Immunohistochemical expression of interferon-γ in different types of granulomatous lesions associated with bovine paratuberculosis

Animals infected with Mycobacterium avium subsp. paratuberculosis (Map) show a variety of lesions, from focal forms, seen in subclinical stages to diffuse lesions in clinical cases. The purpose of this study was to evaluate the local expression of IFN-γ by immunohistochemistry in relation with the type of lesion in naturally Map-infected cows. The number of immunolabelled cells, −the majority morphologically consistent with lymphocytes-, was higher in focal and diffuse paucibacillary forms than in diffuse multibacillary lesions, where they appeared closely related to epithelioid cells. Diffuse multibacillary lesions had the lowest numbers, but higher than controls, and positive cells were intermingled among the macrophages. The peripheral IFN-γ production was higher in all Map infected cows and a positive correlation was found with the number of immunolabelled cells in the intestine. The findings of this study show that IFN-γ would play a role in the development of the different types of lesions in paratuberculosis, and also points out the importance of adequate sampling of lymphoid tissue containing samples when studying the local immune response in which IFN-γ expression may be involved, especially in cases where focal lesions are present.     

比色法测定稻米γ-氨基丁酸含量的体系优化

[目的]优化比色法测定糙米中γ-氨基丁酸(GABA)的方法,建立一套简便、快速、准确测定糙米GABA的测定体系。[方法]基于Berthlot显色反应,从测定波长及测定时间,苯酚、次氯酸钠及糙米用量上优化了比色法测定糙米中γ-氨基丁酸含量的技术体系。[结果]试验得出,比色法测定γ-氨基丁酸的最佳波长为630 nm,5%苯酚溶液的最佳用量为3.4 ml,有效氯为7%的次氯酸钠溶液的最佳用量为1.5 ml,建议最佳的测定时间为60～180 min。以该试验方法处理米样时,米样的最佳用量为1.2 ml。[结论]该体系能简单、快速、经济、准确地测定稻米中γ-氨基丁酸的含量,可为以后筛选和选育富含GABA的稻米品种提供参考。     

γ-聚谷氨酸对土壤结构、养分平衡及菠菜产量的影响

通过盆栽试验研究了土壤增施γ-PGA(0.1%,0.2%,0.3%,0.4%)对土壤团聚结构、土壤养分平衡、菠菜产量与经济效益的影响。结果表明:随γ-PGA施量增加,水稳性团聚体(WR0.25)、平均重量直径(MWD)和几何平均直径(GMD)均显著增大,而团聚体破坏率(PAD)与分形维数(D)均减小,说明γ-PGA对改善土壤团聚结构具有显著的效果;随γ-PGA施量的增加,菠菜对氮、磷、钾的吸收量减少,土壤养分残留量与盈余量增加,养分表观损失量减小;5组处理中,0.1%施量的土壤养分平衡状况最优,0.4%施量对作物养分吸收与土壤养分的平衡产生负效应;菠菜产量及经济效益均随γ-PGA施量增加而显著减小,0.4%施量时较未施γ-PGA减产42.98%;菠菜经济效益拟合结果表明γ-PGA在0～0.1%施量范围内菠菜获利高于不施加γ-PGA处理。综上,0.1%施量对改善土壤结构与养分状况、促进作物增产及作物养分吸收效果较优,这对进一步将γ-PGA应用于实际生产中提供了理论参考。     

Cytokine profiles in peripheral blood mononuclear cells from patients with cystic echinococcosis

IntroductionCystic echinococcosis (CE) is a chronic zoonotic disease caused by the larval stage of Echinococcus granulosus (E. granulosus), which affects domestic and wild carnivores as the definitive host and ungulates as intermediate hosts. In intermediate hosts, both Th1 and Th2 cells are involved in the immune responses to an echinoccocal infection. This study aimed to investigate production of IL-4, IL-10, and IFN-γ cytokines in peripheral blood mononuclear cells (PBMCs) of CE patients before and after surgical treatment.MethodsTo evaluate cytokine production in response to E. granulosus antigens, we investigated IL-4, IL-10, and IFN-γ production in PBMCs of 20 CE patients in response to hydatid cyst fluid antigen (HCF-Ag) before and after surgical treatment using ELISA.ResultsThe mean IL-4 production from HCF-Ag stimulated PBMCs was significantly decreased (p < 0.05), while IFN-γ was significantly increased in HCF-Ag stimulated PBMCs in patients after surgery (p = 0.005).Furthermore, our results showed that there is no significant difference between IL-10 production in patients before and after treatment (p = 0.562).ConclusionsOur data Indicated production of IL-4 in cultured PBMCs of CE patients stimulated with HCF-Ag was decreased significantly. While, production of IFN-γ was increased significantly in responses to HCF Ag after surgery. We concluded that the evaluation of IL-4 and IFN-γ in HCF-Ag stimulated PBMCs of CE patients should be considered as a useful marker in the follow up of patients with cystic echinococcosis.     

Spatiotemporal heterogeneity of PPARγ expression in porcine uteroplacenta for regulating of placental angiogenesis through VEGF-mediated signalling

Non-infectious prenatal mortality severely affects the porcine industry, with pathological placentation as a likely key reason. Previous studies have demonstrated that peroxisome proliferator-activated receptor gamma (PPARγ) deficiency causes defects in the uteroplacental vasculature and induces embryonic losses in mice. However, its role in porcine placental angiogenesis remains unclear. In the present study, PPARγ expression was investigated in porcine uteroplacental tissues at gestational day (GD) 25, GD40 and GD70 via quantitative polymerase chain reaction (qPCR), Western blot and immunohistochemistry (IHC). Moreover, the roles of PPARγ in porcine placental angiogenesis were investigated using a cell model of porcine umbilical vein endothelial cells (PUVECs) to conduct proliferation, migration and tube formation assays in vitro and a mouse xenograft model to assess capillary formation in vivo. The results showed that PPARγ was mainly located in the glandular epithelium, trophoblast, amniotic chorion epithelium and vascular endothelium, as indicated by the higher expression levels at GD25 and GD40 than at GD70 in endometrium and by higher expression levels at GD40 and GD70 than at GD25 in placenta. Moreover, PPARγ expression was significantly downregulated in placenta with dead foetus. In PUVECs, knocking out PPARγ significantly inhibited proliferation, migration and tube formation in vitro and inhibited capillary formation in mouse xenografts in vivo by blocking S-phase, promoting apoptosis and downregulating the angiogenic factors of VEGF and its receptors. Overall, the spatiotemporal heterogeneity of PPARγ expression in porcine uteroplacental tissue suggests its vital role in endometrial remodelling and placental angiogenesis, and PPARγ regulates placental angiogenesis through VEGF-mediated signalling.     

γ-戊内酯/水复合溶剂体系中金属硫酸盐催化半纤维素定向转化制备糠醛

以廉价金属硫酸盐为催化剂,在γ-戊内酯/水复合溶剂中催化半纤维素定向转化制备糠醛,糠醛得率高达50.2%,半纤维素液化转化率达95.5%。在γ-戊内酯/水复合溶剂中,以金属硫酸盐为催化剂进一步研究了直接催化木质纤维生物质原料玉米芯和竹粉定向转化制备糠醛,其中糠醛得率分别达39.5%、29.7%,木质纤维原料液化转化率分别达86.5%、80.5%。     

东北虎γ-干扰素基因的克隆、生物信息学分析及其在毕赤酵母中的表达

本试验旨在通过克隆东北虎γ-干扰素(IFN-γ)基因,研究其分子特征并预测蛋白生物学功能,为后续研究干扰素抗病毒活性做前期准备。通过RT-PCR从ConA诱导过的东北虎血淋巴细胞中扩增东北虎IFN-γ基因并测序,应用生物信息学方法进行序列分析。结果表明:东北虎IFN-γ编码区由504个核苷酸组成,共编码167个氨基酸,蛋白相对分子质量为19.59 ku,等电点为9.03,所编码的蛋白为碱性亲水性蛋白,其中前23个氨基酸可能为信号肽,IFN-γ编码蛋白保守结构域为IFN-γ超家族,且存在跨膜结构,其中1-6位氨基酸为胞内区域,7-28位氨基酸为跨膜区域,29-167位氨基酸为胞外区域;IFN-γ编码蛋白二级结构主要以α-螺旋(58.08%)和无规则卷曲(33.53%)为主,存在5个潜在的B细胞抗原表位,3个潜在的N-糖基化位点;分子进化分析显示,东北虎IFN-γ与GenBank上发表的东北虎、非洲狮、金钱豹、美洲狮、猎豹、家猫、加拿大猞猁、野猪等的核苷酸相似性为80.4%~99.8%,氨基酸相似性为70.5%~100%,东北虎IFN-γ与非洲狮、金钱豹亲缘关系最近,美洲狮、猎豹、家猫、猞猁次之,野猪最远。通过合成改造后的东北虎IFN-γ基因,构建能表达IFN-γ蛋白的重组质粒pPIC9K-IFN-γ,将其导入高效表达系统-毕赤酵母中进行诱导表达,经SDS-PAGE分析,表达蛋白的分子量约17.8 ku,与预期大小相符,表明东北虎IFN-γ成功表达。     

γ-聚谷氨酸对番茄穴盘育苗基质矿质养分供应及幼苗生长发育的影响

【目的】研究聚谷氨酸不同加入量对基质理化性状和蔬菜生长的影响,可以为其在番茄穴盘育苗中的合理应用提供依据。【方法】以草炭、 蛭石、 珍珠岩(3 ∶1 ∶1,v/v)混合物料为育苗基质, 50孔塑料穴盘为育苗容器进行盆栽试验。基质适宜添加量试验: 播种前在基质中混入-聚谷氨酸 (-PGA) 0、 1、 3、 5、 10 kg/m3,调查了添加-聚谷氨酸后番茄穴盘育苗基质理化、 生物学性状及幼苗生长发育参数的动态变化。 顶部灌施适宜用量试验: 在播种后16 d,将-聚谷氨酸0、 1、 3、 5、 10 g/L -PGA溶入水溶性肥料溶液(20-20-20)进行顶部灌施,肥料氮(N)浓度为200 mg/L, 施用量为1 L/tray,测定了播种后36 d番茄幼苗生长发育参数。【结果】 1)基质添加-聚谷氨酸显著提高了基质初始持水孔隙度、 最大持水量、 铵态氮、 硝态氮、 速效磷、 速效钾、 交换性镁含量和EC值,降低了通气孔隙度和pH值,增强了番茄穴盘苗生长发育过程中基质速效氮、 速效磷、 速效钾和交换性镁供应能力,显著提高了番茄穴盘苗生长发育后期基质过氧化氢酶活性和中性磷酸酶活性; 这些有效作用随-PGA添加量的增加而显著增加,不同添加量处理间差异显著。2)基质添加-聚谷氨酸,随着添加量增加,番茄苗叶片叶绿素含量呈逐步增加趋势,根系活力峰值则出现在添加量3 kg/m3水平; 3)-聚谷氨酸无论基质添加或顶部灌施,对番茄穴盘苗茎叶都表现出促进生长的作用,对根系发育却表现出一定的抑制作用; 4)基质添加-聚谷氨酸对番茄穴盘苗生长发育的促进作用还表现出明显的延迟效应,在幼苗生长发育后期的效果显著优于前期。【结论】番茄穴盘育苗施用-聚谷氨酸能增强基质水分、 养分供应能力,促进番茄穴盘苗后期生长发育。     

贯叶连翘提取物对流感病毒感染小鼠肺中IFN-γ·TNF-α的影响

[目的]研究贯叶连翘提取物(HPE)对流感病毒感染小鼠肺组织中肿瘤坏死因子(TNF-α)、γ-干扰素(IFN-γ)等细胞因子的影响。[方法]以流感病毒滴鼻感染小鼠,分别灌胃给药(贯叶连翘提取物高剂量组100 mg/kg、低剂量组50 mg/kg和病毒唑组10 mg/kg),0.2ml/次,2次/d,5 d后摘眼球放血处死动物,摘出全肺,低温条件下制成肺匀浆,离心后取上清采用双抗体夹心ELISA法观察小鼠治疗前后肺组织中IFN-γ、TNF-α的变化。[结果]与模型对照组相比,贯叶连翘提取物流感病毒感染小鼠肺组织中TNF-α显著降低(P<0.05),IFN-γ极显著升高(P<0.01)。[结论]贯叶连翘提取物对机体抗感染免疫有一定调节作用。     

moFcγRⅢ、γ链真核表达载体的构建及其共转染稳定细胞系的建立

将moFeγRⅢ、γ链编码区eDNA分别亚克隆到真核表达栽体pcDNA3的巨细胞病毒启动子下游,构建了重组表达质粒pe3moRⅢ、pe3moγ;用PvuⅠ线性化重组质粒,以线性化重组质粒其转染COS -7细胞,用玫瑰花环试验检测moFeγRⅢ在转染细胞表面的表达,通过G418抗性筛选和单细胞克隆,在转染细胞表面稳定表达...