SARS—CoV N蛋白与病毒mRNA相互作用的初步研究

SARS冠状病毒(SARS-CoV)是一种新型的冠状病毒,其基因组大小约为30,000 nt,为单股正链RNA。病毒基因中的1-72个核甘酸为前导序列。核衣壳(Nucleocapsid,N)蛋白是冠状病毒的主要结构蛋白,它在病毒基因转录,翻译以及病毒颗粒包装中起重要作用。在本研究中,我们通过PCR的方法从SARS-CoV cDNA中克隆N基因,将基因克隆到大肠杆菌表达载体中,经表达纯化获得大量重组蛋白,通过亲和层析和凝胶过滤层析获得高纯度的N蛋白。同时构建前导RNA的转录模板,经体外转录得到地高辛标记的RNA。使用Northwestern分析技术,我们证实纯化的N蛋白在体外可以与RNA发生特异性的结合。N蛋白与病毒RNA的结合特性及其在病毒生活周期中的所起作用的初步研究,为下一步设计出有效的阻断病毒周期从而达到抗病毒目的的药物或疫苗奠定了基础。     

人热休克蛋白gp96基因的克隆及其在大肠杆菌中的表达与纯化

热休克蛋白gp96是热休克蛋白90家族成员,能够引起非特异性和特异性免疫反应。得到大量高纯度的蛋白质是研究开发gp96的关键。然而重组的gp96容易在E．coli中降解,并在一定条件下形成多聚体。实验先将人gp96基因克隆到pET-30a载体上并在E．coli Blstar中表达,再经过亲和层析、阴离子交换、分子筛分别纯化gp96。最终去掉了大部分的降解片段和多聚体,得到一定量的可溶性gp96,为进一步研究其结构和功能打下一定的基础。     

香港嘉道理农场次生林区碰撞诱捕网和黑光灯捕虫器采集所得鞘翅目甲虫多样性比较

本文根据1990-1995年在香港嘉道理农场次生林区采集的昆虫标本,首次分析了鞘翅目及其优势科的种类、数量和季节性变化,以及由碰撞诱捕网和黑光灯捕虫器采集所得甲虫在种类、数量和季节性上的差异。在13260号标本中,已鉴定到科的有13253号,分属45科,231种。其中,朽木甲科(Alleculidae)、毛蕈甲科(Biphyllidae)、丸甲科(Byrrhidae)、坚甲科(Colydiidae)、拟球甲科(Corylophidae)、隐食甲科(Cryptophagidae)、水缨甲科(Hydroscaphidae)、伪叶甲科(Lagliidae)、薪甲科(Lathridiidae)、泽甲科(Limnichidae)、黑蕈甲科(Zopheridae)等11个科为香港地区的首次报道,约占本次调查科总数的25％。分析表明：(1)该次生林区的鞘翅目甲虫以蛀木性为主。天牛科(Cerambycidae)、瓢虫科(Coccinellidae)、象甲科(Curculionidae)、花蚤科(Mordellidae)、金龟甲科(Scarabaeidae)、小蠹科(Scolytidae)等6科均为多样性较高(种类≥15或者个体数量≥200)的优势科。(2)鞘翅目个体数量季节性明显,每年自2月开始数量逐渐增加,6—7月为甲虫发生的高峰期,8月显著减少。各优势科甲虫的季节性也存在一定的差异,庞大的小蠹标本数量(85％)说明在此调查期间该科正处于大发生时期。(3)黑光灯捕虫器所捕的甲虫科类和种类较之碰撞诱捕网所捕不尽相同,黑光灯捕虫器所捕的甲虫数量发生高峰期比碰撞诱捕网所捕的甲虫提前一个月。(4)各项多样性指数对不同捕虫器采集所得鞘翅目的测度差异明显,黑光灯捕虫器所捕甲虫的多样度和均匀度指数高于碰撞诱捕网。     

小球藻病毒PBCV-1特异性溶壁酶(Lysin)的溶壁活性

从PBCV－1感染小球藻NC64A的细胞裂解液中提取了Lysin的粗制剂,酶活底物范围分析表明,几丁质酶、壳聚糖酶的β-1,3-葡萄糖苷酶是Lysin活性的主要组成部分,并与小球藻细胞壁的组成甩分相吻合。其中几丁酯酶和壳聚糖酶,特别是几丁酯酶在裂解小球藻细胞壁的过程中发挥了重要的作用。Lysin粗制剂经FPLC分离纯化得到分子量分别为52kD、56kD的两个几丁质酶（Chil和Chi2）和一个分子量为36kD的壳聚糖酶。     

不受欢迎的生物多样性：香港的外来植物物种

香港早在19世纪中叶开始就有外来植物入侵的记录,迄今为止,已发现多达238种已归化的外来或怀疑为外来的植物,其中又以薇甘菊（Mikania micrantha)、五爪金龙（Ipomoea cairica)、假臭草（Eupatorium catarium)、大黍（Panicum maximum)等最常见,外来植物最常见于受人为干扰的生境,例如荒废农田及路旁等,而较少在天然林地生境及贫瘠的灌草丛中发现,外来植物的对本地生态系统的影响主要局限于低地生境,它们常形成单优种群,减少了生境及贫瘠的灌草丛中发现,外来植物对本地生态系统的影响主要局限于低地生境,它们常形成单优种群,减少少了生境及动植物的多样性,外来动物对香港原生植物影响最大的是于20世纪70年代入侵的松树线虫（Bur-saphelenchus xylophilus)。外来的脊椎动物也有可能对香港的植物被演替产生影响,目前香港的外来植物当中,有些在大陆较少分布或没有记录,作为华南最大的港口,香港对外来物种的引入扮演着重要的角色,因此制定控制外来种在香港及华南地区的引入及传播的政策及措施非常重要。     

Pollen allergies in czechoslovakia

The article discusses the possible utilisation of the palynological and geobotanical observations in immunotherapy of children with pollinosis in Czechoslovakia. Three approaches to this problem exist: 1. Use of the individual pollen mixtures (prepared by the Institute of Sera and Vaccines (USOL), Prague), according to the clinical history and the results of diagnostic tests of each patient. 2. Use of the available standard pollen mixtures. 3. Improvement of the specificity of these mixtures by eliminating the redundant pollen types (i.e. pollen of the species not spread in the area). Intentions for the future, including the establishment of a pollen forecasting service, are mentioned.     

IL-22-Producing RORγt-Dependent Innate Lymphoid Cells Play a Novel Protective Role in Murine Acute Hepatitis

Retinoid-related orphan receptor (ROR) γt is known to be related to the development and function of various immunological compartments in the liver, such as Th17 cells, natural killer T (NKT) cells, and innate lymphoid cells (ILCs). We evaluated the roles of RORγt-expressing cells in mouse acute hepatitis model using RORγt deficient (RORγt^−/−) mice and RAG-2 and RORγt double deficient (RAG-2^−/− × RORγt^−/−) mice. Acute hepatitis was induced in mice by injection with carbon tetrachloride (CCl₄), to investigate the regulation of liver inflammation by RORγt-expressing cells. We detected RORC expression in three compartments, CD4⁺ T cells, NKT cells, and lineage marker-negative SCA-1⁺Thy1^high ILCs, of the liver of wild type (WT) mice. CCl₄-treated RORγt^−/− mice developed liver damage in spite of lack of RORγt-dependent cells, but with reduced infiltration of macrophages compared with WT mice. In this regard, ILCs were significantly decreased in RAG-2^−/− × RORγt^−/− mice that lacked T and NKT cells. Surprisingly, RAG-2^−/− × RORγt^−/− mice developed significantly severer CCl₄-induced hepatitis compared with RAG-2^−/− mice, in accordance with the fact that hepatic ILCs failed to produce IL-22. Lastly, anti-Thy1 monoclonal antibody (mAb), but not anti-NK1.1 mAb or anti-asialo GM1 Ab administration exacerbated liver damage in RAG-2^−/− mice with the depletion of liver ILCs. Collectively, hepatic RORγt-dependent ILCs play a part of protective roles in hepatic immune response in mice.     

Flux boundary measurements for the study of tree stability

The shear strength of soil is an important parameter that affects tree stability and can vary depending on the magnitude of the soil’s negative pore-water pressure (matric suction). The surface flux boundary condition affects the matric suction of soil, and therefore is important for tree stability. Field measurements were performed around a roadside tree for 2 years. The instrumentation results show that the matric suction in the soil fluctuated between 0 and 35 kPa. Matric suction changes in the soil could lead to a decrease in the tree resistive moment of up to 80 %.