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25 January 2011, Volume 42 Issue 1
遗传繁育
Influence of Castration on Porcine Thyroid Transcription Factor-1,2 (TTF-1, 2)Gene Expression
WANG Ying;ZHANG Li-fan;CAI Zhao-wei;CHEN Zhe;JIANG Xiao-ling;ZHOU Hong-mei;HUA Xu-chuan;XU Ning-ying
2011, 42(1):  1-7.  doi:
Abstract ( 1033 )   PDF (1171KB) ( 664 )  
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In order to investigate the effect of castration on expression of TTF-1 and TTF-2 genes, realtime fluorescence quantitative PCR was applied to investigate the developmental expression patterns of TTF-1 and TTF-2 genes from the thyroid tissue of castrated and uncastrated Jinhua pigs on the age of 60, 90 and 120 days. The results showed that the expression of TTF-1 and TTF-2 genes increased slightly in uncastrated pigs, while they were relatively stable in castrated pigs. The expression level of TTF-1 and TTF-2 mRNA in castrated Jinhua pigs on 60 and 90 days were higher than that of uncastrated pigs, while the expression level of TTF-1 and TTF-2 on 120 days were lower than that of uncastrated pigs, but no significant difference (P>0.05). The results indicated that the real-time PCR method was faithful for detecting the expression analysis of TTF-1 and TTF-2 genes, and the expression of TTF-1 and TTF-2 mRNA increased after castration. Furthermore, there was a significant correlation between TTF-1 gene expression and meat quality traits.
The Study of Protein Interaction Network of Cattle Based on the Phylogenetic Profile
SHEN Qing-hang;XU Jia-bao;JIANG Ming-feng;ZHONG Jin-cheng;
2011, 42(1):  8-17.  doi:
Abstract ( 660 )   PDF (1508KB) ( 637 )  
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The objective of this study was to study the protein interaction network of cattle based on the phylogenetic profile and to provide useful information for the post-genome program of cattle, and the results could be used in production of cattle. The known protein-protein interaction (PPI) data set of C.elegans, S.cerevisiae and Homo sapien were used to deduce the PPI of cattle based on phylogenetic profile in this paper. A PPI network of cattle which included 2 953 proteins and 3 034 interactions was obtained. The analysis of PPI network showed that it was a scale-free property network and 23 unknown proteins including ENSBTAP00000011562 were annotated. The results showed that the partial network related to proteins of lipid metabolism was analyzed based on the known protein information. The result provide effective information for improving the quality and production of milk.
Associations of Ghrelin (GHRL) and Its Receptor (GHSR) Genes Polymorphismswith Duck Growth and Carcass Traits
FANG Mei-xia;;LI Ying;XU Hai-ping;XIE Liang;LIAO Xin-di;LIANG Min;NIE Qing-hua;ZHANG Xi-quan
2011, 42(1):  18-24.  doi:
Abstract ( 1080 )   PDF (549KB) ( 755 )  
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The aim of this study was to analyze the association of GHRL and GHSR genes polymorphisms with duck growth and carcass traits. In this study, two SNPs of GHRL gene (C-729T and C+985T) and two SNPs of GHSR gene (T404C and G3427A) were genotyped by PCR-RFLP in a random Sanshui White Duck population to investigate the associations of SNP and their haplotypes with growth and carcass traits. Result showed that C-729T was significantly associated with body weight (BW) at 21 days (BW21), BW28, BW35, BW49, eviscerated weight and Thymus gland weight (P<0.05) Meanwhile, the individuals with CT genotype had better duck body weight gain. T404C was significantly associated with leg muscle weight and spleen weight (P<0.05). And in these traits, the Least-square means of individuals with TT genotype was higher than others. G3427A was significantly associated with BW35, BW49, dressed weight, eviscerated weight with giblet, eviscerated weight and wing weight (P<0.05). And individuals with AA genotype had the highest value of all traits. Otherwise, no significant association of C985T with any growth and carcass traits was found Association analysis of haplotypes indicated similar results with that of SNP It was suggested that C-729T of duck GHRL gene and T404C and G3427A of duck GHSR gene were important molecular markers for duck growth and carcass traits.
Conservation Priorities of Genetic Diversity in Chinese Indigenous Duck Breeds
ZOU Jian-min;LI Hui-fang;CHEN Kuan-wei;HAN Wei;ZHU Yun-fen;SHU Jing-ting;SONG Wei-tao
2011, 42(1):  25-32.  doi:
Abstract ( 1057 )   PDF (383KB) ( 726 )  
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The objective of this study was to analyze conservation priorities of genetic diversity in Chinese indigenous duck breeds, and to supply a theoretical basis for better conservation programs. The DR genetic distance, gene flow, heterozygosity and polymorphism information content were measured by 28 microsatellite markers. Two conservation strategies, Weitzman and Caballero methods were used to assessed the conservation priorities of 24 Chinese indigenous duck breeds. The results showed that all the loci were neutral by Ewens-Watterson test, the mean PIC was 0.494 4. The SS and DY breeds respectively had the highest (0.864 8) and the least (0.720 8) observed heterozygosity. The JXD and JD breeds respectively had the highest (0581 6) and the least (0.434 7) PIC. SC breeds had the farthest mean genetic distance (0.425 7) and the least gene flow (0.471 1) with HZ breeds, XL breeds had the nearest mean genetic distance (0.166 4) and the most gene flow (1.380 8) with JXD. The Weitzman method suggested that the most important breeds to be preserved are LW, SC, GY and BJ, their contribution rate to diversity were higher than 5%. The Caballero method suggested that SX, SC, CH and XY breeds had higher positive contributions to total diversity. The priority order of Caballero method had some difference from that of Weitzman,s. The reason was that the Weitzman method did not take into account the withinpopulation genetic diversity. The full consideration to interpopulations genetic diversity and genetic diversity within populations should be given in the analysis of Chinese indigenous duck resources conservation levels. Sixteen breeds, including SX, SC, CH, XY, GY, LW, HZ, MY, JJ, SS, ES, BJ, HN, YN, YX and WS had higher contributions to overall genetic diversity of Chinese indigenous duck breeds, and should be given preferential protection levels.
Cloning, Sequence Analysis and Specific Expression in Different Tissues of Duck Akirin2 Gene
DAI Fei;HUANG Kai-liang;LIU He-he;HAN Chun-chun;LI Liang;WANG Ji-wen
2011, 42(1):  33-38.  doi:
Abstract ( 1006 )   PDF (2096KB) ( 608 )  
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In order to understand the function of Akirin2 gene in duck, the duck Akirin2 gene was cloned by RT-PCR, and the relative mRNA expression of Akirin2 was detected in different tissues using the qPCR method. The results of the sequence analysis showed that the CDS region of the duck Akirin2 gene included 594 nucleotides, encoding for 197 amino acids, and the amino acid homology of Akirin2 between the duck and the chicken was 99%. The secondary structure of Akirin2 amino acids had a common domain with the mammals Akirin1 and Akirin2.The results of the qRT-PCR indicated that the duck Akirin2 express in the muscle and the immune organ spleen. These results supported the previously views that duck Akirin2 gene had a common function with the Akirin1 and Akirin2 genes in mammals, and these assays made a foundation work for the further research about the Akirin2 gene in myogenesis in birds.
动物营养
The Effects of Injecting pGRF Gene Plasmid on Growth Axis Hormones in the Growing Pigs
ZHANG Jin-xia;;ZHANG Hong-fu;ZHOU Zhan-qin;LIU Yu-tian;LIU Sheng-jun;WEI Xing-hui;DING Guo-qing
2011, 42(1):  39-43.  doi:
Abstract ( 639 )   PDF (363KB) ( 640 )  
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This study was conducted to investigate the effect of pig growth hormonereleasing factor gene plasmid (pGRF) on the change of growth axis-hormones in growing pigs by injecting. Eight crossbred castrated boar pigs (Landrace ×Large white) with similar weight((15±0.43)kg) were allocated into two treatments, the experimental and control groups. The experimental pigs was injected with 2 mL (4.5 mg) pGRF at neck, and the control ones were done in the same way with 2 mL normal saline. Serum samples were taken from all pigs at days 1, 4, 7, 11, 16, 21, 26 and 31 after injecting, and the levels of serum GH,IGF-Ⅰ,GRF and SS were determined. The results showed that the serum GH level of pigs in the experimental group increased significantly by 113.47% (P<0.01) compared with the control pigs at day 7 after injecting. Then the values declined gradually, it increased by 47.09% (P<0.05) at day 11 after injecting, and there was no significant difference at day 31. The serum IGF-Ⅰand GRF levels of the experimental pigs had no obvious increase, but they correlated significantly with serum GH level. The correlation coefficient were respectively 0.589(P<0.05) and 0.678 (P <0.05). Their peaks appeared also at day 7 and then declined gradually Compared to the control pigs, the serum SS concentrations in experimental pigs shown a downward trend before day 11 (they reached the lowest point at day 11), but they ascended at day 16 and declined slowly after that, they declined to the same level as that of control pigs at day 31. There were significant negative correlation among the serum SS and GH and GRF levels,the correlation coefficient were respectively -0.777(P<0.01) and -0.689(P<0.05). It can be seen, pGRF gene plasmid could express in vivo and promote the secretion of GH by improving the serum content of GRF. The secretion of SS was inhibited by the injection, which meant the inhibition of GRF and GH by SS was decreased to a certain degree. It suggested that pGRF gene plasmid could play a growth-promoting role in the pigs. In the same time, we found that although the blood samples were taken from pigs by venous fistula, the variances of the determined data on hormones were still big, so increasing the number of experimental pigs was necessary for the better objective results
Comparative Study on Fatty Acid Composition between Normal Milk and Subclinical Mastitis Milk of Dairy Cow
CHANG Ling-ling;YANG Zhang-ping;WU Hai-tao;CHEN Ying;SHI Xue-kui;MAO Yong-jiang;CEN Ning;LIANG Xiang-huan;YIN Zhao-hua
2011, 42(1):  44-47.  doi:
Abstract ( 682 )   PDF (926KB) ( 793 )  
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The aim of the study was to investigate that subclinical mastitis of dairy cow causes the decline of milk yield and the change of milk composition. Detect milk fatty acids of 57 Holstein cows (32 of them suffer from subclinical mastitis)with gas chromatography,and compare fatty acid composition of normal milk with subclinical mastitis milk. The results showed that 21 fatty acids which were from C4 to C22 could be found out, the content of palmitoleic(C16∶1) in subclinical mastitis milk was higher than that in normal milk,but other 20 kinds of fatty acids were lower, 9 fatty acids in subclinical mastitis milk were significantly lower than that in nomal milk(P<0.05), and 2 fatty acids in subclinical mastitis milk were great significantly lower than that in nomal milk(P<0.01). Subclinical mastitis reduced the total content of milk fatty acids and the content of saturated fatty acid, monounsaturated fatty acid and polyunsaturated fatty acid. Saturated fatty acid percentage and monounsaturated fatty acid percentage of subclinical mastitis milk were higher than that in normal milk,but polyunsaturated fatty acid percentage of subclinical mastitis milk was lower than that in normal milk. Consequently, subclinical mastitis affects the synthetic process of cow fatty acids and changes the composition of fatty acids.
预防兽医
The Analysis of Antiviral Cytokines Transcriptional Profiles of PK-15 Cell Cultures Following Infection with Porcine Parvovirus
LI Hou-wei;WEI Zhan-yong;YIN Hai-yan;CHEN Hong-ying;LI Jin-lei;HAN Zhi-tao;CUI Bao-an
2011, 42(1):  48-55.  doi:
Abstract ( 973 )   PDF (2123KB) ( 778 )  
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In order to survey the host inflammatory responses, particularly the antiviral cytokines responses of Porcine Parvovirus (PPV) infection, and to investigate the host-PPV interaction,Porcine Kidney-15 cells (PK-15) were infected by PPV. Then the viral DNA was measured and analyzed using real-time PCR. The transcript level of cytokines (IFN-β, IFN-γ, IFNAR-1, IFNAR-2, MHC-Ⅰ, MHC-Ⅱ, MX1, NOS, 2-5AS, RNase L and IRF-3) were detected by real-time PCR too. The results showed that PPV proliferated rapidly in PK-15 cell 12 hours after infection, and reached peak 48 hours after infection; the transcription levels of IFN-β, IFN-γ, IFNAR-1, IFNAR-2, MHC-Ⅰ, MHC-Ⅱ, MX1, iNOS, 2-5AS, RNase L and IRF-3 were increased obviously, and the transcription level of Mx1 gene was increased 8 423 times at 24 hours after infection. The results revealed that the level of antiviral cytokines increased in PPV infected PK-15 cell.
Preparation and Analysis of Homogeneity and Heterogeneity and Recognized Epitopes of Monoclonal Antibodies Against Newcastle Disease Virus Strain WF00C
ZHAO Lei;ZHANG Xun-hai;WANG Xuan;SUN Tao;GONG Zheng;BAO Chun-hui
2011, 42(1):  56-64.  doi:
Abstract ( 638 )   PDF (963KB) ( 629 )  
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To analyze antigenic diversity of envelope protein among NDV strains isolated from different avian, monoclonal antibodies (MAbs) against WF00C strains of NDV isolated from chicken were prepared by whole virus which was purified by ultrafiltration. Then, the homogeneity and heterogeneity of MAbs were further analyzed by homology of antibody variable region and ownership of gene family, and key amino acid sites on variable epitope were determined by amino acid sequence analysis of antigenic site and reaction with expressed peptide, which were on the basis of its general biological characteristics and diversity comparison. Six MAbs were identified in this study, and named as 1B6, 4B4, 4E6, 4E11, 5H4 and 5H9. Except 4B4 and 5H4, the rest MAbs had hemagglutination inhibition (HI) activity, and neutralizing activity to NDV WF00C strain. However, 1B6 and 4E11 which recognized linear epitopes didn’t react with two NDV strains isolated from pigeon in cross HI test. The homogeneity and heterogeneity analysis showed that antibody light and heavy chain variable region of 1B6 and 4E11, 4B4 and 5H4 belonged to the same gene family. 1B6 and 4E11 only had five and one amino acids difference in the light and heavy chain complementarity determining region, respectively, but 4B4 and 5H4 were identical. 4E6 and 5H9 had obvious difference in gene sources and amino acid sequences of complementarity determining regions. Amino acid sequence analysis of NDV HN antigenic site 14 showed that site of aa347 had six kinds of amino acids, where glycine was common amino acid for this site of NDV strains isolated from pigeon (26/27), and site of aa349 had three kinds of amino acids, where glutamate was specific to NDV strains isolated from pigeon. 1B6 and 4E11 recognized amino acid E347 and D349 which were variable sites and crucial for these epitopes. In this study, six MAbs against NDV which belonged to three groups according to biological characteristics could be attributed to four types on the basis of homology. Site of aa349 on HN protein being glutamate was specific to NDV strains isolated from pigeon, and this site was a new key amino acid site on the linear neutralizing epitope.
Development of an Indirect ELISA for Detection of Sheep Antibodies against Echinococcosis granulosa
JIA Hong;LIU Dan;HOU Shao-hua;YU Lin-lin;BAI Li-hua;ZHU Hong-fei
2011, 42(1):  65-70.  doi:
Abstract ( 637 )   PDF (995KB) ( 696 )  
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This study was designed to develop an indirect ELISA for detection of sheep antibodies against Echinococcosis granulosa, which will provide a fast and simple means for detection Echinococcosis granulosa of sheep. The amino acid sequences of EG95 published in GenBank were analyzed using DNAStar software, and the hydrophilic epitopes were selected, named EG95s. Then the gene of EG95s were cloned and expressed. As a result, an indirect ELISA for detection of sheep antibodies against Echinococcosis granulosa were developed using the purified recombinant fusion protein as the coating antigen, therefore a variety of conditions were optimized. The results of SDS-PAGE showed that the recombinant fusion protein, GST-1EG95s and HIS1EG95s, were efficiently expressed, which were soluble and purified easily Then the results of Western blot showed that the expression products were reacted with positive serum of sheep with Echinococcosis granulosa, which indicated they were good immunogenicity. The optimized conditions of indirect ELISA showed that: HIS-1EG95s was better than GST-1EG95s as the coating antigen. The determine criteria of the indirect ELISA were: Samples with an OD450 nm value ≥ 0.235 were judged as positive, and with an OD450 nm value≤ 0.191 were judged as negative, while with an OD450 nm value ranging between them was suspicious. 70 positive and negative serum of sheep collected in Xinjiang with or without Echinococcosis granulosa were detected by the indirect ELISA respectively, and the results showed that this indirect ELISA method was completely consistent with the indirect ELISA provided by Wallaceville Animal Research Centre of New Zealand. There were no cross reactivity detected with other proteins. The variant coefficient in batch and between batches varied from 3.8% to 5.6% and from 5.7% to 8.5%, respectively. All these results showed that the indirect ELISA possessed specificity, sensitivity and good reproducibility. The indirect ELISA for detection of sheep antibodies against Echinococcosis granulosa were successfully developed, which provide a fast and simple means, and can be used for clinical detection.
Mutant Prevention Concentrations of Fluoroquinolones for Clinical Isolates of Streptococcus suis and Screen of Singlestep Resistant Mutants
ZHANG Yu-han;WANG Li-ping;LU Cheng-ping
2011, 42(1):  71-76.  doi:
Abstract ( 584 )   PDF (387KB) ( 619 )  
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The mutant prevention concentration (MPC) represents a threshold above which the selective proliferation of resistant mutants is expected to occur only rarely. Measurement of the minimal inhibitory concentration (MIC) and the MPC for 32 veterinary clinical strains of Streptococcus suis revealed that ofloxacin and enrofloxacin are more potent than ciprofloxacin and pefloxacin mesylate to restrict the selective enrichment of resistant mutants. Based on their potential for restricting the selection of resistant mutants, the four fluoroquinolones, in descending order, were found to be enrofloxacin > ofloxacin > ciprofloxacin > pefloxacin mesylate. For several compounds, all of the clinical isolates lacked a known resistance mutation in the quinolone resistance-determining region (QRDR) of gyrA, gyrB, parC and parE. Additionally, the isolates were tested for CCCP or reserpine-sensitive efflux. The MICs of ciprofloxacin, enrofloxacin, ofloxacin and pefloxacin mesylate for each single-step mutants were determined by agar dilution on MuellerHinton agar plates plus 5% sheep blood in the presence and absence of CCCP (40 mg·L-1) or reserpine (50 mg·L-1). As a result, isolates showed no reduction in MIC in the presence of CCCP and considered negative for CCCPsensitive efflux. Nevertheless, it revealed a four or morefold decrease of ciprofloxacin accumulation in the presence of reserpine in the ciprofloxacin first-step mutants compared to that in the parent resistant strain, and it′s considered that there is a reserpine-sensitive efflux to decrease the level of ciprofloxacin accumulation in single-step mutants.
Molecular Characterization of Integron-gene Cassettes in Multidrug-resistant Escherichia coli Isolates from Food-producing Animals
LIN Ju-chun;CHEN Ya-li;CAO San-jie;SHU Gang;WEN Xin-tian
2011, 42(1):  77-81.  doi:
Abstract ( 651 )   PDF (513KB) ( 639 )  
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To study the distribution and molecular characterization of integron-gene cassettes in multidrug-resistant Escherichia coli isolates from food-producing animals in Sichuan province, multiplex PCR was used to detect 3 different classes of integrase genes in 327 isolates. The sequences of the gene cassettes in integron-positive strains were analyzed by PCR-sequencing. The results of multiplex PCR indicated that 294 (89.91%) of 327 isolates harbored intIⅠ, and neither intIⅡ nor intIⅢ was detected in those isolates. 800-3 000 bp amplicons of gene cassettes were yielded in 67 of 81 intIⅠpositive strains choosed randomly. The main cassettes were those encoding aminoglycoside adenyltransferase A (aadA) and dihydrofolate reductase (dfrA, dhfrI). The most prevalent array of gene cassettes was dfrA17+aadA5. aacA4+catB3+dfrA1 and aadA22 were discoveried in two strains, respectively. aadA22 encoding resistance to aminoglycosides was firstly reported in avian E. coli from Sichuan province. IntIⅠwas extensively existed in E. coli isolates. Although there was not a correlation between resistant patterns and integron-gene cassettes in the study, identical gene cassette fragments were found in different strains, suggesting horizontal transfer among different strains may have occurred by integron.
Risk Approximate Based Studies on the Current Status of Global Pattern of African Swine Fever and Its Import Risk Model
ZHANG Zhi-cheng;HUANG Jiong;BAO Jing-yue;ZHANG Yong-qiang;ZHONG Qi;WANG Li-jian;WANG Zhi-liang;LI Chang-you
2011, 42(1):  82-91.  doi:
Abstract ( 972 )   PDF (2094KB) ( 1018 )  
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Global pattern of current status of African swine fever (ASF) were explored, and its import risk model were constructed and derived. Based on the data mining techniques and probability risk theory, methods of Geography risk analysis, the first law of geography and risk approximate theory were applied in this manuscript Results were as follows: In Global Scale, the swine production were mainly distributed in China, United States, Brazil, Canada, German, Spain, and French,the combination swine inventory of above country account for 76.67% of global proportions,among which most density region were Spain, German, Netherlands and Belgium in EU,and China, Vietnam in Asian. Based on the Toble’s first law of geography, “risk approximate” were introduced in this manuscript for the animal disease risk management and vet epidemiology, it’s means that risk can be more approximated in the neighborhood than in distance. ASF have been recorded reported at least 89 frequency globally up to now, among which the countries like GuineaBissau, Namibia, Russia, Senegal, Benin, Burkina Faso, Cape Verde, Ghana, Italy, Madagascar, Mozambique and Togo have three years consecutive reported cases of ASF since 2007. Based on the Toble’s first law of geography and risk approximated theory, the population above can be regarded as the most risk population country for African swine fever Import swine and its products from countries at risk of ASF can pose risk to the imported countries, the imported risk (one batch of imported products) model can be depicted as follows:PInport=f1·(1-Se)+(1-f1)·f2·(1-Se)+(1-f1)·(1-f2)·f3·(1-Se)+(1-f1)·(1-f2)·(1-f3)·f4·(1-Se)+(1-f1)·(1-f2)·(1-f3)·(1-f4)·f5·(1-Se), among which f1 was the most weighted risk for import, it imply the importance of local quarantine or export quarantine before the movement of animal or animal products. Currently, the most risks of ASF were mainly coming from the countries in Guinea Bissau, Namibia, Russia, Senegal, Benin, Burkina Faso, Cape Verde, Ghana, Italy, Madagascar, Mozambique and Togo, next were Cameroon, Congo, Malawi, Rwanda, Uganda, Angola, Kenya, Nigeria, South Africa, Tanzania, Zambia. Potential risk can be imported if import swine or its products from countries at risks of ASF. Enforcing the efficiency of local quarantine or export quarantine,and initiate the geographical risk classification were most important to the risk management of ASF.
基础兽医
The Study on Pharmacokinetic/Pharmacodynamic Model of Enrofloxacin against Escherichia coli in Serum and Tissue Cage Fluid of Pigs ex Vivo
YANG Yu-hui;;LI Xiao-chun;HAN Xin-chou;WANG Xue-mei;WU Ke-bang;HU Ri-cha
2011, 42(1):  92-98.  doi:
Abstract ( 1078 )   PDF (1151KB) ( 682 )  
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The research was performed for the rational usage of enrofloxacin in treating pigs infected by Escherichia coli. The combined method of pharmacokinetics in vivo and pharmacodynamics in vitro was used to survey and evaluate the pharmacokinetics-pharmacodynamics relationship of enrofloxacin against Escherichia coli. The MICs of enrofloxacin against Escherichia coli in serum or tissue cage fluid were 0.4 μg·mL-1 in vitro. The concentration of enrofloxacin that is more than MIC in serum or tissue cage fluid will be needed to inhibit the bacteria if lots of Escherichia coli was added. EC50 were 77.67±31.12 and 15.78±4.99, respectively in serum and tissue cage fluid of pigs after intramuscular administration at 5 mg·kg-1 body weight. Enrofloxacin produces 50% of the maximal antibacterial effect, when the concentration of enrofloxacin is 1.29 and 0.26 μg·mL-1, respectively in serum and tissue cage fluid. These results indicated that the dosage of enrofloxacin should be increased when it was used to prevent the infection of Escherichia coli, and MIC shouldn’t be the only standard that set interval of administration.
Expression of c-Myc in Pulmonary Vascular Smooth Muscle Cells in the Development of Pulmonary Vascular Remodeling in Broilers Induced by Low Ambient Temperature
WANG Jian-lin;YIN Yan-bo
2011, 42(1):  99-103.  doi:
Abstract ( 596 )   PDF (1457KB) ( 668 )  
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The study was conducted to assess the expression of c-Myc in pulmonary vascular smooth muscle cells in the development of pulmonary vascular remodeling in broilers induced by low ambient temperature. 120 male broiler chicks (Arbor Acre) of commercial strain were randomly allocated to control group (raised at the temperature of (22±1.5) ℃) and low temperature group (raised at the temperature of (11±2) ℃) at 15 days old. Six broilers in each group were sampled every week from 15 to 50 days of age and lungs were paraffin-embedded, sectioned. The percentage of relative medial thickness (MT%)and the percentage of relative lumen area (RLA%),the indexes of pulmonary vascular remodeling,were examined by computer-image analyzing system.Positive indexes (PI) of c-Myc in pulmonary vascular smooth muscle cells were assessed. The result indicated that pulmonary vascular remodeling were significantly elevated in low temperature group from 36 days of age (P<0.01 or P<0.05). PI of c-Myc in pulmonary vascular smooth muscle cells were significantly higher than those in the control group from 29 days of age (P<0.01). The study demonstrated that c-Myc in pulmonary vascular smooth muscle cells were expressed significantly induced by low ambient temperature and have a pivotal role in the development of pulmonary vascular remodeling.
Studies on the Determination of Main Metabolites of Mequindox Excretion in Chicken Feces
LIU Ying-chun;DING Huan-zhong;LIU Yi-ming;YANG Fan;WANG Li-qi;ZENG Zhen-ling
2011, 42(1):  104-109.  doi:
Abstract ( 967 )   PDF (1312KB) ( 664 )  
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In order to investigate the excretion quantitation of main metabolites of mequindox in chicken feces,a high performance liquid chromatography method was developed to detect the concentrations of metabolites M1, M2 of mequindox. Fifteen seven-week-old chickens were administered mequindox orally at a dose of 30 mg·kg-1 BW. After administration, all of their feces were collected at different time intervals and weighed. Then 2 g of feces were extracted using methylene chloride/acetonitrile (1︰1 V/V) solution and the organic phase was separated and evaporated. The residual was redissolved with methanol/water (30︰70 V/V) solution, and then analyzed by HPLC. The results showed that the limit of detection (LOD) and limit of quantitation (LOQ) of metabolites M1 and M2 were 0.01 and 0.05 μg·mL-1, respectively. The range of average recovery of metabolites M1 and M2 were 69.5%-72.4%, 73.5%-80.6% under the concentrations of 0.05, 1.0 and 50 μg·mL-1. The proportions of metabolites M1 and M2 in total amount of mequindox administered were 3.20% and 4.58%, respectively. The excretion peak time of metabolite M1 was 4.8 h with 38.09% of accumulated excretion total amount of metabolite M1. And the excretion peak time of metabolite M2 was 12.24 h with 42.29% of accumulated excretion total amount of metabolite M2 after administration. The present results indicated that the method developed in this study could be used to detect the concentration of metabolites M1 and M2 of mequindox in chicken feces.
Study on Pharmacokinetics of Idazoxan Hydrochloric in Deer
YIN Bai-shuang;HA Da;WANG Hong-bin;GAO Li
2011, 42(1):  110-115.  doi:
Abstract ( 1042 )   PDF (618KB) ( 636 )  
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This experiment was to establish a method for the determination of idazoxan hydrochloric in deer plasma by RP-HPLC, and to discuss pharmacokinetics of idazoxan hydrochloric in deer. Six clinical healthy Cervus nippon Temmincks (breed in a room) were injected with the idazoxan solution at the dose of 044 mg·kg-1. 8 mL blood sample was taken from a jugular vein and plasma was separated for drug analysis using high liquid chromatography with ultraviolet detection. The results showed that the kinetics of idazoxan hydrochloric was fitted to one compartment model with first order absorption by route of administration. The main pharmacokinetic parameters were as follows: the halflives of absorption (t1/2ka) and elimination (t1/2ke) were (2.09±0.34) and (13.18±0.24) min, respectively, area under the plasma drug concent ration-time curve from 0 to ∞(AUC) was (70±3.50) (μg·mL-1)·min, the peak plasma concent ration (Cmax ) was (4.70±0.50) μg·mL-1, peaking at (12.46±0.12) min after dosing. The results indicated that idazoxan hydrochloric is characterized by rapid drug action,fast metabolism with little residue in the blood.
Elimination of Residues of Thiamphenicol in Eggs of Laying Hens
XIE Kai-zhou;YAO Yi-lin;XU Dong;CHEN Shu-qin; XIE Xing;PEI Yan;DAI Guo-jun;WANG Jin-yu;LIU Zong-ping
2011, 42(1):  116-123.  doi:
Abstract ( 745 )   PDF (1027KB) ( 613 )  
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A study on elimination of residues of thiamphenicol (TAP) was conducted in egg white, egg yolk and whole egg of laying hens. TAP was extracted from eggs of laying hens with alkalized ethyl acetate. The extract solution was degreased in n-hexane saturated with acetonitrile, dried in nitrogen evaporator and residues were dissolved in mobile phase. TAP was determinated by high performance liquid chromatography (HPLC) with fluorescence detector. The lowest limit of detection(LOD) was all 1.5 μg·kg-1(S/N=3) for TAP in egg white, egg yolk and whole egg and the lowest limit of quantitation(LOQ) was all 5 μg·kg-1(S/N=10) for TAP. After the laying hens orally administered successively Tap capsules of 10.0, 20.0 and 50.0 mg·kg1 of body weight once every day for 5 days, respectively, residues of TAP in egg white, egg yolk and whole egg have a rise trend. The maximum residue of TAP was detected at the first day after withdrawal time in egg white, egg yolk and whole egg. At the early days after withdrawal time, the residues of TAP in egg white, egg yolk and whole egg were eliminated faster, but they were eliminated slowly at the later period. Residues of TAP in whole egg and egg white were all lower than 50 μg·kg-1, and the lowest LOD (1.5 μg·kg-1)at 6th withdrawal day, respectively. Residues of TAP in egg yolk and whole egg were all lower than the lowest LOD (1.5 μg·kg-1)at 8th withdrawal day. Residues of TAP in egg white, egg yolk and whole egg were positively correlated with TAP orally administered doses
Effects of Betulinic Acid on Lymphocyte and Macrophage in Mice
YI Jin-e;YUAN Li-yun;WEN Li-xin;B Obminska-Mrukowicz;YUAN Hui
2011, 42(1):  124-130.  doi:
Abstract ( 632 )   PDF (1271KB) ( 823 )  
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This study aimed to observe the effects of betulinic acid (BA) on the lymphocyte and macrophage in mice. BA was synthesized by oxidizing betulin from the bark of the white birth with Jone′s reagent, followed by selective oxidation of the resulting betulonic acid with NaBH4. 168 KM mice (6 weeks old) were orally administered the BA for 14 days with the dose of 0, 0.25, 0.5 and 1 mg·kg-1 body weight. The effect of BA on the lymphocytes proliferation, thymocyte and splenocyte subsets, the number of plaque forming cells (PFC) and the phagocytic activity of the macrophages were evaluated. Oral administration of BA was found to significantly increase the thymus and spleen indices, lymphocytes proliferation induced by Concanavalin A or lipopolysaccharide, the percentage of CD4+ cells in thymus as well as the percentage of CD19+ B cells and the ratios of CD4+/CD8+ cells in spleens The number of PFC and the macrophage phagocytic activity was also enhanced. The results suggest that BA enhances mouse cellular immunity, humoral immunity, and activity of macrophages.
The Localization of Ghrelin and Its mRNA in the Rat Hypothalamus and Pituitary
WANG Lin;FANG Fu-gui;ZHANG Xiao-rong;ZHANG Yun-hai;WANG Suo-lu;PU Yong;LI Yun-sheng
2011, 42(1):  131-135.  doi:
Abstract ( 610 )   PDF (2965KB) ( 455 )  
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To investigate the regulation of Ghrelin on the hypothalamus and pituitary of rat. The localization of Ghrelin and its mRNA in hypothalamus and pituitary tissues were detected by immunohistochemical PV-9000 2-step and in situ hybridization, respectively. Results showed that Ghrelin and its mRNA was mainly distributed in the arcuate nuclei, ventromedial nucleus, median eminence, periventricular nucleus of hypothalamus and anterior pituitary, the colour of cells in nuclei were different. A large number of positive cells were found in anterior pituitary, no positive cells were distributed in neurohypophysis. Ghrelin and its mRNA were both presented in the hypothalamus and pituitary tissues of rat, which suggested that the Ghrelin may play a role in the secretion of gonadotropin-releasing hormone and gonadotropin.
研究简报
Analysis on the IL-8 Receptor and Lactoferrin Genes in Subclinical Mastiticsdairy Cows by PCR-SSCP
HE Gao-ming;ZHANG Su-hua;ZHAO Zong-sheng;LI Da-quan
2011, 42(1):  136-140.  doi:
Abstract ( 594 )   PDF (926KB) ( 660 )  
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The aim of this study was to detect the relationships between the genotypes of IL-8R and Lactoferrin genes and subclinical mastitics. The genotypes of IL8R and Lactoferrin genes of 80 healthy and 80 subclinical mastitics-dairy cows were detected by using PCRSSCP technique, and the relationships with the number of somatic cells in the milk were analysed. The results showed that the frequencies of genotype SSCP-5 and SSCP-7 of IL-8R CXCR1 were high in the healthy dairy cows and indicated that individuals with genotype AB performed resistance to subclinical-mastitics.
Sexing of in vitro-produced Yak Embryos by Nested-PCR
ZHANG Da-wei;ZI Xiang-dong;HUANG Lei;MA Li;CHEN Da-wen;XU Hua-wei;LIANG Guan-nan
2011, 42(1):  141-144.  doi:
Abstract ( 926 )   PDF (738KB) ( 642 )  
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The objective of this study was to obtain an accurate and reliable method for determining the sex of yak embryos through amplification of SRY gene of the yaks, and evaluate embryonic development after biopsies. Two pairs of SRY gene nested primers for sex determination and two pairs of HSL gene primers as internal standard and optimized PCR reaction conditions were designed. The accuracy of PCR amplification for sex determination was verified by 24 blood samples. The result showed that in every case, the results were as expected for both female and male, i.e., accuracy was 100%. Using this optimized procedure, clear signals following PCR amplification were obtained from all samples of IVFderived yak embryos, eight cells were sampled from each embryo, 45.8% (11/24) embryos for males and 54.2% (13/24) for females. 56.7% (17/30) of biopsied embryos could further develop. It was concluded that this PCR system was highly reliable for sex determination of yak embryos.
Two-dimentional Gel Electrophoresis Analysis of Differential Protein Spots from BHK-21 Cell Infected FMDV Serotype Asia 1
LIU Yong-jie;ZHANG Ke-shan;SHANG You-jun;GUO Jian-hong;ZHENG Hai-xue;TIAN Hong;LU Guo-dong;LIU Xiang-tao
2011, 42(1):  145-149.  doi:
Abstract ( 678 )   PDF (889KB) ( 637 )  
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To examine the differences and changes of proteome of BHK-21 cells after infection with foot-and-mouth disease virus (FMDV) serotype Asia 1, the monolayer BHK-21 cells infected with FMDV and normal cells were prepared as experimental materials. The total protein from the BHK-21 cells were isolated and used to analyze the proteome differences between the infected and the control cells by 2-dimentional gel electrophoresis (2-DE). Results showed that there were 1 457 protein spots visible in the infected cells, and 1 427 visible protein spots in the control cells. Compared with control cells, there were significance of statistics (P<0.05) at 472 differential protein spots in infection cells, including 251 up-expressed protein spots, 221 downexpressed protein spots and 30 newly proteins spots. These results indicate that the expression pattern of cells are changed after infection with FMDV. It maybe results from inter-actions between viral and cellular components. This is the first time to use proteomics technologies to study the proteome differences after FMDV infection and this study is helpful to further research about molecular mechanisms of interactions between FMDV serotype Asia 1 and BHK-21 cells.
Visualized Detection of Avian Leukosis Virus by Reverse Transcription Loop-mediated Isothermal Amplification Assay
LIU Ye-bing;ZHANG Lei;SUN Yue-hui;MAO Ya-qing;LI Jun-ping;Ning Yi-bao
2011, 42(1):  150-156.  doi:
Abstract ( 997 )   PDF (1334KB) ( 709 )  
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We established a rapid detection of avian leukosis virus (ALV) by reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP) in this study. According to the published ALV sequences in GenBank, multiple pairs of primers were designed targeting the conserved region of ALV. The primer, reaction system and processes of LAMP were detected and analyzed by the LAMP Real-time Turbidimeter so as to evaluate its specificity and sensitiveness. Meanwhile, the amplified products were colored by SYBR GreenⅠ after completion of the reaction, so that the amplification can be detected with naked eyes and the results were compared to the former detected by LAMP Realtime Turbidimeter. RT-LAMP showed a highly efficient amplification for ALV nucleic acid which performed at 63 ℃ for 36 min. The results detected with naked eyes by the addition of SYBR GreenⅠ at the end of reaction were consistent with the results detected by Turbidimeter. The RT-LAMP assay showed a strong specificity that the results were all negative towards other nosazontology detection in chicken; and it also showed a high sensitivity with a detection limit of 14.2 pg·μL-1. The results of 24 clinical samples detected by LAMP developed in this study were consistent with the ones tested before by ELISA and COFAL test. A visualized RTLAMP method which can detect ALV rapidly was established and the results can also be read with naked eyes. The method was easy to operation, which was suitable for rapid detection of ALV.