正交实验法在PCR反应条件优化中的应用

本研究以小肠结肠炎耶尔森氏菌为研究材料,将纯理论数学方法一正交设计实验法。应用于分子生物学技术PCR体系的优化之中．首先采用搜索性正交实验,在此实验结果基础上。进一步设计细调性正交实验来寻求PCR反应的最适条件．所得优化扩增体系稳定,重复性好,表明正交实验设计方法高效快速,科学性强,是PCR反应条件优化中值得推广使用的有效方法。     

Effect of Physical Factors on Bacterial Distribution in the Jordan River Mouth Area in Israel

The Jordan River mouth is an area of complicated hydrography and variable and vulnerable ecology. It has changeable water and sediment regimes, for example, large spatial gradients of biological, chemical, physical, and hydrological characteristics. The Jordan River enters Lake Kinneret in the north and carries with it sewage from the Upper Galilee; thus the river is the major source of enteric bacteria in the lake. This research is based on theoretical analysis as well as on biological-hydrodynamical measurements along a discharging free, turbulent jet flow. Three components of velocities were recorded downstream from the exit cross-section for a distance of 700 m, and the mean streamwise velocity turbulence intensity and the turbulence scale were calculated. The measuring devices were an original three-dimensional velocity-fluctuation meter. Results of these measurements show that the Jordan River flows through the whole transect until it reaches the crest of a bar. After this, there is a separated exponential flow from the bottom and upper layers for ~100m after the bar. Fecal coliforms, Escherichia coli, and Klebsiella spp. were enumerated as colony-forming units from samples taken along the Jordan River path as it entered Lake Kinneret. Bacterial numbers were similar in surface and bottom waters in front of the bar. Behind the bar, however, there was a sharp decrease of bacterial numbers in the surface water, probably because of photooxidative stress. Despite the more protective environment of the bottom waters, the numbers here also decreased, indicating that the bar is a physical barrier for bacterial distribution. Furthermore, we found a significant effect of the flow velocity of Jordan River water on the bacterial distribution in Lake Kinneret. When the velocity is high, bacteria are distributed over a longer distance, regardless of their numbers in Jordan River mouth.     

Spherical Boundary Conditions: A Finite and System Size Independent Geometry for Simulations of Electrolytic Liquids

The system size dependence of the thermodynamic properties of electrolytic systems in "Spherical Boundary Conditions" (SBC) have been examined in this work. Coulombic systems were simulated with different system sizes (N ) for a wide range of concentrations, different ionic charges and solvent permittivities. The effects of system size upon the mean internal energy ?U? and mean ionic activity coefficient ?γ _±? values were determined. Our results indicated that there was no dependence of the thermodynamic properties upon system size in the non-Euclidean geometry. Different methods of extrapolating the thermodynamic properties to infinite numbers were studied in detail. In contrast to the Euclidean geometry, the method of extrapolating ?U? or ?γ _±? versus N ^-1, N ^-2/3 and N ^-1/3 was not statistically justifiable nor advantageous. Therefore, SBC is well suited to simulating systems involving long-range electrostatic interactions because the results do not dependent upon system size.     

Induction and Analysis of Epithelial to Mesenchymal Transition

Epithelial to mesenchymal transition (EMT) is essential for proper morphogenesis during development. Misregulation of this process has been implicated as a key event in fibrosis and the progression of carcinomas to a metastatic state. Understanding the processes that underlie EMT is imperative for the early diagnosis and clinical control of these disease states. Reliable induction of EMT in vitro is a useful tool for drug discovery as well as to identify common gene expression signatures for diagnostic purposes. Here we demonstrate a straightforward method for the induction of EMT in a variety of cell types. Methods for the analysis of cells pre- and post-EMT induction by immunocytochemistry are also included. Additionally, we demonstrate the effectiveness of this method through antibody-based array analysis and migration/invasion assays.     

EMOST: Report about the application of low-frequency and intensity electromagnetic fields in disaster situation and commando training

Recently, we published our results (Bókkon et al., 2011. Electromagn Biol Med.) regarding the effectiveness of the EMOST (Electro-Magnetic-Own-Signal-Treatment) method for the reduction of phantom limb pain under clinical circumstances. However, EMOST treatments not only significantly reduced phantom pain, but that most of the patients also reported about additional benefits such as improvement of their sleep and mood quality after treatments. Here we report some unusual applications of EMOST method under special situations. That is, we report about our effective EMOST treatments of humans under catastrophic conditions and commando training course. This article points out that it is reasonable to apply biophysical electromagnetic management under unique circumstances. We also report some preliminary experiments on 12 members of our BioLabor regarding the effectiveness of single EMOST treatment on some serum parameters and electrocardiogram.     

舌苔细菌PCR-DGGE分析条件的建立与优化

目的针对口腔舌苔细菌16S rDNA序列进行变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)适用序列的筛选及电泳条件的优化。方法以DGGE图谱的高分辨率为指标,选择舌苔细菌DGGE分离最适的16S rDNA高变区、电泳电压和电泳时间,并采用优化的条件分析健康青年人舌苔细菌群落的分布。结果舌苔细菌16S rDNA V3高变区引物序列能获得更加丰富清晰的DGGE条带;基于该区,当变性剂浓度为30%～60%、电泳温度60℃、电压60 V和电泳时间14 h时能得到分辨率最佳的DGGE图谱。运用此优化条件对12个样本舌苔细菌群落的分析表明,舌苔微生物主要由厚壁菌门、梭杆菌门、拟杆菌门和变形菌门等组成。优化后的DGGE技术对舌苔细菌多样性的分析具有准确性、灵敏性和可重复性。结论 DGGE图谱显示,不同分析条件对图谱类型和细菌多样性指数均有所差异。利用优化的DGGE条件能有效分离舌苔细菌16S rDNA V3区序列,为口腔微生物群落结构分析提供可靠的技术支持,也为其他不同生态细菌的多样性分析提供参考。     

The birth of the Acheulian techno-complex in the Caucasus region

The lithic industries of the sites Kermek, Ciscaucasia (2.1–2.0 Ma) and Karakhach, South Caucasus (1.85–1.78 Ma) contain almost all the components of the Acheulian, indicating that this techno-complex appeared in the different areas of the Caucasus region not later than at the end of the Gelasian. Such extremely old ages of these industries as well as a number of their features due to the influence of the local raw material (tabular clasts) suggest that they were formed independently of the Early Acheulian industries known in Africa and the Near East. The very early appearance and further dispersal of Early Acheulian in the Caucasus can be explained by the presence there of raw materials with qualities supporting the production of large tools inherent of Acheulian, and by the very favorable natural conditions of the region in the final Gelazian-initial Calabrian (low or medium altitude terrain with subtropical climate and savanna-like landscape).     

嗜热菌Anoxybacillus sp．DL3产蛋白酶条件及其酶学性质研究

目的：对Anoxybacillussp．DL3的产蛋白酶条件及其酶学性质进行研究,为下一步进行蛋白酶基因的克隆、表达提供依据。方法：应用常规方法液体培养细菌,研究温度、pH、培养基中碳源、氮源对菌株产蛋白酶的影响,硫酸铵盐析的方法提取酶液,并采用Folin法测酶活性。用紫外分光光度计在OD680hi／1测吸光值。并对提取的蛋白酶液进行酶的最适温度、pH以及酶的热稳定性和pH稳定性研究,向酶液中添加金属离子和EDTA、PMSF,研究其对酶活性的影响。结果：在培养基初始pH是6．5,培养温度为40℃时菌株产酶酶活性最大;培养基中以乳糖为碳源,酵母膏和硫酸铵为氮源,碳源与氮源的比例为1：2时,酶活最大。酶学性质研究结果显示：该酶的最适反应温度是55℃,最适反应pH是7．0;在50℃保温20min-80min内,酶活力下降幅度较小。60℃保温60min后,仍保持约60％的酶活。70℃保温60min后,残余酶活为30％。该酶在pH为6．0～8．0范围内,相对酶活差别不是很大,下降趋势大致相同。在强碱条件下,相对酶活下降很明显。Fe2＋、Cu2＋和Hg2＋对酶活性有明显的抑制作用;Ca2＋、Mg^2＋、Mn2＋等金属离子对酶活性有明显的促进作用;乙二胺四乙酸（EDTA）和苯甲基磺酰氟（PMSF）对酶活性也有一定抑制作用。结论：Anoxybacillussp．DL3所产的蛋白酶为嗜热中性蛋白酶,此酶具有较好的热稳定性和pH耐受性,该菌株具有进一步开发、利用的价值。     

云南传统发酵豆豉中产β-半乳糖苷酶乳酸菌的筛选及其产酶条件的研究

目的从云南豆豉样品中筛选产β-半乳糖苷酶的乳酸菌,并对其产酶条件进行研究。方法从云南省元阳、红河、建水、石屏等地采集豆豉样品,并从中分离得到355株微生物。结果经明胶诱导、脱脂乳平板实验,复筛得到87株蛋白酶产生菌,从中筛选产β-半乳糖苷酶的乳酸菌。通过X-Gal平板实验,共获得34株产β-半乳糖苷酶菌株,通过酶活测定,最终筛选得到1株高产β-半乳糖苷酶菌株GJ-1-3L,经16S rDNA序列分析鉴定为短乳杆菌;GJ-1-3L在以葡萄糖为碳源、多聚蛋白胨为氮源、起始pH 6.5的MRS培养基中,接种量为4%,35℃发酵培养12 h,其β-半乳糖苷酶活性高达6.73 U/mL,Cu2＋、Ba2＋对酶活有抑制作用,而K2HPO4、MgSO4则能促进酶活。结论 GJ-1-3L菌株来源于豆豉,能够产生β-半乳糖苷酶发酵乳糖,同时产生乳酸,其在食品与乳品加工等方面具有很好的应用前景。     

Studies on Strains of Trichogramma evanescens Westwood from Different Regions of Eurasia

Strains of Trichogramma evanescens Westwood (Hymenoptera: Trichogrammatidae) collected from six different geographical regions in Eurasia were compared in terms of their biological and morphological characteristics and electrophoretic patterns. Significant differences were observed between the strains for biological characteristics such as the proportion of female progeny, developmental period, longevity, proportion of adults emerging and fecundity. Strains from the Altai region in Russia were more fecund at all hygrothermal conditions in which they were reared (27 C, 80% relative humidity (RH); 27 C, 40% RH; 23 C, 80% RH; and 23 C, 40% RH). Poor adult emergence took place at 27 C and 40% RH in strains from Moldavia and Slovakia. Morphological examination of male antennae showed that strains from Moldavia and Slovakia had the highest ratio of the length of antennal hairs to the maximum width of flagellum. The patterns of esterases after electrophoresis indicated that strains differed in the mobility and intensity of esterase bands. Electrophoretic and morphological relationships among the strains did not correspond with their biological relationships. The possibility of the use of strains from the Altai region in Russia in biocontrol programmes in southern parts of Europe is discussed.