Acta Veterinaria et Zootechnica Sinica

Association of Polymorphism for Porcine FSHβ Subunit Gene with ReproductiveTraits and Placental Traits in Large White

CHEN Lai-hua;;WANG Li-xian;JI Yue-guang;YAN Hua;CHENG Du-xue;ZHANG Long-chao;WANG Li-gang;LI Yong

2010, 41(11): 1365-1370. doi:

Abstract ( 963 )

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FSHβ subunit gene was regarded as a candidate gene for reproductive traits in this study, and the polymorphisms of FSHβ was detected by electrophoresis method in 117 Large White pigs. Meanwhile, the effect of polymorphisms of FSHβ subunit gene on total born number(TNB), number born alive(NBA), birth weight(BW) and placental traits(PT) were analyzed. The results showed, about FSHβ locus, the first parity sows with genotype AA and AB had 3.25 and 2.37 TNB more than those with genotype BB. The first parity sows with genotype AA and AB had 3.31 and 2.49 NBA more than those with genotype BB, respectively. The multiparous sows with genotype AA and AB had 2.71 and 1.60 TNB more than those with genotype BB. The multiparous sows with genotype AA and AB had 2.95 and 1.74 NBA more than those with genotype BB. In multiparous sow population, the placental efficiency(PE) with AA genotype were higher than those with AA and BB genotypes significantly (P<0.05) with 16.28% and 23.09%.The results of combined genotype effective indicated that A allele at FSHβ locus in all population had the positive effective on TNB, NBA and PE.

Molecular Cloning, Sequence Analysis and Expression in E. coli of DECR1 Gene in Pig

LI Bu-gao;GUO Xiao-hong;CAO Guo-qing;YANG Xiao-fen;WANG Xiao-jing;ZHOU Zhong-xiao

2010, 41(11): 1371-1378. doi:

Abstract ( 1058 )

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This experiment was conducted to obtain sequence of 2, 4 -dienoyl -CoA reductase 1 (DECR1), and to study its prokaryotic expression. By Reverse Transcription PCR and Rapid Amplification of cDNA Ends (RACE), the fulllength cDNA of DECR1 was cloned from Mashen pig liver, and inserted into the expression vector pET-32a+. After confirmation by the sequencing and restriction enzyme analysis, the recombinant plasmid was transformed into E. coli BL21. The results indicated that the cDNA of pig DECR1 contained 2 352 nucleotides, including a 987 bp open reading frame (ORF) flanked by a 53 bp 5′-UTR and a 1 312 bp 3′-UTR. Pig DECR1 CDS encoded 328 amino acid residues, which shared 99%, 88%, 88%, 87%, 87%, 87%, 87% and 83% identity with those of Sus scrofa (predicted), Bos taurus, Homo sapiens, Macaca mulatta, Pan troglodytes, Equus caballus, Canis and Mus musculus, respectively. SDS-PAGE results showed that the recombinant protein was expressed and then the expression level reached the highest level after induced for four hours. Western blot analysis indicated that the molecular weight of the expressed protein was the same as predicted size of approximately 35 ku. Collectively these data provided the important information for further studying the physiological functions and molecular mechanism of pig DECR1 gene.

Cloning and Functional Analysis of Serine/arginine-rich Specific Kinase 1 in Pig

E Guang-xin;LIU Di;ZHANG Dong-jie;YANG Xiu-qin;CUI Yu;ZHU Ji-yuan;YANG Shao-cheng;WANG Jia-bo;XIE Yu-tong

2010, 41(11): 1379-1386. doi:

Abstract ( 1076 )

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The objective of this study was to illuminate the molecular features and the expression profile of the porcine SRPK1 gene. The SRPK1 gene from the Yorkshire pig was cloned and sequenced, and then its molecular features were analyzed. Use inverse PCR to clone the partial 5′UTR sequence of SRPK1 in pig. The mRNA distribution profile of the porcine SRPK1 gene in ten tissues (heart, muscles, liver, kidney, lung, stomach, small and large intestine, spleen, brain) of Yorkshire and Duroc (one-day, one-month) were examined by real-time PCR. Construct pig skeletal muscle injury model by subcutaneous injection to study the expression characteristics of SRPK1 gene in repair process of skeletal muscle. A sequence with a length of 2 499 bp was cloned, it contained the CDS (coding sequence) region of 1 968 bp of the porcine SRPK1 gene, encoded a protein composed of 656 amino acids, including two P_Kc domains. The sequence of the porcine SRPK1 protein shared high similarity with human and chimpanzee, and they were closed in the Phylogenetic tree. The result of bioinformatics analysis showed that 5′UTR obtained in the study contained multiple transcription binding sites：HSF1, HSF2, Ik-1, IK-2, SRY, SP1 MyoD, USF, E47, p300, CP2, RREB-1, E2F and AP-1. The study results showed that, by realtime PCR, pigs had the organization- and species-specific in expression, but highly in stomach, small intestine and large intestine. The expression of SRPK1 gene fluctuated in the injury and repair process. There are several transcription binding sites in 5′UTR related to muscle development, it mainly expressed in digestive system and the expression level increased in injury and repair process of skeletal muscle. It was inferred that SRPK1 gene may be related to skeletal muscle cell development, or other growth-related factor expression.

The Effects of Dynamic Addition of FSH on Sheep Ovarian Cortical Tissue in vitro Culture

PENG Xia-yu;YANG Mei;CHEN Tong;WANG Li-qin;GUO Zhi-qin

2010, 41(11): 1387-1393. doi:

Abstract ( 678 )

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The objective of this work was to examine the effect of follicle-stimulating hormone (FSH) on the in vitro activation and viability of sheep primordial follicles. Sheep ovarian cortical fragments were cultured in a steel mesh culture system, various concentration of FSH (1, 10, 50, 100 ng·mL^-1) were either stabilized or dynamically (different days with different concentration of FSH) added in culture medium. Ovarian tissues, both fresh and cultured, were processed for histological and proliferating cell nuclear antigen (PCNA) assays, while tissue activity was evaluated by estrogen (E2) production during culture duration. As results showed, FSH of dynamic addition FSH started with up-grade dose (1-100 ng·mL^-1) followed by down-grade dose (100-1 ng·mL^-1), stimulated primordial follicle activation, survival, growth or granulosa cells proliferating, and also promoted the E2 production in tissues in vitro cultured, compared to other groups. The concentration of FSH with 50 ng·mL^-1 stimulated primordial follicle activation after 1 d culture, and in dose higher than 10 ng·mL^-1 of FSH groups stimulated follicle survival and growth. During culture duration, E2 production in all culture groups were increased constantly, which indicated that tissue activity were well maintained in the present study. In conclusion, this study suggestd that FSH involved in early follicle development and dynamic addition of FSH would benefit to primordial follicle developing and tissue activity maintenance.

Genomic Structure, Polymorphism in Promoter Region and Their Associations with Production Traits of Dkk1 Gene in Goats

CAO Gui-ling;ZHANG Yu-jie;LI Biao;TANG Pei-rong;JIANG Yun-liang

2010, 41(11): 1394-1400. doi:

Abstract ( 650 )

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The objective of this study was to clone goat Dkk1 gene, identify its polymorphisms and find DNA markers associated with cashmere production To clone goat Dkk1 gene, PCR with primers designed within conserved encoding region was performed. Polymorphisms were screened by PCR amplification and sequencing with two genomic DNA pools of goats differing in cashmere production, and detected by PCR-SSCP approach. A fragment of 3 491 bp in size for goat Dkk1 gene was amplified and sequenced, including four exons and three introns and encoding 265 amino acid residues. It has high homology with the corresponding sequences of other mammalian species both at mRNA and amino acid levels. Altogether 33 single nucleotide substitutions and an insertion of 4 bp in intron 2 were revealed. Two polymorphic sites in (T593C and C603A) the promoter region were detected in five goat populations. Allele A was predominant in all of the five goat populations, and all populations were in Hardy-Weinberg equlibrium (P>0.05) at the two sites. The cashmere production, cashmere production per body surface area, birth weight, gaining weight from birth to weanling among different genotypes had no significant difference in White Cashmere goats (P>0.1), although allele A had a trend to increase cashmere production Goat Dkk1 is homologous to other mammal species. Most of the polymorphisms were in noncoding region of goat Dkk1 gene. No significant association was found between the T593C and C603A polymorphisms in Dkk1 promoter region and cashmere production

Immunological Response Analysis of Chinese Merino Sheep with DifferentMHC-DRB1/DQB1 Haplotypes Infected with Echinococcus granulosus

DU Ying-chun;JIA Bin;SHEN Hong;JIANG Song;HUI Wen-qiao;TIAN Yong-zhi;WEN Qing-na;ZHOU Qi-wei;LV Li-min;LI Ren-yan

2010, 41(11): 1401-1406. doi:

Abstract ( 1067 )

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The aim of this study was to analyze the immunological response to Echinococcus granulosus in Chinese Merino (Xinjiang Army Wasteland Reclamation) sheep with different MHC-DRB1/DQB1 haplotypes. 16 two-year-old female healthy Chinese Merino sheep with different haplotypes were artifically infected with Echinococcus granulosus under the same raising condition, which were divided into two groups: the test group (8 resistant haplotyoe sheep ) and the control group (8 nonresistant haplotype individuals ). The percentage of CD4⁺ T, CD8⁺ T and B lymphocyte subpopulations in peripheral blood were analyzed by flow cytometry on 7 day before infection（－7），and on 0(the day of infection)，7，21，30 and 60 day after infection, respectively. During the same period, the content of IgG, IgM, IgE, IFN-γ in the serum were quantitatively assayed by ELISA and the quantity of granulocyte were counted on Wright’s stained blood smears The results showed that: 1) the incidence of Cystic Echinococcosis in the test group was significantly lower than that in the control group (P=0.019).2) The percentage of CD4⁺T cells had no significant difference between the two groups (P＞0.05). The percentage of CD8⁺ T cells in the test group was significantly higher than that in the control group (P＜0.05) at day 7. 3) Serum levels of IgG in the two groups were significantly higher at day 7 when compared to day (－7) (P＜0.05).Compared with the test group, significantly higher IgM and IFN-γ occurred in the control group at day 30 (P＜0.05). 4) Leukocytes and lymphocytes in the test group were significantly more than that in the control group (P＜0.05) at day 7 and 21.Neutrophils and albumin in the test group were obviously lower than that in the control group (P＜0.01) at day 21.The significantly higher resistance to Echinococcus granulosus occurred in the resistant haplotype sheep, when compared with the nonresistant haplotype individuals. In addition, the levels of CD8⁺ T cells, IgM, IFN-γ, leukocytes and lymphocytes in the resistant haplotype sheep were significantly higher than those in nonresistant haplotype individuals during the different periods after infection.

Cloning of Goose ACSL1 Gene, Tissues Expression and the Effect of Overfeeding on Its mRNA Level

PAN Zhi-xiong;LV Jia;LU Li-zhi;WANG Ji-wen

2010, 41(11): 1407-1413. doi:

Abstract ( 1084 )

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This experiment was conducted to study the effect of overfeeding on ACSL1 mRNA level in goose. In this study, the Landes geese and Sichuan White geese were used to clone the gene sequence of ACSL1, and the effect of overfeeding on the transcriptional level of ACSL1 in liver was researched The result indicated that the obtained CDS sequence of ACSL1 gene was 2 100 bp, the sequence analysis revealed that its open reading frame (ORF) encoding 699 amino acids, which contain a ATP/AMP motif and a FACS motif, and has high homology with the corresponding sequences of chicken (93%), cattle (80.4%), human (78.5%) and mouse (77.3%), respectively. The expression of ACSL1 was measured in several tissues, and the effects of overfeeding on the expression of ACSL1 were studied. The results of real time RT-PCR demonstrated that, compared to the other tissues, goose ACSL1 mRNA was more abundant in abdominal adipose tissues, subcutaneous adipose tissues and liver. Overfeeding markedly increased the mRNA expression of ACSL1 gene in the liver of two breeds, and gene expression was markedly higher in Landes geese than that in Sichuan White geese The mRNA abundance of ACSL1 was positively correlated to the relative weight of liver, TG levels and total of fatty acids after overfeeding, and the correlation in Landes geese was stronger than that in Sichuan White geese It was concluded that overfeeding induced the significant increase of ACSL1 mRNA level in goose liver, and the effect of overfeeding was different between the two breeds.

Effect of Different Concentrations of Vitamin D on Growth Performance, Blood Biochemistry and Tibia Parameter for White Pekin Duckings

WANG Shuang;HOU Shui-sheng;XIE Ming;HUANG Wei;YU Jun-ying

2010, 41(11): 1414-1420. doi:

Abstract ( 1137 )

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This experiment was conducted to study the effect of vitamin D on metabolism of calcium and phosphorus in Starter White Pekin Duckings. Seven hundred and twenty one-day-old male White Pekin Duckings with similar body weight were randomly allotted into 12 treatments with 6 repicates of 10 ducks each. Duckings were fed two levels of dietary calcium/phosphorus (0.8/0.4, 0.4/0.2) and six levels of vitamin D (0，250，500，1 000，2 000，3 000 IU·kg^-1) in a 2×6 factorial arrangement for 14 days. The results showed that: vitamin D in diets significantly effected the daily weight gain，daily feed intake and feed/gain (P<0.05). When the vitamin D was 2 000 IU·kg^-1, the plasma calcium reached the most level. With the increasing of vitamin D, plasma phosphorus and Alkline Phosphatase(AKP) activity were decreased signficantly (P<0.05). Tibia ash content increased when vitamin D increased from 500 to 1 000 IU·kg^-1 (P<0.05). More vitamin D was not effect any more (P>0.05). At the 2 000 IU·kg^-1 level of vitamin D, the calcium content in tibia ash reached its most level. Vitamin D in diets did not effect the phosphorus content in tibia ash. The results indicated that the vitamin D and calcium/phosphorus in diets significantly effected the growth performance, blood biochemistry and tibia parameter in White Pekin Duckings. The requirement of vitamin D was between 1 000 and 2 000 IU·kg^-1 for White Pekin Duckings of 14 days old.

A Study on Digestibility in Small Intestine of the Ruminal Undegraded Protein Fractions According to CNCPS Model of 14 Feedstuffs

ZHANG Ying-xue;LIN Xue-yan;SU Peng-cheng;WANG Yun;WANG Zhong-hua

2010, 41(11): 1421-1427. doi:

Abstract ( 1321 )

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The amino acid sub-model of Cornell net carbohydrate and protein system is one of the three published major model to predict ruminant duodenal amino acid flux at present. Previous studies indicated that the ruminal undegraded C protein fractions defined by CNCPS model of non-forage feedstuffs may be digestible in small intestine, which questioned the reliability of setting the digestibility of C protein fraction in small intestine to zero by the CNCPS model. Contents, dynamic degrading parameters in the rumen of each protein fractions, and CP digestibility of ruminal degrading residue at different sampling time point of 14 feedstuffs were measured in the present study. These data were used to estimate small intestine digestibility of each ruminal undegraded protein fraction by least square data fitting according a deduced equation. Reliable estimates were acquired when taking small intestine digestibility of ruminal undegraded C protein fraction was 100% as the restrictive condition. Results of the present study indicated that it was reasonable to set the small intestine digestibility of ruminal undegraded B1, B2, and B3 protein fractions as 100%, 100%, and 80% by the CNCPS model. Small intestine digestibility of ruminal undegraded C protein fraction of 8 feedstuffs, 4 of which were roughage, were high enough and can not be neglected, therefore it was reasonable to further investigate the reliability of setting the digestibility of C protein fraction to aero by the CNCPS model.

Function Analysis of a Conserved Stem-Loop Structure in the 5′ Untranslated Region of the Porcine Reproductive and Respiratory Syndrome Virus Genome

LU Jia-qi;GAO Fei;LIU Ping;YUAN Shi-shan

2010, 41(11): 1428-1434. doi:

Abstract ( 762 )

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The objective of this study was to analyze the secondary structure and function of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) 5′ UTR. Based on the PRRSV type 2 full-length infectious cDNA clone pAPRRS, a series of mutants which can modify a conserved stem structure were obtained through PCR-based mutagenesis. The mutants are transfected into MARC-145 cells for cytopathic effect (CPE) observation. Meanwhile, the indirect immunofluorescence analysis (IFA) and Northern blot analysis show the genomic replication and subgenomic transcription for the rescued viruses. In addition, the growth features are also detected by plaque morphology compared with the parental virus. It was found that the secondary structure of the conserved stem loop in PRRSV 5′ UTR wasn’t essential for virus rescue. Novertheless, the primary sequence of the stem structure is functional for viral replication. In addition, it is demonstrated that a key nucleotide in PRRSV 5′ UTR directly regulate the viral replication. All results demonstrate that the essential sequences in PRRSV 5′ UTR are functional in viral genomic eplication.

Preparation and Application of Swine Hepatitis E Virus Diagnostic ELISA Kit

2010, 41(11): 1435-1441. doi:

Abstract ( 986 )

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The objective of this study was to establish effective diagnostic technique and diagnostic kit for swine hepatitis E virus (HEV). An indirect ELISA assay was developed for detection of swine HEV using synthetic peptide swHEV11 screening from one sequence of the open reading frame (ORF) 3 of swine HEV. This assay was optimized for an antigen coating concentration of 3 μg·mL^-1 and a serum dilution of 1:10, with a standard incubation time of 30 min. The optimal dilution and reaction time of enzyme labeled antibody was 1:12 000 and 60 min, respectively. When the kit was applied to detect HEV antibodies in sera of pigs, the results were quite consistent with the HEV diagnostic kit from Wantai. The conformity rate reached 73.4% (205/279), the sensitivity rate and specificity rate reached 73.3% (200/273) and 83.3% (5/6), respectively. The coefficient of variation (CV) was less than 10% in the same batch, and less than 20% in different batches. The results indicated that the application of indirect ELISA and the HEV ELISA kit is specific, sensitive and stable, and may provide a simple and rapid method for monitoring serum antibodies against HEV infection and seroepidemiological surveys of HEV infection

Establishment and Clinical Application of RT-PCR Assay for Detection of Rabbit Hemorrhagic Disease Virus

HU Bo;WEI Hou-jun;WANG Fang;FAN Zhi-yu;XU Peng;XU Wei-zhong;XUE Jia-bin; HE Kong-wang

2010, 41(11): 1442-1446. doi:

Abstract ( 646 )

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The RT-PCR assay for detection of Rabbit hemorrhagic disease virus (RHDV) in nasal swab specimens collected from rabbits was developed for the research of virus carrying in healthy immunized rabbit. A pair of primers were designed according to the conserved sequences of vp60 from RHDV published in GenBank, and a 591 bp segment was amplified by RT-PCR. The least amount of RHDV could be detected was 2.40 ng·μL^-1, and the sensitivity of the RT-PCR was 8×103 times higher than that of HA method. 168 nasal swabs collected from 5 provinces was detected by the assay, the results showed that the positive samples was 22 and the positive rate was 13.09%. These results indicated that the phenomenon of virus carrying in healthy immunized rabbit and the developed RT-PCR might be a promising approach in clinical screening, detection and carrying research of RHDV in a large number of immunized rabbit nasal swabs.

Development of Real-time Fluorescent Quantitative RT-PCR Assay for Detection of Genotype ⅤⅡ Newcastle Disease Virus （NDV）

SUN Jun-feng;LIU Hua-lei;WANG Zhi-liang;ZHAO Yun-ling;ZHENG Dong-xia;ZHANG wei;LIU Wen-hua;FENG Li-li;CHANG Wei-wei

2010, 41(11): 1447-1452. doi:

Abstract ( 680 )

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In order to establish a differential diagnosis Real-time RT-PCR with highly sensitivity, specificity and repeatability, a pair of primers and TaqMan probe were designed according to characteristics of cleavage site sequences of F gene of genotype ⅤⅡ Newcastle disease virus strains in this study. The standard curve was established by using cRNA standard prepared in vitro transcription as positive template, the related indexes were tested. The results showed that this assay has a good effect of differential detection of genotype ⅤⅡ NDV; 102 copies·μL^-1 of template RNA could be detected at least. The sensitive of this assay was 100 times higher than conventional RT-PCR. The real-time fluorescent quantitative RT-PCR for detection of genotype ⅤⅡ Newcastle disease virus was established. This assay could be used as differential diagnosis method of genotype ⅤⅡ NDV strains, which couldn’t be identified by conventional methods. Our results indicated that the method established in this study can be used as rapid identification assay for the detection of genotype ⅤⅡ NDVs.

Relative Quantification of mRNA Transcription of Toll-like Receptors inDifferent Tissues of Chicken by Real-time Quantitative PCR

ZHOU Zuo-yong;NIE Kui;SONG Zhen-hui;HU Shi-jun;ZHOU Rong-qiong;GUO Zhi-li;LIU Li-qiong;QIN Bo

2010, 41(11): 1453-1459. doi:

Abstract ( 700 )

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According to the sequences of chicken Toll-like receptors （ChTLR）, the realtime PCR primers were designed and synthesized to establish a method for the detection of mRNA transcription of ChTLRs by real-time quantitative PCR- And the transcriptions of ChTLRs in different tissues of Luo-man chicken were analyzed. The result showed that the ChTLRs mRNA were transcripted in spleen, bursa, thymus and intestinal tract. The ChTLR1 is transcripted at high level in bursa, kidney and cecum, in contrast, ChTLR2 mRNA is restricted in the partial tissues, and at high level in spleen, bursa and liver, and not detected in kidney, lung and skin. ChTLR4 is transcripted at approximately the same level in all tissues tested. The ChTLR5 is transcripted at high level in kidney, spleen and cecum, and ChTLR15 is transcripted at high level in bursa, spleen and cecum. These results indicated that the real-time quantitative PCR was established successfully for the detection of ChTLRs mRNA transcription, and there were comparatively large differences on the transcription level of ChTLRs mRNA in different tissues, which may be associated with the ability of chicken to recognize pathogens.

Distribution of Orexin B in Porcine Hypothalamo-Pituitary-Gonadal Axis and the Regulation of Reproduction in Vitro

SU Juan;YAO Yuan;YANG Gui-hong;LI Xun;LIU Yan-peng;KOU Rui;LIU Jing;LEI Zhi-hai

2010, 41(11): 1460-1466. doi:

Abstract ( 689 )

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The experiment was conducted to investigate the distribution of Orexin B in the hypothalamo-pituitary-gonadal （HPG） axis of 1-month-old Suzhong swine and the sexual maturity sow and its direct biological actions in vitro by immunohistochemical technique and radioimmunoassay. The result showed that Orexin B- immunoreactive neurons localized within the hypothalamus, mainly in the perifornical nucleus (PeF), the lateral hypothalamic area (LHA), dorsomedial hypothalamic nucleus (DMH), and posterior hypothalamic area (PH) and with a sparser distribution in the preoptic, anterior hypothalamic area(AH) and suprachiasmatic nucleus. In pituitary, Orexin B localized in endocrine cell of adenohypophysis pars distalis and intermedia. In gonad, Orexin B distributed in granular cell of primary and secondary follicle and cytoplasm of oocyte, leydig cell of the testis and epididymis. The hypothalamic plants, anterior pituitary cells and granular cell were challenged with 1, 10,100 and 1 000 nmol·L^-1 Orexin B and GnRH, LH and E2 were measured after treatment. Compared to control, 1 000 nmol·L^-1 dose Orexin B significantly increased（P<0.01）GnRH secretion, 100 and 1 000 nmol·L^-1 doses of Orexin B significantly stimulated （P<0.05）LH and E2 secretion These results demonstrated that Orexin B regulated reproduction of porcine in every level of HPG axis.

Effect of Danshen Root on Morphologic Change of Spleen and Kidney in Diabetic Rats Induced by STZ

LI Jing;LIU De-yi

2010, 41(11): 1467-1471. doi:

Abstract ( 897 )

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To study the effect of Danshen Root on the glucose, the structure of spleen and kidney in diabetic rats induced by STZ. And the diabetes model was established by Streptozotocin injected 60 mg·kg^-1. The SD rats were randomly divided into two groups as diabetes group and Danshen treated group, 15 in each group.There were 20 rats in Normal control group. Danshen treated group were orally administered by Danshen Root boiled liquid (5 g·kg^-1·d^-1)， and the diabetes group， normal control group were administered by the similar dosage of distilled water The period of experiment was 28 days. At the 7th, 14th, 21th and 28th day, glucose was measured in serum that is prepared by taking blood through orbital venous plexus and centrifuging. At the end of the experiment, histological section of all rats′ spleen and kidney were made and observed after Bouin liquid fixing and HE staining. Glucose of Danshen treated group were significantly lower than that of the model control group (P<0.01) and higher than that of the normal control group (P<0.01). For the model control group, the following results of histological observation showed that there were less splenic cords, invisible germinal centers and thinner periarterial lymphatic sheath, larger renal glomerulus, smaller renal capsule, protein exudates and exfoliative epithelial cells in renal tubule. For the Danshen treated group, the observation results showed that splenic cords with clear demarcation, visible germinal centers and thicker periarterial lymphatic sheath, closely aligned lymphocytes, normally aligned renal glomerulus, renal capsule and renal tubule, normal epithelial cells but a small quantity of homogeneously redstained protein exudates. The results showed that Danshen Root could improve the abnormal glycometabolism of diabetic rats and help prevent the histological change of the spleen and kidney for diabetes model rats.

Expression of Leptin Receptor in Neuroendocrine and Reproductive Organs of Small Tail Han Sheep

XIAO Huan;CUI Ya-li;CHENG Yu-bao

2010, 41(11): 1472-1477. doi:

Abstract ( 615 )

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To study the localization and expression of the leptin receptor(OB-R) in various tissues of Small Tail Han Sheep. In the study, we analyzed the expression of leptin receptor in neuroendocrine and reproductive system of Small Tail Han Sheep with immunohistochemistry and Western Blot. The results demonstrate: positive brown granules were located in some of the cells of hypothalamus, pituitary, ovary and uterus. Three bands with a molecular mass close to 120,110 and 98 ku were identified by Western Blot There were different amount of expression in different organization. There were a plenty of expression in many tissues, which showed that OB-R plays a key role in the reproductive function of Small Tail Han Sheep.

Expression and Immunolocalization of Endothelin Receptor B in Alpaca Skin of Different Colors

GENG Jian-jun;MU Xiao-li;SUN Le-tian;JIANG Jun-bing;ZHANG Jie;LI Hong-quan;ZHANG Ying;DONG Chang-sheng

2010, 41(11): 1478-1484. doi:

Abstract ( 699 )

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The aim of this experiment is to study the expression and immunolocalization of Endothelin receptor B in alpaca skin, to compare the expression quantity in different colors, and to explore the function of EDNRB in the process of coat color formation. The whole testes were obtained from adult alpaca of white and brown hair colors. The mRNA and protein expression level of EDNRB in alpaca skin of different colors were examined by immunohistochemistry, Real-time quantitative PCR and Western blot. The results showed that the EDNRB gene was expressed in alpaca skin reliably. The expressive quantity of EDNRB gene in the brown alpaca skin tissue was 2.165 1 times than that in white one. The distribution of EDNRB in alpaca skin were demonstrated, the expression was notable difference between the white and brown alpaca based on the average optical density. Our findings showed that EDNRB was closely related to the activity of alpaca melanocytes, and was involved in the formation of alpaca hair and skin color.

Effect of Dietary Se-enriched Yeast on Blood Biochemical Indicator and Endocrine Dysfunction of Chai Hens in Summer

GUO Yun-xia;HAO Qing-hong;HUANG Ren-lu

2010, 41(11): 1485-1489. doi:

Abstract ( 1006 )

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The experiment was conducted to study the effects of dietary Se-enriched yeast on blood biochemical parameters and endocrine capacity of Chai Hens in summer. 360 thirty week-old Chai laying-hens were randomly divided into five groups, with three replication per group. The control group fed with basal diets containing 0.08 mg·kg^-1 Se, the testing group fed with basal diets adding 0.15, 0.3, 05, 1 mg·kg^-1 Se, respectively. Preliminary trial period was 7 days and experimental time was 50 days. The results showed as follows: With the increasing of diet selenium content, Feeding diet with 0.3 mg·kg^-1 selenium had increased the ratio of albumen and globulin (P>0.05) ; but the blood sugar had decreased(P>0.05); The activity of GPT and CK had decreased (P<0.05), and the activity of LDH and ALP had increased (P<0.05).Feeding diets of Se-yeast had decreased the activity of T3,T4 and CORT(P<0.05). The results suggested that adding 0.3-0.5 mg·kg^-1 of Se could lessen the animal body heat stress response, increase the body′s immune system. At the same time, its could improve the metabolism of material and then increase the production performance of Chai Hens.

Study on Changes of Blood Biochemical Metabolites for Subclinical Ketosis in Dairy Cows

JIA Zong-fei;SHI Jian-peng;HAN Yi-chao;PANG Quan-hai

2010, 41(11): 1490-1496. doi:

Abstract ( 1072 )

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In this study, 24 cows with similar age, birth and body condition were chosen and grouped by serum β-hydroxybutyric acid (BHBA) content. The cows with greater BHBA content (>1.2 mmol·L^-1) were assigned as the experiment group, and cows with less BHBA content (≤1.2 mmol·L^-1) were assigned as the control group. The concentration of biochemical indicators in the serum which taken at the 14th, 7th, 1st day of prenatal and the 1st, 7th, 14th, 21th, 28th day of postpartum, were tested and the changing trends of them during the experimental stage were analyzed. The results shown that the concentration of serum BHBA in the experiment group was very significantly higher than that in the control group (P<0.01); nonesterified fatty acid (NEFA) was significantly higher than that in the control group (P<0.05); glucose (GLU) was very significantly lower than that in the control group (P<0.01); blood urea nitrogen (BUN) was very significantly higher than that in the control group (P<0.01); Other indices were difference in different stages. In the two groups, the concentration of serum BHBA was in a high level at the 7th to the 21st day of postpartum, NEFA was highest at the 21st day of postpartum, the activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were highest at the 7th day of postpartum. This test proved that subclinical ketosis can occur at the 7th day of postpartum, and occur easily at the 21st day of postpartum.

Genotype Frequencies of the Fishy Taint Susceptible Gene FMO3 in 11 Chinese Local Chicken Breeds

WANG Xiao-liang;XU Gui-yun;ZHENG Jiang-xia;YAO Jun-feng;QU Lu-jiang;YANG Ning

2010, 41(11): 1497-1501. doi:

Abstract ( 672 )

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The objective of the study was to detect the distribution of T329S mutation in 11 Chinese local chicken breeds. The FMO3 genotype frequency in 11 Chinese local chicken breeds including Beijing You, Hebei Chai, Gushi, Liyang, Rugao, Taihu, Huainan Mahuang, Silky, Dongxiang Blue-shelled, Wenchang and Tibetan chicken, were detected by PRC-RFLP method. The results showed that Dongxiang Blue-shelled chicken only had TT genotype, suggesting that no individual would lay fishy taint eggs in this breed. Fishy taint susceptible individuals with genotype SS were detected in all other 10 local breeds. Of these breeds, Beijing You chicken had the highest frequency of the SS genotype (8.8%) and Tibetan chicken present the lowest (0.7%). In conclusion, the frequencies of fishy taint genotype in Chinese local chicken breeds were generally low and the susceptible individuals could be easily detected by PCR-RFLP method

Preparation, Affinity Purification and Identification of Polyclonal Antibody Against Pig Annexin A2

CHEN Jiang-tao;WANG Guang-xiang;JI Yan-hong;SHANG You-jun;LIU Yan-hong;LIU Xiang-tao

2010, 41(11): 1502-1509. doi:

Abstract ( 645 )

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The study was aimed at preparing and purifying polyclonal antibody of rabbit anti-pig ANNXA2 by GST-ANNXA2-coupled affinity chromatography. The fragment of pig Annexin A2 and EGFP fusion gene was amplified from pA2EGFP plasmid by PCR. And then the fragment was subcloned into the prokaryotic expression vector pET-30a(+) to construct the recombinant plasmid p30a/AE, and expressed in inclusion body of E. coli. Rosetta (DE3) by IPTG induction. The fusion protein ANNXA2-EGFP which was purified by NiResin HP affinity chromatography was used as immunogen to inject male rabbit to prepare polyclonal antibody anti-pig Annexin A2. The soluble fusion protein GST-ANNXA2 which has been prepared was coupled on NHS-activated Sepharose Fast Flow column, and then the polyclonal antibody was purified by this column- Purified antiserum titer amounted to 1:12 800 by indirect ELISA, and the purified antiserum was also demonstrated high specificity by Western blot and immunohistochemistry staining. The polyclonal antibody of rabbit anti-pig ANNXA2 was prepared and possessed high titer and high specificity, which laid the groundwork for the investigation of relationship between virus infection and pig Annexin A2.

The Effect of DES on the Testis Cell of Xiang Pig

TIAN Xing-gui;LI Jiang-sen;ZHU Xing;FENG Hui-li;LIU Ruo-yu

2010, 41(11): 1510-1514. doi:

Abstract ( 660 )

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The objective of this study was to explore the effects of diethylstilbestrol daily exposure on testis Sertoli cells and germ cells of immaturity Guizhou Xiang-Pig. Twenty male Xiang pig, 20-25 days old, were divided into four groups (one control group and three test groups) of 5 each. Test group 1, 2 and 3 was received different doses, 0.03, 0.3 and 3 mg·(kg^-1·d^-1), of diethylstilbestrol for 9 days by intraperitoneal respectively. 24 h after the last injection, the left testis was removed and fixed in 40 g·L^-1 paraformaldehyde for 24 h, and then was processed to paraffin wax Fivemicronthick sections were cut. We used an immunohistochemical method with antibody against GATA-4, a special protein of Sertoli cell, to determine the Sertoli cells, the number of Sertoli and germ cells were counted under microscope. The result showed that DES significantly (P<0.05) increased the germ cell number, but significantly (P<0.05) decreased Sertoli cell number at 0.3, 3 （mg·kg^-1·d^-1） dose，no significance was found at 0.03 mg·（kg^-1·d^-1） dose of DES （P>0.05）, compared with that in control group. DES could promote the germ cell proliferation of immature xiang pig testes, while decrease the Sertoli cell proliferation. It was concluded that the DES had different effects on different cell types.

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