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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。

Theory of motion and motionlessness and view on changes inphilosophy of Taoism are further inquiries into the situation of the world(i.e.,"how"is the world)on the basis of the inquiries into the essence of the world(i.e.,"what"isthe world)by ontology and view on deities.

道教哲学的动静论和变化观是在本体论和形神观探讨了世界本质的基础上，进一步探讨世界的状况，道教哲学在此问题上主张本体静止而现象变动，并探讨了变化的原因及其规律。

Puncture into Attain long sheath of CS through left infraclavicular vein, guided by CS pipe, take sheath into CS, make reverse contrast of heart vein with Attain saccule contrast pipe, put the left ventricle electric pole into the vein through sheath, then bring it into right ventricle apex and right cardiac ear through the electric pole of right ventricle and atrium, connect it with 3-cavity pulse generator, which is buried under left chest skin.

方法］经左锁骨下静脉穿刺送入Attain冠状静脉窦长鞘，在CS导管的导引下，将鞘送人CS，用Attain球囊造影导管进行心脏静脉逆行造影，将左心室电极经长鞘放入选择的静脉，送人右心室及右心房电极至右室心尖和右心耳。与三腔脉冲发生器连接，脉冲发生器埋藏于左上胸皮下。

Method］ Puncture into Attain long sheath of CS through left infraclavicular vein,guided by CS pipe,take sheath into CS,make reverse contrast of heart vein with Attain saccule contrast pipe,put the left ventricle electric pole into the vein through sheath,then bring it into right ventricle apex and right cardiac ear through the electric pole of right ventricle and atrium,connect it with 3-cavity pulse generator,which is buried under left chest skin.

双心室起搏治疗心力衰竭的临床观察"版权属于原作者所有！请勿将"双心室起搏治疗心力衰竭的临床观察用于商业用途！［目的］观察双心室起搏治疗心力衰竭临床疗效。［方法］经左锁骨下静脉穿刺送入Attain冠状静脉窦长鞘，在CS导管的导引下，将鞘送人CS，用Attain球囊造影导管进行心脏静脉逆行造影，将左心室电极经长鞘放入选择的静脉，送人右心室及右心房电极至右室心尖和右心耳。

The thesis is based on income question, forest coverage rate and grop production.to beging with, date envelopment analysis is proved that it can be applicated into grain for green in shan-bei district, and then in view of the fact, seven esential factors which have influence on the project are found out: expense of dam, expense of crop,expense of cash tree ,expense of defense tree,other economic crop,expense of grass and graziery;and the output factors include:income of gdp,pure income per captia,the area of decreasing land loss,graziery income ,crop production,income of tree,the totle income of economic crop.after the date of each facts are puted into dea model,unefficiencial decision making units found.the data that are got through adjusting unefficiencial dmus dy dea can offer guide in shanbei district upgrading of an industrial structure.at the same time ,taking into account some possible problems in or after grain for green in shan-bei district,the thesis bring out some propesal to improve the circumstance,enhance the life level and putthe relation of population,resource and circumstance into a healthy orbit.

论文以解决陕北地区的收入问题，植被问题以及粮食产量问题为出发点，首先从理论上分析了包络分析方法在陕北地区退耕还林中运用的可行性，然后结合陕北地区目前的实际情况，综合分析出影响陕北地区退耕还林的关键的七个因素作为输入指标：水利水保设施投入，农作物投入，经济林种支出，防护林支出，其它经济作物，草类支出，畜牧业投入；以退耕还林所要解决的最终问题作为输出指标：GDP收入、农民人均纯收入、水土流失减少量、牧业总产值、粮食产量、林业总产值、其他经济作物总产值。将各指标所对应的数据代入模型后，通过分析求解得出非有效的决策单元，再运用包络分析方法的一些基本原理对非有效的决策单元进行调整，由此所得的数据对陕北地区的实际投入具有很好的指导意义。同时，又针对陕北地区在退耕还林过程中以及之后可能面临的困境，提出了一些建议和对策。最终使得陕北地区的投资环境尽快得以改善，使农民尽快脱贫致富，使人口、资源、环境的关系尽快纳入良性循环的轨道上。

A country's development, must be internal to the people bring happiness to the world outside to bring economic take-off, this is that China should do.

一个国家的发展，必须对内给人民带来幸福，对外给世界带来经济腾飞，这才是中国应该要做的。

This is merely apprearance. Flowers have soul. Their beauty, nobleness and high-hearted are larruping in numerous static life . Except they bring admiration and enjoyment , Moreover , They can expatiate meaning of life . A flower seems a soul. Flowers can bring serenity and day dream in insatiate world. There are flowers , There will be hopes. Who can say wild lily hasn't spring ?

花是有魂灵的，在众多静态生命中，花的美丽、高洁与清幽与众不同，除了带给人们欣赏与享受外，花还能阐释生命的意义，一朵花如同一个心灵，在物欲横流的世界里，花能带给人宁静、遐想，有花就有希望，谁能说野百合没有春天呢？

In 2005,coordinating with city construction,the company buy 40000m2 land in Shanghai Jiading Malu high-tech Park,build factory 18000m2,15000000RMB assets,more than 400 employees, has a scientific research group with 70 technologists and 20 senior engineer.Many years experiences bring up a well-trained riveter and welder team. The company have a long-term cooperation with many research institutions in China and aboard,bring in advanced technology of the world.

公司以生产大、中型系列矿山机械、冶金机械、建材设备为主，集研发、生产、销售为一体的股份制企业。2005年，为配合市政建设，公司在上海嘉定马陆高科技园区置地60余亩，新建厂房18000平方米，资产1.5亿，员工400多人，拥有专业技术人员70多人，中，高级工程师20多人的专业科研队伍，历年的创业造就了一支训练有素的铆工和焊工队伍，公司长期与国内外科研单位合作，引进了具有国内外先进的技术、工艺及检测设备。

I could not only bring the Western-style democracy thinking to China, but also could bring Chinese civilization to the world.

我可以把西方的民主思想传递给中国，也可以把中国的悠久文明展示给世界。

Its real-time performance of 3D space capabilities, the operating environment involved in man and computer and bring the " personal experience " feeling, bring a change in the past interestless, rigid and passive conditions between man and computer , and make human explorate and study the micro - macro world greatly facilitated.

其实时的三维空间表现能力、人机交互的操作环境以及给人带来的&身临其境&的感觉，一改过去人与计算机之间枯燥、生硬和被动的状况，为人类探索和研究宏观世界与微观世界提供了极大的便利。

The shaping method of noncircular part and the tool holder's radial motion characters in noncircular turning process are discussed in detail in the thesis.

论文详细研究了非圆零件的成型方法和加工过程中刀架的径向运动规律。

I have not really liked him,I do not like his this kind of disposition.

我没有真的喜欢他，我不喜欢他的这种性格。