Characterization of bovine ileal epithelial cell line for lectin binding,susceptibility to enteric pathogens,and TLR mediated immune responses

In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.     

鸡源株与人源株H9N2流感病毒血凝素受体结合位点氨基酸比较与分析

H19N2病毒可以感染多种禽类和哺乳动物，包括人类。流感病毒血凝素受体结合位点的氨基酸可以影响受体结合特性。为了解鸡源株与人源株H19N2病毒受体结合部氨基酸的差异，作者对二者进行了比较与分析，结果表明，鸡源株与人源株血凝素受体结合部位氨基酸的第137、183、190、226位点存在差异，鸡源株在这些位点分别为K、N、AorVorT、LorQ而人源株则分别为R、H、E、L。虽然尚不清楚这些位点的改变对病毒与细胞亲和力及宿主范围的影响，但由于H9N2在我国养鸡业的普遍存在，加强H9N2病毒的分子流行病学监测有重要意义。     

棉花黄萎病菌毒素结合位点初探

以纯化的棉花黄萎病菌毒素(PLPC)免疫新西兰白兔制备了PLPC特异性抗血清。将PLPC(15μg·ml 1)用不同浓度的抗体(1～20μg·ml 1IgG)吸附后处理泗棉3号的切根苗,毒素所引起的症状都有不同程度的减轻。表明所制备的抗体在与毒素发生特异性免疫学反应的同时,可部分封闭毒素分子上与毒素受体结合的位点。利用竞争ELISA测定了泗棉3号幼苗子叶的质膜制剂与PLPC的结合活性。结果表明,质膜制剂与毒素结合后能部分阻断毒素与其抗体的免疫学反应,即质膜制剂中含有毒素的结合位点。分别用胰蛋白酶和煮沸处理质膜制剂后,质膜制剂对毒素与其抗体的反应的抑制作用消失,初步表明质膜制剂中与毒素结合的是蛋白质。     

淡紫拟青霉发酵滤液对大豆胞囊线虫趋化性的影响

 实验室条件下,研究了大豆胞囊线虫幼虫对大豆根和根浸出液的趋化性以及淡紫拟青霉代谢产物对其趋化性的影响。结果表明,大豆胞囊线虫二龄幼虫(J₂)对大豆根存在着一定的趋化性,而淡紫拟青霉发酵滤液对J₂存在明显的驱避性。在根浸出液、真菌滤液及其混合液剂处理下,大豆胞囊线虫J₂在WA平板上靠近处理液的0~1cm区间中分布率存在极显著差异(P <0.0 1),而加样时间(液剂与线虫同时放置或提前2 4h处理)对其分布率没有显著影响。大豆幼根蘸取滤液后对线虫J₂在该区间的分布与不经处理的幼根差异显著(P <0.0 5),证明淡紫拟青霉发酵滤液可以明显地降低线虫与大豆根的亲和力,并强烈抑制线虫对大豆根的侵染。     

不同温度下TMV侵染枯斑三生烟的LncRNA差异表达

【目的】 筛选不同温度下烟草花叶病毒（Tobacco mosaic virus,TMV）侵染后枯斑三生烟（Nicotiana tabacum var. Samsun NN）差异表达的长链非编码RNA（long non-coding RNA,lncRNA）,研究lncRNA在枯斑三生烟抗性反应中的作用。【方法】 N基因的温度敏感性使枯斑三生烟在25℃时具备对TMV的抗性、在31℃抗性丧失,在这两个温度条件下对枯斑三生烟接种TMV和磷酸盐缓冲盐水（phosphate buffered saline,PBS）,48 h后提取系统叶总RNA,构建链特异性文库后进行深度测序。对测序结果进行过滤后利用HTSeq将有效数据与近缘品种TN90（N. tabacum var. TN90）基因组比对,筛选得到lncRNA后利用FPKM法估计lncRNA的表达水平。通过edgeR筛选差异表达lncRNA（differentially expressed lncRNA,DElncRNA）,并利用qRT-PCR技术对这一结果进行验证。通过共定位及共表达分析预测DElncRNA的靶基因,通过参考基因组注释、GO和KEGG富集分析研究靶基因的功能。【结果】 4个处理共12个样本经lncRNA-seq各测得约8 000万条clean reads,共获得4 737条已知lncRNA、40 169条新lncRNA。其中64个lncRNA在不同温度条件下TMV侵染后存在差异表达,qRT-PCR测定结果显示这些lncRNA的测序正确率在80%左右,表明本研究所得测序数据具备较高的可信度。对DElncRNA进行靶基因预测,发现一些基因同时被25℃下调和31℃上调的DElncRNA靶向。靶基因注释功能丰富,主要参与植物抗病、激素和代谢等生理过程。部分可能与激素通路相关的lncRNA,在25℃下TMV侵染时呈现下调趋势,而在31℃下TMV侵染则呈现上调趋势。GO富集分析显示靶基因主要参与构成膜、囊泡等组分,具备钙、钾离子通道抑制剂活性等分子功能,使相应离子得以转运引发随后的反应,同时也参与发病、抗原加工和呈现、细胞分裂素代谢等生理过程。KEGG分析发现靶基因显著富集在植物激素信号转导通路,25℃下调和31℃上调的DElncRNA靶基因同时富集在激素信号传导、ABC运输蛋白、苯丙烷类生物合成等通路。【结论】 不同温度（25℃和31℃）条件下TMV侵染枯斑三生烟后,长链非编码RNA差异表达,DElncRNA通过作用于激素信号传导、物质转运等过程参与寄主系统获得性抗性反应。研究结果可为揭示植物系统获得性抗性中lncRNA的调控功能以及新型抗病毒技术开发提供依据。     

三江源退化高寒草原植物种间亲和性和土壤团聚体特征

为研究三江源地区不同退化程度的高寒草原土壤团聚体和种间亲和性特征,以轻度、中度、重度和极度退化草地为研究对象,对土壤特征和群落特征进行测定。结果表明：随着退化程度的加剧,同一土层相同粒径土壤颗粒的分形维数逐渐减小,>0.5 mm粒径土壤团聚体占比下降、平均重量直径减小,<0.5 mm粒径土壤颗粒的平均重量直径增大,各粒径团聚体分布受土层深度影响较小;中度退化草地物种多样性最高,其种间相关性也最为复杂;随退化程度加剧,正联结种对占比增大,而负联结种对占比下降,种对间的相关性和性质也发生显著变化;退化过程中土壤沙化明显,群落建群种发生变化、杂类草比例显著增加,种对间相关性趋于简单,种间竞争强度逐渐降低。因此,在草地退化过程中维持群落种间亲和性,可有效遏制土壤沙化和草原进一步退化。     

Adrenomedullin (AM) and adrenomedullin binding protein (AM-BP) in the bovine mammary gland and milk: Effects of stage of lactation and experimental intramammary E. coli infection

Adrenomedullin (AM) has been characterized as an endogenous tissue survival factor and modulator of many inflammatory processes. Because of the increased susceptibility of the mammary gland to infection during the time surrounding parturition in the cow, we investigated how milk and tissue content of AM and its binding protein (AM-BP) might be affected by the stage of lactation and the udder health status. Milk and mammary biopsy samples were obtained from Holstein cows 21 days prior to and at various times after calving to represent the dry period and early and mid-stages of lactation. Additional cows received an intramammary challenge with Escherichia coli for immunohistochemical characterization of AM and AM-BP. Milk AM concentrations were relatively constant across the stages of lactation while AM-BP increased two-fold (P<0.04) between early and mid-lactation. Milk AM (P<0.04) and AM-BP (P<0.03) increased as somatic cell counts (SCCs) increased within a given stage of lactation. Tissue content of both (AM and AM-BP) were significantly affected by stage of lactation, lowest in the dry period and progressively increasing to peak at mid-lactation as well as increasing in association with higher levels of SCCs. Following E. coli challenge, AM increased in epithelial cells surrounding mammary alveoli presenting high levels of SCCs. The data suggest that AM and AM-BP are cooperatively regulated in the mammary gland during lactation; changes in localized tissue AM and AM-BP content reflect a dynamic regulation of these tissue factors in the bovine mammary gland consistent with their protective effects within inflamed tissue.     

Comparative pharmacokinetics of marbofloxacin in healthy and Mannheimia haemolytica infected calves

The pharmacokinetics of marbofloxacin were investigated in healthy (n=8) and Mannheimia haemolytica naturally infected (n=8) Simmental ruminant calves following intravenous (i.v.) and intramuscular (i.m.) administration of 2 mg kg(-1) body weight. The concentration of marbofloxacin in plasma was measured using high performance liquid chromatography with ultraviolet detection. Following i.v. administration of the drug, the elimination half-life (t(1/2 beta)) and mean residence time (MRT) were significantly longer in diseased calves (8.2h; 11.13 h) than in healthy ones (4.6 h; 6.1 h), respectively. The value of total body clearance (CL(B)) was larger in healthy calves (3 ml min(-1) kg(-1)) than in diseased ones (1.3 ml min(-1) kg(-1)). After single intramuscular (i.m.) administration of the drug, the elimination half-life, mean residence time (MRT) and maximum plasma concentration (C(max)) were higher in diseased calves (8.0, 12 h, 2.32 microg ml(-1)) than in healthy ones (4.7, 7.4 h, 1.4 microg ml(-1)), respectively. The plasma concentrations and AUC following administration of the drug by both routes were significantly higher in diseased calves than in healthy ones. Protein binding of Marbofloxacin was not significantly different in healthy and diseased calves. The mean value for MIC of marbofloxacin for M. haemolytica was 0.1+/-0.06 microg ml(-1). The C(max)/MIC and AUC(24)/MIC ratios were significantly higher in diseased calves (13.0-64.4 and 125-618 h) than in healthy calves (8-38.33 and 66.34-328 h). The obtained results for surrogate markers of antimicrobial activity (C(max)/MIC, AUC/MIC and T > or = MIC) indicate the excellent pharmacodynamic characteristics of the drug in diseased calves with M. haemolytica, which can be expected to optimize the clinical efficacy and minimize the development of resistance.     

高亲和钾转运体在植物抗盐中的作用

土壤盐渍化是导致作物减产的一项重要原因,Na+对大多数作物是有害的,而钾是植物不可缺少的营养成分,植物根系在外界低钾条件下主要通过高亲和性系统吸收K+.介绍了高亲和钾转运体Trk/HKT和KUP/HAK/KT 2大家族的结构和功能.主要讨论了当植物处于K+饥饿及盐胁迫下,2大家族在改善植物钾营养、提高植物耐盐性方面的作用.