电泳的

Methods The DNA damage in CHL cells processed by salted aconite root extractive and aconitine by SCGE was detected.

方法采用单细胞凝胶电泳法检测盐附子、乌头碱对CHL细胞DNA即刻损伤的影响。

Actomyosin were purified from human heart, and further isolated into CMLC with urea degradation and ammonium sulfate precipitation, CMLCⅠ and CMLCⅡ were purified later from this CMLC by the improved PAGE method, both CMLCⅠ and CMLCⅡ were without any protein contamination.

从人心肌提取肌动球蛋白，用尿素降解法和硫酸铵沉淀法从肌动球蛋白中分离、纯化CMLC，应用改进的聚丙烯酰胺凝胶电泳法从总CMLC中分离CMLCⅠ和CMLC Ⅱ，二者均不含有任何杂蛋白。

Three methods,the electrophoresis aggradation,brushed coating and water cooking method were presented to deposit MoS2 coatings in the material surfaces,and the deposited mechanisms were analyzed.

选用电泳沉积法、刷涂法和水煮法3种方法在材料表面制备了二硫化钼涂层，并探讨了3种方法形成涂层的机制。

An amidase from a strain Bacillus cereus ZJB-07112 was purified to homogeneity by using sonication, anion-exchange chromatography, phenyl-sepharose chromatography.

用一株蜡状芽孢杆菌新菌株ZJB-07112 ( Bacillus cereus ZJB-07112)发酵生产酰胺酶，经超声波细胞破碎、High Q阴离子色谱、Phenyl-Sepharose疏水色谱等步骤获得了凝胶电泳均一的酰胺酶。

A amidase from a strain Bacillus cereus ZJB-07112 was purified to homogeneity by using of the sonication, anion-exchange chromatography, Pheny1-Sepharose chromatography.

用一株蜡状芽孢杆菌新菌株ZJB-07112 (Bacillus cereus ZJB-07112)发酵生产酰胺酶，经超声波细胞破碎、High Q阴离子层析、Phenyl-Sepharose疏水层析等步骤获得了凝胶电泳均一的酰胺酶。

A basic knowledge of microbiology, will be detected with a microscope observation of micro-organisms, medium Preparation, disinfection and sterilization, separation and purification of micro-organisms, bacteria counting method A chemical analysis of the theoretical knowledge and can use chemical analysis methods and equipment of the components of material analysis Master fermentation process principles, understanding of beer, liquor, fermentation production technology Master protein related knowledge, PAGE gel electrophoresis technology, high-speed centrifugal Genetic engineering master the basic theory of knowledge, the extraction and purification of genes, PCR principle, gene recombination technology Master of the basic theoretical knowledge, the immobilized enzyme, the enzyme preparation Application Master of animal and plant tissue culture techniques of experimental steps studied experimental apparatus: Balance (scales, electronic scales, Analytical Balance), or acidity, spectrophotometer, liquid gas chromatography, acid-base titration apparatus, microscopes, centrifuges, ultrasonic cleaning, constant temperature oscillation incubator, super-clean table, steam sterilization pot, the pot constant temperature water bath, PAGE device apparatus, oven, fermenter, such as PCR instrument.

具备微生物理论基础知识，会用显微镜检测观察微生物，培养基的制备、消毒与灭菌，分离与纯化微生物，细菌的计数法具备分析化学的理论知识，能运用化学分析和仪器分析方法对物质的组分进行分析掌握发酵工艺原理，了解啤酒、白酒、发酵生产工艺掌握蛋白质相关知识，PAGE凝胶电泳技术，高速离心掌握基因工程的基本理论知识，基因的提取和纯化，PCR技术原理，基因重组技术掌握酶的基本理论知识，酶的固定化方法，酶制剂的应用掌握动植物组织培养技术的实验步骤学过的实验仪器：天平（台天平，电子天平，分析天平）、酸度计、分光光度计、液气相色谱仪，酸碱滴定仪器，显微镜，离心机，超声波清洗器、恒温振荡培养箱、超净工作台，蒸气灭菌锅、恒温水浴锅，PAGE电泳装置仪器、干燥箱、发酵罐、PCR扩增仪等。

Through designing of composition and structure of the bioactive graded coating,innerstress and its distribution in the coating were analyzed and calculated, the resultsshowed that when composition distribution coefficient n was 1.5, a reasonable stressdistribution could be got, that was at the beginning of deposition the suspension containingrichly BG granules was used so that a rich BG granules layer, a good transitional layerbetween BG layer at the bottom and the coating could be obtained at the titanium alloy side,the bottom of the coating; the stress value near the interface and surface and its character,pressure stress or tensile stress, were decided by the character of its composition itself.Changing composition distribution coefficient n could only change the variation tendency ofstress in the coating, but did not change the stress distribution rule in the coating. Thethinner the coating is, the sharper stress variation in the coating is, which does not mean thatthicker coating is better because the thicker the coating is, the little the permitteddeformation of coating is, so the coating thickness should be thinner, for example, about50μm for bending applications, but for applications only bearing pure shear stress, such asroot of tooth implant, the coating can be thicker little, for instance, about 80～100μm. The study on electrification characteristic and electrophoresis deposition of HAand BG granules in aqueous and non-aqueous solution system found that EPD almost didn'toccur in aqueous solution system. However, because HA granules take position charges inabsolute alcohol, a homogeneous EPD be carried out on the cathode titanium alloy slice, but taking negative charge in absolute alcohol the BG granules not be deposited on the cathode. A guided HA crystallizing, 100～300nm, on surface of the BG granules be realized by metathetical reaction, which cover BG granules with HA microcrystals and make the covered BG granules taking position charges in absolute alcohol, sequentially realize the EPCD of the BG and HA granules on the cathode, so it is feasible to make a titanium alloy/BG/HA bioactive graded coating by making use of EPCD technology. The corrosion experiment of rich boron bioglass coating and plasma spray coating showed that split phase, rich boron and rich silicon phase, occurred during its preparation. In basic medium the corrosion behavior of 〓 BG coating showed uniformity corrosion, the corrosion mostly occurred at rich boron phase area, therefore batch formula design of BGshould avoid the occurring of split phase. The corrosion appearance of plasma spray coatingappeared a non-uniform corrosion, mostly occurred at the edge of the laminated HA moltendrops, and emerged an accelerated corrosion tendency, which will easyly lead to corrosioncrackles extending to the interface and the happening of osmotic interfacial corrosion, thatmay be one of the major reasons leading to the coating cracking-off in the later period. Thetesting results of thermal expansion coefficient of 〓 and 〓BG showed the thermalexpansion coefficient of 〓 BG matched with that of titanium alloy better, and 〓 BG couldsinter with titanium alloy into densification enamel layer at low temperature (720℃).

将Ti6Al4V合金在1000℃下进行真空热处理会降低其力学性能，且合金内的V元素会向表面富集，因此，钛合金真空热处理和表面涂层的烧结温度不能过高，即应低于其相转变点；通过对生物活性梯度涂层的组成和结构的设计，分析和计算了梯度涂层内的应力大小和分布，结果表明：对于本研究，当成分分布系数n=1.5时，可以获得较合理的涂层力学性能，即在沉积开始时，采用富含BG颗粒的悬浮液，以便在钛合金侧获得同底层BG有良好过渡的富BG涂层；梯度涂层界面和表面的应力大小、性质由材料组成本身的性质决定，改变成分分布系数，只能改变涂层内应力变化的趋势；涂层的厚薄不影响涂层内的应力分布规律，但涂层越薄，涂层内的应力变化越快，但这并不意味着涂层越厚越好，因为涂层越厚，涂层允许的变形越小，对于应用于弯曲受力部位的涂层而言，涂层应薄一点为好(50μm)；而对于仅纯受剪切应力的部位，如牙根种植体，涂层可适当加厚(80～100μm)；通过对HA和BG颗粒在水溶液体系和非水溶液体系中的带电特性和电泳沉积的研究发现，它们在水溶液体系中很难发生电沉积；在无水乙醇溶液中，HA颗粒带正电，可在阴极钛合金片上发生均匀的电泳沉积，而BG颗粒则带负电荷；利用复分解反应法，可以制得100～300nm的HA，通过诱导HA在BG颗粒表面结晶，可对BG颗粒进行表面包覆，获得了被HA包覆的BG颗粒，改变了BG颗粒表面的带电特性，使BG和HA颗粒在无水乙醇中均带上正电荷，从而实现了HA和BG颗粒在阴极上的共沉积。

After the cell growth curves was recorded, RPE cells of the 3-5th passages were utilized. 2、Three different siRNA (siRNAl,siRNA2,siRNA3) targeting against human cx43 gene and one negative control siRNA were designed and transfected into cultured human RPE cells via liposome reagent. The most effective siRNA can be determined by semi-quantitative reverse transcription PCRRT-PCR. 3、To the most effective siRNA, after transfected into human RPEs with different concentration, the cellular proliferate activities were messured by MTT colorimetry ; the percentages of RPE in different cell circle phase was assayed by FCM; the changes of phenotypical properities were observed with SCM; the protein expression of cx43 was studied through immunocytochemistry stain and Weston blot; the communication intercellular was calculated with FRAP; and the ability of recovery was assessed by using an in vitro wound healing model.4、The total proteins of siRNA1 and RPE were seperated by two-dimensional gel electrophoresis and visualized by silver staining. Proteins with significant expression alterations were selected and their peptide mass fingerprints (PMFs were obtained by matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS).The PMFs were used to search NCBInr database by Auto MS-Fit software.

实验方法：1、培养原代的人RPE细胞，经过细胞角蛋白、S-100和神经胶质原纤维酸性蛋白免疫细胞化学鉴定后，通过AO/PI染色技术确定培养细胞的存活率，描记其生长曲线，第3-5代用于以下细胞实验2、生物合成针对人cx43基因的三条小干扰RNA和一条阴性RNA通过脂质体转染RPE细胞后，通过RT-PCR的方法确定抑制效率最高的干扰片断3、将该片段以不同浓度通过阳离子脂质体转染培养的人RPE细胞后，采用MTT法观察其对细胞的增殖力的作用；通过流式细胞仪观察其对细胞周期的影响；通过扫描电镜观察其对细胞形态的影响；通过免疫细胞化学和Weston blot观察其对cx43蛋白表达的作用；采用激光共聚焦和荧光淬灭恢复技术观察荧光恢复速率平均百分率，评价其对细胞间通讯功能的影响；通过制作RPE细胞损伤模型，观察其对损伤修复能力的作用4、分离纯化转染siRNA的RPE组和正常对照组RPE细胞的全部蛋白质，应用等电聚焦电泳和SDS-PAGE双向电泳技术，银染显示分离出的蛋白质斑点，经凝胶图像分析软件对两个样本进行胶图分析，寻找差异蛋白点。

Results of non-reducing sodium dodecyl sulphate polyacrylamide gel electrophoresis of the supernatant and aggregate precipitate formed in refolding process show that except being refolded to native egg white lysozymes, the reduced-denatured lysozymes can also form the aggregates with molecular weights being separately about 30.0 and 35.0 kD, while the reducing SDS-PAGE and the refolding results in the presence of sodium dodecyl sulphate show that these aggregates are formed chiefly through the misconnection of disulfide bonds between the reduced-denatured lysozymes, and the aggregate precipitates are formed through the non-covalent interactions between the aggregates with molecular weight being about 30.0 kD.

复性过程形成的上清和集聚体沉淀的非还原电泳结果表明，除了能复性成天然态的溶菌酶分子外，还原变性蛋白溶菌酶同时还能形成分子量分别约为30.0KD和35.0KD的蛋白溶菌酶分子集聚体；而它们的还原电泳和在SDS存在时的复性结果表明，这些集聚体主要是通过还原变性蛋白溶菌酶分子间的二硫键错配而形成，而集聚体沉淀则是通过分子量约为30.0KD的集聚体分子之间的非共价相互作用而形成。

In our research, through derivatization of stable calix[4]pyrroles, active substituting units were jointed on the calixpyrrolic skelecton. After acylamide and free radical transfer reaction, the preparation of new calix[4]pyrrole-bonded stationary phases were fulfilled. the separation abilities of them to inorganic and organic anions and medicines were exploited and new analytical methods were established. The separation mechanisms were discussed. At the same time, through using calix[4]pyrroles as additives in CZE to influence the electrophoresis of inorganic anions, the recognition mechanisms and influencing factors in supramolecular chemistry were also studied.

本项目以新型杯吡咯大环化合物分子识别化学研究为基础，设计合成了含活泼取代基的官能化杯[4]吡咯化合物，通过酰胺化反应和原位自由基链转移反应制备了新型的杯[4]吡咯色谱固定相，系统研究了具有阴离子识别功能的杯[4]吡咯大环化合物作为离子色谱的分离介质对无机、有机离子型化合物和药物分子的色谱分离性能，建立了新的分析方法，对分离机理进行了探索和验证；同时将杯[4]吡咯化合物用作毛细管电泳流动相添加剂，利用其对无机阴离子电泳行为的影响探讨超分子化学中分子识别的机理和影响因素。

Breath, muscle contraction of the buttocks; arch body, as far as possible to hold his head, right leg straight towards the ceiling (peg-leg knee in order to avoid muscle tension).

呼气，收缩臀部肌肉；拱起身体，尽量抬起头来，右腿伸直朝向天花板（膝微屈，以避免肌肉紧张）。

The cost of moving grain food products was unchanged from May, but year over year are up 8%.

粮食产品的运输费用与5月份相比没有变化，但却比去年同期高8％。