miRNA-132增强人类免疫缺陷病毒1型复制

小分子RNA（miRNA）在CD4＋T细胞活化期间上调可能有助于人类免疫缺陷病毒1型（HIV-1）在静息期扰动后复制效率的提高。该研究发现,miRNA-132（miR-132）在CD4＋T细胞活化后高度上调,且能增强病毒在JurkatCD4＋T细胞系中的复制。     

人单核细胞系中HIV-1前病毒转录调节新型研究模型的建立

【目的】抗反转录病毒疗法(ART)能够有效控制人免疫缺陷病毒(Human immunodeficiency virus-1,HIV-1)的复制,但是不能将其完全清除。至2012年底,全球仍有3 500万HIV-1感染者,同年约160万人死于艾滋病(Acquired immune deficiency syndrome,AIDS)及其相关疾病。HIV-1感染难以根治的主要原因之一是机体内HIV-1潜伏储存库(Reservoir)的存在。HIV-1潜伏储存库主要由CD4+T细胞和单核巨噬细胞构成,与CD4+T细胞相比,目前研究者对单核巨噬系细胞中HIV-1病毒复制机制尚不明了,且缺乏适宜的研究体系。因此,为探讨单核细胞活化或分化信号对HIV-1复制的影响,我们建立了旨在研究HIV-1前病毒转录调控机制的人单核巨噬细胞系模型。【方法】构建env区域移码突变和nef区域携带EGFP或Nano Luc报告基因的HIV-1 NLn GFP-Kp或NLn Nano Luc-Kp重组病毒,分别感染2种人单核细胞系THP-1或U937细胞。通过有限稀释法制备单克隆细胞系,利用流式细胞术或Nano Luc荧光素酶活性分析检测报告基因的表达。筛选EGFP或Nano Luc阴性表达的细胞克隆,经激活剂佛波酯(Phorbol-12-myristate-13-acetate,PMA)刺激后鉴定潜伏感染的细胞克隆。【结果】研究中鉴定了4个HIV-1潜伏感染的细胞克隆。其中2个是表达EGFP的THP-1克隆,2个是以Nano Luc为报告基因的U937克隆。这些克隆在PMA刺激处理后皆有报告基因的表达,而在恒态条件下未检测到报告基因的表达。【结论】成功建立了4个HIV-1潜伏感染的人单核细胞系克隆,该模型有助于理解单核巨噬系细胞的HIV-1病毒复制机制,可能成为进一步研究HIV-1前病毒转录调控机制的有力工具。     

人白血病细胞系KG-1a中肿瘤干细胞样亚群细胞的初步研究

探讨人白血病细胞系KG-1a中是否存在具有肿瘤干细胞样生物学特性的亚群细胞.瑞氏染色和吖啶橙染色分别观察白血病细胞系KG-1a细胞形态和RNA含量:流式细胞术检测细胞周期分布;免疫组化和流式细胞术检测KG-1a细胞CD34的表达;流式细胞术检测CD34 CD38-亚群细胞;烟酸己可碱Hoechst33342染色后用荧光显微镜观察KG-1a细胞中侧群(side population,SP)样细胞所占比例.结果显示,白血病细胞系KG-1a细胞核仁易见,形态原始;部分细胞RNA含量低,细胞处于G0/G1期的比例占15.5%.绝大多数细胞的胞浆和胞膜皆表达CD34抗原,KG-1a中CD34 细胞占96.3%,CD34 CD38-亚群细胞占7.02%;SP样细胞大致比例为7.60%.本研究表明.人白血病细胞系KG-1a中存在具有肿瘤干细胞样生物学特性的亚群细胞.     

急性病毒性肝炎患者外周血T 细胞亚群的检测和分析

目的:检测急性甲、乙型病毒性肝炎患者外周血T淋巴细胞亚群变化,探讨其对疗效和预后意义。方法:采用APAAP桥联酶标法检测115例急性甲、乙型病毒型肝炎患者外周血CD3+、CD4+、CD8+T细胞亚群比例,计算CD4+/CD8+值。并检测了34例患者治疗前后T淋巴细胞亚群变化。结果:115例急性甲、乙型病毒性肝炎患者外周血CD3+、CD4+T细胞比例及CD4+/CD8+值均低于正常对照组(P<0.05),而CD8+T细胞比例均高于正常对照组(P<0.01)。6例无明显疗效者,各亚群比例在治疗前后无显著差异(P>0.05)。28例有明显疗效者,治疗后各亚群比例恢复正常水平,与治疗前相比差别具有统计学意义(P<0.05)。结论:急性甲、乙型病毒性肝炎患者外周血CD4+/CD8+T细胞比值可在一定程度上反映疗效及预后。     

HIV-1潜伏感染体外实验模型研究进展

潜伏感染的静息记忆CD4+T细胞是清除HIV-1病毒的一个重要障碍。处于潜伏状态的病毒多以原病毒c DNA的形式整合至宿主基因组中,但是病毒基因表达处于沉默状态,因此潜伏感染的细胞难以受到病毒的致细胞病变效应或机体特异性细胞毒性T细胞的杀伤,也不易受到抗反转录病毒治疗药物的作用。如何减少潜伏感染的细胞储存库是艾滋病治疗中亟需解决的一个问题。体内及体外HIV-1潜伏感染模型有助于深入了解HIV-1潜伏感染的建立、维持或打破机制,评价潜伏感染再激活剂的活性。在此侧重于介绍采用永生化细胞系、原代静息CD4~+T细胞或活化的CD4+T细胞建立的HIV-1潜伏感染体外实验模型。     

人类免疫缺陷病毒与细胞凋亡

人类免疫缺陷病毒1型(HIV-1)感染可使被感染者体内CD4细胞数量减少,最终导致艾滋病。关于HIV-1如何杀死免疫细胞的精确机制仍是1个争论的问题。现已知道,细胞凋亡为HIV-1诱导细胞死亡的一个重要机制。HIV可直接诱导细胞凋亡,也可以通过活化作用,同源被感染的细胞的介导,以及CD^8 T细胞诱导细胞凋亡。且细胞因子在HIV诱导细胞凋亡的过程中发挥着重要的作用。本综述主要从以上几个方面总结HIV-1诱导细胞凋亡的机制。     

多发性骨髓瘤患者外周血淋巴细胞亚群分析及与预后因子的相关性

目的通过对多发性骨髓瘤（MM）患者外周血淋巴细胞亚群的检测以评价MM患者机体的免疫功能状态。方法采用流式细胞术检测36例MM患者和25例健康志愿者外周血T、B淋巴细胞、NK细胞及CD4＋CD25＋T细胞的表达。结果与正常对照组相比,MM患者外周血的CD4、CD19细胞的表达显著下调,CD8细胞的表达显著上调,CD4/CD8比值则显著降低（P〈0.05或〈0.01）;MM患者外周血的CD4＋CD25＋T细胞占CD3＋T细胞的比例明显增高（P〈0.01）,且与血清中的β2-MG浓度成正相关（γ=0.56,P〈0.05）。结论 MM患者体内存在淋巴细胞亚群的异常表达、CD4＋CD25＋Treg细胞的异常扩增,可能是MM患者体内广泛存在免疫缺陷的一个主要原因。     

009 细胞内染色检测人类免疫缺陷病毒特异性γ干扰素的产生

细胞免疫反应在控制人类免疫缺陷病毒Ⅰ型(HIV-1)感染的传播中发挥着重要的作用,它是通过裂解感染的细胞以及产生强有力的抗病毒细胞因子(如γ干扰素(IFN-γ)]起作用的,而流式细胞技术已被广泛用于检测HIV感染引起的免疫反应中T细胞亚群CD4和CD8细胞经抗原刺激后产生的IFN—     

HIV-1感染者中全血miRNA表达谱的变化

初步了解HIV-1感染者全血中miRNA表达谱的变化。使用Taqman低密度miRNA表达谱芯片,分别检测10例HIV-1感染者和10例未感染者全血样本中754条miRNA的表达情况。通过BRB分析软件对样本中miR-NAs的表达情况进行比较,筛选出差异表达的miRNA。使用DIANA在线工具预测差异表达miRNAs的靶基因和细胞功能。筛选结果显示有56条miRNAs显著差异表达(P<0.001),其中有49条miRNAs在感染者中下调,7条miRNAs的表达上调。差异表达miRNAs的靶基因涉及的生物学功能相对集中,主要富集在MAPK、TGF-be-ta、Wnt等信号通路。提示:HIV-1感染引起全血中部分miRNAs表达情况的改变。56条显著差异表达的miRNA可能有着重要的作用,对这些miRNA的进一步研究有望发现新的与致病机制相关的关键分子和HIV-1感染诊断潜在的标志物。     

BST-2抑制HIV-1病毒样颗粒释放的研究

BST-2是最近发现的可以抑制成熟HIV-1(human immunodeficiency virus,HIV)病毒颗粒从哺乳动物细胞表面释放的宿主因子,随之发现其也可以抑制多种包膜病毒的释放。本研究采用密码子优化的表达HIV-1 gag和gag-pol蛋白的质粒所形成的病毒样颗粒作为研究对象,观测BST-2对这两种病毒样颗粒(Virus-like particle,VLP)的释放抑制情况及其作用机制。结果发现,瞬时表达和稳定表达的BST-2均可以显著抑制病毒样颗粒从哺乳动物细胞释放,同时发现这两种病毒样颗粒(gag/gag-pol)的释放都可以被BST-2抑制;而且,HIV-1中Vpu蛋白可以拮抗BST-2抑制HIV病毒样颗粒释放的作用,另外,通过化学试剂和酶学方法处理,确证BST-2可以被包装进病毒样颗粒中。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

医疗机构合理用药评价模型的构建研究

目的 针对医疗机构的合理用药水平进行评价研究。方法 根据医疗机构合理用药的具体要求,构建医疗机构合理用药评价指标体系,采用基于模糊群决策的方法和多指标评价分析法构建医疗机构合理用药评价模型。结果 构建了基于模糊群决策的医疗机构合理用药评价模型,并通过实例分析证明了评价模型的可行性。结论 建立的基于模糊群决策的医疗机构合理用药评价模型能够对医疗机构的合理用药水平进行科学评价,为提高医疗机构合理用药水平奠定基础。