胞外酶

The chondrocytes of different generation were observed with light-microscope and transmission electron microscope for cellular growth and ultromicrostructure, with the method of MTT assay for grow curve and proliferation, with alcian blue test for GAG of ECM, with immuocytochemistry and RT-PCR (reverse transcript -polymerase chain reaction)for type Ⅱcollagen, with flow cytometry for cell life cycle ,histochemistry for S-A-β-galand so on by which to identify cataplasia and senescence of chondrocytes cultured in vitro.

对软骨细胞退变老化进行观察，采用相差显微镜观察其生长情况，透射电镜观察细胞结构，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，免疫细胞化学法和RT-PCR方法检测Ⅱ型胶原鉴定软骨细胞，以MTT比色法描绘生长曲线、检测生长状态，组化法检测老化相关β-半乳糖苷酶，流式细胞仪分析细胞周期和增殖指数。3。

This bacterium is resided in xylem be used to, short staff state, sheet is unripe, provide the fasciculus outside the cellular wall of several winding and afterbirth, micron of 1.0-4.0 of size 0.25-0.50 ×, without flagellum, not move about, oxidation enzymatic negative, cross oxidation enzymatic positive, good oxygen, blame barmy, of the salt that be not be addicted to, colorless element is deposit, education is difficult, the requirement has special education base, if the ability such as BCEY, JD-3 grows.

该菌在木质部习居，短杆状，单生，具数层卷绕的细胞壁和胞外纤维束，大小0.25-0.50×1.0-4.0微米，无鞭毛，不游动，氧化酶阴性，过氧化酶阳性，好氧，非发酵的，非嗜盐的，无色素沉积，培养困难，要求有非凡培养基，如BCEY.JD-3等才能生长。

The results showed:among the 7 isolates, five isolates of HY3、GY1－3、ZJ1－1、HP1、FC3 had same colony shape, irregular shape, liquidlike, slimy, opacity with smooth surface;the other two isolates had same shape, irregular shape, dry, opacity with coarse surface. By inoculating eucalyptus with the 7 isolates, the plants were infected apparently, and the young plants of eucalyptus in control experiment with tap water were not infected. By cultivating eucalyptus cuttings with the bacterial suspensions without EPS, the incidence of disease was very distinct,but compared with the former bacteria suspension,the incidence of disease has decreased at different degrees. By screening out two isolates of strong pathogenicity and two isolates of weak pathogenicity from the 7 isolates,making the bacterial suspensions with them to inoculate the young plants of eucalyptus, two treatments of cutlings and ramets with rats were set with 5 repetitions in every treatment, the results of data analysis showed: for the cutlings, the bacterial contents in upper and middle parts、upper and lower had significant difference;for ramets with roots, the bacterial contents in upper, middle parts, lower had significant difference between each other; For both the cutlings and ramets with roof, the bacterial contents in xylem and phloem had significant difference. The interaction between vertical and horizontal parts for the bacterial content had significant difference. For the two isolates of HY3 and 93B which were screened out at last,their activities of the cellulase were: 1.955ug/ and 1.288ug/ respectively, and had significant difference; the activities of pectase were: 1.325 ug/and 1.24ug/ respectively, and had no significant difference. The content of EPS extracted from the two isolates of HY3 and 93B was very different: 7.08x10-8ug/cell and 5.17x10-8ug/cell.

结果显示：7个菌株中，其中5个菌株HY3、GY1－3、ZJ1－1、HP1、FC3的菌落形态相同：不规则形状、流体、粘性、不透明、表面光滑；另外2个菌株93B、GN1菌落形态相同：不规则形状、干燥、不透明、表面粗糙；用7个菌株接种剪根桉树苗，发病情况非常明显，而自来水对照实验中桉树苗却不发病；无EPS菌悬液培养桉树剪根苗，发病率也很明显，但是相比原菌液，则发病率有不同程度的下降；从7个菌株中间筛选出来2个强致病性菌株和2个弱致病性菌株，用它们配制菌悬液培养桉树苗，设置剪根和不剪根两个处理，每个处理设置五个重复，数据分析结果显示：对于剪根苗，上部和中部、上部和下部的含菌量有显著的差异，中部和下部含菌量差异不显著；带根苗，上部、中部、下部含菌量彼此之间差异显著；不管是剪根苗还是带根苗，木质部和韧皮部含菌量之间的差异都非常显著；上中下与木韧交互作用中，含菌量差异非常显著；最后筛选出来的强弱2个菌株HY3和93B，它们的纤维素酶活性分别为：1.955ug/和1.288ug/，具有显著的差别；果胶酶的活性分别为：1.325 ug/和1.24ug/，没有显著的差别，而且HY3和93B两个菌株细胞分泌的胞外多糖的含量差异很显著，分别为：7.08×10-8ug/cell和5.17×10-8ug/cell。

The lysis efficiency was about 90% for heat-inducible vector or around 30% forUV inducible vectors. This set of autolytic units should be transferable to other vectors.

本体系能够通过简单的物理方法，将具有生物活性的胞内酶释放到胞外的，解决了定向进化的一个瓶颈问题。

METHODS: The BEC were preincubated with crocetin (1, 0.1 and 0.01 μmol/L) for 12 h, then exposed to AGE (100 mg/L). The RAGE mRNA expression was detected by RT-PCR analysis, the intercellular adhesion molecule-1(ICAM-1) was measured by ELISA. The extracellular superoxide ion and thiobarbituric acid reactive substances were assessed with superoxide ion kit and colorimetric assay, respectively The intracellular I2O2 was also detected using the probe 2,7-dichlorofluorescein. The mitochondrial membrane potential and mitochondrial Succinate dehydrogenase were analyzed by the retention of rhodamine 123 (Rh123) and MTT.

不同浓度的西红花酸(1、0.1、0.01μmol/L)预孵BEC细胞12h后，用AGE(100mg/L)刺激细胞12h，RT-PCR法测定RAGEmRNA的表达水平；ELISA法测定细胞间黏附分子－(ICAM-1)的表达；试剂盒分别检测胞外超氧阴离子和硫代巴比妥酸反应产物浓度；同时，还用2，7-二氯荧光素测定了胞内H2O2的浓度，并用罗丹明123(Rh123)荧光法及MTT法分别检测细胞线粒体膜电位水平和其琥珀酸脱氢酶的活性。

The former stimulates the cells and transfer the signals into the cells through the ligation with receptors on the cell membrane. The latter, a derivative of nucleotide, causes a cytotoxic effect to suppress thymidylate synthetase activity, and, therefore, to damnify RNA and DNA synthesis.

前者通过与细胞表面的受体结合，把胞外刺激传递到胞内，诱导免疫受体介导的信号级联放大作用，引起一系列免疫效应；后者是是小分子的核苷酸衍生物，可直接进入细胞内，通过抑制胸苷酸合成酶及损伤RNA、DNA等对基因转录和核苷酸代谢产生影响，最后产生细胞毒效应。

Moreover, no significant changes in flagellar mobility, biofilm formation and production of extracellular cellulase and xylanase in vitro were observed in △rbfCxoo compared to PXO99A. Most importantly, the deletion mutation of rbfCxoo resulted in enhanced virulence and gene expression.

此外，△rbfCxoo鞭毛运动性、生物膜形成和胞外纤维素酶和木聚糖酶活性都无明显改变，但对水稻的致病性显著增强，毒性相关基因的表达也有所增加。

Na-K-Exchanging ATPase is a universal cell membrane protein in higher eukaryotic cells which mediates the anti-concentration gradient exchange of intracellular Na+ for extracellular K+. It plays essential metabolic roles such as maintaining sodium and potassium ion gradients across the plasma membrane. The targeted sequences were amplified by PCR to determine the genome structure of the hithertofore unsequenced portion of the alpha 1 subunit of the human Na-KExchanging ATPase gene which encodes 80-130 aa of its extracellular domain using two different templates, human genomic DNA and a human muscle cDNA library. The PCR products were analyzed by restriction endonuclease digestion and then cloned into a plasmid vector for chemiluminescence sequencing and further analysis.

Na-K-Exchanging ATPase是一种普遍存在于高等生物体内的细胞膜蛋白，主要参与介导K+和Na+在细胞内外之间的逆浓度梯度的转运，并维持一定细胞内外的离子梯度，我们采用聚合酶链式反应方法分别以人基因组DNA及cDNA文库为模板对人Na-K-Exchanging ATPaseα1亚单位基因胞外区约80-130位氨基酸编码序列进行扩增，限制性酶切分析扩增产物，并进行荧光测序，对测序结果进行同源性分析及剪接位点的搜索并对得到的核苷酸序列进行进一步的分析，发现人基因组DNA和cDNA经过扩增后分别得到833bp和195bp两种不同大小的片段Fg，Fc。

A part of microbial cells would autolyzed to release the intercellular alkanedegrading enzymes when they were cultured on alkane. Thus alkane could be degraded and synthesized alkane derivative with surface activity extracellularly. This surface-active agent would improve alkane emulsifying, and those unautolyzed cells could utilize the emulsified alkane sequentially.

微生物细胞在烃类基质中培养时，为了适应其疏水性环境以更好地摄取烷烃，会通过一部分细胞的自溶解体而使得胞内烃类代谢酶等释放，这样烃类在胞外被代谢生成具有表面活性的烷烃衍生物，该表面活性产物将促进烃类的乳化，使得剩余的未自溶解体的细胞可以借助生物表面活性剂而摄取降解烃类。

To better understand the make-up of the enzyme producing strains, API 20E identification system were employed and results indicated that majority of the enzyme producing strains were vibrios. They made up 46%, 42% and 40% of the protease, lipase and cellulase producing populations respectively.

API鉴定结果表明，在有能力分泌胞外蛋白酶、脂肪酶和纤维素酶的菌株中，弧菌菌群所占的比例分别为46％、42％和40％，而其中河流弧菌又占各弧菌菌群比例的83.3％、62.5％和100％。

However, as the name（read－only memory）implies, CD disks cannot be written onorchanged in any way.

然而，正如其名字所指出的那样，CD盘不能写，也不能用任何方式改变其内容。

Galvanizes steel pallet is mainly export which suits standard packing of European Union, the North America. galvanizes steel pallet is suitable to heavy rack. Pallet surface can design plate type, corrugated and the gap form, satisfies the different requirements.

镀锌钢托盘多用于出口，替代木托盘，免薰蒸，符合欧盟、北美各国对出口货物包装材料的法令要求；喷涂钢托盘适用于重载上货架之用，托盘表面根据需要制作成平板状、波纹状及间隔形式，满足不同的使用要求。