精子膜抗原提取、纯化及SDS—PAGE分析

目的：分析正常生育男性精细胞膜抗原，以便进一步应用精子抗原于其抗体的检测。方法：1．精子膜抗原粗制方法，按“全国临床检验操作规程”(第二版，1998年，P398)描述的方法进行，即收集20名正常生育男性精子样品，用含0．5％NP—40的Tris—Hcl缓冲液抽提。2．粗提液蛋白含量测定采用紫外分光光度法。3．蛋白抗原纯化方法，采用冰冷乙醇沉淀法，以粗提液：冰冷95％乙醇(—20℃)＝1：9(v/v)混合，并按1／50容量加入饱和乙酸钠混匀，于—20℃放置48h。取出后于—10℃，1800g离心30分钟，弃上清，例放试管，沥干沉淀，—20℃保存备用。4、SDS—PAGE分析，步骤3所获样品，以NS溶解，通过紫外分光光度计测定含量，并调节到6mg/ml。SDS—PAGE分析条件的点样量25μl，分离胶17．5％，隔层胶5％，电泳缓冲液用甘氨酸—Tris，PH18．6，电压20v过夜，次日再150V4h。电泳后凝胶经考麦斯亮兰染色、乙酸—甲醇洗脱，直至显示清晰电泳图谱。结果：1、粗提液经紫外分光光度计测定，其原液含蛋白质为24mg/ml，最大吸收峰值280nm。2．纯化的精子膜抗原溶液经SDS—PAGE分析，图谱显示2个蛋白斑，与标准蛋白比较，其分子量一个为35—45KD，另一个为15—25KD；75—85KD处出现微量痕迹。结论：我们成功地进行了精子膜抗原提取、纯化及SDS—PAGE分析实验，结果有清晰记录，为进一步应用精子抗原检测精子抗体的研究提供物质基础。     

棘球蚴抗原纯化对免疫球蛋白IgG免疫应答的影响

以纯化的棘球蚴抗原和未经纯化的粗抗原用于棘球蚴感染者血清中抗棘球蚴抗原的特异性免疫球蛋白IgG水平的检测试验,结果表明,当血清稀释度为1∶200时,使用纯化抗原可以获得较高的特异性 (98.9%),并可保持在使用粗抗原及血清稀释度为1∶400时获得的较高的敏感性（93.7%）,提示在棘球蚴感染者血清中抗棘球蚴抗原特异性免疫球蛋白IgG水平的检测中,使用纯化棘球蚴抗原和粗抗原均可获得较高的敏感性(93.7%)和较高的特异性 (98.4%,98.9%),使用纯化抗原时的特异性略高,但在统计学上并无显著差异.     

抗Sa抗体和RF对RA诊断的比较研究

目的：比较抗Sa抗体和FR对RA诊断的效果。方法：用Wextenn－blot方法检测抗Sa抗体,用胶乳凝集法检测RF。结果：抗Sa抗体对RA诊断的敏感度为42．7％,特异度为100％,阳性确诊率为100％,阴性确诊率为71．1％。RF对RA的诊断敏感度为72O％,特异度为79．2％,阳性确诊率为71．2％,阴性确诊率为80.0％。结论：两种方法比较表明,抗Sa抗体诊断RA的特异性和阳性确诊率均优于RF.     

水牛梭形住肉孢子虫缓殖子抗原的纯化及SDS—PAGE分析

应用葡聚糖凝胶柱层纯化了水牛梭形住内孢子虫缓殖子抗原，并应用ＳＤＳ－ＰＡＧＥ分析了缓殖子可溶性粗抗原及纯化抗原的蛋白组分，结果表明：缓殖子可溶性粗抗原含有１４种蛋白成分，分子量范围为４５．９－７１．２ＫＤ，经过ＳｅｐｈａｄｅｘＧ－１００柱层析纯化扣的抗原在ＳＤＳ－ＰＡＧＥ图谱上显示５条蛋白带，分子量分别为５８ＫＤ，６１ＫＤ，６４ＫＤ，６６ＫＤ，７０ＫＤ。     

重金属铅多克隆抗体的制备及纯化

制备重金属铅完全抗原,进而制备和纯化重金属铅的多克隆抗体。使用双功能螯合剂p-SCN-Bn-DTPA、Pb2+标准溶液和牛血清白蛋白(BSA)制备重金属铅的完全抗原Pb-DTPA-BSA;运用聚丙烯酰胺凝胶电泳(SDSPAGE)方法判断完全抗原Pb-DTPA-BSA是否合成成功,并应用紫外分光光度计测定免疫抗原Pb-DTPA-BSA的浓度;取0.5 mg免疫抗原Pb-DTPA-BSA加入弗氏佐剂乳化后免疫家兔,制备抗重金属铅多克隆抗体,并运用凝胶柱层析法纯化抗重金属铅多克隆抗体,SDS-PAGE方法判定抗重金属铅多克隆抗体的纯化结果。SDS-PAGE结果显示成功制备重金属铅的完全抗原Pb-DTPA-BSA,其浓度为3.4mg/mL;应用免疫抗原Pb-DTPA-BSA免疫家兔,成功制备抗重金属铅多克隆抗体;应用凝胶柱层析法纯化获得高纯度抗重金属铅多克隆抗体。成功制备重金属铅的完全抗原PbDTPA-BSA,并成功制备和纯化抗重金属铅的多克隆抗体。     

从对流免疫扩散电泳免疫复合物沉淀线提取抗原小核RNA的方法

可提取性核抗原(ENA,包括Sm、RNP、Ro和La等多种抗原成分)是小核RNA和蛋白质组成的复合物。本文介绍从对流免疫扩散电泳(CIDE)形成的免疫复合物沉淀线提取单一抗原小核RNA的方法。其结果与纯化抗原的小核RNA组成相同。此法仅需约100微升的单价特异性抗体,抗原不需纯化,操作简单易行。     

口服耐受增效剂对抗原诱导性葡萄膜视网膜炎的作用

目的：观察口服牛视网膜S抗原与脂多糖(LPS)对Wistar大鼠抗原诱导性葡萄膜视网膜炎(AIU)的影响。方法：用纯化牛视网膜S抗原和福氏完全佐剂的混合乳剂致敏大鼠，14d后接种S抗原于致敏鼠眼玻璃体腔复制AIU模型。观察致敏前口服S抗原及LPS、单独口服S抗原或牛血清白蛋白(BSA)对AIU眼部表现和迟发型超敏反应(DTH)的影响。结果：与口服BSA组比较，口服S抗原及LPS组在AIU第1、3、5d的临床分级参数显降低，炎症持续时间显缩短，DTH显降低。与单独口服S抗原组比较，口服S抗原及LPS组在AIU的炎症持续时间显缩短，DTH显降低。结论：口服S抗原及LPS明显抑制AIU的炎性反应和DTH，口服LPS可加强S抗原对AIU的免疫抑制作用。     

琼脂微球偶联氨基化合物及其效果分析

琼脂微球表面偶联了氨基这一易被氧化的亲和性基团,制备了对口蹄疫病毒抗原的亲和作用。以琼脂微球为基质,通过正交试验筛选、凯氏定氮法测定,选取最优偶联条件下的琼脂微球介质,用其亲和吸附口蹄疫病毒灭活抗原,检测其亲和吸附效果。结果显示:琼脂微球15g,环氧氯丙烷15mL,3 M NaOH 30mL,40℃活化反应3.5h;6mL苯乙胺,2M NaOH 20mL,50℃偶联反应2.5h。该条件偶联后的含氮量是0.4486%,得到的该介质对口蹄疫灭活病毒抗原的层析效果和某生产疫苗的药厂的分离纯化效果相似,表明该自制介质对分离纯化口蹄疫灭活病毒抗原具有一定的应用潜力。     

带His-tag的乙肝病毒融合抗原在毕赤氏酵母中的表达

乙肝病毒融合抗原SpreS1（简称SS1抗原）较S抗原有更强的免疫原性,可望有更好的临床应用前景．为了便于表达产物的纯化,利用化学合成的寡聚核苷酸链和PCR扩增,在SS1基因的5’端和3’端分别融合了6His-EK和6His的编码序列,将其重组质粒pPIC3.5k-6His-EK-SS1,pPIC3．5k-SS1-6His转化毕赤氏酵母菌GS115,筛选鉴定得到的重组酵母工程菌进行诱导培养,然后采用镍离子柱（Ni-NTA Basin）纯化表达产物,纯化的蛋白样品经ELISA效价测定、SDS-PAGE和Western Blot分析,结果表明融合抗原基因6His-EK-SS1和SS1-6His在毕赤氏酵母中均能表达,纯化的产物仍有免疫原性,所用的纯化方法简便有效．     

日本血吸虫天然抗病分子疫苗的结构与功能分析 Ⅱ.SIEA 28kDa分子的高效液相色谱分离及薄层等电聚焦分析

采用高效液相离子交换法对电泳纯SIEA28kDa抗原组分进一步分离纯化。对获得的色谱纯SIEA28kDa抗原分子进行薄层等电聚焦分析,测定其等电点。结果证明电泳纯SIEA28kDa抗原分子经高效液相离子交换法纯化后,280nm,220nm波长色谱图均为一个两侧对称,光滑的高峰曲线,色谱洗脱液对其免疫活性光密度图也为一个两侧对称,光滑的高峰曲线,色谱纯化证明SIEA28kDa抗原组分可能为单一分子,其等电点范围在pi5-pI7之间。     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

A computer generator for randomly layered structures

AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…     

海南岛地体及其毗邻陆缘晚中生代—新生代古地磁研究和构造演化

本文叙述了对海南岛及其毗邻大陆边缘白垩纪到第四纪地层岩石进行古地磁研究的全部工作过程。通过分析岩石中剩余磁矢量的磁偏角及磁倾角的变化,提出海南岛白垩纪以来经历的构造演化模式如下:早期伴随顺时针旋转而向南迁移,后期伴随逆时针转动并向北运移。联系该地区及邻区的地质、地球物理资料,对海南岛上述的构造地体运动提出以下认识:北部湾内早期有一拉张作用,主要是该作用使湾内地壳显著伸长减薄,形成北部湾盆地。从而导致了海南岛的早期构造运动,而海南岛后期的构造运动则主要是受南海海底扩张的影响。海南地体运动规律的阐明对于了解北部湾油气盆地的形成演化有重要的理论和实际意义。     

Exciton Seebeck in Molecular Systems

Various applications relevant to the exciton dynamics,such as the organic solar cell,the large-area organic light-emitting diodes and the thermoelectricity,are operating under temperature gradient.The potential abnormal behavior of the exicton dynamics driven by the temperature difference may affect the efficiency and performance of the corresponding devices.In the above situations,the exciton dynamics under temperature difference is mixed with     

Achieving Unusual States of Matter Under High Pressure

The elongation method,originally proposed by Imamura was further developed for many years in our group.As a method towards O(N)with high efficiency and high accuracy for any dimensional systems.This treatment designed for one-dimensional(ID)polymers is now available for three-dimensional(3D)systems,but geometry optimization is now possible only for 1D-systems.As an approach toward post-Hartree-Fock,it was also extended to     

《Chinese Science Bulletin》SUBJECT INDEX TO VOLUME 59(2014)

正~~     

An on-line scaling method for improving scalability of a database cluster

The explosive growth of the Internet and database applications has driven database to be more scalable and available, and able to support on-line scaling without interrupting service. To support more client's queries without downtime and degrading the response time, more nodes have to be scaled up while the database is running. This paper presents the overview of scalable and available database that satisfies the above characteristics. And we propose a novel on-line scaling method. Our method improves the existing on-line scaling method for fast response time and higher throughputs. Our proposed method reduces unnecessary network use, i.e. , we decrease the number of data copy by reusing the backup data. Also, our on-line scaling operation can be processed parallel by selecting adequate nodes as new node. Our performance study shows that our method results in significant reduction in data copy time.     

An improved R-tree based on childnode''s probability

R-Tree is a good structure for spatial searching. But in this indexing structure,either the sequence of nodes in the same level or sequence of traveling these nodes when queries are made is random. Since the possibility that the object appears in different MBR which have the same parents node is different, if we make the subnode who has the most possibility be traveled first, the time cost will be decreased in most of the cases. In some case, the possibility of a point belong to a rectangle will shows direct proportion with the size of the rectangle. But this conclusion is based on an assumption that the objects are symmetrically distributing in the area and this assumption is not always coming into existence. Now we found a more direct parameter to scale the possibility and made a little change on the structure of R-tree, to increase the possibility of founding the satisfying answer in the front sub trees. We names this structure probability based arranged R-tree (PBAR-tree).     

Integrating GIS Web services based on mediating architecture

The geographic information service is enabled by the advancements in general Web service technology and the focused efforts of the OGC in defining XML-based Web GIS service. Based on these models, this paper addresses the issue of services chaining,the process of combining or pipelining results from several interoperable GIS Web Services to create a customized solution. This paper presents a mediated chaining architecture in which a specific service takes responsibility for performing the process that describes a service chain. We designed the Spatial Information Process Language (SIPL) for dynamic modeling and describing the service chain, also a prototype of the Spatial Information Process Execution Engine (SIPEE) is implemented for executing processes written in SIPL. Discussion of measures to improve the functionality and performance of such system will be included.     

Tracking data management using topology of moving object database system for moving object constrained trajectory in LBS application

Advances in wireless technologies and positioning technologies and spread of wireless devices, an interest in LBS (Location Based Service) is arising. To provide location based service, tracking data should have been stored in moving object database management system (called MODBMS) with proper policies and managed efficiently. So the methods which acquire the location information at regular time intervals then, store and manage have been studied. In this paper, we suggest tracking data management techniques using topology that is corresponding to the moving path of moving object. In our techniques, we update the MODBMS when moving object arrived at a street intersection or a curved road which is represented as the node in topology and predict the location at past and future with attribute of topology and linear function. In this technique, location data that are corresponding to the node in topology are stored, thus reduce the number of update and amount of data. Also in case predicting the location,because topology are used as well as existing location information, accuracy for prediction is increased than applying linear function or spline function.