细胞内的

Abrin is one of the potent toxins isolated from the beans of plant abrus precatorius.the toxic protein consists of two disulfide-bonded subunits,the a and b chains.the b chain is a galactose-specific lectin that facilitates the binding of abrin to the cell membrane that precedes endocytosis.after entering the cells,the a chain catalytically inactivates 60s ribosomal subunits by removing adenine position 4324 of 28s r–rna,and thereby inhibits protein biosynthesis.

相思子毒素是从豆科植物(abrus precatorius l。)种子中分离的一种细胞毒性蛋白。它由a，b两条链组成，由一个二硫键相联，b链具有半乳糖凝集活性，可与细胞膜上受体结合，帮助a链进入细胞内，a链进入细胞催化失活60s核糖体亚基，从而使细胞蛋白合成被抑制。

Results demonstrated that the level of ROS was greater in embryo than in aleurone layer, and only very little ROS could be detected in starch. Furthermore, the ROS produced in embryo were mainly localized inside cells but not on cell membrane.

结果表明，胚细胞产生的活性氧最多，糊粉层细胞次之，淀粉细胞几乎不产生活性氧；而且，胚细胞产生的活性氧主要定位在细胞内，细胞膜上不产生。

There is now strong evidence that chloroplasts and other cell organelles, such as mitochondria, represent prokaryotic organisms that invaded heterotroph ic eukaryotic cells early in evolution and are now part of an indispensable symbiotic union see endosy

这恰恰是现在有关叶绿体及一些其他的细胞器如线粒体是被原始的真核细胞吞噬进细胞内，与宿主长期共生而逐渐演化出独立单元的内共生起源学说的重要证据。

These ingredients includes epidermal growth factor, hepatocyte growth factor, dimethyl sulfoxide, phenobarbital sodium and all trans-retinoic acid. We isolate and culture primary 10-weeks-old human fetal hepatocytes, observe hepatocytes form and its function markers. Inoculate hepatocytes with HBV Dane particles to observe if HBV susceptibility of fetal hepatocytes can be induced in vitro. To detect HBV covalently closed circular DNA within hepatocytes, we improve the existent PCR method.

基于上述假设，我们设计了10种培养条件，在自行设计的无血清培养基中添加不同的细胞分化诱导物，包括表皮生长因子、肝细胞生长因子、二甲基亚砜、苯巴比妥钠、全反式维甲酸等，分离孕10周龄人胎肝细胞进行原代培养，观测肝细胞形态及功能指标变化，并定时给予HBV感染，观察能否在体外诱导胎肝细胞出现HBV易感性；为检测肝细胞内HBV共价闭合环状DNA(covalently closed circular DNA，cccDNA)，我们改进了现有的PCR检测法。

After Bastard halibutwas poured in Edwardsiella tarda through mouth, the immunocyte' s immunocompetent of pronephros and spleen increased because the number of monocytes added and large granule' s inchon driosis strengthened, also many small granules produced in granulocytes.

当经口腔灌注大量迟缓爱德华氏菌后，牙鲆的头肾和脾脏内的免疫细胞活性增强，表现为单核细胞数量增加，颗粒细胞内产生大量的颗粒以及颗粒吞噬作用的增强。

Get high purity DCs by Cultured plastic-adherent monocytes isolated from healthy human peripheral blood with GM-CSF and IL-4 for 7 days. To observe the morphology of DCs by inverted phase contrast microscope ,electron microscope and laser confocal microscope. Analyse phenotype of DCs with flow cytometry. Investigate the endocytosis ability of DCs as a group by Horseradish peroxidase endocytosis assay. To appraise allogeneic mixed lymphocytes reaction of DCs by MTT reduction assay. Analyse the levels of IL-12 and TNF in liquids of cultured medium by ELISA and MTT reduction assay respectively. Soluble antigens of HCCs was obtained by 3 freeze-and-thaw cycles. Biological characteristics of HC soluble antigens pulsed DCs were monitored by flow cytometry. According to MTT reduction assay estimated the cell proliferation of self lymphocytes activated by HC antigens pulsed DCs. Get high purity BCG HSP 70 protein by SDS-PAGE electrophoresis and determined its biological activity with ELISA. Analyse phenotype of antigen pulsed DCs primed by BCG HSP70 with flow cytometry. By MTT reduction assay estimated the cell proliferation of self lymphocytes and the MLR of DC based vaccine. Analyse expression of HLA-DR molecule on surface of HCC lines. The IFN-γ mRNA in lymphocytes after actived by DC vaccine and the Fas-L expression on DC and DC vaccine primed lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes and nonspecific inhibition of liquids in culture medium against HCC lines were also tested. Detect expression of hAFP on four HCC lines with Cell-ELISA. Induce apoptosis of HCCs with actinomycin-D. Interaction of DCs and apoptotic cells was observed under transmission electron microscope. Growth inhibition test of DC against HCC lines was also performed. Establish the nude mouse model bearing human HC xenografts and indentify the characteristic of tumour by histochemistry and immunohistochemistry techniques. Prevent and treat transplanted human HC on nude mouse with Freezing and anabiotic HC specific lymphocytes.

用GM-CSF和IL-4从健康人外周血诱导DC；分别用倒置相差显微镜、电子显微镜及激光共聚焦显微镜观察DC形态；流式细胞术检测DC表型；HRP吞噬实验测定DC的群体内吞能力；MTT法检测同种异体混合淋巴细胞反应；ELISA法和MTT法分别测定DC培养上清液中IL-12和TNF水平；冻融法制备肝癌细胞可溶性抗原；流式细胞术检测负载肝癌可溶性抗原后DC的生物学特性；MTT法检测DC负载肝癌抗原后对自身淋巴细胞增殖的影响；SDS-PAGE制备电泳纯化BCG HSP70并鉴定纯度，ELISA测定活性；流式细胞术检测负载抗原DC经BCGHSP 70活化后的表型；MTT法检测肝癌DC疫苗对自身淋巴细胞增殖的影响和混合淋巴细胞反应；流式细胞术检测肝癌细胞表面HLA-DR表达；MTT法检测肝癌DC疫苗对自身淋巴细胞的活化；原位杂交法检测肝癌DC疫苗活化后的淋巴细胞IFN-γmRNA表达；流式细胞术检测DC和肝癌DC疫苗活化后淋巴细胞表面Fas-L；MTT法分别检测肝癌DC疫苗活化的淋巴细胞和其培养上清对肝癌细胞的特异性杀伤和非特异性抑制作用；Cell-ELISA检测人肝癌细胞hAFP表达；MTT法检测负载AFP表位肽和凋亡肝癌细胞DC对自身淋巴细胞增殖的影响；ELISA法和MTT法分别测定活化后淋巴细胞培养上清中TNF和IL-12水平；肝癌细胞凋亡的诱导和检测；DC吞噬凋亡肝癌细胞后的电子显微镜观察；DC对肝癌细胞的生长抑制试验；人肝癌裸鼠皮下移植瘤动物模型的建立及其组织学和免疫组织化学鉴定；DC及肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤；冻存和复苏后的肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤。

Grew on the normal temperature, it was shown that the deposits of calcium antimonate being the indicator for Ca(superscript 2+) localization mainly concentrated within the vacuoles and intercellular spaces and there was also some Ca(superscript 2+) deposits in cell walls. But when Garyota urens L. was treated by the temperature of 2℃ for 48 h, the level of Ca(superscript 2+) increased in cytoplasm and plasma membrane, but decreased in vacuoles and intercellular spaces considerably. At the same time, the ultrastructure of chloroplasts suffered from chilling: the membrane of chloroplasts had been damaged, the layer of thylakoids was exiguous and unclear, the photosynthetic rate decreased evidently. And when Garyota urens L. was treated by the temperature of 2℃ for 120 h, the deposits of Ca(superscript 2+) mainly concentrated within the cytoplasm, nucleus and plasma membrane and there was also some Ca(superscript 2+) deposits in vacuoles, and the ultrastructure of some cells was simultaneously damaged severely: Chloroplasts structure, vacuole membrane and nuclear membrane had been damaged fully, the structure within the cell had become unclear, and the cell only have respiration.

研究结果表明，未经低温处理的董棕幼苗叶肉细胞，焦锑酸钙沉淀颗粒大量出现在液泡和细胞间隙中，细胞壁中也可见少量沉淀，而细胞基质中则看不到焦锑酸钙沉淀；经2℃ 48 h低温处理后，细胞基质和细胞膜上焦锑酸钙沉淀增加，而液泡和细胞间隙中的焦锑酸钙沉淀则显著减少，并且超微结构已初步显示出寒害的特徵，叶绿体外膜部分破损，类囊体片层稀疏且排列不规则，光合速率明显下降等；经2℃ 120 h低温处理后，细胞间隙内的焦锑酸钙沉淀极少，有的也紧贴在细胞外壁上，而细胞基质和细胞膜上则分布有非常多的焦锑酸钙沉淀，在核基质和液泡中也可见到少量的焦锑酸钙沉淀，并且超微结构遭到了显著破坏，叶绿体结构完全被破坏，核膜与液泡膜严重破损，内部结构模糊，细胞只表现为呼吸作用，不进行光合作用。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank，选择包含人主要T细胞抗原表位序列的人PSCA基因片段，应用异种化抗原设计技术，保留人T细胞抗原表位，设计异种化PSCA融合抗原片段；(2)根据核酸序列按中心模板法设计引物，应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因，以PCR、限制性酶切和DNA序列测定法进行鉴定：(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点，采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4)，并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中，转染293T细胞，借助免疫荧光+流式细胞术考察插入片段表达效率，最终选定PSCA_3片段进行下一步研究；(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下，插入pVAX1-neo-IRES—GM/B7载体中，构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB，并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况；(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞，制备小鼠前列腺癌模型，并采用股四头肌肌肉注射+电脉冲法(Electroporation，EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB，同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照，通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率，来评价该DNA疫苗在小鼠体内的抑瘤效果。

Protein disulfide isomerase is one of main retention proteins in ER, which is an abundant protein in luman of the ER especially in secretory cells.

蛋白质二硫键异构酶是内质网腔内的主要驻留蛋白之一，在分泌细胞的内质网腔中含量非常丰富，其主要功能是催化细胞内分泌蛋白和膜蛋白新生多肽的二硫键形成。

There is now strong evidence that chloroplasts and other cell organelles, such as mitochondria, represent prokaryotic organisms that invaded heterotrophic eukaryotic cells early in evolution and are now part of an indispensable symbiotic union.

这恰恰是现在有关叶绿体及一些其他的细胞器如线粒体是被原始的真核细胞吞噬进细胞内，与宿主长期共生而逐渐演化出独立单元的内共生起源学说的重要证据。

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。