筛选法

Use constituent piece to stick mural way depart to foster Niu Er to become fiber cell above all, after classics generation is rarefied, fat simple way will be contained double the skin of the 4th acting ox that outside conveying carrier to turn into body, the particularity of person bacteriolysis enzymatic mammary gland that chooses number fosters becomes fiber cell, g/mLG418 chooses 500 μ 2 weeks, g418 halve continues to develop 3 generation of 2 ～, catch the cell after choosing to undertake PCR detects to turning next reach nucleus analysis.

首先采用组织块贴壁法分离培养牛耳成纤维细胞，经传代纯化后，脂质法将含有双重筛选标记的人溶菌酶乳腺特异性表达载体转入体外培养的第4代牛皮肤成纤维细胞，500μg/mLG418筛选2周，G418减半继续培养2～3代，然后对转染筛选后的细胞进行PCR检测及核型分析。

Sample data were selected by standard deviation law and factor variables were selected by the method of principal components analysis, stepwise and enter regression analysis. The forecast precision of the model is 87.35%.

样本数据筛选采用标准差法，因子变量筛选采用主成分因子分析法、多元线性回归的逐步回归和强行进入法等方法，建立的多元回归模型预测总体精度达到87.35%。

To select five blood samples for cross match with mixed field agglutination in order to screen the best disposal methods by CaCl2 centrifugalization method and EDTA-Na2 method which were designed by ourselves, To select another cryoprecipitate which were diluted 1: 10 and 1: 15 after mimesis do cross match and antibody screening with CaCl2 centrifugalization method and EDTA-Na2 method.

用自行设计的CaCl2离心法和EDTA-Na2法对5份交叉配血中有混合外观凝集的血样本分别进行处理，筛选出最佳处理条件；另选取一份冷沉淀，按1：5和1：10稀释后用CaCl2离心法和EDTA-Na2法分别模拟进行交叉配血和抗体筛选。

Results:thrity-one drugs were sfited by germinative test,twenty-one by agar diffusion method,fourteen by both method.

结果 ：应用芽管试验法筛选的有效药物有 31种，应用琼脂扩散法筛选的有效药物有 2 1种，其中两种方法均有效的有 1 4种。

Six experimental stages were designed in our procedure, those are:(1) metabolite recovery and tested sample preparation: the metabolites were recovered by Amberlite XAD-2 absorption, followed by MeOH elution and solvent concentration;(2) antioxidant detection and strain selection: samples were quantitatively analyzed by the inhibition effects on formation of lipid peroxides and TBARS to screen the strains able to produce antioxidants. According to the established screening methods, we chose out a strain of actinomycetes, designed as AMBL-029C;(3) antioxidant purification: the fermentation broth was recovered by a series of separation techniques including centrifugation, Amberlite XAD-2 absorption, followed by MeOH elution and a successive TLC purification. The resulting primary purified compound [temperately designed as AMBL-029C-TS] was further analyzed by HPLC to monitor its purity;(4) physical-chemical characteristics: judging from the acid-base fractionation experiments, and the pH and temperature stability tests, the compound was deduced to be a acidic compound with the properties of low polarity and highly pH and temperature stable;(5) mechanism of the antioxidant: in comparison with some other known antioxidants, TS was subjected to investigate its antioxidant mechanism, together with BHT,-tocopherol, as well as two streptomyces metabolites, homogentisic acid and -phenylpyruvic acid, which were previously isolated as the natural antioxidants in our laboratory.

针对本实验目的，我们设计了以下的实验步骤﹔（1）二次代谢物回收及检测样本处理：我们将发酵所得的培养上清液，利用疏水吸附性树酯Amberlite XAD-2吸附回收，并以甲醇溶离及真空减压浓缩脱水等方式处理，以取得提供抗氧化活性筛选之检测样本；（2）抗氧化活性检测及菌种筛选：以「过氧化脂质」和「硫丙二醯尿」的生成量进行定性定量分析以作为抗氧化物质生产菌筛选之用；经此筛选程序，我们选获了具有抗氧化物质高生产力的菌株，命名为AMBL-029C；（3）抗氧化物质的分离纯化：针对生产菌株的发酵回收处理液，以矽胶薄层色层分离法经物质层析纯化后，并以高效能液相层析法(High performance liquid chromatography；HPLC)分析物质可得一初级纯化物质，命名为AMBL-029C-TS；（4）抗氧化物质的物理化学性质分析：由酸碱转溶（acid-base fractionation）实验得知，此抗氧化物质属於中低极性的强酸性物质，对温度（37℃-100℃）及酸碱度（pH3.0-13.0）均表现出高稳定性；（5）在抗氧化机制探讨方面，我们针对数种不同的抗氧化机制进行探讨，即： 1。

Methods:(1) Silica column chromatography and LH-20 were used in phytochemistry study.(2) Based on the extraction ratio of arctiin, the best extraction procedure was acheived under the direction of L9(34) orthogonal experimental design. From 10 types of resins, the one with the maximal binding ability for Arctii was selected and silica-column choramatograpy was used in its seperation and purification procedure.(3) The animal diabetic experimental model in Wistar rats was established with streptozotocin and many biochemical factors were used to indicate the pharmacological effects of arctiin, including glycosylated hemoglobin, serum creatinine, seralbumin, monoxide nitrogen, glycerin trilaurate, cholesterol, HDL-ch, LDL-ch, Blood urea nitrogen, creatinine, SOD, ET, MDA, total Urine protein and albuminuria.

(1)采用硅胶柱层析、聚酰胺层析、LH-20等方法对牛蒡子的化学成分进行研究；(2)采用正交实验设计方法，以牛蒡子苷为指标，确定了牛蒡子乙醇回流提取的最佳工艺；对10种类型大孔吸附树脂进行筛选，筛选出对牛蒡子苷吸附效果最佳的树脂；采用硅胶柱层析法，对牛蒡子苷进一步分离纯化；(3)建立实验性糖尿病大鼠模型，以GLU、TC、TG、HDL-ch、LDL-ch、TB、ALB、BUN、CREA、GHbAlc、NO、SOD、ET、MDA和尿液中TB、ALB为指标，研究牛蒡子苷药理活性；(4)以差速消化法纯化Wistar大鼠的主动脉内皮细胞并进行传代培养，测定牛蒡子苷对高糖条件下RAECs活性、RAECs释放LDH、MDA和NO的变化以及RAECs表达内皮型一氧化氮合酶表达的影响。

TF primary extracts condensate used alone or used with the general stored grain chemistry protect pharmaceutical to get the experimental treatments. Adopt filter paper medicine membrane method and grain mixed medicine method to seeking TF plant single pharmaceutical and the extender, screen out two kinds of emulsion Basuling-1, Basuling-2 and one powder Basuling powder have better prevent and cure result to target pest Sitophilus zeamais Motschusky and Rhizopertha dominica Fabricius.

TF植物性物质单用或与常用的化学防护剂混配，采用滤纸药膜法和小麦拌药法对优选TF植物单剂和混配剂的筛选，筛选出了对靶标害虫玉米象和谷蠹有较好防治效果的乳油两种巴素灵1号、巴素灵2号和粉剂巴素灵粉。

According to the test data, quality evaluation indicators for rotating sprinklers were sieved by means of cluster analysis and principal component analysis. Effectiveness and applicability of two methods were given in comparative analysis.

该文依据室内性能测试数据，借助聚类分析法和主成分分析法对旋转式喷头质量评价指标进行定量筛选，对比分析了两种方法的筛选效果和适用性。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法，从人组织细胞总RNA中扩增可溶性TWEAK胞外区（sTWEAK1和sTWEAK2）的cDNA序列及全长编码序列，用琼脂糖凝胶电泳分析PCR产物，胶回收目的基因片段，连接到pMD18-T克隆载体中，转化大肠杆菌K802，PCR和酶切筛选阳性克隆，全自动DNA测序验证序列；(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中，分别转化大肠杆菌BL21和TB1，PCR筛选和酶切鉴定，阳性克隆用IPTG诱导表达，表达产物用SDS-PAGE分析和Western blot验证融合蛋白；(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白；(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测，贴壁细胞用结晶紫染色法，悬浮细胞用磺酰罗丹明B染色法，酶标仪检测OD值；(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量；(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况；(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响；(8)用双荧光素酶报告基因检测法，测定表达产物对敏感细胞NF-κB的影响；(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上，用脂质体转染法转染HEK293细胞，PCR鉴定重组质粒。

In this paper a new contingency screening and ranking method is formed by combiningiterative filtering screening technology with continuation power flow analysis. Researchingpreventive control policy in reactive power compensation and load control when themargin of stability is serious, and reciprocal math model is given;through the exempleof IEEE39 typical system , the availability of the discussed methods in this dissertationwas approved. Aimed at the real grid, the contingency screening analysis and preventivecontrol policy based on Beijing power grid is discussed;the static voltage stabilityanalysis software based on figure draw of Beijing grid is realized, and the software hasthe junction to BPA .

论文提出了将迭代过滤筛选技术和连续潮流法相结合的事故筛选分析方法；从无功补偿控制和负荷控制两方面研究了系统裕度紧张时的预防控制策略，给出了相应的数学模型；采用IEEE39节点的标准系统算例，验证了本文提出的静态电压稳定事故筛选分析方法的可行性；针对实际电网讨论了基于北京电网的事故筛选分析和预防控制策略；通过与BPA接口实现了基于北京电网图形平台的静态电压稳定分析软件。

What are your goals and strategies for growth?

你的成长目标和策略是什么？

And unto the angel of the church in Sardis write; These things saith he that hath the seven irits of God, and the seven star I know thy works, that thou hast a name that thou livest, and art dead.

3：1 你要写信给撒狄教会的使者，说，那有神的七灵和七星的，说，我知道你的行为，按名你是活的，其实是死的。