使纯化

Morphological conversion from spiralto coccoid forms occurred when HP was cultured with selenium and most of coccoid forms had fragmentary flagella and pilus; the effection of selenium to activity of urease, catalase, alkaline phosphatase is not exist or not evidence; selenium enable pET32a-vacA-E.coli BL21 which can express VacA produce yellow lipochrome, and the color become deeper with increasing of concentration of selenium; selenium can decrease expression of recombined VacA by restrain the growth of pET32a-vacA-E.coli,but can not influence activity and gene sequence of VacA; selenium can enhance the suppression effect of recombined VacA for gastric cancer cell, depress the index of cell multiplication and the forming rate of colony, decrease the number of cell in S period, increase total of cell in G period; After purified, the recombined VacA can be identified by VacA antibody.

体外实验结果表明：一定浓度的硒作用HP后，HP菌落数量随着硒浓度的增加而减少，菌落颜色由灰白色变为砖红色，溶血现象随硒浓度的增加和作用时间的延长而逐渐消失，HP的形态由弯曲形变为球形或椭圆形，鞭毛及菌毛出现脱落现象；硒对尿素酶、过氧化氢酶、碱性磷酸酶活性无影响或影响不明显；一定浓度的硒能使表达VacA的pET32a-vacA-E.coli BL21工程菌产生脂溶性砖红色色素，颜色随硒浓度增加而加深；一定浓度的硒通过抑制pET32a-vacA-E.coli BL21工程菌的生长，减少重组VacA表达量，但不影响其空泡毒活性及编码基因序列；纯化后的VacA重组蛋白可被VacA抗体识别，具有良好的抗原性。

We also discovered that selenium can decrease hematolysis and effects function of vacuole toxin of HP. morphological conversion from spiralto coccoid forms occurredwhen HP was caltured with selenium and most of coccoid forms had fragmentary flagella and pilus; the effection of selenium to activity of urease, catalase, alkaline phosphatase is not exist or not evidence; selenium enable pET32a-vacA-E. coli BL21 which can express VacA produce yellow lipochrome, and the color become deeper with increasing of concentration of selenium; selenium can decrease expression of recombined VacA by restrain the growth of pET32a-vacA-E. coli, but can not influence activity and gene equence of VacA; selenium can enhance the suppression effect of recombined VacA for gastric cancer cell, depress the index of cell multiplication and the forming rate of colony, decrease the number of cell in S period, increase total of cell in G period; After purified, the recombined VacA can be identified by VacA antibody.

结果：体外实验结果表明：一定浓度的硒作用HP后，HP菌落数量随着硒浓度的增加而减少，菌落颜色由灰白色变为砖红色，溶血现象随硒浓度的增加和作用时间的延长而逐渐消失，HP的形态由弯曲形变为球形或椭圆形，鞭毛及菌毛出现脱落现象；硒对尿素酶、过氧化氢酶、碱性磷酸酶活性无影响或影响不明显；一定浓度的硒能使表达VacA的pET32a-vacA-E.coli BL21工程菌产生脂溶性砖红色色素，颜色随硒浓度增加而加深；一定浓度的硒通过抑制pET32a-vacA-E.coli BL21工程菌的生长，减少重组VacA表达量，但不影响其空泡毒活性及编码基因序列；硒能增强重组VacA对胃癌细胞的抑制作用，使细胞分裂指数和集落形成率降低、S期细胞数量减少，G1期细胞总数增加；纯化后的VacA重组蛋白可被VacA抗体识别，具有良好的抗原性。

A set of isolation and purification methods will be established, so it can be used for the separation of the trace amount compound.

同时建立一套分离纯化方法，使其能普遍应用于难于分离的微量化合物的分离纯化。

If you thought they didn't make albums like Neil Young's "Everybody Knows This is Nowhere" anymore, cue up "Ghosts of the Great Highway," and marvel over the fact that Kozelek and company are able to cross-pollinate folk, country, punk, and psychedelic influences without sounding the least bit retro, stealing the purifying flame of Crazy Horse meltdowns like "Cortez the Killer" while sounding like no one but themselves.

你认为，他们不象尼尔杨一样使集子"，每人知道这无处"再，向上的暗示是"大的公路的鬼线"，并且越过那些事实的奇迹那Kozelek 和公司能够穿过受粉人们，国家，朽木和引起幻觉影响没有发出那些重新流行一点儿最小，在只听上去是他们自己时象"科尔特斯消除剂"一样偷使燃烧纯化不含的发疯的马熔化。

First of all, this study will be multi-walled carbon nanotubes by chemical oxidation process purified so it can be modified to deal with than the purification of purity before the modified 5 wt%, more than 1.89 times the surface area to increase the surface potential decrease of about 10 ~ 20 mV and the surface functional base (-COOH and-OH) to increase 1.45 times, and then the control technology of electroless preparation parameters include: analysis of plating time, plating analysis of temperature, metal ions in solution than the (Fe2+/ Ni2+), bath pH and the added value of dispersion Agent and other research towards Fe-Ni particles to increase the iron content and Fe-Ni particles spread in the MWCNT upper fixed targets,the results found that when the parameters for the preparation of 50 ℃, pH10, do not add dispersant, metal ions than the solution (Fe2 +/ Ni2 +) 7, the highest iron content can be Fe = 40.55 at%(flat iron content 111.17 mg / g) of nanocomposites Fe-Ni/CNT, then for a series of the nature of its analysis, the final evaluation Fe-Ni/CNT nanocomposites were processed on the effectiveness of the application of mixed pollutants, the results showed that in 120 minutes at the same time when Adsorption of heavy metal ions lead nitrate 10 ppm, selenium heavy metal ion degradation of 1 ppm and orange azo dye AO7 50 ppm standard of effluents, and after a total Fe-Ni nano-particles to SEM / EDS analysis of more than 50% still remaining.

本研究首先将多壁奈米碳管以化学氧化法做纯化改质处理能使其纯度较纯化改质前提高5 wt％、比表面积提高1.89倍、表面电位下降约10~20 mV以及表面官能基提高1.45倍，再控制无电镀技术之制备参数包括：析镀时间、析镀温度，镀液中金属离子比(Fe2+/Ni2+)、镀液pH值及是否添加分散剂等，研究朝提高Fe-Ni粒子的含铁量与Fe-Ni粒子散布固定在MWCNT上等目标进行，研究结果发现当制备参数为50 ℃、pH10、不添加分散剂、镀液金属离子比(Fe2+/Ni2+)7时，可得到最高铁含量Fe = 40.55 at％(单位铁含量111.17 mg/g)之奈米复合材料Fe-Ni/CNT，接著对其作一系列性质分析，最后评估奈米复合材料Fe-Ni/CNT对共处理混合污染物之应用效益，结果显示其在120分钟时能够同时吸附重金属硝酸铅离子10 ppm、重金属硒酸根离子1 ppm及降解偶氮橘色染料AO7 50 ppm达放流水标准，且共处理后奈米Fe-Ni粒子以SEM/EDS分析仍剩余50 %以上。

Chemiluminescent immunoassay is one of the advanced immunoassay ofnon-radioisotopic immunoassay because of its high sensitivity, wide dynamic range,high accuracy, stable labeled protein, full automation and extensive application field.CLIA using acridinium ester derivatives as chemiluminescent label has advantages oflow background, high signal-to-noise ratio, no need of catalyst and simpleluminescence system. Different aspects associating the DMAE·NHS-based CLIA were studied in thisthesis, including synthesis of acridinium ester, chemiluminescentcharacteristics of DMAE·NHS, labelling antibody or streptavidin with DMAE·NHS,two-site sandwich chemiluminescent immunoassay for TSH, two-sitesandwich chemiluminescent immunoassay involved biotin-streptavidinsystem for TSH, and competitive chemiluminescent immunoassay usingbiotin-streptavidin system for TT〓. The desired acridinium ester, DMAE·NHS, was synthesized according to thereference method with some modifications. The products were identified by IR, NMR,MS and elemental analysis. In our method, KOH was used in place of NaOH tosynthesize benzyl ester of 3,5-dimethyl-4-hydroxybenzoic acid,2',6'-dimethyl-4'-(n-succinimidyloxycarbonyl) phenyl-acridinium-9-carboxylate waspurified on a silica gel column with chloroform/ethylacetate(4:1, v/v) as eluent andfurther purified by triturating with hexane/acetone(2:1, v/v).

DMAE·NHS的合成是本论文的关键和难点，我们对文献方法进行改进：文献方法用氢氧化钠与3，5-二甲基-4-羟基苯甲酸反应制得3，5-二甲基-4-羟基苯甲酸钠，再用3，5-二甲基-4-羟基苯甲酸钠与苄氯作用制备3，5-二甲基-4-羟基苯甲酸苄酯，我们用氢氧化钾代替氢氧化钠，使合成取得成功；在合成2'，6'-二甲基-4'-苯基-吖啶-9-甲酸酯时，文献方法对粗产品进行两次硅胶柱层析纯化，第一次柱层析用氯仿／乙酸乙酯(4∶1，V∶V)作溶剂和淋洗剂，第二次柱层析用己烷／丙酮(2∶1，V∶V)作溶剂和淋洗剂，按照文献方法得到的不是所需要的化合物，我们只进行第一次柱层析纯化，然后用己烷／丙酮(2∶1，V∶V)进行研磨，过滤，洗涤，除去溶于己烷／丙酮(2∶1，V∶V)的部分，得到所需要的产品。

This is achieved by first crystallizing the purified biomolecule into ordered arrays and then using X-ray diffraction to analyze the crystals.

这种方法的运用是基于首先使纯化的生物分子结晶为有序排列然后用X射线分析结晶体。

The products were identified by IR,NMR,MS and elemental analysis. In our method,K.OH was used in place of NaOH to synthesize benzyl ester of 3,5-dimethyl-4-hydroxybenzoic acid,2',6'-dimethyl-4'-(n-succinimidyloxycarbonyl) phenyl-acridinium-9-carboxylate was purified on a silica gel column with chloroform/ethylacetate(4:l,v/v) as eluent and further purified by triturating with hexane/acetone(2:l,v/v).The luminescence produced by DMAE-NHS is a flash light with maximumemission at 0.4s and decay half-time of 0.9s. The luminescence intensity is 6.11x10 cps/mol,which is affected by the composition of trigger and surfactant.

DMAE·NHS的合成是本论文的关键和难点，我们对文献方法进行改进：文献方法用氢氧化钠与3，5-二甲基-4-羟基苯甲酸反应制得3，5-甲基-4-羟基苯甲酸钠，再用3，5-二甲基-4-羟基苯甲酸钠与苄氯作用制备3，5-二甲基-4-羟基苯甲酸苄酯，我们用氢氧化钾代替氢氧化钠，使合成取得成功；在合成2'，6'-二甲基-4'-苯基-吖啶-9-甲酸酯时，文献方法对粗产品进行两次硅胶柱层析纯化，第一次柱层析用氯仿/乙酸乙酯(4：1，Ⅴ：Ⅴ)作溶剂和淋洗剂，第二次柱层析用己烷/丙酮(2：1，Ⅴ：Ⅴ)作溶剂和淋洗剂，按照文献方法得到的不是所需要的化合物，我们只进行第一次柱层析纯化，然后用己烷/丙酮(2：1，Ⅴ：Ⅴ)进行研磨，过滤，洗涤，除去溶于己烷/丙酮(2：1，Ⅴ：Ⅴ)的部分，得到所需要的产品。

The unfolded protein correctly refolded to the native conformation through the column, and was partly purified during the elution process.

其原理是将层析技术应用于蛋白质复性和纯化，使变性蛋白质在层析柱上折叠为正确的空间构象，在洗脱的同时实现部分纯化。

The results showed: when TLC solvent system was: chloroform - methanol - diethyl amine = 90:9:1, alkaloids from lotus leaves were well isolated, and five components were isolated and identified by spraying Dragendorff reagent. The optimal conditions of HPLC solvent system were: methanol- water - diethyl amine = 75:25:0.0125. Under the conditions, components of alkaloids from lotus leaves were separated very well, and ideal RP-HPLC peaks were obtained.5、Methods of High Speed Counter Current Chromatography and preparative High Performance Liquid Chromatography applied to identify and purify alkaloids from lotus leaves were set up. When solvent system of HSCCC was: chloroform - methanol - water (pH=4.00)= 4:3:2, speed of chromatogram was 700rpm, flow speed of the mobile phase was 2mL/min, four pure components relatively were attained by isolation and purification.

结果表明：薄层层析溶剂系统为氯仿：甲醇：二乙胺=90：9：1，使荷叶中的生物碱达到了较好的分离效果，并用改良的碘化铋钾试剂喷雾显色，共分离鉴别出了5种荷叶生物碱；分析型HPLC分析检测的较优溶剂系统为甲醇：水：二乙胺=75：25：0.0125，使荷叶生物碱的各个组分达到了基线分离，并获得了较好的峰形。5、建立了高速逆流色谱和制备型HPLC技术分离纯化荷叶生物碱的具体方法。H(来源：ABC46论文网www.abclunwen.com)SCCC法在溶剂系统为氯仿：甲醇：pH4.00的水=4：3：2，色谱仪转速为700rpm，流动相流速为2mL／min的条件下，分离纯化得到了4个纯度较高的化合物。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失