腺病毒载体介导的GFP在鸡胚成纤维细胞中的表达及RNA干涉

鸡胚成纤维细胞（CEF）是多种动物及人类病毒的易感细胞;也是研究鸡的基因组功能的理想材料．本试验的目的是研究非复制型腺病毒载体对CEF的转染效率及安全性;并建立在CEF细胞中进行RNAi的技术平台．采用GFP重组非复制型腺病毒载体（Ade-GFP）转染CEF细胞：结果证明0．1．2000 MOI Adv-GFP均可转染CEF细胞并表达GFP;转染后16h开始出现GFP阳性细胞：胞核与胞浆可见明亮的荧光：荧光细胞数量及荧光强度在24～36h达到高峰：以后逐渐衰减;约180h全部消失;转染效率最高为22.3％;形态学观察及流式细胞仪测定结果显示：Adv-GFP转染的CEF细胞未见细胞病变;多数转染组细胞活力不受影响：说明用Adv-GFP携带外源转染CEF细胞是安全可行的;用脂质体转染试剂转染化学合成的GFP-siRNA;成功地干涉了腺病毒载体介导的GFP基因在鸡胚成纤维细胞的表达：干涉效率为85％：证明了鸡胚成纤维细胞中存在RNAi机制：为进一步利用非复制型腺病毒载体递送siRNA在CEF细胞内进行RNAi的研究奠定了基础．     

空斑滴定中甲纤复盖物的简易配制方法

在研究病毒的过程中,测定病毒的效价是一项重要的工作。病毒效价的测定,视病毒的种类不同其方法各异。如流感病毒有凝集红血球的能力,可用血凝试验测定其效价;单纯疱疹病毒可使细胞产生病变用TCID_(30)测定效价;单纯疱疹病毒还能在致密的单层细胞中形成一个个空斑,利用空斑测定其效价,是一种比较准确的测定效价的方法,所以目前应用较广泛。     

含狂犬病毒糖蛋白基因的重组痘苗病毒(RTTVV-3)传代后生物学特性研究

将重组病毒在原代鸡胚细胞上连续传至30代(P30),其结果P30与第11代(P11)一样,在含中性红和X-gal的琼脂糖平板上为蓝色蚀斑;空斑法测定P11至P30,各代病毒滴度均稳定在3×10~7PFU/mL～4×10~7PFU/mL;血凝试验表明。不同代次的血凝滴度基本稳定在相同水平;Western blot检测P30与P11在同一位置出现相同蛋白带;血清中和试验证实传代后的重组病毒抗原活性及保护性稳定。重组病毒经传30代后,在生物学特性和表达水平上都具有良好的遗传稳定性。     

猫疱疹病毒Ⅰ型的分离与鉴定

目的从临床疑似猫病毒性鼻气管炎的病例中分离猫疱疹病毒Ⅰ型。方法用猫肾细胞(FK-81)对临床疑似猫病毒性鼻气管炎病猫的眼鼻分泌物进行分离培养,连续传4代,对获得的培养物进行了电子显微镜观察、血凝试验、病毒核酸型试验、理化特性试验以及PCR扩增和扩增产物基因测序鉴定。结果该病毒粒子呈球形,直径为160 nm左右,有囊膜。凝集猫红细胞,不凝集猪、兔、犬、小鼠和鸡的红细胞。病毒对乙醚、酸、热、胰蛋白酶敏感,其核酸型属DNA。PCR扩增为猫疱疹病毒Ⅰ型阳性,扩增产物基因序列与Nunberg等报道猫疱疹病毒Ⅰ型的基因序列同源性为100%(登录号:M26660)。结论在国内首次成功分离一株猫疱疹病毒Ⅰ型,其形态、血凝性、理化特性、基因序列等与国外文献报道一致,为猫病毒性鼻气管炎病原学、疫苗免疫、诊断及分子生物学研究奠定基础。     

大黄素体外抗柯萨奇病毒B_4的实验研究

为研究大黄素的体外抗柯萨奇病毒B4(CVB4)作用,以病毒唑为阳性对照药,用MTT法从药物抑制病毒增殖、直接杀灭和抗吸附3种方式评价了大黄素抗CVB4效果,并用实时定量PCR检测了大黄素对CVB4基因mRNA表达的影响,用空斑法测定了大黄素对病毒感染细胞不同时间空斑形成率的影响.结果表明:大黄素不能直接杀灭病毒或阻断病毒吸附细胞,但能显著抑制病毒在细胞内的生物合成,其IC50=(14.10±0.74)μM,TI=(4.35±0.60),能显著降低细胞内CVB4基因mRNA表达(p0.05),其抑制作用发生在病毒进入细胞0~4 h.故大黄素主要通过抑制CVB4穿入细胞后复制环节发挥抗病毒作用,并具有时间依赖性和剂量依赖性.     

间接血凝试验诊断日本血吸虫病的研究

本文对间接血凝试验诊断日本血吸虫病的实验方法和影响因素进行了研究.实验证明:不论采用1％或2.5％鞣化红细胞,加入1％正常兔血清(Normal rabbit serum,简称1％NRS)后,可以消除鞣化红细胞的自凝现象.比较鞣化红细胞加入1％NRS后再致敏与致敏红细胞加入1％NRS后对间接血凝试验的影响,结果表明:二者对间接血凝试验的敏感性无显著差异(P>0.05).不同致敏红细胞浓度对间接血凝试验的影响,实验结果显示:采用1％致敏红细胞试液可提高间接血凝试验的敏感性;采用2.5％浓度则可提高间接血凝试验的特异性.故在血吸虫病重流行区,为了提高间接血凝试验的特异性,可考虑用2.5％浓度的致敏红细胞试液,而在非流行区及轻流行区,为了提高间接血凝试验的敏感性,可采用1％浓度,以节省致敏红细胞的用量.温度与间接血凝试验反应速度的关系是:随着温度的升高,间接血凝试验反应速度加快,观察结果的时间可以相应缩短.     

1株新型小球藻病毒的分离

以常见淡水普通小球藻为材料,用反冲超滤法从自然水体中分离到1株能裂解普通小球藻的病毒,并用空斑法对病毒进行了纯化,命名为WH-1株.电镜观察显示,WH-1株病毒为球形多面体结构,直径约为21 nm;噬藻斑显示,64 h后所形成的空斑直径约为3 mm.由于与已报道的小球藻病毒株具有明显的差异,认为WH-1株是一种新型的小球藻病毒.     

Marc-145细胞库的建立及其生物学特性研究

从中国典型培养物保藏中心(CCTCC)引进Marc-145细胞,建立了原始细胞库、主细胞库和工作细胞库,并对工作细胞库的细胞复苏后进行了活力、形态、生长特性、微生物污染、核型、乳酸脱氢酶同工酶和对猪繁殖与呼吸综合征病毒(PRRSV)敏感性等特性进行了鉴定.结果显示,Marc-145细胞为上皮型细胞,生长状态良好,生长曲线呈"S"型,最大增殖浓度为4.71×105/mL,倍增时间为26.1h,细菌、真菌、支原体和红细胞吸附检测均为阴性,染色体核型为2n=60,对PRRSV敏感.建立的细胞,为开展猪繁殖与呼吸综合征病毒及其疫苗研究提供了细胞资源.     

预防流感蜡烛对流感病毒抑制杀灭作用做试验

为测定预防流感蜡烛对流感病毒的抑制和杀灭作用,本试验采用了对流感病毒敏感的鸡胚细胞和鸡胚最适部位接种病毒。以细胞病变(CPE)、红细胞吸附试验和红细胞凝集试验做检测指标。结果表明:该蜡烛对流感病毒具有较强的抑杀作用。故用于群体预防具有流行病学意义。     

MDV中meq基因对CEF细胞chTERT,chTR表达影响的研究

利用TaqMan探针技术,采用实时荧光相对定量RT-PCR方法检测了MDV中meq基因对鸡胚成纤维细胞(CEF)的端粒酶催化亚单位基因(chTERT)、端粒酶RNA亚基(chTR)表达水平的影响,并用TRAP法测定了端粒酶活性,用流式细胞仪检测了细胞周期的变化.实验结果表明,转染48 h后chTERT表达水平为转染后72 h的16倍,chTR表达水平在转染前后基本不变;转染48 h后CEF细胞的相对端粒酶活性是转染72 h后的12倍;在转染后72 h S期细胞的百分比较未转染细胞显著增加.     

An Index mechanism for multi-scale view in spatial databases

There are numerous geometric objects stored in the spatial databases. An importance function in a spatial database is that users can browse the geometric objects as a map efficiently. Thus the spatial database should display the geometric objects users concern about swiftly onto the display window. This process includes two operations:retrieve data from database and then draw them onto screen. Accordingly, to improve the efficiency, we should try to reduce time of both retrieving object and displaying them. The former can be achieved with the aid of spatial index such as R-tree, the latter require to simplify the objects. Simplification means that objects are shown with sufficient but not with unnecessary detail which depend on the scale of browse. So the major problem is how to retrieve data at different detail level efficiently. This paper introduces the implementation of a multi-scale index in the spatial database SISP (Spatial Information Shared Platform) which is generalized from R-tree. The difference between the generalization and the R-tree lies on two facets: One is that every node and geometric object in the generalization is assigned with a importance value which denote the importance of them, and every vertex in the objects are assigned with a importance value,too. The importance value can be use to decide which data should be retrieve from disk in a query. The other difference is that geometric objects in the generalization are divided into one or more sub-blocks, and vertexes are total ordered by their importance value. With the help of the generalized R-tree, one can easily retrieve data at different detail levels.Some experiments are performed on real-life data to evaluate the performance of solutions that separately use normal spatial index and multi-scale spatial index. The results show that the solution using multi-scale index in SISP is satisfying.     

Wuhan University Journal of Natural Sciences TOTAL CONTENTS

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Advances in broadband wireless communications under high-mobility scenarios

Future mobile communication systems aim at providing very high data transmission rates, even in high-mobility scenarios such as high-speed wheel-track trains, maglev trains, highway vehicles, airplanes, guided missiles or spacecraft. A particularly important commercial application is the strong and increasing worldwide demand for high- speed broadband wireless communications （up to 574.8 km/ h test speeds or 380 km/h commercial speeds） in railways, providing data, voice and video services for applications such as onboard entertainment services to passengers, train control, train dispatch, train sensor status handling and sur- veillance. In such high-mobility scenarios, there are a number of communication challenges, including fast hand- over, location updating, high-speed channel modeling, estimation and equalization, anti-Doppler spreading tech- niques, fast power control, and dedicated network architec- ture. Because signal transmission in very high-speed scenarios will inevitably experience serious deterioration, it is imperative to develop key broadband mobile communi- cation techniques for such high-speed vehicles.     

Perspherical Approaches for Electrons in Atoms and Charged Excitons in Quantum Wells

Instead of following Fock’s expansion,we solve the Schrodinger equation for some quantum mechanical manybody systems such as electrons in atoms and charged excitons in quantum wells in a similar way in hyperspherical coordinates by expanding the wave functions into orthonormal complete basis sets of the hyperspherical hannonics(HHs)of hyperangles and generalized Laguerre polynomials(GLPs)of the hyperradius.This leads the equation to     

Scale-based spatial data model for GIS

Being the primary media of geographical information and the elementary objects manipulated, almost all of maps adopt the layer-based model to represent geographic information in the existent GIS. However, it is difficult to extend the map represented in layer-based model. Furthermore, in Web-Based GIS, It is slow to transmit the spatial data for map viewing. In this paper, for solving the questions above, we have proposed a new method for representing the spatial data. That is scale-based model. In this model we represent maps in three levels: scale-view, block, and spatial object, and organize the maps in a set of map layers, named Scale-View, which associates some given scales.Lastly, a prototype Web-Based GIS using the proposed spatial data representation is described briefly.     

英文诗歌中的喻类修辞

为了有助于提高英文爱好者的个人情操、文化素养和学习兴趣,以及专业人员对英语学习和研究进行多途径的探索,本文通过一些精选诗歌的引证和分析,着重论述了喻类修辞法在英文诗歌中的运用其及效果。     

Man Is Unable to Change the Miserable Life——Analysis of the Arrangement of Four Characters in The Glass Menagerie

Tennessee Williams is considered as one of the most important American playwrights since World War II.The Glass Menagerie is his first successful drama,which describes a tragic situation of family and means to say that Man is unable to change the miserable life,no matter whatever means he try,This essay focuses on the analysis of the arrangement of the four main characters:Laura.Amanda,Jim and Tom to reveal the theme.Laura is fragile.Amanda is brave.Jim is vital.Tom is sensible.And all of them develop and try the different means to struggle against life,but fail tragically.With the evidence,the paper comes to conclusion naturally that Man is unable to change the miserable life,and he dooms to fail.     

Characterization of root-nodulating bacteria on Retama raetam in arid Tunisian soils

The aim of this study is to investigate the diversity of Retama raetam root-nodule bacteria isolated from arid regions of Tunisia. Twelve isolates, chosen as representative for different 16S rRNA gene patterns, were characterized by 16S rRNA gene sequencing and phenotypic analysis. Isolates were assigned to Sinorhizobium, Rhizobium and Agrobacterium. Symbiotic properties of Sinorhizobium and Rhizobium isolates showed a large diversity in their capacity to infect their host plant and fix atmospheric nitrogen. Strain RK 22 identified as Rhizobium was the most effective isolate.     

An Affordable Accurate Evaluation of the Binding Conformations Predicted by Molecular Docking Studies in Bio-System

正Recently,docking has been widely used to predict the binding-modes of protein-inhibitors,when the crystal complexes structure was absent.Most docking algorithms are able to generate a large number of probable conformations,it,however,is difficult to effectively evaluate these docking poses and identify the most reasonable bindingmode.In the present study,on the basis of the crystallographic data of human 3-hydroxy-3-methylglutaryl coenzyme     

WIMAX与WI-FI技术分析及联合组网

介绍了WiMAX与Wi-Fi两种无线宽带接入技术,并对两者之间的关系及相互之间的影响做了对比及分析,并对WiMAX的关键技术进行了详细说明,最后对两者的联合组网方式做了简单的探讨。