表达性

Src l was an acidic cytolysin, which was reported forthe first time. The cDNA encoding the mature Src l was cloned into non-fusion expression vectorpBV220 and expressed successfully in E.coli DH5α in inclusion body. After washing anddenaturing-renaturing, the recombinant Src l protein was purified by Q Sepharose FastFlow ion exchange chromatograph and Phenyl Sepharose hydrophobic interactionchromatograph.

首先构建了Src I基因的非融合蛋白表达质粒pBV220-Src I，并成功地在原核细胞E.coli进行了重组表达，筛选出稳定表达的工程菌株DH5α，摸索了稳定表达的条件，摸索了包涵体的洗涤、变性和复性条件，摸索出了复性蛋白Src I的纯化工艺，纯化过程简单，经QSepharose Fast Flow阴离子交换层析和Phenyl Sepharose疏水层析两步纯化，就可获得了高纯度的重组蛋白样品(纯度达99％以上)，该纯化方法适合重组蛋白的大量纯化，为Src I的性质、结构和功能研究奠定了良好的基础。

The result showed that twocDNA fragments which expressed at high level both in shoot and radicle representedthe gene encoding beta-D-glucosidase; one cDNA fragment expressed specifically inshoot represented the gene encoding mitochondria HSP60; most clones of MF12 andMF17 fragments respectively represented the chloroplast genes encoding prp22 andprp19 proteins which are two components of ribosomal small subunit; while thededuced amino acid sequence from each exceptional one clone was respectivelyhomologous to CDC5 proteins and vesicle-associated membrane proteins; otherthree cDNA fragments expressed preferentially in shoot had no homologue inGenBank.

结果发现2个在胚芽和胚根中表达量都很高的cDNA片段代表的是编码玉米β-D-葡糖苷酶的基因；一个在胚芽中表达而在胚根中不表达的片段代表的是编码线粒体分子伴侣HSP60的基因；片段MFl2的大部分克隆测序结果是与叶绿体基因组中编码核糖体小亚基蛋白prp22的基因同源，但其中有一个克隆测定的cDNA片段序列推测的氨基酸序列与CDC5家族成员有较高的同源性；片段MF17的大部分克隆测序结果与叶绿体基因组中编码核糖体蛋白prp19的基因同源，而有一克隆测定的cDNA片段序列推测的氨基酸序列与参与信号转导的膜结合蛋白VAP27和VAMP有较高的同源性；另有3个优先在胚芽中表达的cDNA片段未查询到同源序列。

24H treatment group compared with 24h model group: the density of light of HSP70 of the group increased and which of TGF-β〓, is not different with the model group; the density of the light the HSP70 of the Baicalin group increased and which of it the TGF-β〓, increased with the model group; the density of light of the Concha Margatitifera Usta is not different and which of TGF-β〓, increased with the model group; the density of light of the Cholic Acid group is not different and the density of light of it of TGF-β〓 increased with the model group; the density of light of the Hefang Group is not different and which of TGF-β〓 is not different than the model group.

合方治疗脑缺血12小时组缺血脑组织TGF-β1光密度值较缺血12小时模型组无显著性差异（p＞0.05）。本实验结果发现，与单纯缺血12小时、24小时大鼠相比，药物干预组缺血侧大脑皮层TGF-β1免疫反应明显增强，计算机辅助图像半定量分析两组间缺血侧大脑皮层TGF-β1免疫阳性细胞数差异非常显著。说明药物对局灶性脑缺血损伤可能有一定保护作用，其机制可能与诱导脑细胞TGF-β1合成增加有关。然而，药物调控TGF-β1表达，其促TGF-β1表达增强的机制可能是这样的：脑缺血时TGF-β1mRNA表达增加可能与缺氧诱导神经和胶质细胞刺激生长因子表达有关。

Results We obtained a lysogenic bacteriophage MZTP01with clear plaque and 1 mm diameter. Fragment D with 2362bp (Genebank No. AY639599) was obtained after the phage DNA hydrolyzed by HindⅢ/EcoRⅠ. Among fragment D, the pep gene with molecular weight of 47 kDa and length of 1101bp was cloned and expressed. Recombinant M15 (pQE30pep) was built and overexpressed in Escherichia coli with a 47kDa clear band. At the same place a clear band was observed by Western blot. Judging from the time course expression, we could conclude that PEP protein produced at 1 hour after induction and then increased gradually. PEP protein was mainly in the form of inclusion body in the recombinant and slowed the growth speed of host. Homologous comparison of PEP protein from phage MZTP01 with other PEPs from BLAST were that phage MZTP01 PEP protein had 100% homologe with that of Escherichia coli K12, and most of others took the similarity in the range between 37%~84%.

诱导获得的溶原性噬菌体MZTP01斑点清晰，直径约1mm，成斑时间12h；从噬菌体基因组DNA双酶切(HindⅢ/EcoRⅠ)片段中回收长度为2362bp的D片段(Genbank登录号： AY639599)，又从D片段中克隆了长度为1101bp、编码367aa、分子量为47kDa的pep基因，表达载体M15(pQE30pep)在大肠杆菌(Escherichia coli， E.coli)中表达获得了47kDa的清晰表达带，在1h 时开始产生蛋白并有逐步上升的趋势； Western blot 也在47kDa处得到一条清晰的条带；可溶性分析表明PEP蛋白在重组菌株中是以不可溶的包含体形式存在的，该蛋白的产生明显地抑制了宿主的生长速度；噬菌体PEP氨基酸序列之间的同源性比较表明，噬菌体MZTP01 PEP蛋白与来自E.coli K12噬菌体的PEP蛋白的同源性程度最大。

In Hp positive group, the expression of IL-10 in active gastritis is lower than that in inactive gastritis, which are different with the expression of COX-2 and iNOS, indicating IL-10 is a protect factor in gastritis, but COX-2 and iNOS may relate to pathopoiesis.

Hp阳性组，IL-10在活动性胃炎粘膜组织中的表达低于非活动性胃炎，COX-2、iNOS在活动性胃炎中的表达显著高于非活动性胃炎，说明IL-10在活动性胃炎的发病中可能起保护作用，而COX-2、iNOS可能与致病过程有关。

The results showed that the expression of calm3 and trm112l genes had no remarkable differences in oocytes and 8-cells stage embryos, but were significantly different in blastocysts. It was supposed that these two genes were possibly maternal genes, and they might be involved in oocyte maturation and zygotic genome activation.

结果显示，在卵母细胞和8细胞期胚胎，基因calm3和trm112l的表达量无显著性差异，而在囊胚期的表达量与前两个时期的表达量存在显著性差异，推测这两个基因在牛胚胎发育过程中的表达可能属于母型调控，提示该基因在卵母细胞成熟和合子基因激活中起一定作用。

Baculoviruses have emerged as a popular system for overproducing recombinant proteins in eukaryotic cells. Several reasons lead to this popularity. First, the rod shaped capsid can extend to accommodate large DNA inserts.

昆虫杆状病毒载体表达系统由于对人畜安全无害，外源基因容量大，表达水平高，表达产物的抗原性、免疫原性与功能均类似天然产物，因而在诸多表达系统中日益引人注目。

Methods Samples from 78 patients with pituitary adenomas, including 37 invasive and 41 noninvasive, and 5 normal pituitary tissue specimens from autopsied patients were examined. The expressions of OPN mRNA and OPN were examined by real-time fluorescence quantitative PCR and immunohistochemical SP method, respectively.

分别对78例垂体腺瘤手术标本（其中侵袭性37例，非侵袭性41例）和5例来源于尸检的正常垂体组织进行检测，采用实时荧光PCR技术检测组织中OPN mRNA表达，免疫组织化学SP法检测OPN蛋白表达，结合临床资料分析OPN mRNA及其蛋白的表达与垂体腺瘤侵袭性的关系。

Methods samples from 78 patients with pituitary adenomas, including 37 invasive and 41 noninvasive, and 5 normal pituitary tissue specimens from autopsied patients were examined. the expressions of opn mrna and opn were examined by real-time fluorescence quantitative pcr and immunohistochemical sp method, respectively. the relation of opn and opn mrna expressions to invasiveness of pituitary adenomas was analyzed.

分别对78例垂体腺瘤手术标本（其中侵袭性37例，非侵袭性41例）和5例来源于尸检的正常垂体组织进行检测，采用实时荧光pcr技术检测组织中opn mrna表达，免疫组织化学sp法检测opn蛋白表达，结合临床资料分析opn mrna及其蛋白的表达与垂体腺瘤侵袭性的关系。

Results were shown as followings:(1) Sodium selenite at 0～2.5 μmol/L significantly increased the antioxidative capacity of L-02 cells without having remarkable impact on SMMC-7721 cells;(2) Sodium selenite at concentrations above significantly increased telomerase activity, hTERT gene expression and telomere length of L-02 cells without significant impact on SMMC-7721 cells;(3) Sodium selenite at higher concentrations (larger than 5 μmol/L) resulted in peroxidation of L-02 cells, while scutellarin significantly counteracted its effect;(4) Selenium-rich amino acids from silkworm pupas in the range of 0.5～2.5 μmol/L Se significantly inhibited SMMC-7721 cell growth, induced apoptosis and cell cycle change, and the generation of reactive oxygen species. In contrast, sodium selenite and selenomethionine only had weak impact on them at the same concentrations;(5) A new selenium-containing protein was found from selenium-rich silkworm pupas, which is worthy to be study further;(6) An expression vector containing ansense RNA of hTERT gene were constructed and used to transfect SMMC-7721 cells. They were observed to inhibit hepatoma cells.

结果如下：（1）0～2.5μmol/L亚硒酸钠显著性增强L-02细胞的抗氧化能力；而对SMMC-7721细胞的作用不显著；（2）该浓度硒显著性提高L-02细胞端粒酶活性、增强hTERT基因表达和延长细胞端粒长度；而对SMMC-7721细胞的作用均不显著；（3）高浓度硒（5μmol/L以上）显著性抑制L-02细胞生长、致细胞过氧化，灯盏花素能拮抗硒所致过氧化、降低硒毒性；（4）0.5～2.5μmol/L富硒蚕蛹氨基酸显著性抑制肝癌细胞SMMC-7721生长、导致细胞凋亡和周期改变、诱导细胞产生活性氧，同浓度亚硒酸钠和硒代蛋氨酸对其抑制不显著；（5）富硒蚕蛹蛋白经分离纯化和鉴定后发现存在一新含硒蛋白，其结构和功能有待研究；（6）通过已有的含hTERT基因质粒，成功构建hTERT反义RNA表达质粒，转染SMMC-7721细胞后对其生长具有抑制作用。

I'm strongly against the death penalty — it's an eye for an eye.

我不赞成死刑——这是以牙还牙的报复行为。

And to get you the support you need, we're enlisting all elements of our national power: our diplomacy and development, our economic might and our moral suasion, so that you and the rest of our military do not bear the burden of our security alone.

并给你们所须的支援，我们正徵召国家所有各种的力量：我们的外交及发展，我们的经济力量与道德劝说，所以你们与其他军人不须要孤独地负起国家安全的责任。