Feng;CHEN Hongbo;WANG Xiaowei;CHENG Ziqiang;
2011, 42(11):  1643-1648.  doi:
Abstract ( 420 )   PDF (1223KB) ( 448 )  
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An outbreak of productive diseases occurred in a commercial layer breeding company in Yuci, Shanxi Province during August 2009. Lymphocytoma presented in lung, liver, kidney, heart, etc. ELISA, PCR and IFA results indicated that the positive rate of serum antigen to ALVJ was 8/8 and the viremia rate was 7/8. The coinfection rate with REV was 2/8, and found no single REV infection. The pathogenic observation indicated that the most serious lymphocytoma presented in coinfected viremia chicken, but no myelocytomas were observed. Phylogenitic analysis showed that the homology was over 96.9% of three strains of ALVJ with the prototype HPRS103; The REV strain showed the highest homology with suvenv strain (93.8%). The case revealed that viremia state induced by ALVJ is necessary for lymphocytomas formation induced by REV, and it is the reason why lymphocytomas showed earlier than myelocytomas. So, it is critical to eliminate vertical transmission of ALVJ for prevention these diseases. The synergistic effect mechanism of ALVJ and REV need to be further study.
Prokaryotic Expression and Immunogenicity of E2 Gene of CSFV Guizhou Epidemic Strain
TANG Deyuan;LI Chunyan;LUO Xianfeng;HUANG Tao;ZENG Zhiyong;GAN Zhenlei;WANG Feng
2011, 42(11):  1649-1653.  doi:
Abstract ( 413 )   PDF (718KB) ( 566 )  
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According to published cDNA sequence of E2 gene in GenBank of CSFV Shimen and C strain, the synthesized specific primers was designed. E2 gene fragments of classical swine fever virus Guizhou strain were amplified by extraction tissue RNA of the suspected CSF dead and RTPCR. The PCR products were sequenced and comparatively analyzed. The products of RTPCR were cloned into pMD18T vector to form recombinants pMD18TE2 and subcloned into prokaryotic expressing vector pET32a(+). The pET32aE2 was transformed into the E. coli strain BL21(DE3) and the expression of the E2 protein was induced. After analysis by SDSPAGE electrophoresis of the induced expressed protein and Western blot, we get about 58 kDa protein in line with the expected size. Through further extracting and purifying the target protein, the expressed protein was used to immunize mice by adding adjuvant, and then the antibody titers of mice were detected. The results showed that the target protein induced antiCSF antibody in mice. This study laid the foundation for the research of subunit vaccine of swine fever.