Effect of dipeptide on intestinal peptide transporter 1 gene expression: An evaluation using primary cultured chicken intestinal epithelial cells

Peptide transporter 1 (PepT1) is a transporter responsible for absorbing dipeptide and tripeptide in enterocytes and is upregulated by dipeptide in mammals. It has not been certain whether intestinal PepT1 expression is responsive to dipeptides in chickens because of the lack of in vitro study using the cultured enterocytes. This study established a primary culture model of chicken intestinal epithelial cells (IECs) in two-dimensional monolayer culture using collagen gel by which the response of chicken PepT1 gene expression to dipeptide stimuli was evaluated. The cultured chicken IECs showed the epithelial-like morphology attached in a patch-manner and exhibited positive expression of cytokeratin and epithelial cadherin, specific marker proteins of epithelial cells. Moreover, the chicken IECs exhibited the gene expression of intestinal cell type-specific marker, villin1, mucin 2, and chromogranin A, suggesting that the cultured IECs were composed of enterocytes as well as goblet and enteroendocrine cells. PepT1 gene expression was significantly upregulated by synthetic dipeptide, glycyl-l-glutamine, in the cultured IECs. From the results, we herein suggested that dipeptide is a factor upregulating PepT1 gene expression in chicken IECs.     

中国地方绵羊群体TYRP1基因遗传变异研究

本研究旨在了解酪氨酸酶相关蛋白1（tyrosinase related protein 1,TYRP1）基因在中国地方绵羊群体内的遗传变异,以及TYRP1基因突变与不同毛色表型绵羊群体的相关性。通过直接测序法和PCR-RFLP技术对10个中国地方绵羊群体进行单核苷酸多态性（SNP）检测,利用Beagle、PLINK和POPGENE等软件对突变位点数据进行单倍型构建、连锁不平衡分析和遗传变异研究。突变位点检测结果表明,在绵羊TYRP1基因内识别了13个SNPs,其中位于TYRP1基因外显子上的10个SNPs位点,除个别位点在大尾寒羊、中国美利奴羊和岷县黑裘皮羊中没有发生突变外,其他突变位点在所有绵羊品种中均出现不同程度变异,说明中国地方绵羊群体具有较高的遗传多样性。单倍型分析结果表明,所有样本中共有42个单倍型,优势单倍型0000000000（245/918）、0100000001（91/918）在所有绵羊群体中均存在,除单倍型0101100000（93/918）在中国美利奴羊中没有出现,单倍型0001000001（69/918）在岷县黑裘皮羊、哈萨克羊群体中没有出现外,在其他群体中均存在。连锁分析结果表明,10个SNPs在所有样本中均存在2个连锁模块。群体遗传变异分析表明,中国地方绵羊群体具有较高水平的群体内遗传变异,各绵羊品种间存在明显的遗传分化模式,且各品种遗传关系与其品种传统分类结果基本一致。本研究为进一步研究TYRP1基因对绵羊毛色遗传性状的影响提供了参考依据。     

干扰素诱导跨膜蛋白1对猪源冠状病毒吸附宿主细胞的影响

为探索干扰素诱导跨膜蛋白1（interferon-inducible transmembrane protein 1,IFITM1）对猪源冠状病毒复制的影响,本研究选用猪传染性胃肠炎病毒（Transmissible gastroenteritis virus,TGEV）高致病毒株及其宿主细胞PK15作为研究对象。正常PK15细胞接种TGEV后,实时荧光定量PCR检测感染后不同时间点PK15细胞中一些干扰素刺激基因（interferon-stimulated genes,ISGs）的mRNA表达水平;利用慢病毒表达系统构建稳定表达和稳定干扰IFITM1表达的PK15细胞系。将一段靶向干扰猪源IFITM1的shRNA序列及猪源IFITM1全长,分别插入pLKO.1-EGFP-Puro载体及pLVML-Myc-MCS-IRES-Puro载体中,分别构建出pLKO.1-IFITM1shRNA-EGFP-Puro及pLVML-Myc-IFITM1-IRES-Puro重组质粒。将重组质粒与慢病毒包装质粒共转染293FT细胞后获得带有目的基因的重组慢病毒,慢病毒侵染PK15细胞后用嘌呤霉素进行筛选,获得稳定表达及稳定干扰IFITM1表达的PK15细胞系,分别命名为PK15-IFITM1及PK15-IFITM1^-/-,并分别用实时荧光定量PCR、间接免疫荧光试验（IFA）及Western blotting检测IFITM1干扰效率及表达情况;TGEV接种PK15-IFITM1^-/-和PK15-IFITM1,实时荧光定量PCR测定细胞中TGEV的拷贝数。结果显示,PK15细胞接种TGEV后的48 h内,一些ISGs的mRNA水平均有所上升;PK15-IFITM1^-/-细胞系的干扰效率为70%,PK15-IFITM1细胞系表达成功;在PK15-IFITM1^-/-细胞系中,IFITM1的mRNA水平显著下调,促进了TGEV的复制。反之,在PK15-IFITM1细胞系中,TGEV的复制受到了抑制。但IFITM1的表达或缺失却不影响TGEV对PK15的吸附作用。总之,IFITM1对TGEV有显著的抗病毒作用,IFITM1不影响TGEV对PK15细胞的早期吸附,这为后续IFITM1抗冠状病毒机制的研究奠定了基础。     

奶牛α1-抗胰蛋白酶及基因多态性研究进展

α1-抗胰蛋白酶（α1-antitrypsin，α1AT）是一种多功能糖蛋白。奶牛α1AT编码基因多态性很高，并对奶牛经济性状具有重要意义。作者主要综述了α1-抗胰蛋白酶和其编码基因在奶牛上的研究进展。     

“三三法”快速检测沙门氏菌的研究

将DHL平板上产H_2S和不产H_2S的菌落分别用TSI、LMA、甘露醇或TSI、LMA、ONPG三管判定,辅以肠杆菌科诊断噬菌体0—1、C多价、E多价三滴法快速检测沙门氏菌的方法简称“三三法”。近两年来我们用此法检测了125批、341份样品中236株可疑菌株,检出沙门氏菌27株、11个血清型,经系统生化验证符合率100％。表明该法具有快速、简便、准确、实用的优点。     

中国禽(番鸭)呼肠孤病毒分离株S1基因全序列分析

采用RT-PCR技术,分别以DRV-YH、DRV-YJL两株中国禽(番鸭)呼肠孤病毒RNA为模板,扩增了S1全基因的cDNA片段.将S1 cDNA克隆到T载体后进行序列测定,测序结果表明所扩增的cDNA片段长1643个核苷酸,包含了完整的S1基因的三个开放阅读框架(ORF1,ORF2和ORF3)和基因两端的非编码区.核苷酸序列比较分析结果表明:DRV-YH与ARV-S1133,176分别有6个,10个核苷酸的差异,DRV-YJL与ARV-S1133,176分别有8个,12个核苷酸的差异,DRV-YH与DRV-YJL有4个核苷酸的差异.     

Immunopathology of <Emphasis Type="Italic">Dirofilaria immitis</Emphasis> Infection

Heartworm disease caused by Dirofilaria immitis affects canine and feline hosts, with infections occasionally being reported in humans. Studies have shown that both dirofilarial antigens and those derived from its bacterial endosymbiont Wolbachia, interact with the host organism during canine, feline and human infections and participate in the development of the pathology and in the regulation of the host’s immune response. Both innate and acquired immune responses are observed and the development of the acquired response may depend on the host and, or on its parasitological status. This review aims at illustrating current research on the role of both D. immitis and Wolbachia, in the immunology and immunopathology of dirofilariosis.     

Antigen-nonspecific macrophage factors modulating the antibody response in vitro

Antibody response to an antigen involves the co-operation between three types of cells: macrophages, T cells and B cells. The cognate interactions between these cells play a fundamental role in the expression of a specific antibody response, but the last is modulated by antigen-nonspecific soluble factors produced either by macrophages or by T cells. Macrophages elaborate a spectrum of molecules modulating the function of lymphoid cells; among them are IL1 and prostaglandins of the E series, which are respectively enhancer and inhibitor of the antibody response in vitro. These molecules alter T cell and B cell activities through different mechanisms involving activation or inhibition of IL2 production, or alteration of cells surface antigens. However, the cellular events following the fixation of soluble factor on its receptors are not known.     

BRCA1基因多态性及其与奶牛乳房炎抗性的相关性研究

为了研究BRCA1基因突变与荷斯坦奶牛体细胞数和体细胞评分的关系,试验通过对BRCA1基因外显子13、14进行克隆、序列比对和挖掘已有突变的方法确定该基因的多态位点,采用SNaPshot技术检测了BRCA1基因25025 T>A和46126 G>T突变位点在北京郊区荷斯坦奶牛群体中的分布,并对突变位点与体细胞数和体细胞评分进行了关联分析。结果表明,荷斯坦奶牛BRCA1基因2个位点均检测到3种基因型,其中25025 bp位点TT基因型为优势基因型,46126 bp位点GT基因型为优势基因型。25025 bp位点AA基因型个体体细胞数(P<0.05)和体细胞评分(P<0.01)都显著低于TT和TA基因型;46126 bp位点TT基因型个体体细胞数显著低于GG和GT基因型个体(P<0.05),但3种基因型个体体细胞评分无显著差异(P>0.05)。本研究结果初步表明,BRCA1基因25025和46126 bp位点可作为中国荷斯坦牛乳房炎抗性的标记辅助选择。     

鲁梅克斯对草甸盐土的改土效应研究

在草甸盐土上种植鲁梅克斯Rumex patientia×R.tianschanicus cv.Rumex K-1,脱盐改土效果十分明显.种植鲁梅克斯3～4年,鲜草产量达100.51 t/hm2,pH值由8.43降为7.99,耕作层土壤总孔隙度增加5.29%～7.06%,土壤容重降低0.14～0.21 g/cm3,水稳性团粒结构增加23.14%～32.34%,自然含水量增加72.12～89.44 g/kg,土壤有机质、速效氮、磷、钾亦随之增加.