Acta Veterinaria et Zootechnica Sinica

Study on Polymorphism and Genetic Differentiation of pGH Gene in 11 Pig Breeds

SHUAI Su-rong;LI Xue-wei;ZHU Li;LI Hui;ZHAO Zhong-quan

2007, 38(8): 753-759. doi:

Abstract ( 2101 )

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By applying PCR and sequencing technique, the polymorphism and genetic differentiation of porcine growth hormone gene in 65 individuals form 11 pig breeds were studied.43 SNPs loci and 7 bp lack segment within intron 3 were found. Analysis results showed that the variance is different at different function region The percentage of SNPs loci in exon region, intron region and 5'UTR and 3'UTR region are 1860%, 74.42% and 6.98%. There are 8 SNPs loci in exon region which bring about 4 amino acid loci variation, exon 2 is the high variation region.The variance of SNPs loci belongs to neutral mutation The pGH gene haplotype among 11 breeds is unique. Results of AMOVA analysis showed that the variation of pGH gene sequence is higher in interbreeds (Percentage of variation is 73.45%) than interbreeds （Percentage of variation is 26.55%）, the diversity among breeds is significant at 0.01, and the genetic differentiation of pGH gene is significant The main favorable factor affecting pGH gene evolution and breeds genetic differentiation is intron region. The genetic differentiation among breeds is related to gene flow and geographic distribution

Analysis on Associations of Myopalladin Gene Polymorphisms with Carcass Traits in Pigs

WANG Chong;HUANG Zhi-hong;ZHANG Xi-quan;QIU Zhi-peng;LI Jia-qi;FANG Wei;CHEN Yao-sheng

2007, 38(8): 760-764. doi:

Abstract ( 2267 )

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Single nucleotide polymorphisms (SNPs) were detected by RTPCR amplification and sequencing on the EST (GenBank accession No. BM445313)which was expressed in porcine muscle. Two new SNPs were found and located in the 3′UTR region of Myopalladin gene by homologous comparative analysis in the nonredundant sequence database of NCBI through BLAST search. One of the SNPs was genotyped by RFLP with restriction enzyme Csp6I and used for association analysis with carcass traits in the SCAU-LL F₂ pig resource population. The results showed that the average carcass lean meat percentage of CT and CC genotype were 46% and 45% respectively, and the average carcass skin & fat percentage of the two genotype were 42% and 44% respectively, these differences were significant (P<0.05). Therefore, it could be seen that porcine Myopalladin genotypes had significant influence on carcass traits, and might be considered as a potential genetic marker for selection.

Polymorphism of FcRn Gene and Its Effects on the Sow Colostrum IgG Concentration

WANG Bing-yan;WANG Li-xian;WANG Ting;ZHAI Li-wei;WANG Chu-duan;LI Zhen-kuan;ZHOU Wei-liang

2007, 38(8): 765-768. doi:

Abstract ( 1515 )

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The polymorphism of the FcRn gene was analyzed, and two SNPs were found by the technique of single strand conformation polymorphism (SSCP) and finally confirmed by sequencing. The frequencies of different genotypes in 207 sows of the three breeds were calculated and the relationship between three genotypes and IgG Concentration in Colostrum of the Sow were analyzed based on liner model. The results showed that the mutation in the Site1 did not have effect on IgG concentration in colostrum of the sows( P>0.05) , the mutation in the Site6 had effect on IgG concentration in colostrum of the sows ( P<0.05); Multiple t comparisons in site6 indicated that IgG concentration in colostrum of the sows between CC,CD and DD genotype were significant(P<0.05)：IgG concentration in colostrum of the sows of CC is the highest, while that of the sows of DD is the lowest. The result indicated FcRn gene could be a candidate gene of IgG concentration in colostrum of the sows.

Expression Rule of TGH Gene in Adipose Tissue and Primary Cultured Adipocytes of Pig

ZHANG Li-jie;WANG Qi;ZHANG Li-hong;YANG Gong-she

2007, 38(8): 769-775. doi:

Abstract ( 828 )

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Triacylglycerol hydrolase (TGH) is an intracellular neutral TG lipase identified in murine adipocytes. To explore the transcriptional expression rule of TGH gene in porcine adipose tissue, mRNA levels of TGH and HSL genes in variously located adipose tissue from pigs in different types, growth stage were assessed by semiquantitative RT-PCR. The results showed that TGH mRNA level was significantly higher in adipose tissue of lean pigs than those of obese and crossbreed ones, and also higher in piglets at birth than mature pigs. On the other hand, the abundance of TGH mRNA increased in the order as subcutaneous, peritoneal and visceral fat. All these variances of TGH gene expression were in accordance with that of HSL gene. To elucidate the time sequence variety of TGH gene expression during the adipocytes differentiation, preadipocytes isolated from adipose tissue of 1-3 day-old piglets were induced to differentiate and the differentiation state was confirmed by Oil Red O staining. The results of TGH gene expression determination showed that instead of expressing in preadipocytes, TGH mRNA expressed in differentiating adipocytes and the level reached its peak at 4 d after differentiation induction, which was earlier than that of HSL mRNA did. These data indicated that TGH gene expression was related to the site of adipose tissue, economic type, and age of pig as well as differentiating state of adipocyte, implying that TGH might play a major role in the basic lipolysis of adipose.

ERα mRNA Expression in Reproductive Tissues of Meishan Sows during Estrus Period

DING Li-jun;LIU Yan-jie;DING Jia-tong

2007, 38(8): 776-780. doi:

Abstract ( 741 )

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To investigate localization and quantum of ERα mRNA in ovary and uterus of porcine on D 2, 7, 12, 17 throughout menstrual cycle, nonradioactive in situ hybridization using oligonucleotide probes labeled by DIG was applied In the porcine ovary, positive signals for ERα mRNA were limited exclusively to the granulose cells of all types of follicles and were present in a few cells of early corpora lutea. Level of ERα mRNA in ovary remained low during the whole oestrus cycle but increased during proestrus on D 17. ERα mRNA positive signals were mainly detected in glandular epithelial cells and a few of stroma cells in the uterus Abundance of ERα mRNA in porcine endometrium on D 17 was significantly higher than that of D 12 (P<0.01). E₂ concentrations of serum on D 2, 7, 12, 17 during the menstrual cycle were measured by radioimmunoassay. The levels of ERα mRNA in porcine endometrium synchronized the changes of estrogen with positively relations (r=0.918).

SNP Detection and Haplotype Analysis on Exon 16 of Chicken lmbr1

HUANG Yan-qun;CHEN Wen;DENG Xue-mei;QIU Xiang-pin;LI Ning;WU Chang-xin

2007, 38(8): 781-785. doi:

Abstract ( 811 )

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Exon16 is the biggest exon of chicken lmbr1,here we conduct the SNP detection and haplotype analysis on Silkies and White Plymouth Rock broiler. It shows that the distribution of exon 16 PCR-SSCP genotypes had clear difference between Silkies and White Plymouth Rock broiler ,4 variations were detected by sequencing from 24 individuals, the distribution of allele at T32C site had clear difference between Silkies and White Plymouth Rock broiler too, all the individuals of Silky have 32T allele(in homologous TT or heterozygous TC),while all White Plymouth Rock are homologous CC at T32C site. 5 haplotypes were detected from 24 individuals, the distribution of haplotypes between two breeds also had clear difference,hap1 and hap2 were the special haplotypes of Silkies, while hap5 was the special haplotype of White Plymouth Rock broiler, the hap3 and hap4 were mainly detected from White Plymouth Rock broiler, which were rare in Silkies.

The Effect of ADSL and GARS-AIRS-GART Genes on Inosine Monophosphate Content in Chicken Meat

SHU Jing-ting;JI Wen-lin;BAO Wen-bin;CHEN Guo-hong;ZHANG Xue-yu;JI Cong-liang

2007, 38(8): 786-791. doi:

Abstract ( 836 )

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This study was designed to investigate the effect of ADSL, GARS-AIRS-GART and combination genotypes of ADSL and GARSAIRSGART on IMP content in chicken meat. Five chicken breeds, including Recessive White, Silkies, Baier, Tibetan and Xiaoshan chickens were used. The primers for the exon2 in ADSL gene and 5′UTR region in GARS-AIRS-GART gene were designed and the SNPs were detected by PCR-SSCP method and DNA sequencing. Two SNPs were detected in exon2 of ADSL gene and 5′UTR region of GARS-AIRS-GART gene separately. The least square analysis showed that TT genotype birds had greatly significant higher IMP content than CC (P＜0.01) and significant higher than CT (P＜0.05) genotype birds, TC genotype birds also had a little higher IMP content than CC genotype birds, but the difference was not significant in ADSL gene TT genotype birds had significant higher IMP content than CC and CT genotype birds (P＜0.05), TC genotype birds also had a little higher IMP content than CC genotype birds, but the difference was not significant in GARSAIRSGART gene. The combination genotypes of ADSL and GARS-AIRS-GART genes also had significant effect on IMP content. The IMP content of the individuals with TTTT combination genotype was 1.584 mg/g higher than the individuals with CCCC combination genotype. Therefore we putatively drew the conclusion that the combination genotypes of ADSL and GARSAIRSGART gene could be used as the molecular genetic marker to select the chicken for meat quality traits.

Relationship between the Polymorphisms of Microsatellites DNA and ongissimus dorsi Traits in Two Meat Sheep Populations

REN Hang-xing;LIU Guo-qing;ZHANG Xing-guo;DAI Rong;SHI Guo-qing;LV Gao-rui;GUI Dong-cheng

2007, 38(8): 792-799. doi:

Abstract ( 1267 )

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To investigate the relationship between the polymorphisms of the microsatellites and the Longissimus dorsi (L. dorsi) traits, ten highly polymorphic microsatellite loci which probably be linked with the Carwell gene were selected from the genetic linkage map of chromosome 18 in sheep. The polymorphisms of these microsatellites were detected, and the genetic character was analysed in two grading crossing sheep group(F2 from Dorset♂and Suffolk♂×Xinjiang fine wool sheep♀,respectively ) with nondenaturing polyacrylamide gel electrophoresis by multiplex silver staining. Results showed that there were differences for genetic parameters between the two meat sheep groups among these loci . The effect of these loci on L. dorsi traits were analyzed in a generalized linear model(GLM). Results indicated that : (1) The effect of BM3413 on thickness of L. dorsi reached a significant level at P<0.05 , and TGLA122 had a significant effect on width of L. dorsi (P<0.05) , respectively; The locus MCMA26 had a significant impact on thickness, width and area of L dorsi traits (P<0.05) in the Dorset group, respectively. While the other loci(ILSTS54, TGLA337, HH47, TGLA122, BP33, MCM38 and CSSM18 ) had no impact on the L dorsi traits (P>0.05) in the same group; (2) In the Suffolk population, the loci of MCM38, CSSM18, ILSTS54 and BM3413 each had a significant impact on all the three L. dorsi traits simultaneously (P<0.05 or P<0.01 ); And the other 6 loci( TGLA337, HH47, TGLA122, BP33, OB2 and MCMA26 ) had no effect (P>0.05) in the same group. We also found that, 196 bp/222 bp and 192 bp/220 bp at MCMA26 were the most favourable genotypes for area of L. dorsi in the Dorset group; 124 bp/134 bp at CSSM18, 130 bp/148 bp and 134 bp/148 bp at ILSTS54 , 190 bp/232 bp at BM3413 were the most favourable genotypes for area of L. dorsi in the Suffolk group. Two conclusions were drew : (1) The Carwell gene probably located in the region of MCMA26- CSSM18, and there may be a new gene or QTLs (quantitative trait loci) flanking the locus BM3413; (2) The most favourable genotypes we found may be regarded as the candidateloci for the marker -assisted selection or markerassisted breeding in the merit meat sheep on early period.

Genetic Variation in GDF9 and BMP15 Genes and Their Relationships with Twinning Trait in Cattle

ZHANG Lu-pei;ZHANG Xiao-hui;XU Shang-zhong;GAO Xue;REN Hong-yan;CHEN Jin-bao

2007, 38(8): 800-805. doi:

Abstract ( 801 )

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Growth differentiation factor 9 (GDF9) gene and bone morphogenetic protein 15 (BMP15) gene were studied as candidate genes on the twinning trait in cattle Genetic variation of GDF9 and BMP15 genes in Luxi cattle, Qinchuan cattle, Nanyang cattle and Chinese Holstein cattle was analyzed. The relationship between genetic polymorphisms and twinning trait was studied in Luxi cattle The results showed that the 2-base deletion at 3′UTR of GDF9 gene was only detected in Luxi cattle. The χ2 test of association between genotypic distribution and twinning or monotocous trait in Luxi cattle showed significant difference (P=0.006). Frequency of B allele in Luxi twinning cows was higher than that in Luxi monotocous cows. According to bioinformatics analysis, the total free energy of the secondary structure of the mutation type mRNA was almost the same as the wild type mRNA But the stability of the secondary structure in translation initiation region (TIR) of mutation type mRNA was significantly higher than that of wild type mRNA.4-base deletion in coding sequence of BMP15 gene was detected in Luxi, Nanyang and Qinchuan cattle Homozygous mutation was not detected in these groups. The χ2 test of association between genotypic distribution and twinning or monotocous trait in Luxi cattle showed no significant difference (P=0.947).

Effects of L-carnitine and CoQ₁₀Supplementation on Performance and Some Immune Functions in Ascites-Susceptible Broilers

GENG Ai-lian;LI Bao-ming;GUO Yu-ming

2007, 38(8): 806-813. doi:

Abstract ( 730 )

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To investigate the effects of L-carnitine and CoQ10 (CoQ) supplementation on performance and immune functions in ascites-susceptible broilers. L-carnitine supplemental levels were 0, 75, 150 mg/kg, CoQ supplemental levels were 0 and 40 mg/kg, respectively. Four hundred and eighty 1 day-old Arbor Acre male broiler chicks were randomly allocated into 12 groups with 5 replicates each, 8 birds per replicate. The results showed that performance was not significantly influenced by Lcarnitine and CoQ supplementation alone, but heart index of broilers was significantly increased during 0-3 week (P＜0.05),and the development of immune crgans was not significantly affeded.PCV was significantly decreased by L-carnitine supplementation alone (P<0.05), and was not affected by CoQ supplementation alone AHI and ascites mortality were decreased by L-carnitine, CoQ supplementation alone, and L-carnitine + CoQ supplementation together (P<0.05). Serum IgG content was improved by Lcarnitine supplementation alone (P<0.05), and was not affected by CoQ supplementatim alone and L-carnitine+CoQ supplementatim together. Lysozyme activity was increased by L-carnitine + CoQ supplementation together (P<0.05). Supplemental CoQ alone and L-carnitine + CoQ together decreased the peripheral blood lymphocyte (PBL) proliferation in response to concanavalin A (ConA) (P<0.05). The study suggested that the reduction of broilers′ susceptibility to ascites might be relevant with L-carnitine + CoQ supplementation’s improving some immune functions of broilers.

Regional and Ontogenetic Expression of PepT1 mRNA in Intestine of Broiler

WANG Xiu-qi;ZOU Shi-geng;ZUO Jian-jun;SU Hai-lin;HUANG Zhi-yi;FENG Ding-yuan

2007, 38(8): 814-821. doi:

Abstract ( 1122 )

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120 1-day-old parental male Arbor Acre (AA) broiler and 120 1-day-old parental male Lingnan yellow broiler were randomly divided into 4 replicates, respectively. Regional expression of Peptide transporter 1 (PepT1) mRNA in different intestinal tract segments of Lingnan yellow broiler on 30 d and ontogeny of PepT1 mRNA in duodenum and jejunum of AA broiler and yellow broiler were determined by relative quantitative RT-PCR. The results showed that: (1) Regional expression of PepT1 mRNA in duodenum, jejunum, ileum and colorectum was declined gradually. The abundance of PepT1 mRNA in duodenum was higher than in colorectum (P<0.05)； (2)Ontogeny patterns of PepT1 mRNA in duodenum and jejunum were consistent between AA broiler and Lingnan yellow broiler. The abundance of PepT1 mRNA in two strains was the highest on day 16, decreased from day 16 to day 44 and slightly increased again on day 58. While compared two breeds directly, the abundance of PepT1 mRNA in AA broiler was not significant difference than that in Lingnan yellow broiler from day 2 to day 58(P>0.05). These results indicated that： (1) Regional expression of PepT1 mRNA in intestine of Lingnan yellow broiler was declined gradually along the downward intestine from duodenum, jejunum, and ileum to colorectum. The abundance of PepT1 mRNA in duodenum was higher than in colorectum (P<0.05)；（2） Ontogeny of PepT1 mRNA in two breeds had the same pattern in duodenum and jejunum, and there was no difference at same development stage between AA broiler and Lingnan yellow broiler, indicating that PeptT1 mRNA expression could be regulated by developmental stage but not breeds.

Cloning and Analysis of Replicase Polyprotein-Encoding ORF1 of Transmissible Gastroenteritis Virus SC-Y Strain

SONG Zhen-hui;GUO Wan-zhu;YIN Hua-ping;ZENG Zhi-yong;ZHU Ling;LUO Ai-fang;ZHANG Ying-jun

2007, 38(8): 822-826. doi:

Abstract ( 1544 )

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In this study, the complete sequence of the replicase polyproteinencoding ORF1 of TGEV SC-Y strain was obtained by RT-PCR, the entire nucleotide sequence was determined(GenBank Accession No. DQ390461).Results showed that there were two partially over lapping open reading frames in replicase polyprotein-encoding ORF1 of SC-Y strain , the ORF1a contains 12 053 nucleotides encoding a polypeptide of 4 018 amino acids, the ORF1b contains 8 036 nucleotides encoding 2 678 amino acids. The sequences were compared among different TGEV strains and other coronary viruses. The results showed that the SC-Y has the highest homology with PUR46MAD strain,the ORF1a and ORF1b nucleotide sequences had 995% and 998%, amino acid sequences had 99.2% and 99.8% respectively. The ORF1b is highly conserved among the different coronaviruses.

Construction and Expression in IBRS-2 Cells of the Eukaryotic Expressed Plasmid of MIC3 from Toxoplasma gondii

JIANG Tao;ZHANG Dong-lin;NIE Hao;YAO Bao-an;ZHAO Jun-long

2007, 38(8): 827-831. doi:

Abstract ( 828 )

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The sequence encoding MIC3 was obtained by amplification from genomic DNA of Toxoplasma gondii RH strain and cloned into the vector pMD18-T. The target gene was subcloned into the eukaryotic vector pcDNA3.1 after the identification of MIC3 in pMD18-T by enzyme digesting, PCR amplification and sequencing. Then the target recombinant plasmids pcMIC3 were transfected into IBRS-2 cells, and the positive cells containing pcMIC3 plasmids were obtained under the selection of G418. The expressed proteins from the positive cells were detected by SDS-PAGE, Western blot and ELISA.The results showed that the DNA sequence identity was 99.9% between amplified MIC3 and that from GenBank. The molecular weight of the recombinant MIC3 protein with good immuno-activity was about 39.2 ku. These available data would lay the foundation for further studying on DNA vaccine against Toxoplasma gondii.

Construction of a Subtractive cDNA Library for Infective Larvae of Ascaris suum

HUANG Cui-qin;CHEN Ning;ZHANG Xin-gao;LIN Rui-qing;SONG Hui-qun;ZHU Xing-quan

2007, 38(8): 832-836. doi:

Abstract ( 1569 )

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The objective of present study was to construct a subtractive cDNA library for infective larvae of Ascaris suum, for the identification and cloning of differentially expressed genes in infective larvae of A. suum. The subtractive cDNA library for infective larvae of A. suum was constructed with the technique of suppression subtractive hybridization (SSH) using the PCR-Select cDNA Subtraction kit (Clontech) after subtraction with cDNAs representing other developmental stages (adults, eggs, L3 and L4). The specificity of the constructed archives was determined using Southern dot blot, followed by sequencing of 45 random clones showing significant expression in the infective stage. The results showed that the subtractive cDNA library for infective larvae of A. suum was specific highly as confirmed by Southern dot blot. 40 of the 41 valid expression sequence targets (ESTs) had significant similarities with that of other known genes, mainly representing L3 genes and female head gene of A. suum. One EST may represent a new gene. The successful construction of the subtractive cDNA library for infective larvae of A. suum provides a foundation for further studies of genes related to the infectivity and migration of A. suum larvae.

A Report on New Causative Agent (Gasterophilus spp.) of the Myiasis of Przewalski’s Horse Occurred in China

LI Kai;WU Zhuan;HU De-fu;CAO Jie;WANG Chen

2007, 38(8): 837-840. doi:

Abstract ( 1255 )

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A new causative agent attracts us in present control of the myiasis of Przewalski′s horse. The species, which is attributed to Gasterophilus, Gasterophilidae, Diptera according to the collected larva specimen, has been unrecorded in national literatures by now. Features of the 3rd instar larvae were as follows: fully-grown larvae (maggots) are 14-17 mm long (double of the width) and have green, tough skin; the body is spindly and blunt at the back end; the front part of each body segment (from the 3rd thoracic segment to the 6th abdominal segment) is ringed with single line of strong, yellow, longitudinal-narrow spines. The line of spines at the backside become incomplete since the 4th abdominal segment, and there is no spine at the backside of the 7th abdominal segment.

Cloning and Expression of Porcine Interleukin-15

JIANG Yun-bo;FANG Liu-rong;XIAO Shao-bo;ZHANG Hui;PAN Yong-fei;LUO Rui;LI Bin;CHEN Huan-chun

2007, 38(8): 841-845. doi:

Abstract ( 744 )

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Interleukin-15 (IL-15) is a multi-functional cytokine of IL-2-like cytokine family and exerts profound effects on the proliferation and differentiation of lymphocytes and NK cells and the secretion of immunoglobulin. In this study, the porcine IL-15 (pIL-15) full-length gene was cloned from porcine peripheral blood molecular cells (PBMCs) using RT-PCR and ligated into pMD18-T vector Sequence analysis showed that the cDNA of pIL-15 is 486 base pairs in length and encodes a protein consisting of 162 amino acid (aa). The gene of mature pIL-15,which was deleting a 48 aa leader sequence at N-terminus,was cloned by PCR and then was introduced into downstream of 6 His of pET-28a, resulting in the prokaryotic expression plasmid pET-pIL15. After transforming into E．coli BL21 (DE3) and inducing by IPTG, the results of SDS-PAGE showed that the 6×His-pIL15 fusion protein about 15ku was acquired. Gel scanning analysis.revealed expression quantity of the fusion expression in all tropina was 17.5%. The result of Western blot demonstrated that the recombinant fusion protein 6 His-pIL15 can be recognized by anti-6 His monoclonal antibody. The cloning and expression of pIL-15 made a foundation for the study of function and application.

Preparation of Protein Samples from Mammary Tissues and Its 2-DE Analysis in Dairy Cow

YANG Yong-xin;TAO Jin-zhong;ZHANG Yong;CAO Sui-zhong;ZHAO Xing-xu

2007, 38(8): 846-850. doi:

Abstract ( 1256 )

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The mammary gland is a complex organ, and associated with immune factors and secretion that play a vital role in protecting the mammary gland from infection or inflammation. Centrifugation, ultracentrifugation, 2-D clean-up kit and TCA/acetone methods were used to prepare samples and corresponding 2-DE maps were compared by PDQuest7.4 software. The results showed that samples prepared by ultracentrifugation resulted in an efficient concentration, desalting and removing non-protein, its 2-DE map has showed clear spots with less background than the others. Precipitation with 2-D clean-up kit and TCA/acetone were also efficient desalting and removing nonprotein, these gel maps have strong spots, but the protein spots less than that of ultracentrifugation method. The results indicated that ultracentrifugation method was a efficient sample preparation of mammary gland for proteomic analysis.

Morphological Study on the Reproductive Organs of the Male Ostrich Chicks

WEI Lan;PENG Ke-mei;LUO Lai-qiang;SONG Hui;WANG Yan;LI Sheng-he;TANG Li;DU An-na;JIN Er-hui;WANG Jia-xiang

2007, 38(8): 851-855. doi:

Abstract ( 786 )

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In order to investigate the morphological feature of the reproductive organs of the male ostrich chicks, 6 male ostrich chicks were studied by paraffin section and the HE staining method, and compared with that of some domestic fowls The morphological feature of the reproductive organs of the male ostrich chicks were observed under the light microscope. The results of the present study were as follows:(1)There are not mediastinum testis and lobules of testis in ostrich testis. Spermatogonia cells in the convoluted seminiferous tubules arrange regularly and tight, and there is less interstitial tissue of testis in ostrich testis. (2) The ostrich epididymis is composed of rete testis, efferent duct and ductus epididymidis. (3) The tubal wall of spermatic duct is composed of mucosa, muscular layer and adventitia. (4) The penis is composed of 2 round fibrolymphcorpuscles, lymph space and micrangium together.

Effects of Sodium Selenite on Morphology, Proliferation and Viability of Mice Osteoblast Cultured in vitro

FENG Li-fang;WU Pei-fu;SUN Li-ting;HAN Bo

2007, 38(8): 856-860. doi:

Abstract ( 1238 )

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To observe the effect of the selenium with different concentrations on the morphology, proliferation and viability of mice osteoblast in vitro. The 24 hourold mice osteoblasts, which were obtained from skull with complete enzyme digestion, were cultured in DMEM solution seperatively containing various concentrations of sodium selenite（0.00，0.05，0.1，0.2，0.4，0.6，0.75，1.0 mg/L）. The morphological changes were viewed; The proliferation rate and cell viability were measured by MTT and crystal violet staining, respectively. The results indicated that the proliferative capacity and viability of osteoblasts exposed to Na2SeO3, was increased at the concentration of 0.2 mg/L in 48 h, however they were inhibited over 0.4 mg/L Na2SeO3. Selenium has two-side effects for proliferation and viability of osteoblasts. It is time-dossage dependent.

Effects of Chronic Exposure to Phoxim on Oxidative Stress in the Liver of Rats

LI Hui-min;WANG Fu-min;GU Jian-hong;CAO Xue-zhi;FENG Ya-jie;LIU Zong-ping

2007, 38(8): 861-865. doi:

Abstract ( 1503 )

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In order to study the toxicity of phoxim on rats, and explore the role of oxidative stress in the mechanism of phoxim intoxication, phoxim was administrated intragastrically to SD rats with the concentrations of 30 and 300 μmol/kg body weight. After 15 and 30 days, the content of malondialdehyde (MDA) and the activity of cholinesterase（ChE）,superoxide dismutase（SOD）,glutathione peroxidase（GSH-Px）in liver tissue and plasma were determined and histopathological changes were detected. Results showed that activity of ChE in the plasma and liver were sighificant lower than that in the control group（P＜0.01）. Especially , the activity of ChE in high dosage group decreased to 22％ in plasma and 75％ in liver compared to the control. The activity of SOD and GSH-Px both decreased in plasma, the content of MDA increased in plasma and liver along with the exposed time extended Histopathological changes showed that fatty degeneration in hepatocytes of rats. The results indicated that phoxim can induce enhanced lipid peroxidation on SD rats and lesions of structure in the liver. The oxidative stress plays an important role in the mechanism of hepatotoxicity

Cloning and Sequence Analysis of DRD4 Gene in Six Horse Breeds

FAN Cai-yun;MANG Lai;CHENG Jian-bo;Mori Yuji

2007, 38(8): 866-871. doi:

Abstract ( 1267 )

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The genome DNA was extracted from the blood of six horse breeds including SH, BH, XN, WZ, TB, AI. Four pair of primers were designed based on the sequence of human, mouse and ferret DRD4 gene cDNA to amplify the corresponding horse gene. The PCR products were ligated into a linear plasmid pMD17, and sequenced respectively. These sequences were aligned to a conting to get most sequence of the horse DRD4 gene. The horse DRD4 gene has four exons and three introns, and share above 75% similarity with human, mouse and dog. And by comparing the DRD4 gene sequence of six breeds, some variations among the sequence of six breeds horse DRD4 gene were detected in intron1 and exon3.

Regularity of IgA, IgM and IgG Antibodies in Local Mucosal and Systemic Immunity of Ducks Induced by Attenuated Duck Plague Virus Vaccine Strain Vaccinated Orally

QI Xue-feng;CHENG An-chun;WANG Ming-shu;YANG Xiao-yan;ZHOU Xia;CHEN Xiao-yue

2007, 38(8): 872-876. doi:

Abstract ( 780 )

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The regularity of IgA, IgG and IgM antibodies in local mucosal and systemic immunity of ducks induced by attenuated duck plague virus vaccine strain vaccinated orally were studied in this paper. Twenty-day-old Cherry Valley ducklings were vaccinated orally with attenuated duck plague virus Cha strain and every five vaccinated ducklings were killed punctually in 60 days, and DPVspecific antibodies were detected in serum, bile and the secretion from trachea and enteron (oesophagus, duodenum, jejunum, ileum, caecum and rectum) by indirect ELISA(results were presents as the log10 geometric mean antibody titer±SD).The results showed that the order of antibody titers from high to low was:① IgG, IgM and IgA in serum, and could be detected at 9-60, 3-15 and 9-36 days respectively;②IgA, IgG and IgM in bile,and could be detected at 6-15, 12-36 and 9-12 days respectively;③IgA, IgM and IgG in trachea, and could be detected at 6-60, 3-12 and 9-27 days respectively;④IgA, IgM and IgG in oesophagus and could be detected at 3-60, 3-27 and 9-60 days respectively; ⑤The antibody titers in intestines from high to low and the period of time when the antibodys could be detected were respectively as follows: IgA(3-60 days),IgM(3-15 days)and IgG(9-27 days) in duodenum; IgA(6-60 days),IgM(6-12 days)and IgG(9-36 days) in jejunum; IgA(9-60 days),IgM(6-9 days)and IgG(15-21 days) in ileum; IgA(6-60 days),IgG(12-36 days)and IgM(6 days) in caecum; IgA(3-60 days),IgG(12-21 days) and IgM(6 days) in rectum. Data presented here indicated that IgM was produced firstly and IgG was the most important antibody in systemic immunity, and IgA, IgG and IgM preserved in the bile to be short, and IgA was the most important antibody in trachea and enteron after the duckling were vaccinated orally with attenuated duck plague virus strain.

Common Antigen Epitopes on Eimeria acervulina Sporozoites and Duodenum Epithelium of Chickens

WANG Cheng-min;WEI Gang-cai;QIN Jian-hua;HE Hong-xuan;LIU Li-yan;QI Yong-hua;WU Yan-yun

2007, 38(8): 877-880. doi:

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An monoclonal antibody(EASP-3G3) produced previously against Eimeria acervulina sporozoites was used as tool to detect common antigen both on duodenum epithelium and sporozoites. The weight of soluble antigen recognized by MAb were determined by Western blot analysis. Immuno histochemistry and immunofluorescence techniques were used to determine whether common epitopes exist on sporozoites and duodenum epithelium The results showed that the anti-sporozoites MAb was found to react specifically with duodenum epithelium，in addition to the other parts of the digestive tract. In Western blot after sodium dodecyl sulfatepolyacrylamide gel electrophoresis，the monoclonal antibody recognized an 35-48 ku protein The results above suggested that the interrelationship between specific site invaded by Eimeria acervulina sporozoites and certain antigen existed on the surface of duodenum epithelium, can exist during the infection.

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