一种用磷酸钙法高效转染HEK 293T细胞方法的建立

利用磷酸钙转染法将lacZ-pShuttle质粒导入HEK 293T细胞中,实验证明,pH值、DNA纯度、转染后培养时间和甘油休克时间是影响转染效率的重要因素.转染后培养6h进行甘油休克效果最佳,比培养0h直接进行休克转染效率升高584.4%;甘油休克时间为4.5min时转染效果最佳,比不用甘油休克转染效率升高130.8%.通过对影响转染的几个条件进行优化,建立了1种可以高效转染HEK 293T细胞的方法,且简便易行.     

Rapid detection of virulent protease secreted by Vibrio anguillarum by dot enzyme-linked immunosorbent assay

Dot enzyme-linked immunosorbent assay （dot-ELISA）, indirect ELISA and Westem blot were performed to detect the virulent protease secreted by Vibrio anguillarum which was isolated from the diseased left-eyed flounder, Paralichthys olivaceous. Sensitivity results showed that dot-ELISA is a more sensitive, rapid and simple technique for the protease detection. The minimal detectable amount of protease is about 7 pg in the dot-ELISA test, while 7.8 ng in the indirect ELISA and 6.25 ng in the Westem blot respectively. Protease could be detected 2 h after incubation of V. anguillarum in the 2216E liquid medium but enzyme activity was very low at that period. From 6 to 12 h, the amount and enzyme activity of protease increased markedly and reached maximum at stationary phase. Analysis of serum samples periodically collected from the infected flounders showed that after 2 h of infection by V. anguillarum, the pathogenic bacteria could be detected in the blood of the infected flounders but no protease was found. It was 5-6 h after infection that the protease was detected in blood and then the amount increased as infection advanced. Quantitative detection of protease either incubation in the medium or from the blood of infected flounders could be accomplished in virtue of positive controls of quantificational protease standards （＂marker＂） so that the alterations ofprotease secretion both in vitro and in vivo could be understood generally. In addition, the indirect ELISA and dot-ELISA were also performed to detect V. anguillarum cells. Results indicated that the sensitivity of indirect ELISA to bacteria cells is higher than that of the dot-ELISA, and that the minimal detectable amount is approximately 10^4 cell/mL in the indirect ELISA, while 10^5 cell/mL in the dot-ELISA.     

天然热释光技术在海洋油气田勘查中的应用

沉积物热释光能有效地反映海洋环境下深部油气田引起的微弱放射性异常。热释光在不同的测试粒级、不同的岩性中的丰度不同，结果为粗粒级＞细粒级、砂岩＞粉砂质砂岩＞泥质粉砂岩＞泥岩。热释光也与部分金属元素含量有关，Ti、Ba、Zn、Fe^3 /Fe^2 与热释光呈正相关关系。热释光与烃类也呈较强的正相关性。不同采用深度热释光测量结果表明，热释光剂量随沉积物的沉积深度和沉积时间的增加而增加，深层样的热释光对油气藏的指示效果更好。     

Fully nonlinear numerical wave tank (NWT) simulations and wave run-up prediction around 3-D structures

A finite-difference scheme and a modified marker-and-cell (MAC) algorithm have been developed to investigate the interactions of fully nonlinear waves with two- or three-dimensional structures of arbitrary shape. The Navier–Stokes (NS) and continuity equations are solved in the computational domain and the boundary values are updated at each time step by the finite-difference time-marching scheme in the framework of a rectangular coordinate system. The fully nonlinear kinematic free-surface condition is implemented by the marker-density function (MDF) technique developed for two fluid layers.To demonstrate the capability and accuracy of the present method, the numerical simulation of backstep flows with free-surface, and the numerical tests of the MDF technique with limit functions are conducted. The 3D program was then applied to nonlinear wave interactions with conical gravity platforms of circular and octagonal cross-sections. The numerical prediction of maximum wave run-up on arctic structures is compared with the prediction of the Shore Protection Manual (SPM) method and those of linear and second-order diffraction analyses based on potential theory and boundary element method (BEM). Through this comparison, the effects of non-linearity and viscosity on wave loading and run-up are discussed.     

黄河流域2002年麦收期间一次强降水过程分析

本文通过对陕西南部的佛坪县2002年6月8日——lO日的降水达到了489mm的特大暴雨降水过程的初步分析，对进一步了解大尺度环流背境场下的强降水过程的成因，提高对暴雨天气过程的预报能力是非常有益的；本文收集的降水过程期间的天气图和传真图资料对预报员和海洋工程技术人员有参考价值。     

Experimental Investigation into the performance of the Axial-Flow-Type Waterjet according to the Variation of Impeller Tip Clearance

The tip clearance inside the duct from the tip of the impeller is very important to the performance of waterjet systems, which fact has been proven in the pump field. The tip clearance is especially important on the model scale because it is very difficult in manufacture to keep the tip clearance constant and minimally small along the inside of the duct. In the present study, a flush-type waterjet propulsion unit (duct, impeller, stator, and nozzle) was designed for an amphibious tracked vehicle. Two impellers of different inner diameter were designed and manufactured in order to investigate the gap effect. Resistance and self-propulsion tests with a 1/5-scale model were conducted in PNU towing tank. The flow rate at the nozzle exit, the static pressure at the various sections along the duct and also the nozzle, the revolution of the impeller, and the torque, thrust, and towing forces at various advanced speeds were measured. Based on these measurements, the performance was analyzed according to the ITTC 96 standard analysis method. Based on this analysis method, the full-scale effective and delivered power of the tracked vehicle was estimated according to the variation of tip clearance.     

青岛近海地区SO2和SO42-干沉降速率的季节变化

用小麦岛和团岛观测站提供的青岛近海气象和水温资料,以适于SO2气体和硫酸盐(SO42-)气溶胶的干沉降模型研究了模型中的输送阻尼对两种污染物质干沉降速率的贡献,并计算了2003年青岛近海地区两种污染物质干沉降速率的季节变化。结果表明:对SO2而言,粘性副层传输系数对沉降速率的贡献大于空气动力学传输系数;对于SO42-,空气动力学传输系数对沉降速率的贡献要大于表面传输系数。SO2和SO42-的干沉降速率变化范围分别为0.187~0.868cm/s和0.188~0.532cm/s。两种污染物质干沉降速率的四季变化有相似的规律,即冬季>秋季>春季>夏季。     

黄茅海河口湾现代动力地貌体系和冲淤过程分析

1980—1993年对黄茅海河口湾进行沉积物采样和水流测定及水深测量。根据水动力和地形条件，冲淤分析及Mclaren模型研究河口湾的动力地貌体系、冲淤特征和现代沉积物运移。结果表明：（1）水下地形主要为下泄流或上溯流控制的“深槽－槽沟－浅滩－湾口”的动力地貌体系，反映了河口湾“东进西出”的水流格局；（2）整个河口湾以淤积为主，只有崖门深槽有较明显的优势冲刷特征，并随着崖门深槽向海推移和河口湾“东进西出”水动力作用，黄茅海落潮三角洲相应向西南进积；（3）应用Mclaren模型揭示了黄茅海河口湾现代沉积物运移规律，同样反映了河口湾具有“东进西出”的运移趋势。     

Application of the first internal transcribed spacer(ITS-1) of ribosomal DNA as a molecular marker to population analysis in farrer''s scallop Chlamys farreri

Sequence variation of the first internal transcribed spacer of ribosomal DNA （ ITS - 1 ） was examined and its application to the study of genetic variation was explored in four populations of farter＇ s scallop Chlamys farreri. ITS - 1 fragments, with a length of about 300 bp,of 78 individuals collected from Dalian, Qingdao, Yantai in China and Korea respectively were amplified via PCR, cloned and sequenced. Intra-genomic variation was examined by sequencing several clones of single individuals. Alignment and polymorphism analysis detected 44 haplotypes and 50 polymorphic sites which consist of 30 substitutions and 20 indels, indicating a high level of polymorphisms. Sequence analysis also showed a very low level of intra-individual variation. All these features validated the feasibility of application of ITS - 1 fragment to population analysis. Polymorphism analysis showed that the Korea sample has the richest genetic variation, followed by Yantai and Qingdao samples. AMOVA （analysis of molecular variance） showed that the majority （96.26%） of genetic variation was distributed within populations and 3.74% resulted from among populations, but with P 〈 0.05 （ = 0.042）, indicating that the populations in this study have significant divergence. This output was basically concordant with the result arising from RAPD data and different from that from mitochondrial 16S rDNA sequence data. Discussion on this inconsistency was made accordingly.     

3种蛏类线粒体16SrRNA和COI基因片段的序列比较及其系统学初步研究

对3种蛏类大竹蛏（Solen grandis），长竹蛏（Solen strictus）和小刀蛏（Cultellus attenuatus）的线粒体16SrRNA和COI基因片段序列进行了比较并对其系统学进行了初步研究。得到的序列总长度分别为472-481bp（16S）和658bp（COI）。3种蛏序列的碱基组成均显示出较高的A＋T比例（16SrRNA基因62．1％；COI基因62．8％）。对位排序比较表明，16SrRNA片段种内个体间变异较小，3种类间存在128个碱基变异位点（其中包括127个简约信息位点）和5个插入／缺失位点，总共12个碱基长；COI片段有200个碱基存在变异，其中包括191个简约信息位点，不存在任何插入／缺失位点。数据分析结果表明16SrRNA和COI基因片段大竹蛏与长竹蛏两片段的遗传距离分别为0．0856和0．1712，两竹蛏类与小刀蛏的遗传距离分别为0．3054，0．2798和0．2662，0．2933。作者认为小刀蛏与竹蛏之间的遗传距离已达到科之间的水平，结果支持将其提升为刀蛏科的分类观点。3种蛏类线粒体16SrRNA和COI基因在种间明显的多态性，证实了16SrRNA和COI基因序列均普遍适用于蛏类种及以上阶元的系统学分析。