Acta Veterinaria et Zootechnica Sinica

Mapping, Cloning and Expression Profile Analysis of Porcine NDUFS2 Gene

TANG Wen-hua;LI Yong;WANG Heng;TANG Zhong-lin;ZHU Zheng-mao;YANG Shu-lin;JIN Er-hui;GONG Wei-hua;LI Kui

2007, 38(11): 1137-1142. doi:

Abstract ( 805 )

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In this study, we completed the chromosomal mapping of porcine NDUFS2 （NADH dehydrogenase (ubiquinone) Fe-S protein 2）gene using a radiation hybrid panel, cloned the coding sequence, predicted the structure and function of NDUFS2 gene with some software tools, and analyzed the expression profile of NDUFS2 gene in 11 different porcine tissues with semiquantitative RT PCR. The NDUFS2 gene was physically mapped on porcine chromosome 4q and closely linked to marker SW589. The coding sequence of porcine NDUFS2 gene is 1 392 nt, coding for 463 amino acids. The results of phylogenetic analysis among different species indicated that the porcine NDUFS2 gene is more homologous to bovine NDUFS2 gene. According to the structural prediction, porcine NDUFS2 gene has a conserved NuoD domain involved in generating and transforming of energy. The results of tissue expression profile showed that the expressive levels of NDUFS2 gene were different in 11 porcine tissues.

Partial cDNA Cloning and Tissue Expression of FoxO1 in Pig

YANG Yan-jun;BAI Liang;PANG Wei-jun;YANG Gong-she

2007, 38(11): 1143-1148. doi:

Abstract ( 727 )

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The partial cDNA of FoxO1 was cloned from pig’s liver by RT-PCR, and primer was designed according to the homology blast among human, chimpanzee and rat. Analysis of tissue expression showed that FoxO1 was widely expressed in the liver, lung, kidney, spleen, heart, stomach, subcutaneous adipose, visceral adipose, m.longissimus dorsi and m.quadriceps femoris. Moreover, FoxO1 expressional profile was different between in 1-day-old and 9-month-old pigs following the different developmental stage and tissues. FoxO1 was highest expressed in visceral adipose of 1-day-old pigs while lower in heart and skeletal muscle. Furthermore, FoxO1 had the highest expression in spleen of 9-month-old pigs and significantly higher than piglet that didn’t have complete immunity ability, showing FoxO1 may be involved in body immunomodulation. In addition, the expression profile of FoxO1 not only was significantly different from smooth muscle and skeletal muscle, but also was significantly different from various skeletal muscle of 9-month-old pig. It implied that the expression of FoxO1 is intimately related to skeletal muscle types and exercise intensity.

Analysis of Polymorphism of MHC-DRB1 Gene and Resistance of Hydatidosis in Dolang Sheep

YU Zhi-yong;LI Hai;JIA Bin;JIANG Wen-sheng;PENG Lin-ze;SHEN Hong;ZENG Xian-cun;DU Ying-chun

2007, 38(11): 1149-1153. doi:

Abstract ( 1418 )

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The major histocompatibility complex (MHC) DRB1 exon2 was amplified by polymerase chain reaction (PCR) from 122 healthy and 70 infectious with Dolang sheep. PCR products were characterized by the RFLP technique using three restriction enzymes, SacⅠ, Hin1Ⅰ and HaeⅢ. The results showed that rich polymorphism was observed in the exon2 of MHC-DRB1, which 2 ,2 and 6 RFLP patterns were found with enzymes SacⅠ,Hin1Ⅰ and HaeⅢ, respectively , and 3，3 and 18 kinds of genotypes were identified in Dolang sheep. By analyzing allele and genotype frequencies of healthy and infectious Dolang sheep, it was found that allele HaeⅢa was associated with susceptibility of hydatidosis (P＜0.05), and genotypes SacⅠab and Hin1Ⅰaa were resistant to hydatidosis (P＜0.05), while the genotypes of HaeⅢbe and HaeⅢ ef were rather highly susceptible to hydatidosis (P＜0.01).

Study on SNPs of H-FABP Gene and Its Relationship with Growth and Carcass Traits in Shanbei White Cashmere Goats

YU Gang;LUO Jun;HAN Xue-feng;QUAN Song-an;XUE Rui;WANG Wei;SHENG He-jun;ZHOU Zhan-qin

2007, 38(11): 1154-1159. doi:

Abstract ( 740 )

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PCR-SSCP technique was used to analyze the polymorphisms at the exon 2 of the HFABP gene in 262 yearling Shanbei White Cashmere goats to further investigate the correlation between polymorphism and growth and carcass traits. The results showed that there was one mutation (G to C) at this gene locus with the gene frequency of the population in HardyWeinberg disequilibrium. The backfat thickness of AB genotype of H-PFBP gene was significantly different from that of AA and BB (P < 0.05), significant difference was found between the genotypes of AA and BB for the backfat thickness (P < 0.01); there was a significant difference between the genotypes of AB and BB for the heart girth and circumference of cannon (P < 0.05) as well as between the genotypes of AA and BB for the loin eye area, the significant differences between genotypes were not noted for other traits. Moreover, the values of birth weight, loin eye area, backfat thickness and other 3 traits ranked as AA>AB>BB, the values of body height, heart girth and circumference of cannon ranked as AB>AA>BB, which indicated that HFABP gene may be a candidate gene for the selection of growth and carcass traits in Shanbei White Cashmere goats and a possible molecular marker for meat performance of cashmere goat.

Correlation Analysis of Growth Traits in Native Sheep Using Microsatellite DNA Markers

WANG Jian-min;YUE Wen-bin;QIN Zi-juan;QU Xu-xian;MA Yue-hui

2007, 38(11): 1160-1167. doi:

Abstract ( 764 )

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Eleven polymorphic microsatellite loci in sheep 1st, 2nd, 3rd, 4th, 6th, 7th and 8th chromosome were selected from GenBank. The relating effects between genotypes and growth traits were analyzed in 133 animals of four native sheep breeds from sympatric geography regions, by using least squares linear model and DNA analysis technique. The results indicated that，compared with age and marker factors, the breed factor had large effect on growth traits（P<0.000 1）. Eight microsatellite loci had significant effect on some growth traits of sheep（P<0.05 or P<0.01）. The most favourable genotypes for interrelated traits are 136/136 at BMS1248 and 136/143 at BM3033 for height at withers, 117/121 at BM6444 for circumference of cannon bone, 178/198 at BM3501 for heart girth, 118/130 at BMS710 and 178/168 at BMS1724 for body length, 142/144 at MAF70 for dody weight, 108/116 at BM1227 for thurl width, respestively.We also inferred that some alleles above loci had positive/ negative effect on interrelated growght traits.

Analysis on the Genetic Polymorphism and Systematic Evolution in Domestic Ducks by Mitochondrial DNA D-loop Region

ZHANG Tang-jie;LI Hui-fang;CHEN Kuan-wei;CHANG hong;TANG Qing-ping;ZHANG Jing-xin

2007, 38(11): 1168-1175. doi:

Abstract ( 1140 )

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Investigation of the genetic polymorphism and systematic evolution in domestic ducks by the structure and polymorphism of mitochondrial DNA. The 667 bp control region on mitochondrial DNA from 106 domestic ducks of 9 breeds was sequenced. The result showed that content of nucleotide A,C,G,T were 25.6%,33.3%,15.2% and 25.9%, respectively. There were 34 polymorphic sites (5.1% in total analyzed sites).The transition, transversion, and insertion/deletion were found in this region. 31 haplotypes were defined in this study, in which A7 was major haplotype in ducks and shared 9 haplotypes among breeds. The haplotype diversity(Hd) and average nucleotide diversity(Pi) were 0.798 and 0.28%, respectively. Hd was the highest in Jingjiang Sheldrake, followed then by Youxian Sheldrake and Enshi Sheldrake, and lowest in Wendeng Black duck. Kimura 2-parameter distance between the breeds is 0.001 3-0.004 4. A Phylogenetic analysis of the 31 haplotypes identified that there was only one distinct maternal lineage in domestic duck breeds and revealed that no evidence of contribution of Anas zonorhyncha to the maternal origin of domestic duck breeds.

Differentially Expressed Gene in Chicken Sperm Storage Tubules

ZHU Jin-jin;LIU Gui-qiong;WANG Zhi-yue;GUO Chun-yan;HUANG Jun;LAI Yu-yan

2007, 38(11): 1176-1179. doi:

Abstract ( 733 )

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The difference of gene expression in chicken sperm storage tubules (SST) was investigated by differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). One specific fragment, named SST01, was found. After secquencing and BLAST analysis, SST01 displayed high homology (98%) to neurexophilin gene of chicken. Neurexophilin is a glycoprotein which is detected only from neurons, which indicated that nervous system be related to the storage or release of sperm in SST. Neurexophilin functions as the ligand of neuridine which can cause cytoadherence, which indicated that neurexophilin be related to the storage of sperm in SST.

Effects of Effective Temperature and Dietary ME on Performance and Carcass Composition in Broiler

GU Xian-hong;LI Sheng-sheng;ZHAO Xiang-hong

2007, 38(11): 1180-1187. doi:

Abstract ( 1109 )

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In a 4×3 factorial design trial, 216 21-day-old Arbor Acre (AA) broilers were randomly allocated into 12 groups, each with 3 replicates of 6 birds, reared in 4 chambers which effective temperature (ET) was controlled at 15.0, 21.2, 35.3 and 37.6℃, respectively, and fed on one of the trial diets containing 11.71, 12.55 and 13.39 MJ/kg ME, respectively, for three weeks. Body weight and feed consumption were measured at d 21 and d 42. After feeding trial, two birds from each replicate were slaughtered for carcass and meat measurement. The results showed that: (1) Under high temperature (35.3℃ and 37.6℃), average daily feed intake (ADFI) and average daily gain (ADG) significantly decreased (P<0.05), and feed conversion rate (FCR) markedly rose (P<0.05). Trial dietary ME level and the interaction between ET and ME had no remarkable effect on performance. (2) High temperature (35.3℃ or 37.6℃) increased significantly the ratio of carcass yield to live weight, percentage of halfeviscerated yield, percentage of eviscerated yield and the ratio of meat yield in left thigh and drumstick to uneviscerated carcass yield (P<0.05), but decreased significantly carcass yield, meat yield in right breast, meat yield in left thigh and drumstick, the ratio of meat yield in right breast to uneviscerated carcass yield, subcutaneous fat thickness and fat width between muscles(P<0.05). Under low temperature the ratio of carcass yield to live weight increased and subcutaneous fat thickness decreased significantly (P<0.05). Carcass yield increased, and the ratio of meat yield in right breast to uneviscerated carcass yield and subcutaneous fat thickness decreased significantly (P<005), as the dietary ME level rose. (3) High temperature (37.6℃) decreased the protein of breast meat and thigh meat significantly (P<0.05); Both high and low temperature significantly increased the fat deposition of breast and thigh meat (P<0.05); High temperature (35.3℃ and 37.6℃) increased the moisture of breast meat significantly (P<0.05); Low temperature decreased the meat ash significantly (P<0.05). The protein of thigh meat in the broilers reared under high dietary ME was significantly lower than that under proper dietary ME (P<0.05). It was concluded that ET had more significant effect on performance and carcass composition in broiler than dietary ME level, and that there were lower performance, carcass composition and crude protein content in meat in broiler reared under high temperature.

Effects of Dietary Daidzein on Egg Quality and Meat Quality of the Offspring of Broiler Breed Hens during the Late Period of Laying Cycle

NI Ying-dong;HONG Wen-jie;REN Ling-zhi;HU Yan;DAI Jing;RAO Kai-qing;CHEN Jie;ZHAO Ru-qian

2007, 38(11): 1188-1194. doi:

Abstract ( 765 )

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The effects of dietary daidzein on broiler breeders during the late periods of the laying cycle on egg quality, hatching rate and meat quality of the offspring chickens were studied. Broiler breeders feed a basal-diet (n=240, 8 replicates, control) or the basal diet supplemented with 10 mg/kg daidzein (n=240, 8 replicates, daidzein). The laying rate was increased over the whole investigation with the daidzein treatment. Egg quality analysis showed that eggshell thickness and relative weight of eggshell to the whole egg increased 0.03 mm (P < 0.01) and 0.45% (P < 0.05),respectively, in daidzein group. Compared with the control group, egg Haugh unit increased significantly in daidzein group by 4 units(P < 0.05). The hatching rate of fecundated eggs and hatched eggs were decreased by 11.5% and 8.3% after daidzein supplementation, respectively (P < 0.05). Although postnatal growth of the offspring chickens from 1 to 63 days of age was not significantly affected, maternal daidzein supplementation tended to reduce the muscle mass of female chicken. As the breast muscle and leg muscle, the rate of cooking loss24 and drip loss24 were significantly decreased, and the yellowness of breast muscle was significantly increased by daidzein treatment (P < 0.05), while the ratio of myofiber types were not changed. The results suggested that maternal diet supplemented with 10 mg/kg daidzein during late periods of the laying cycle of broiler breeders affect egg quality and meat quality of the offspring chickens.

Effects of Diet Energy on FSH, LH, P4 in Plasma and Laying Rates of Layers

YANG Yu;HUANG Ying-xiang;LI Qing-hong;NING Guan-bao;HE Xiao-yan

2007, 38(11): 1195-1203. doi:

Abstract ( 943 )

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The objectives of this study were to investigate the effects of diet energy on plasma folliclestimulating hormone (FSH), luteinizing hormone (LH), progesterone (P4) and laying rates of layers and to relate these hormone levels with the laying performance. Radiation immunity analysis was used to determine the plasma hormone concentration during prepubertal development and laying period. The results demonstrated that high energy level led to ahead of the day to first egg. Laying rates of high energy I and low energy Ⅱincreased slowly after first egg. With the decrease of energy level, the time to laying peak was delayed. The order of laying peak value from high to low was control group, low energy group and high energy group. Compared with other groups, laying rates in high energy groups descended rapidly. During sexual maturity, plasma FSH concentration increased with age，FSH concentrations were higher in high energy groups than that in low energy groups,plasma LH concentration increased with age，LH concentration was higher in high energy group than that in low energy groups. High energy I reached FSH level peak at 28 weeks, whereas other groups reached the peaks at 3340 weeks. High energy I，high energyⅡand control groups reached LH level peak at 27，34 and 30 weeks, respectively, whereas low energy I and low energy Ⅱ reached the peaks at 30 and 42 weeks; During laying period, control group had higher FSH level than other groups and higher LH level than low energy groups. Control group had higher laying rate than other groups. When FSH concentrations at peaks and during laying period were related to mean laying rates, a significant positive correlation and a significant positive correlation were obtained respectively. During sexual maturity, P4 concentration increased with age in all groups, P4 concentration was low before 18 weeks and increased at 22 weeks sharply, P4 concentrations in high energy groups were higher than that of low energy groups. During laying period, P4 concentrations and mean laying rates were higher in control and low energy groups than that in high energy groups. Significant positive correlations of P4 concentration at peak, after peak and during laying period with mean laying rate were obtained, respectively.

Dynamic Distribution of GPV-VP3 Gene Vaccine in Geese Vaccinated with Genegun Bombardment

LI Min;CHENG An-chun;WANG Ming-shu;HAN Xin-feng;LIU Xiao-dong;LU Fei;CHE Qian;CHEN Xiao-yue

2007, 38(11): 1204-1210. doi:

Abstract ( 1191 )

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This paper concentrates on developing the Realtime PCR method for detecting GPV-VP3 gene and studying the dynamic distribution of GPV-VP3 gene vaccine （pcDNA-GPV-VP3）in geese (duodenum, liver, spleen, kidney, bursa of Fabricius, thymus, ileum, cecum, pancreas, heart, blood, jejunum, lung, gland of Harder, skin of injected spot, rectum and brain). Ninety 30-day-old geese were divided into 5 groups. Animals in pcDNA-GPV-VP3 inoculation groups were injected via gene-gun with different doses（1, 3, 6 μg）,respectively. Results showed that: ①This assay was specific, highly sensitive and rapid for detecting pcDNA-GPV-VP3.The standard curve showed a good linear relationship between Ct and template concentrations, and the correlation coefficient was 0.999. ②The pcDNA-GPV-VP3 could be detected in all tissues and peripheral blood 1 h post immunization. The copy numbers were most at the skin of injected spots, then liver, kidney, lymphoid organ (spleen, bursa of Fabricius, thymus, gland of Harder). ③After 217 d post-administration, pcDNA-GPV-VP3 could still be detected in tissues of 1 μg dose group. But the copy numbers decreased about 104 copies in the most tissues than that at 1 h post administration, the obvious decrease of pcDNA-GPV-VP3 was found at the injection sites (it dropped about 107 copies ).④The copy numbers in the serum were always little and changed not obvious with the time ( P≥0.05 ).⑤Different dosage groups of pcDNA-GPV-VP3 distributed in the geese tissues did not significantly differ from each other(P≥0.05), the 6 μg group had the highest copy numbers, then the 3 μg group, and the 1 μg group had the lowest. The results demonstrated that FQ-PCR was a reliable method for detecting dynamic distribution of pcDNA-GPV-VP3,and pcDNA-GPV-VP3 could distribute in all tissues of geese 1 h post immunization, and exist in those at least 217 days post immunization.

Study on BP Neural Network in the Prediction of Newcastle Disease

GONG Du-qiang;GAO Li;XIAO Jian-hua;LIU Yun;SUN Xin;SUN Xi-juan;WANG Hong-bin

2007, 38(11): 1211-1216. doi:

Abstract ( 698 )

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The purpose of this paper is to investigate the correlation between meteorological factors and Newcastle disease morbidity, and to determine the key factors that affect Newcastle disease. Having built BP neural network forecasting model by Matlab 7.0 software, we tested the performance of the model according to the coefficient of determination (R2) and absolute values of the difference between predictive value and practical morbidity. The results showed that 6 kinds of meteorological factors were determined,and the model′s coefficient of determination was 0.760, and the performance of the model was very good. Finally, we built Newcastle disease forecasting model, and applied BP neural network theory in animal disease forecasting research.

Molecular Characterization and Identification of A Velogenic Newcastle Disease Virus Isolate

2007, 38(11): 1217-1223. doi:

Abstract ( 728 )

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A Newcastle disease virus (NDV) field strain named SRZ/03 was isolated from a layer flock with typical symptoms and lesions and identified by the HI test, and cloned by plaquepurification of three times. SRZ/03 was determined as a virulent strain with MDT of 55.2h and 51h , ICPI of 1.7 and 2.0, IVPI of 1.91 and 2.79 respectively before and after plaque-purification. Analysis of F gene indicated that SRZ/03 belonged to genotype Ⅱ, and the amino acid sequence deduced from the F gene at the cleavage site was 112R-R-Q-K-R-F117 which was the same as the velogenic virus, and the F protein amino acid sequence had 93.1％, 93.0％, 94.4％ and 94.4％ homologies with that belong to published genotype Ⅱvaccine strains LaSota/46, B1/47, virulent strain Bingham/87 and Texas/48, and very similar homologies of 92.2％, 92.8％ ,92.8％ or 91.2％ were found when comparing with Taiwan/95, SGM/01, SKY/03 and F48E9/48 that belong to published genotypes Ⅶ and Ⅲ. However, the former 210 amino acids of F protein of SRZ/03 shared 96.7％ and 98.1％ homology with that of LaSota/46 and Texas/48， respectively, and the highest homology of the latter 343 amino acids of F protein was 95.7％ and 97.4％ compared with Taiwan/95 and SGM/01, which indicated that SRZ/03 may be a recombinant strain. Furthermore, when HN genes were compared, SRZ/03 demonstrated higher homologies of 95.8％-97.6％ with the known velogenic strains such as Taiwan/95 or SGM/01 , but lower homologies of 87.2％-89.3％ with LaSota/46, B1/47, Bingham/87 and Texas/48.

Establishment and Preliminary Application of Detection of Mycobacterium bovis by Phage Amplified Biologically Assay

ZHONG Zhi-jun;PENG Guang-neng;TANG Ming-xia;BAI Yong-ping;SHI Mei;MA Xiao-ping;HU Zhong-yi;WANG Jie;QIN Lian-hua;YANG Hua

2007, 38(11): 1224-1229. doi:

Abstract ( 1098 )

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A technique for detection of Mycobacterium bovis by PhaB assay was developed, and the method was applied to detect 22 clinical isolates of Mycobacterium bovis,20 non-Mtb strains of Mycobacteria and 5 other bacteria.203 cow milk samples were detected by PhaB assay and the results were compared with skin test，smear method and Lǒwenstein-Jensen culture. It was demonstrated that the optimal working concentration of mycobacteriophage was 1×10⁹PFU/mL and the optimal condition for detection was infection at 37℃ for 2 hours. Meanwhile, the concentration of virucidal agent to inactivate the mycobacteriophage completely was 100 mmol/L at room temperature for 10 min,and 4 mg/mL concentration of the logarithmic phage of mycobacteriophage was selected as the working concentration. Under these conditions, the optimal results could be obtained, and the heatinactivated or indicated bacteria were not infected with mycobacteriophage. The positive results could be obtained by using this method of detection for control strain of M.bovis，M.smegmatis and 22 clinical isolates of M. bovis,but the negative results were demonstrated in 16 reference strains of nonMtb strains of Mycobacteria and 5 other bacteria. The 4 strains of NTM（M.fortuitum, M.intrcellulare, M.aurum, M.phlei）can be positive reaction at concentration of high level (>10⁵ CFU/mL). By using this method 60-120 CFU/mL of M.bovis could be detected. The intra and interbatch variation coefficies were all below 15%, indicating a good repeatability. In all milk samples from 203 cows, 14，17，21 and 12 were detected positive by PhaB assay，smear method，skin test and LǒwensteinJensen culture respectively. The correspondence rate and specificity of phaB method compared to other three methods were all above 96%, the sensitivity was between 71%100%. Rapidity, simplicity, sensitivity and high specificity enable PhaB assay can be used for detecting Mycobacterium bovis.

Phylogenetic Analysis of the Cryptosporidia Isolates Based on CPN60 Gene Sequence

WANG Jin-chan;ZHANG Long-xian;LOU Fei;NING Chang-shen;JIAN Fu-chun;LU Qing-bin

2007, 38(11): 1230-1234. doi:

Abstract ( 1289 )

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The mitochondrial functional protein Chaperonin 60（CPN60）gene was chose as the target to research the phylogenetic relationship between Cryptosporidium strains.The gene of all Cryptosporidium strains that preserved in our lab were amplified and sequenced. Then the sequences were compared to other related reference sequences with Clustal X1.81.Three methods of phylogenetic analysis were used to assess genetic relationships among Cryptosporidia isolates and the related sequences downloaded from GenBank. The distance-based neighbor-joining(NJ) analysis, parsimony(MP) analysis in PAUP4.0 were used to construct NJ tree and MP tree, while ML tree was made by using PUZZLE Version4.1.The phylogenetic relationship of different Cryptosporidium species or genotypes were established and then these phylogenetic trees were contrasted with that drew from 18S rRNA and HSP70 gene sequences, to determine whether the CPN60 gene is suitable for phylogenetic analysis of Cryptosporidium. Cryptosporidium species were divided into two groups in those phylogenetic trees, one group contained C. baileyi and C. meleagridis, the other group contained C. hominis，C. suis，C. parvum cattle genotype and C. parvum mouse genotype. The sequence identity of different Cryptosporidium species ranged from 96% to 100%. The results showed that CPN60 gene is equally suitable for the phylogentic analysis of Cryptosporidium.

Study on the Cytoarchitecture of 9 Nuclei in Pons of Taihe Silky Fowl

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

2007, 38(11): 1235-1241. doi:

Abstract ( 704 )

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In order to investigate the cytoarchitecture of the nuclei in central nervous system of silky fowl, the pons of ten silky fowl brainstems were studied by paraffin continual sections and haematoxylin-eosine (HE) staining. The morphologic characteristics of cochlear nuclei, abducens nuclei, abducens accessorius nucleus, facial nucleus, sensory principal nucleus of trigeminal nerve, motor nucleus of trigeminal nerve, locus ceruleus and nucleus subceruleus, and rapheal nucleus of pons were observed under the light microscope. The results of the present study showed that the cochlear nuclei were also divided into cochlear magnocellular nucleus, laminar nucleus and angular nucleus, but the neurons in this nuclear group were largely mediumsized. The cochlear magnocellular nuclei were relatively developed. The circumscription of the angular nucleus were not clear. The anterior poles of the abducens nuclei and the facial nucleus were mostly parallel. The abducens accessorius nucleus were remarkably close to the abducens nuclei. Facial intermedia nuclei were not developed fully. The motor nucleus of trigeminal nerve were not divided into three subnuclei. The neurons in locus ceruleus are relatively denseness. Most neurons of the locus ceruleus are mediumsized, round, oval. The nucleus subceruleus locates at the inferior, its verge was not obvious, the cells of which distribute sparsely, the neurons are mainly ellipse and spindle, and mostly are mediumsized. The nucleus raphes of pons situates at the lower part of the raphes area in the posterior segment of pons. The neurons are mainly triangle, starlike and ellipse, and mostly are giant neurons.The processes of the neurons are well developed.The distribution of all the nuclei were similar to Pekin duck and fowl.

The Separation and Cultivation of Rabbit Endometrial Cells and Smooth Muscle Cells in vitro

CHEN Xiu-li;ZHAO Yong-zhen;JIN Ya-ping;ZHANG Yan-ming

2007, 38(11): 1242-1247. doi:

Abstract ( 669 )

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In this paper, rabbit endometrial cells(epithelial cells and stromal cells) and smooth muscle cells were successfully isolated and cultured, and the ways used in purifying endometrial epithelial cells and stromal cells were discussed. The ways of differential centrifugal speed and differential sticking on the wall were applied to obtain the purified epithelial cells and stromal cells, the way of sieve was chosen to harvest smooth muscle cells, separately. The cells were cultured in the medium that consist of a 1∶1 mixture of DMEM∶F12 media supplemented with 20% FBS, 100 μg/mL insulin, 63.5 nmol/L estrogen, 7.14 nmol/L progesterone,at 37℃ humidified atmosphere of 5% CO2 in air. Immunohistochemistry and immunofluorescence assay were used to identify the three kinds of cells and their purification. Results showed that rabbit endometrial epithelial cells and stromal cells were successfully separated and cultured, the purification of the two kinds of cells reached about 98%. Decidualization was observed in stromal cells. The purification of smooth muscle cells reached about 97%, cells showed lateral-cut growth. The research indicated that endometrial cells and smooth muscle cell could be cultured successfully in vitro; The DMEM/F12(1∶1) culture medium supplemented with 20% FBS,100 μg/mL insulin,63.5 nmol/L estrogen,7.14 nmol/L progesterone and the incubation under condition of 37℃ humidified atmosphere of 5%CO2 were optimal to support the growth of endometrial cells and smooth muscle cells.

Proteome Maps of Mammary Tissues in Healthy Dairy Cows and Those with Clinical Mastitis

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

2007, 38(11): 1248-1252. doi:

Abstract ( 823 )

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In the mammary glands suffered from invading pathogens that produced and released inflammatory mediators might be accompanied by changes in protein expression profiles. This project presents the protein changes of the mammary gland from healthy dairy cows and clinical mastitic cows that were investigated using two-dimensional gel electrophoresis (2-DE). Acquired gel maps were matched and analyzed using PDQuest 7.4 image analysis software. The results showed that the protein profiles in healthy dairy cows and those with clinical mastitis were shown to be very similar, 480±27 and 453±31 protein spots were statistically detected, respectively. Take "more or less" as the determination criterion, 17 protein spots were detected to be differentially expressed changes in the stain density of threefold or more. That is, 10 protein spots were up or downregulated proteins in clinical mastitic cows, 7 protein spots were unique in healthy dairy cows or clinical mastitic cows. The results indicated significant difference in expression protein between healthy dairy cows and those with clinical mastitis, and may provide valuable information to find new regulation markers of mastitis and potential protein targets for treatment.

Study on the Single Nucleotide Polymorphism in Porcine SHP Gene Exon 1

LI Jian-hua;CHU Xiao-hong;ZHAO Xiao-feng;XU Ningying

2007, 38(11): 1253-1256. doi:

Abstract ( 1129 )

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Small heterodimer partner gene is expressed in liver, heart, pancreas and adipose tissue and influences the lipid homeostasis. Three genotypes (GG, GA and AA) of SHP gene exon 1 were identified by PCRSSCP and their distribution was significant different (P<0.05) in five pig breeds (Large White, Landrace, Pietrain, Jinhua and Wild boar) by Chisquare analysis. The homozygotes were sequenced and one polymorphic locus (G374A，DQ002896) was found which changed the amino acid from Arg to His. Moreover, association analysis of SHP polymorphism with backfat thickness and intramuscular fat contents was conducted in the F2 generation from the Jinhua×Pietrain resource family. The results showed that the pigs with genotype AA had 27.0%, 30.6%, 19.5% and 40.9% lower backfat thickness at three positions (should, thorax-waist and buttock ) and intramuscular fat contents respectively than those with genotype GG (P < 0.05); At 6th-7th rib backfat thickness, the pigs with genotype AA was significantly lower than those with genotype GA by 23.0% (P <0.05).

Ruminal Degradation and Intestinal Release Rate of Protected Methionine and Lysine

GUO Yu-qin;WANG Jia-qi;BU Deng-pan;WEI Hong-yang;ZHOU Ling-yun

2007, 38(11): 1257-1261. doi:

Abstract ( 749 )

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Rumen-protected methionines(RPMet) were obtained from polyoxylate Ⅳ(M1), polyoxylate Ⅳ and ethyl-cellulose(M2), polyoxylate Ⅳ and ethylcellulose and hydroxyprpoyl methylcellulose(M3), ethyl-cellulose(M4), polyoxylate Ⅳ and hydrogenated vegetable oil(M5),and hydrogenated vegetable tallow(M6).Rumen-protected lysines(RPLys) were obtained with polyoxylate Ⅳ and ethyl-cellulose(L1), polyoxylate Ⅳ and hydroxyprpoyl methylcellulose(L2),polyoxylate Ⅳ and hydrogenated vegetable oil(L3),ethyl-cellulose and hydrogenated vegetable oil(L4). The stability of RPMet and RPLys was tested by in vitro and in situ.After incubating for 4, 8, 24 and 48 h in the rumen ,the ruminal release rate of nitrogen was lower and ranged from 11.72% -50.67% and 35.70%-72.09% in M5 and L3, respectively . However ,when incubated in the buffer solution(pH= 6.6) for 4, 8, 24 and 48 h, nitrogen release rate ranged from 24.67%-38.44% and 42.05%-75.02% in M5 and L3, respectively. There was a significant positive linear relationship (RM2=0.867 5、RL2=0.963 0) of the degradability of RPMet and RPLys between in vitro and in situ.Nitrogen release rate was 100 % in M5 and L3 in the small intestine and the buffer solution(pH=2.4).

Influence of Shuanghuanglian Injection on Phosphodiesterase 4B Gene Expression in Neutrophils

JIANG Dai-xun;CHEN Wu;GU Jin-ni;YU Tong-quan;LU Ping;MU Xiang

2007, 38(11): 1262-1266. doi:

Abstract ( 1305 )

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The influence of Shuanghuanglian Injection on phosphodiesterase 4B gene expression in neutrophils and it’s anti-inflammatory mechanism in neutrophils were studied. The quantity of phosphodiesterase 4B gene expression was detected by semi-quantitative RT-PCR in different time to evaluated the influence of Shuanghuanglian Injection. Expression of phosphodiesterase 4B gene in neatrophils had no notable change after incubating with Shuanghuanglian Injection for 1 h, but significantly reduced (P<0.01) after incubating with Shuanghuanglian Injection for 3 h. The results suggested that Shuanghuanglian Injection can inhibit phosphodiesterase 4B gene expression in neutrophils in certain time, making cAMP content rise and inhibiting activation of inflammatory cells such as neutrophils, consequently, to exert the antiinflammation.

Molecular Cloning and Characteristics of cDNA Encoding Porcineβ8 Subunit for FMDV Receptor

DU Jun-zheng;GAO Shan-dian;CHANG Hui-yun;CONG Guo-zheng;SHAO Jun-jun;LIN Tong;LIU Xiang-tao;CAI Xue-peng;XIE Qing-ge

2007, 38(11): 1267-1272. doi:

Abstract ( 1524 )

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Field isolates of foot-and-mouth disease virus (FMDV) are believed to use integrins as cellular receptors for virus internalization in vivo. In this study, we first cloned and sequenced the cDNA encoding porcine integrin β8 from the tongue keratinocytes of pig infected experimentally with FMDV. The 2 304 bp cDNA of porcine integrin β8 encodes a polypeptide of 767 amino acids consisting of a 42-residue putative signal peptide, a 637-residue ectodomain, a 30residue transmembrane domain, and a 58-residue cytoplasmic domain. The ectodomain contains 7 potential N-linked glycosylation sites（NXT/NXS）and 49 cysteine residues. The nucleotide sequence similarity of integrin β8 between pig and cattle, human, dog, rhesus monkey, house mouse, Norway rat are 94.3%，91.4%，91.7%，91.4%，83.5%，83.5%, and the amino acid sequence similarity are 95.0%，91.8%，92.8%，91.5%，84.0%，84.8%,respectively. The secondary structure of porcine β8 contains α-helix, β-sheet, β-turn and coil regions.The hydropathic analysis of the polypeptide revealed two hydrophobic domains,the signal peptide(1-42aa) and transmembrane domain(680-709aa). This study will lay a foundation for understanding the interactions of FMDV with receptors.

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