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MTT assay FAK signaling pathway inhibitor genistein on human corneal epithelial cell cytotoxicity;RT-PCR detection of human corneal epithelial cells adhesion to fungus at different times,extracellular matrix protein including laminin,fibronectin,FN glass,Ⅳcollagen,transmembrane protein integrinαⅤ,integrinβ1(ITGβ1),as well as the FAK signaling pathway FAK1,FAK2 and Paxillin gene expression;Western blot detection of the signal transduction pathway adhesion-associated protein ITGβ1,FAK and PAX expression and the inhibition of genistein. Immunocytochemical method was used to observe the LN,FN and FAK expression in human corneal epithelial cells during interaction with the fungues;Laser scanning confocal microscope had a cell positioning on FN,FAK and PAX,observed the changing of the human corneal epithelial cytoskeleton after stimulated by fungues;Quantitatived by flow cytometry to detect of human corneal epithelial cells with PAX at ITGβ1 fungal expression after adhesion;Optical microscopy quantitied the fungues and human corneal epithelial cell adhesion and recorded to determination the integral optical density afrer adhesion;Scanning and transmitted electron microscope observed the changing of cell ultrastructure after fungues and human corneal epithelial cell adhesion.

第一部分真菌激活人角膜上皮细胞FAK信号转导通路的体外实验研究将三种常见致病真菌(白色念珠菌、烟曲霉菌和茄病镰刀菌)分别与人角膜上皮细胞共孵育,MTT法检测FAK信号通路抑制剂染料木黄酮的对人角膜上皮细胞的细胞毒性作用;RT-PCR检测真菌黏附人角膜上皮细胞后不同时间细胞外基质层连蛋白、纤连蛋白、玻连蛋白、Ⅳ型胶原、跨膜蛋白整合素αV、整合素β1(ITGβ1),以及FAK信号通路中FAK1、FAK2和桩蛋白基因的表达情况;Western blot的方法检测黏附信号转导途径相关蛋白ITGβ1、FAK和PAX的表达,以及染料木黄酮对真菌刺激人角膜上皮细胞FAK信息通路活化的抑制作用;免疫细胞化学方法观察人角膜上皮细胞与真菌相互作用过程中LN、FN和FAK的表达;激光共聚焦显微镜对FN、FAK和PAX进行了细胞定位,并观察真菌刺激后人角膜上皮细胞骨架的变化;流式细胞仪定量检测人角膜上皮细胞ITGβ1与PAX在真菌黏附后表达的改变;光学显微镜观察真菌与人角膜上皮细胞黏附数量,记录并测定了黏附后积分光密度值OD扫描及投射电镜观察了真菌与人角膜上皮细胞黏附后,细胞超微结构的改变。

Objectives:Through the detection of the expression level of interleukins-1β(IL-1β), matrix metalloproteinase 9(MMP-9) and nuclear factor kappa B in Synovium and the detection of the concentration of IL-6 and TNF-αin serum of collagen induced arthritis Wister rat, to investigate the relationship between IL-1β、MMP-9、NF-κB and arthritis index in the development of arthritis,and to understand the pathology of synoviocyte.This may provide an experimented proof for the diagnosis of clinical rheumatical arthritis. Methods: Thirty male Wistar rats were randomly divided into two groups,namely the model controlled group and the normal controlled group,three subgroups were randomly divided in model group and controlled group separately.

目的:通过对白细胞介素1β,基质金属蛋白酶-9,转录因子核因子κB在胶原性关节炎大鼠滑膜细胞表达水平的观察,同时监测同期血清细胞因子IL-6和TNF-α的浓度,旨在探讨白细胞介素1β,基质金属蛋白酶-9,转录因子核因子κB与RA发生发展及其与关节指数之间的相关性,了解炎性病变关节滑膜细胞的病理生理状态,进一步为类风湿关节炎的临床治疗提供实验依据。

The reactions that isocyanate reacted to cellulose, semicellulose, lignin, and water in timber to form the urethane and urea are basic knowledge for the adhesion of isocyanate-terminated polyurethane resins to wood. It has significant meaning to understand that different quantity of urethane and urea affect the strength of adhesive bond.

异氰酸酯基与木材中的纤维素、木素、半纤维素和水在一定条件下生成氨基甲酸酯和取代脲的反应是聚氨酯胶粘剂与木材胶接的基础反应,研究氨基甲酸酯和取代脲的生成量对胶接强度的影响具有积极的理论意义和实践指导价值。

SF was helpful in keeping moisture and could prevent cake from staling. The quality of the cake was not acceptable at higher concentration than 0.5%.

当丝素蛋白的添加量小于0.5%时,丝素营养强化面条添加0.3%(w/w瓜尔豆胶和蛋糕产品的综合评价结果都为良,即可被消费者接受。

The steady state kinetics of the lignin peroxidase catalyzed oxidation of veratryl alcohol by H2O2 in a sodium bis(2-ethylhexyl) sulfosuccinate/isooctane/toluene/water reverse micellar medium was studied and a comparison with the corresponding aqueous medium was made to understand the effect of the reverse micellar medium on the catalytic mechanism and kinetic parameters.

为了理解木素过氧化物酶在反胶束介质中的催化性能与动力学行为,本文以LiP催化过氧化氢氧化藜芦醇反应为模型反应,研究了2-乙基己基琥珀酸磺酸钠/异辛烷/甲苯/水反胶束中上述模型反应的稳态动力学并与水介质做了比较。

Breast cancer commonly metastasizes to bones, producing both osteolytic and osteoblastic deposits. Different markers for quantitative determination of bone turnover have been developed to evaluate bone metastases of breast cancer. The urinary deoxypyridionline, a crosslink product of collagen molecules found in bone and excreted in urine during bone degradation, and bone specific alkaline phosphatase, an isoenzyme localized in the membrane of osteoblasts and released in circulation during bone formation, were recently described as a group of markers of bone turnover in metastatic cancer.

乳癌患者常见骨髓转移造成溶骨性及成骨性沈积物,而利用不同标记来定量骨质转换形态,可以评估乳癌骨骼转移的变化,而尿液中Deoxypyridinoline是骨质内胶原分子在骨骼破坏分解时至尿中的产物,骨特殊碱性磷酸酵素是一种同形酵素,位於成骨细胞的外膜且在骨骼形成时释放至血液中,利用此两种生化标记,可以来预测乳癌转移至骨骼的转换情形。

On the basis of the wholly balanced nutrition and optimization of homeostasis of human body ,fairness element especially designed for ladies in accordance with physiological characteristic of females, contains collagen、papa in 、tea polyphenols、grape seed extract 、soy isoflavone、Honey freezing flour、barley Green、lecithin、oligosaccharides、resveratrol、dietary fiber and so on.

在全面均衡营养和优化人体动态平衡的基础上,根据女性生理特征专为女士设计的靓丽素,内含胶原蛋白、木瓜蛋白酶、茶多酚、葡萄籽提取液、大豆异黄酮、蜂王浆冻干粉、麦绿素、卵磷酸、低聚脂、白藜芦醇、膳食纤维等多种营养因子。

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。

The invention provides a method for preparing a tube-shaped cathode of a direct methanol fuel cell, belonging to the fuel cell field and comprising the steps of: dispensing a single solution for gel injection molding at first, then adding a mesophase bituminous coal micro ball into the single solution and stirring evenly to obtain stable slurry, then casting the slurry into a die, preserving heat until the single solution reacts with a cross-linker completely to obtain a carbon-biscuit, after drying and sintering the biscuit in vacuum, preparing a diffusion layer and a catalyst layer on the surface by dipping process, and then coating polyester electrolyte membrane with a burette, rolling the carbon on a warm table to solidify the electrolyte membrane.

一种制备直接醇燃料电池管状阴极的方法,属于燃料电池领域。本发明首先配制凝胶注模用单体溶液,之后将中间相沥青炭微球加入到单体溶液中,搅拌均匀得到稳定的浆料,然后将浆料浇注到模具中,保温至单体与交联剂完全反应,得到炭管素坯,将素坯烘干、真空烧结后,在其表面采用浸渍工艺制备扩散层、催化剂层,然后采用滴管涂覆聚合物电解质膜,在加热台上对炭管进行滚压,使电解质膜固化。

In the second part, the glycosylation site and glycan structures of pectin methylesterase in jelly fig achenes are determined to provide a better understanding on the relation between glycosylation and enzymatic activity.

第二部分是要鉴定爱玉凝胶酵素上的醣基修饰位置以及醣基结构,目的在於进一步了解之后,对於未来在研究醣基化与酵素活性的关系有所帮助。

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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

But because of its youthful corporate culture—most people are hustled out of the door in their mid-40s—it had no one to send.

但是因为该公司年轻的企业文化——大多数员工在40来岁的时候都被请出公司——一时间没有好的人选。