蛋白的

The main collagens in skin are type I and III. In this research, total collagens were separated from rat skin by enzymatic digest and acetum methods. The collagens were denatured at 60℃ and digested with trypsin. Characteristic peptides typical for collagen type I and III were identified with high performance liquid chromatography/mass spectrometry.

哺乳动物皮肤真皮中胶原蛋白含量约为70%，主要为是I型、III型胶原蛋白，本实验利用稀酸溶解和酶法提取了大鼠皮肤中的总胶原蛋白，将胶原蛋白粗提品在60℃变性后用胰蛋白酶进行降解，液相色谱/质谱联用法分析了两种胶原蛋白的特征多肽，利用特征多肽比较了不同生长期大鼠皮肤中I型和III型胶原蛋白相对含量。

The female salivary gland proteins were influenced by JH Ⅲ. Treatment on d2 after feeding, 10μg dose increased SG protein level and restrained 136kD protein gene expression. Treatment on d0 after engorgement, 10μg, 50μg and 100μg doses significantly increased the d2 SG protein and 100μg dose made 35kD protein gene expression, 200μg treatment made the 136kD and 192kD proteins absent. This is the first found that JHs regulate arthropoda SG activity.

保幼激素对雌蜱唾液腺蛋白及成分有影响，吸血后2天处理，10 μg剂量使唾液腺蛋白含量明显增加，抑制136kD蛋白的基因表达；饱血当天处理，10μg、50μg和100μg显著提高饱血后2天唾液腺蛋白含量，100μg使35kD蛋白表达，200μg使136kD和192kD的蛋白缺矢；饱血当天处理，1μg和10μg剂量均能显著提高饱血后4天雌蜱唾液腺蛋白含量，高剂量（100μg）抑制136kD蛋白的表达，该结果首次报道了保幼激素对节肢动物唾液腺的调控作用。

Up-regulated proteins under salt stress include an ABC transporter permease, glycerol-3-phosphate permease, pyrimidine nucleotide transporter, formate dehydrogenase, and down-regulated proteins include succinate dehydrogenase iron-sulfur subunit, flavoprotein subunit, cytochrome b-556 subunit, and a hypothetical membrane protein similar to charperone DnaJ. Some of these changes were further verified by enzyme activity assay.

盐胁迫诱导上调表达的蛋白包括ABC型转运蛋白，3-磷酸甘油透性酶，嘧啶核苷转运蛋白和甲酸脱氢酶；下调表达的蛋白包括琥珀酸脱氢酶铁硫亚基、黄素蛋白亚基、细胞色素b556亚基，以及分子伴侣DnaJ的同源蛋白，酶活力测定结果表明胁迫条件下上述蛋白的活性变化与表达量变化相一致。

The positive expression both of p14ARF and mtp53 protein in NSCLC tissues were significantly different from that in nonmal lung tissues and tissues adjacent to the cancer (P.01).The expression of p14ARF protein was correlated with pathological degree（P＜0.01）,but it was uncorrelated with pathological staging,lymphaden metabasis and vessel invasion(P＞0.05).The expression of mtp53 protein was correlated with clinical pathology staging,pathological degree and vessel invasion(P＜0.05 or P＜0.01),but it was uncorrelated with lymphaden metabasis(P＞0.05).(2) The coexpression of p14ARF and mtp53 was 36.7％,their consistency was 38.8％,but the expression of p14ARF protein were negatively correlated with the expression of mtp53 protein(r=-0.3553,P＜0.001).Conclusion The results suggest that the decrease of p14ARF and the increase of mtp53 protein may play an important role in the genesis and development of NSCLC.

结果 (1)在正常肺组织、癌旁组织和NSCLC中p14ARF蛋白的阳性表达率分别为93.3%、83.3%、55.1%，mtp53蛋白的阳性表达率分别为6.7%、44.4%、79.6%；p14ARF蛋白和mtp53蛋白在NSCLC中的表达与正常肺组织和癌旁组织比较均有显著性差异(P＜0.01)；p14ARF蛋白表达与临床病理分期、有无淋巴结转移、有无脉管浸润无关(P＞0.05)，而与病理分化程度有关（P＜0.01）；mtp53蛋白表达与临床病理分期、病理分化程度(P＜0.01)及有无脉管浸润(P＜0.05)有关，而与有无淋巴结转移无关(P＞0.05)；(2)在NSCLC中，pl4ARF与mtp53蛋白共表达为36.7％，一致性为38.8％，二者表达呈负相关(r=-0.3553，P＜0.001)。

The results showed that the water retentiveness of soybean separation protein was the best, followed by casein, and pH value, metal ions such as Ca2+, Mg2+, Zn2+, and Na+, etc. had some effects on the water retentiveness, with effects differed greatly among these factors; the emul...

结果表明：大豆分离蛋白的持水性能较好，其次为酪蛋白，pH值、金属离子Ca2+、Mg2+、Zn2+、Na+等对持水性有一定的影响，呈现较大差异；大豆分离蛋白的乳化性能较好，其次为酪蛋白、乳清蛋白，动物蛋白较差，在影响乳化性能中金属离子以多价离子为主要因素；大豆蛋白的热稳定性较差，酪蛋白和动物蛋白较好。

The study showed the recombinant wt/mCREG protein depressed the VSMC proliferation depending on dose and the optimal concentration was 400nM;2biologic function of CREG protein and the membrance receptor mechanism:①effect on VSMC migration: the wound healing experiment showed the OB2 cells migration was slower significantly after added wt/mCREG(400Nm) in supernatant. The HITASY cells migration were very slowly and no remarkable change. The gelatinase digestion and Western blot analysis showed the matrix metalloproteinase was decreased and TIMPs was increased;②effect on differentiation: after added wt/mCREG(400nM), the expression of myocardin, SMα-actin, MHC and caldesmin were increased and that of LM-1 and FN were decreased in OB2 cells. These effects were more significant when adding wtCREG.;③effect on VSMC proliferation: Cell cycle assay and BrDU stain showed: after added the wtCREG and mCREG protein, the ratio of cell in G0/G1 phase increased to 0.5773 and 0.5572 from 0.5308 respectively in OB2 group, which increased to 0.7369 and 0.7034 respectively from 0.6297 in HITASY group;3Role of M6P/IGF2R in CREG biologic function:①ELISA and co-immunoprecipitation showed the wt/mCREG binding to M6P/IGF2R directly.②antibody blocking test: when the anti-IGF2R was added to medium at the same time with wt/mCREG at different concentration(2μg/mL、4μg/mL、8μg/mL),the effects of CREG protein which depressing proliferation, migration, secretion and promoting differentiation were blocked, which had the positive correlation to the concentration of added anti body. The studies showed two combinant CREG promoted VSMC switch to differentiation phaenotype, at the same time, depress VSMC proliferation, migration and secreting extracellular matrix.

上述实验结果证实：两种重组CREG蛋白对VSMC增殖均有剂量依赖性的抑制作用，并且相同浓度的糖基化的CREG蛋白对细胞增殖的抑制效应更为显著，最佳效应浓度为400nM；2两种重组CREG蛋白添加后对HITASY和OB2细胞生物学行为的影响：①CREG蛋白对VSMC迁移的影响：刮伤实验发现，加入最佳效应浓度的wtCREG和mCREG蛋白24h后，OB2组迁移能力下降，HITASY组无明显变化；细胞外基质金属蛋白酶-2，9(Matrix metallo-proteinase 2，9，MMP2 ，9)明胶酶电泳检测和Western blot检测结果证实，两种CREG蛋白均可以使OB2细胞合成细胞外基质MMP2，9减少，而组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases，TIMPs)增加；②CREG蛋白对VSMC分化的影响：加入400nM的wtCREG和mCREG蛋白12h后，OB2细胞myocardin、SMα-actin、MHC、caldesmin表达增加，LM-1、FN表达减少；③流式细胞仪分析细胞周期和BrDU染色分析证实，加入400nM的wtCREG和mCREG蛋白后，OB2组G0/G1期细胞由0.5308分别增加至0.5773和0.5572，HITASY组G0/G1期细胞由0.6297分别增加至0.7369和0.7034；3M6P/IGF2R在重组CREG蛋白的生物学功能中的调控作用：①免疫共沉淀和免疫荧光双染色分析结果显示，CREG蛋白与M6P/IGF2R存在直接结合；②应用抗体阻断实验：将不同浓度的anti- M6P/IGF2R(2、4、8μg /mL)与两种CREG蛋白同时加入培养液中，CREG蛋白抑制VSMC增值、迁移和合成细胞外基质、促进分化的效应减弱，而且与加入anti- M6P/IGF2R浓度正相关。

Expression of Caspase-3 and bcl-2 protein In E20 cerebral cortex, the expression of cleaved Caspase-3 in the encephalocoele group are higher than that in control group, and the expression of bcl-2 in the encephalocoele group are lower than that in control group. These findings were consistent with the results of the western blot analysis in E14 Brain.

四、Western-blot检测E14、E20胎鼠大脑皮质中caspase-3、bcl-2蛋白的表达 1、用Western-blot方法检测不同组E20胎鼠大脑皮质中caspase-3、bcl-2蛋白表达水平，结果表明对照组、给药无畸形组、脊柱裂组胎鼠大脑皮质中caspase-3、bcl-2蛋白表达水平无明显改变，而脑膨出组胎鼠大脑皮质中caspase-3蛋白活化片段表达水平明显增加，bcl-2蛋白表达水平明显减少。

By using multi-dimensional nuclear magnetic resonance method we have studied the folding mechanism of staphylococcal nuclease in vitro; the tertiary interactions for folding of SNase fragments into native-like conformation; the interaction between SNase N- and C-terminal subunits; the relationship of enzyme activity with folding and dynamic states of SNase; the structural properties of enzyme protein while exert its function. We have studied the internal motions of thermophilic Archaea protein Ssh10b and mechanism of its heat-resistance using the NMR 1H-15N relaxation and H/D exchange methods. We have determined the 3D solution structure of human translationally controlled tumor protein TCTP and the Ca2+-binding site; determined the 3D crystal structure of human mitoNEET, a novel protein from distinct groups of iron-sulfur proteins; determined the 3D solution structure of a novel chromatin protein Cren7. Determination of SNase-DNA and Archaea protein-DNA complex structures are in progress.

运用异核多维核磁共振方法研究了金黄色葡萄球菌酶体外折叠机制，酶蛋白片段体外折叠成类天然溶液三维构象的三级相互作用力，酶蛋白亚基间的相互作用，酶蛋白的折叠以及内运动状态与酶活力的关系，酶蛋白发挥功能时的结构特性；运用NMR的1H-15N 驰豫和H/D交换方法研究了嗜热古菌蛋白质Ssh10b双体结构内运动特性，热稳定性机制；确定了人翻译控制的肿瘤蛋白TCTP蛋白的溶液三维结构及其钙离子的结合部位；确定了一类新的铁硫蛋白家族蛋白人线粒体膜上mitoNEET蛋白的晶体结构；确定了一个新型的染色质蛋白Cren7的溶液三维结构；正在研究金黄色葡萄球菌酶及嗜热古菌蛋白质与DNA复合体的溶液三维结构。

The main components in soy protein isolate are 7S and 11S globulins. Studiesof papain on 7S, 11S and mixture of the two proteins suggested that 11S is themain component in SPI to form coagulation. 7S coagulation required higher papainaddition than 11S coagulation, while 11S gel's elastic modular was higher than7S gel. The elasticity of mixture depended on the proportion of 11S. Two proteinases were separated from papain by SP-sepharose cation-exchangechromatography.

首先分离出大豆蛋白中的主要组分7S和11S蛋白，将木瓜蛋白酶分别作用于两种蛋白的溶液和不同7S/11S比例的蛋白溶液，结果表明11S蛋白是大豆蛋白中形成酶促凝胶的主要组分；7S蛋白形成凝胶所用的酶浓度高于11S蛋白所用的酶浓度，但11S蛋白形成的凝胶的弹性强度却远高于7S蛋白；在两种蛋白的混合溶液中11S蛋白的比例对凝胶的弹性强度起决定作用。

In this experiment, a new method was used to purify the toxin protein produced by Verticillium dahliae. The supernatant of fungus culture was frozen and dried with Lyophilizer first, and then dialysed by Dialysis Membranes (MWCO 1000) after dissolved in distilled water. This method can eliminate the salt and sucrose in culture medium and reserve the protein component almost completely. VD-toxin were analysed using sodium dodecyl sulfate polyacrylamide gel analysis. The results indicated that the protein components were very complex, and included glycoprotein within 35.8kDa-83.2kDa. Furthermore, treatments on glycoproteins with high temperature, conA and proteinase alleviated, not abolished, their activities.NO and H_2O_2 production were assayed in two cultivars of cotton cells which were treated with VD-toxin.

本实验首先确定了用冷冻浓缩后透析的方法初步提纯棉花黄萎病菌毒素，该方法能有效去除粗提毒素中的蔗糖和盐离子并能最大程度的保留黄萎病菌分泌的蛋白成分；对毒素蛋白成分鉴定结果表明蛋白条带多，每种蛋白的含量少，小分子量蛋白含量多，糖蛋白染色结果显示，毒素蛋白中分子量在35.8kDa-83.2kDa之间的蛋白大多数为糖蛋白，分子量小于35.8kDa的蛋白多为非糖蛋白；性质鉴定结果显示，毒素中有活性的蛋白成分具有部分可耐高温高压，且毒素蛋白中的蛋白成分和糖基成分都具有致萎活性。

I didn't watch TV last night, because it .

昨晚我没有看电视，因为电视机坏了。

Since this year, in a lot of villages of Beijing, TV of elevator liquid crystal was removed.

今年以来，在北京的很多小区里，电梯液晶电视被撤了下来。