Acta Veterinaria et Zootechnica Sinica

Molecular Cloning, Sequence Analysis and Prokaryotic Expression of the Porcine Sar1b Gene

WANG Xue-min;LIU Bang;ZHAO Shu-hong;FAN Bin;ZHU Meng-jin;YU Mei;XIONG Tong-an;LI Kui

2007, 38(7): 625-629. doi:

Abstract ( 2098 )

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A pair of primers was designed based on the published nudeotide sequence of the porcine Sar1b gene (GenBank accession number: AY819557). Total RNA was isolated from the pig liver tissue and the first-strand cDNA was generated using reverse transcription reactions. The full-length coding sequence of the porcine Sar1b cDNA was amplified by RT-PCR. The product was double-digested by EcoRI and SalI, and then directionally cloned into the pET28a vector. The recombinant expression plasmid in the E. coli BL21 (DE3) was induced with 1 mmol/L ITPG. SDS-PAGE analysis showed that the induced expressed protein was about 26 ku. Western blot revealed that the expressed protein was the His tag fusion protein, which indicated that the recombinant prokaryotic expression vector expressed the fusion protein successfully. The cloning and expression of the porcine Sar1b gene established the foundation for further study on its biological function.

Cloning and Expression Analysis of Porcine RPS3 Gene

YANG Xiu-qin;GUO Li-juan;GUAN Qing-zhi;LIU Hui;LIU Di

2007, 38(7): 630-635. doi:

Abstract ( 2007 )

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In order to further elucidate the structurefunction relationship of RPS3 which has multifunction in vivo and to reveal the possibility of its CDS as a phylogenetic marker, the RPS3 cDNA (GenBank accession No: DQ660373) was cloned using bioinformatics, RT-PCR and 3′-RACE methods; and its expression profile was researched with semiquantitative RT-PCR. Sequence analyses showed that the full ORF is 732 bp, encoding a polypeptide of 243 amino acids which includes the ribosomal protein S3 signature. The phylogenetic tree demonstrated similar topology with the zoology. Semiquantitative RT-PCR analyses showed that the expression level is the highest in heart and the lowest in liver, and there are no significant difference among other tissues. With the age increasing, the expression level gradually decreases and becomes steady in pigs older than three months. The results made clear that the coding sequence of RPS3 fits to construct phylogenetic tree among species and its expression at mRNA level is related to growth and metabolism

Polymorphism of the Exon2 of SLADQB Gene and Its Relationship with Reproductive Performance in Sutai Pigs

BAO Wen-bin;WU Sheng-long;CHEN Guo-hong;JU Hui-ping;HUA Jin-di;HUANG Xue-gen

2007, 38(7): 636-640. doi:

Abstract ( 892 )

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The exon 2 of SLA-DQB gene in Sutai pigs was analyzed by PCR-RFLP method, the results showed that the exon 2 of SLA-DQB gene digested by restriction endonuclease RsaⅠ and HaeⅢ could be divided into four kinds of genotypes and three kinds of genotypes in Sutai pigs respectively. The results of χ2 indicated that the frequencies of patterns digested by RsaⅠand HaeⅢ fit with Hardy-Weinberg equilibrium in Sutai population. Four kinds of combined patterns were observed in Sutai pigs. The correlation between these combined patterns and the reproductive performances of Sutai pigs was analyzed, the results showed that reproductive performances (the total litter, number of viable pigs, litter weight at birth, number of ablactation pigs and litter weight after ablactation in the third litter) of Sutai pigs with BBEE combined pattern were significant higher than those with AADD combined pattern, also higher than the pigs with ABDE combined pattern and ACDE combined pattern, but were no significant difference in reproductive performances between pigs with AADD combined pattern and the pigs with ABDE and ACDE combined patterns.

Mitochondrial DNA D-loop Genetic Diversity and Origin of Chinese Domestic Donkeys

GE Qing-lan;LEI Chu-zhao;JIANG Yong-qing;CHEN Hong;ZHANG Wei;DANG Rui-hua;ZHENG Hui-ling;ZHANG Ai-ling;LI Tian-peng

2007, 38(7): 641-645. doi:

Abstract ( 1524 )

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All 126 mtDNA D-loop sequences 399 bp in length from 12 Chinese domestic donkey breeds were analyzed, including 26 sequences published before. The results revealed 36 different haplotypes with 37 variable sites.The haplotype diversity and the nucleotide diversity were 0.466 7-0.977 8 and 0.001 2-0.028 5, respectively, indicating high level of genetic diversity in Chinese domestic donkeys. A Neighbor-joining tree constructed using 36 haplotypes of Chinese domestic donkeys, 3 sequences from Nubian wild ass, 3 sequences from Somali wild ass and 6 sequences belonging to Asian wild ass firstly showed that the maternal origin of Chinese domestic donkeys was from Somali and Nubian wild ass in Africa but not from Asian wild ass We also discussed transport of donkeys among different regions in China

Yak Myoglobin: Gene Cloning and Sequencing, Purification, Contents and Their Relation to Activities of Lactate Dehydrogenase and Malate Dehydrogenase

ZHENG Yu-cai;SU Yong-jie;WEN Yong-li;JIN Su-yu;CHEN Wei;ZHOU Jing;PIAO Ying

2007, 38(7): 646-650. doi:

Abstract ( 880 )

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To investigate the molecular mechanism for yak to adapt the hypoxia environment of plateau, experiments were carried out to measure myoglobin concentration, activities of lactate dehydrogenase(LDH) and malate dehydrogenase(MDH) in heart and skeletal muscles of yak. Also, yak myoglobin was purified and its gene was cloned and sequenced. Yak heart muscle and skeletal muscle contained significantly higher concentration of myoglobin than buffalo and yellow cattles. Heart tissues had significantly higher content of myoglobin and MDH/LDH ratio than skeletal muscles (P<0.01) in yak, yellow cattle and buffalo. Heart MDH/LDH ratio in yak was significantly higher than those of buffalo and yellow cattle, however, skeletal muscle MDH/LDH ratio showed no difference among the species tested. Significant positive relationship between myoglobin content and MDH/LDH ratio was observed in heart and skeletal muscles. The full-length coding sequence of yak myoglobin was cloned by RT-PCR using total RNA extracted from heart muscle of Maiwa yak.The amplified yak myoglobin gene shares 995% homology with that of bovine and the same amino acid sequence. Myoglobin was purified from yak heart muscle by salting-out, CM-Sephadex cation exchange chromatography, and Sephadex G-50 gel chromatography.The molecular weight of purified yak myoglobin was approximately 17 ku as shown on SDS-gel.

Construction of Expression Vector of Chicken Peroxisome Proliferator-Activated Receptor-Gamma Gene and the Preparation of Its Antiserum

WANG Ping;WANG Qi-gui;LI Hui;ZHANG Fu-chun

2007, 38(7): 651-656. doi:

Abstract ( 883 )

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To construct the prokaryotic and eukaryotic expression vector and prepare the antiserum of peroxisome proliferator-activated receptor-gamma (PPAR-γ) gene in chicken. The sequence of PPAR-γ gene was amplified by reverse transcriptase-PCR(RT-PCR) from the total RNA of chicken fatty acid and insert into pGEX-4T-1 vector and pcDNA3 vector. Recombinant plasmid pGEX-4T-1/PPAR-γ was expressed in E.coli BL21; Mice were immunized with purified recombinant plasmid pcDNA3/PPAR-γ. SDS-PAGE showed that recombinant pGEX-4T-1/PPAR-γ expressed abundantly a fusion protein in E.coli BL21. ELISA and Western blot showed that antibody obtained from the serum of mice by injecting recombinant pcDNA3/PPAR-γ was high effective and strong specific. The recombinant pcDNA3/PPAR-γ in the current study would provide a powerful tool for overexpression research of chicken PPAR-γ gene at cell level; The recombinant protein and antiserum in the current study would also establish a basis for function research of chicken PPAR-γ at protein level.

Cryopreservation Capacity of Chicken Spermatogonial Stem Cells

LI Bi-chun;ZHOU Guan-yue;CHEN Guo-hong;SUN Guo-bo;SUN Peng-xiang;XU Qi;LIU Tie-zheng

2007, 38(7): 657-662. doi:

Abstract ( 1404 )

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To isolate spermotogonia using two combinatorial enzyme three step wises digest the testicular tissues from 19 d-old-embryo after hatching, the cryopreserve effect of dimethylsulphoxide (DMSO), glycerol (GLY) and ethylene glycol (EG) cryoprotectants, each at (5%, 10%, and 15%) concentration on the 19 d chicken embryonic SSCs was compared with fast freeze method. The results indicated that： (1) When SSCs frozen in 5%,10% and 15% DMSO medium, the viability rate of after thawed SSCs were 73.1%,88.6% and 74.8% respectively, the difference of three concentration were significant(P<005) When SSCs frozen in 5%,10% and 15% EG medium, the viability rate of after thawed SSCs were 69.4%,83.1% and 65.2% respectively, and the difference of three concentration were significant(P<0.05). When SSCs frozen in 5%,10% and 15% glycerin medium, the viability rate were less than 15%, and difference of three concentration were not significant (P>0.05); (2) The SSCs were cryopreservated in 10% DMSO for 1-2 weeks, then thawed and seeded on the feed cells to culturing, the chicken embryonic fibroblast cells as feed cells. The results found that the SSCs had formed the colonies which were positive after AKP staining. On the contrary, when the SSCs were cultured without feed cells, the colonies couldn’t form The results showed that the 10% DMSO was suitable freezing media for the 19 d SSCs from chicken embryo

Effect of Linoleic and Linolenic Acid on Milk Fat Conjugated Linoleic Acid from Dairy Cows

BU Deng-pan;WANG Jia-qi;Dhiman T R;LIU Shi-jun

2007, 38(7): 663-671. doi:

Abstract ( 1559 )

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Objective of this study was to determine the comparative effectiveness of linoleic and linolenic acid in enhancing CLA in milk when fed at the same levels to mid lactation dairy cows. Forty dairy cows were randomly assigned to 4 treatments. Cows in four treatments were fed a basal diet that was supplemented oils differed in their linoleic and linolenic acid contents. The four treatments were Low linoleic and linolenic acid(LLALEA), high linoleic(HLA), High linolenic acid(HLEA) or High linoleic and linolenic acid(HLALEA). Experimental duration was 9 weeks and measurements were made during the last 6 weeks. Dry matter intake, energy corrected milk (ECM) , milk fat and protein contents were not differ among treatments. The proportions of C18∶1 trans-11 (TVA) and C18∶2 cis-9, trans-11 (CLA) were 1.48, 6.19, 3.04, and 4.53 (P=0.01), 0.64, 2.39, 1.60, and 1.81% of total fatty acid methyl esters in LLALEA, HLA, HLEA, and HLALEA, respectively. The proportions of unsaturated fatty acids were 27.9, 33.6, 36.4, and 33.8 (P=0.001) in LLALEA, HLA, HLEA, and HLALEA, respectively. Feeding free oil rich in linoleic acid is 100 and 50% more effective in enhancing TVA and CLA in milk fat, respectively, than feeding free oil rich in linolenic acid fed at the similar levels in the diet.The milk fat content of CLA c-9,t11 across the whole treatment increased linearly with increased intake of dietary LA. Milk fat concentration of TVA in HLEA,HLALEA and HLA were 1.56,3.05 and 4.71 g/100 g higher than LLALEA respectively ,while CLA c-9,t11 were 2.5,2.8 and 3.7 times respectively. Therefore, dietary LA ,which was used as the main substrate for CLA endogenous synthesis via SCD in the mammary gland, is more effective than LEA in enhancing milk CLA c-9,t11 concentration.

Influence of Exogenous Alphaamylase Supplementation on Development of Digestive Organs and Intestinal Enzyme Activities of 21-day-old Broilers

JIANG Zheng-yu;ZHOU Yan-min;WANG Tian

2007, 38(7): 672-677. doi:

Abstract ( 1364 )

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Four hundred and forty 1-day-old AA broiler chickens were randomly allocated to four groups for 5 replicates, and fed a commercial starter diet and the same diets supplemented with 1 000 U/kg (250 mg/kg), 3 000 U/kg (750 mg/kg), 9 000 U/kg (2 250 mg/kg) of α-amylase preparation up to 21 day of age to study the effects of supplementary αamylase of different levels on the development of digestive organs, intestinal enzyme activities. The results showed that relative weights of liver, gizzard, anterior intestine and relative length of anterior intestine tended to decrease (P>0.05).The increased activities of amylase, protease (P<0.05), and trypsin (P<0.05) in anterior intestinal and high-dose-depression responses with the amylase supplementation of 2 250 mg/kg were observed.Lipase activity, however, was unaffected by amylase supplementation of all levels (P>0.05). DNA, RNA concentration and the activities of maltase and sucrase within jejunal mucosa were not affected by supplementary amylase of 250 mg/kg and 750 mg/kg. High-dose-depression responses were also observed for the activities of maltase and sucrase (P<0.05)

Endogenous Outputs and True Digestibility of Phosphorus Associated with Peanut Meal and Soybean Meal in Growing Pigs by Multiple Linear Regression Analysis Technique

ZUO Jian-jun;WANG Jing;ZHANG Tie-ying;ZHANG Chang-ming;FENG Ding-yuan;YIN Yu-long;FAN Ming-zhe

2007, 38(7): 678-684. doi:

Abstract ( 1306 )

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Regression analysis technique was used to measure the fecal phosphorus (P) endogenous outputs and true P digestibility values with peanut meal and soybean meal in this study with growing pigs. Six Yorkshire×Landrace barrows, with average initial body weight of (24.8±1.42) kg, were fed in the metabolic cage. According to a 6×6 Latin square design, the six diets with soybean-cornstarch-glucose based experimental diet with peanut meal containing six P level (0.09%，0.17%，0.26%，0.35%，0.43%，0.53%) were formulated, in which the peanut meal and soybean meal were the only P source. Chromic oxide (0.35%) was included in the diets as digestibility marker. Each experimental period lasted 8 d with 6 d adaptation and 2 d collection of the feces.The results showed that the P endogenous outputs were 0.526 6 g/kg DMI, true P digestibility with peanut meal was 28.00%, and true P digestibility with soybean was 38.87% with regression analysis technique.

Detection of Classical Swine Fever Virus in Experimentally Infected Pigs Using a Newly Developed Real-time Fluorescent Quantitative RT-PCR

ZHAO Jian-jun;CHENG Dan;LI Na;SHI Zi-xue;SUN Yuan;ZHAO He-ping;ZHU Qing-hu;TU Chang-chun;QIU Hua-ji

2007, 38(7): 685-693. doi:

Abstract ( 937 )

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A real-time fluorescent quantitative RT-PCR assay for detecting classical swine fever virus (CSFV) was established and evaluated in experimentally infected pigs.The sensitivity of the assay was 10 copies/μL viral RNA, with an agreement of 94.7% with RT-nested PCR in detecting 152 different samples. The assay was used to detect CSFV in the whole blood samples of the pigs infected with 106 TCID50 Shimen strain and contact pigs. The results showed that there was an excellent correlation between clinical symptoms and viral RNA loads in the blood.Viral RNA can be detected in the blood samples of contact pigs 3 to 4 days prior to the onset of clinical signs.

Construction of Co-expressing Plasmids of AgB from Taenia Solium with Porcine CD58 or IFN-γ Gene and Expression in BHK-21 Cells

HOU Jun-ling;JING Zhi-zhong;ZHENG Ya-dong;HU Zhi-min;LUO Xue-nong;MENG Xue-lian;DOU Yong-xi;LIU Jun-long;CAI Xue-peng

2007, 38(7): 694-699. doi:

Abstract ( 1283 )

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Cysticercosis is an important zoonosis.The development of efficient, safety and cheap vaccines is one of contemporary major directions. In this study, AgB, CD58 and IFN-γ gene were amplified from the former constructed recombinant plasmids named pGEM-TEasy-AgB, pGEM-TEasy-CD58 and pGEM-TEasy-IFN-γ by PCR and cloned into pBudCE4.1.The acquired recombinant plasmids were transfected into BHK-21 cell with liposome. 48 hours after transfection, the expression of AgB, CD58 and IFN-γ were tested by indirect immunofluorescence assay (IFA) or direct immunofluorescence assay.The results showed that AgB, CD58 and IFN-γ were successfully expressed in BHK-21 cells, providing a solid basis for development of DNA vaccine against cysticercosis

Cloning and Bioinformatics Analyzing for Capsid Protein (VP60) Gene of Rabbit Hemorrhagic Disease Virus YL Strain

LI Chuan-shan;YANG Ying;ZHANG Shou-tao;YANG Zeng-qi;GUO Ai-guang

2007, 38(7): 700-707. doi:

Abstract ( 1334 )

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The capsid protein VP60 gene of rabbit hemorrhagic disease virus（RHDV）YL Strain was cloned by RT-PCR and then sequenced. YL Strain was the first RHDV strain being sequenced in the Northwest of China. The results showed that the nucleotide sequence of VP60 gene was 1 740nt in length and encoding a protein of 580 aa.The sequence had been submitted to GenBank (the accession number is DQ530363)We downloaded all other 39 sequences of published RHDV isolates from GenBank and then constructed phylogenetic tree and drawn homology and divergence form by bioinformatics software. The results indicated that the VP60 gene of RHDV was very conservative, and the homology of the nucleotide among different isolates exceeded 90%, but they can be divided into 5 gene groups. The gene group and region had negative connection.All Chinese RHDV isolates belonged to the Ⅴ gene group eccept WX84 Strain. In this group, the hereditary distance between the strains isolated in recent years and NJ85 Strain was very close. The RHDV strains of China isolated in recent years probably come from the same strain NJ85. Meanwhile, the secondary structure and antigenic index of four strains from different areas of China were predicted and analyzed. The results revealed that there were certain differences in secondary structure, but the antigenicity was identical

Establishment and Initial Application of Low Density DNAarray for Diagnosing the Subtypes of SIV

CHEN Hong-jun;HOU Yi-hong;BAI Hua;CHEN Gui-lai;WU Shi-you;YIN Yan-bo;QIAN Yong-hua;TIAN Zhen-yu

2007, 38(7): 708-712. doi:

Abstract ( 917 )

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The hemagglutinin(HA) and neuraminidase(NA) gene of five subtypes influenza A virus were amplified by RT-PCR with primers based on the diversity of HA and NA gene. Meanwhile conserved cDNA fragments of M gene was amplified by RT-PCR with universal primer of influenza A virus. Each fragments were cloned and inserted into pMD18-T to construct recombinant plasmids for DNA probe preparation. Recovered DNA probes were spotted on located sites on nitrocellulose filter to prepare DNAarrays. 217 different samples of area was detected by this DNAarray. The results showed that this DNAarray could identify five subtypes of Influenza A virus in samples and show high sensitivity and specificity. This DNAarray provids a quick, sensitive and highflux method for largescale epidemiology investigation and quarantine of swine influenza virus.

Immune Response and Gene Expression in Rats after Inhibin Gene Immunization

MAO Da-gan;ZHANG Zhi-jie;YANG Li-guo;HE Xiao-hong;ZHANG Hong-lin

2007, 38(7): 713-717. doi:

Abstract ( 1352 )

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To study the immune response, estrus and tissue distribution of inhibin after inhibin gene immunization, 60 rats were divided into five groups and immunized (i.m.) with 10（T1）,50（T2）,100 μg pCIS（T3）, 50 μg pcDNA3.1（V）and 0.85%NaCl (S) respectively. The antibody against inhibin was detected by ELISA, the estrus was detected by vagina smear and the tissue distribution of inhibin was detected by immunohistochemistry. The results showed that the P/N value of inhibin antibody went up significantly at day10 after 1st booster（P<0.05）and the P/N value in T2 and T3 group raised significantly at day 20 after 2nd booster（P<0.05）. The P/N value in T3 group was slightly higher than those in the T1 and T2 group（P>0.05）. The estrus was not affected. The immunohistochemistry results showed that there was no positive staining in the tissues such as heart, liver and muscles at inoculated sites, however, positive staining occurred in ovary, kidney and pituitary for both the experimental and control groups 2 weeks after 2nd booster. In the spleen, positive staining appeared in the experimental groups, but not in the control groups. These results suggested that the increased immunization time and dosage of inhibin DNA did not cause significant immunologic tolerance. The inhibin gene is safe to immunize rats and the tissue distribution of inhibin also provides important evidence to elucidate the mechanism of gene immunization.

Study on the Pharmacokinetic of Sulfamonomethoxine Suspension Injection in Pigs

WU Jun-wei;RAO Yong;JIANG Bo

2007, 38(7): 718-722. doi:

Abstract ( 945 )

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The aim was to obtain the basic data for development and registration the sulfamonomethoxine (SMM) suspension preparation.The pharmacokinetics of SMM and SMM-Na suspension injections and SMMNa injection were studied in healthy pigs. 18 pigs of 3 groups were administered muscularly with SMM at 100 mg/kg body weight in single.The SMM concentrations in plasma were analyzed by high performance liquid chromatography (HPLC).The pharmacokinetic parameters were calculated by MCPKP program. The pharmacokinetic processes of 3 preparations were all fitted to twocompartment open model with first-order absorption. The main pharmacokinetic parameters were as follows: t1/2α (2.86±0.85) h，t1/2β (45.57±8.06) h，AUC (712.04±108.20) mg·L-1·h，Cmax (12.16±0.52) μg/mL for the SMM suspension; t1/2α (7.90±1.21) h, t1/2β (24.87±12.92) h, AUC (1 489.78±164.63) mg·L-1·h, Cmax (97.86±10.24) μg/mL for the SMM-Na suspension; t1/2α (0.10±0.04) h， t1/2β(6.20±0.57) h，AUC (1 080.83±93.78) mg·L-1·h，Cmax (84.30±4.26) mg/L for the SMM-Na injection. Though SMM suspension injection was eliminated very slowly, its plasma-drug concentrations were low with a short effective plasma-drug concentration time, and were difficult to meet the clinic treatment requirement. SMM-Na suspension injection showed a long effective plasma drugconcentration time with high plasma-drug concentrations and it was an excellent delayed-release preparation.

Effects of Achyranthes Bidentata Polysaccharide on Immune Efficacy of Vaccine in Chickens

QIU Yan;HU Yuan-liang;CUI Bao-an;ZHANG Hong-ying;WANG Yuan-ge

2007, 38(7): 723-727. doi:

Abstract ( 1469 )

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150 14-day-old chickens were randomly assigned to three groups and vaccinated with ND-IB live virus vaccine. At the same time, the chickens in two experimental groups were intramuscularly injected with achyranthes bidentata polysaccharide at high and low dosages, control group were injected with physiological saline, once a day for three successive days. On day 7, 14, 21, 28, 35 and 42 after vaccination, the dynamic changes of serum Newcastle disease (ND) hemagglutination inhibition (HI) antibody titer were determined by Micromethod. On day 10, 20, 30, 40 and 50 after vaccination, blood samples from 5 chickens per group were collected to separate lymphocyte and to determine the peripheral T lymphocyte proliferation with MTT method. The content of CD4+ and CD8+ T cells were tested by using flow cytometry with doublecolorstaining method, and the weights of whole body, spleen, bursa and thymus were weighted for calculation of immune organ index. The results showed that achyranthes bidentata polysaccharide at high dosage significantly enhanced the antibody titers, promoted the proliferation of peripheral T lymphocyte, and raised the ratio of CD4+ to CD8+T lymphocyte (P < 0.05) and immune organ indexes. These results indicated that achyranthes bidentata polysaccharide had significant immune enhancement role on immunity of ND-IB vaccine in chickens.

Pathological Study on Anti-experimental Atherosclerotic Effects of Rhubarb Ethanolextract in Rabbits

CHEN Juan-hua;LU Zhan-jun;XU Zai-pin;DENG Xiao-yan;MAO Yi-zhi;HUO Xin

2007, 38(7): 728-734. doi:

Abstract ( 792 )

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Thirty healthy male white rabbits were divided randomly into five groups, six rabbits per group. The rabbits in control group were fed with common forage. The rabbits in model group were fed with high lipid forage. The rabbits in three rhubarb groups were fed with high lipid forage and administrated intragastrically with different dosage of rhubarb ethanolextract (REE). The rabbits were killed at the end of the tenth week, the atherosclerosis (AS) plaque areas of aorta were measured and the percentage of AS plaques covering aortic intima (PA) was calculated. The pathological changes of coronary arteries were observed with microscope and the incidence of atherosclerosis of small coronary arteries was calculated. The results indicated that REE could inhibit the progression of aortic and abdominal aortic intimal AS plaques and decrease the incidence of AS in artery branch. REE also could reduce AS formation in coronary arteries.

Maternal Origins of Six Indigenous Chicken Breeds in China

SONG Chun-hong;CHEN Hong-ju;MA Yue-hui;TANG Hui;CAO Ding-guo;FAN Xin-zhong;JIANG Yun-liang

2007, 38(7): 735-740. doi:

Abstract ( 998 )

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The maternal origins of six Chinese indigenous chicken breeds were investigated by sequencing the D-loop region of mtDNA. Of the six chicken breeds, Wenchang, Luxi game, Shouguang, Jining Bairi and Laiwu Black have five, five, five, six and seven haplotypes respectively, whereas Langya breed has only three haplotypes. Six Chinese indigenous chicken breeds were clustered in three clades, of which clade A is dominant. Clade A includes 17 haplotypes which are shared by ten Shouguang, nine Luxi game, five Wenchang, six Laiwu Black, six Jining Bairi and two Langya individuals, accounting for 100％, 90%, 84％, 75％, 60％ and 20％ of the total number of each breed, respectively. These indigenous chickens have close relationship with the G .g .gallus, G. g. jabouile and G. g. spadiceus distributed in Laos and Yunnan Province in China. Clade B contained eight Langya (80%) and one Jining Bairi individuals(10%) . Clade C contained one Luxi game (10%), one Wenchang (16.7%), two Laiwu Black (25%) and three Jining Bairi individuals(30%). Domestic chickens of Clade B and C were clustered with H19 and H32, H33 of G. g. gallus. The cluster results indicated that the six Chinese indigenous chicken breeds may originated from the continental subspecies of Gallus gallus of Yunnan, Laos and Vietnam or its surrounding areas. Gene flow is the major factor affecting genetic diversity of these chicken breeds. Mismatch distribution and Fu’s Fs test suggested that the five Shandong indigenous chicken breeds did not undergo population expansion in the past .

Biological Identification of Newcastle Disease Viruses Isolated from Eggs of a Parent Breeder Farm

SUN Shu-hong;CUI Zhi-zhong

2007, 38(7): 741-743. doi:

Abstract ( 883 )

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Among 24 eggs from a meattype parent breeder chicken flocks with eggdropping, 2 embryos were dead at 8-9 d after incubation. The allantoic fluid from them demonstrated hemagglutination activity. By hemagglutination inhibition (HI) and viral neutralization test in cell culture with monoantiserum against Newcastle disease virus (NDV), H5- and H9- avian influenza virus (AIV), the agent for HA activity in allantoic fluid of dead embryos were identified as NDV and designated as strains Tengz060104 and Tengz060107. The MDT, ICPI and IVPI for Tentz060104 were 56.4 h, 1.88 and 2.67 respectively, and they were 40.8 h, 1.88 and 2.79 for Tengz060107. Both two isolated strains can cause symptoms and lesions typical to Newcastle disease in 6 weeks old SPF chickens. The results indicated that two NDV isolates were velogenic virulent strains. In cross HI, Tengz060104 and tengz060107 had 100% homology, but they have only 667% and 577% antigenic homology with F48E8 strain . This is the first report of isolation of virulent NDV from eggs in China.

Effects of Monochromatic Light on Immune Function of Broilers

XIE Dian;CHEN Yao-xing;WANG Zi-xu;LI Jun-ying;CAO Jing;JIA Liu-jun

2007, 38(7): 744-747. doi:

Abstract ( 848 )

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A total of 260 day-old AA male broilers were randomly assigned to four light treatments to investigate the effect of monochromatic light on immune function. Chicks were exposed to blue (480 nm, B), green (560 nm, G), red (660 nm, R) and white (400-700 nm, W) light by using lightemitting diodes (LED) lamps. All light sources were equalized on the light intensity of 15 lx and applied for 23 h daily. There were five replication pens for each light treatment and 13 birds per pen, and the experiment lasted 7 weeks. On day 49, spleen weights were determined. T lymphocyte proliferation and serum levels of cortisol were analysized on the 21- and 49-day old. Antibody titers to NDV were measured on day 14, 28, 42 and 49. The results were as follows: (1) Blue light significantly increased spleen weight compared to RL groups (42.2%), and no significant difference was found between WL, GL and BL groups at 49 d of age (P<0.05);(2) Blue light significantly decreased cortisol concentrations in serum compared to white light at 21d and 49 d (25.2％and 26.2％ respectively,P＜0.05);(3) A significant elevation in serum anti-NDV level was observed in broilers reared under GL compared to RL at 28 d of age (32.9%), but no significantly difference was detected between BL, RL and WL groups (P>0.05). By 49 d of age, the antibody titers from BL was higher than that of RL (62.8%, P<0.05), but not significant difference among BL, GL and WL groups (P>0.05); (4) A significantly increase of peripheral blood T lymphocytes proliferation was observed in birds reared under WL and GL groups compared to RL at 21 d of age(P<0.05, 101.1％ and 80.8％ respectively). In addition, broilers reared under WL and BL had significantly elevated proliferation response compared to RL at 49 d of age by 41.5％ and 26.9%, respectively (P＜0.05). These results showed that immune function would be increased in broilers when who was illuminated with either green light in early stage of broiler growth stage or blue light in latter growth stage under 15 lx light intensity. Moreover, blue light could, to an extent, show an action of alleviating immunologic stress.

Expression of PTEN Gene and Its Clinical Value in Canine Mammary Tumors

QIU Chang-wei;WANG Heng;QIAO Ming-ming;LIN De-gui

2007, 38(7): 748-752. doi:

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Elevated levels of phosphatase and tensin homolog deleted on chromosometen (PTEN ) have been reported in mammary gland tumors in humans. This study was designed to study the expression of PTEN mRNA in canine mammary tumor tissues. We investigated 38 canine mammary gland tumors(including 15 benign mammary tumors and 23 malignant mammary tumors), 4 normal mammary gland by real-time RT-quantitative PCR- PTEN gene expression levels in canine malignant mammary tumors were much lower than that in benign mammary tumors and normal mammary tissues (P < 0. 001) ; The expression of PTEN in benign mammary tumors vary insignificantly than that in normal mammary tissues(P>0.05); The expression of PTEN in canine mammary cancer without lymph node metastasis vary insignificantly contrast to mammary cancer with lymph node metastasis(P>0.05) .The expression of PTEN was not correlated with tumor size and patient' age. In conclusion, the abnormal expression of PTEN gene protein plays an important role in the development of canine mammary tumors. It may be a predictor for biological behaviors and prognosis of breast cancer

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