细胞

The aim are:lTo examine the proliferation ability and potential chondroblast differentiation ;2To find an ideal condition stimulated BMSC differentiate into chondroblast;3To examine the chondroblast proliferation in porous scaffolds and to explore the interaction between cells and materials;4To examine the release of cytokine in vitro.

取健康成人略后上棘处骨髓 5ml，经 percoll液分离后密度梯度离心，10'砌l密度接种培养，观察原代细胞的贴壁、增殖状况，细胞长满瓶底后进行传代培养；传至第三代细胞，重新消化后以500转／分钟轻度离心 5分钟，10V加 l接种，改用化学限定培养基代替完全培养基培养，倒置显微镜观察细胞生长情况及形态变化，甲苯胺蓝染色观察诱导细胞合成细胞外基质QCM）中的蛋白多糖，免疫细胞化学染色检测ECM中＊胶原的蛋白合成，RT干CR鉴定诱导细胞＊胶原mRNA的表达。

In resent year, several in vitro models for committed differentiation of ES cells have been described which give rise to a variety of cell types, including hematopoietic, neuronal, chondroitic, osteocytic, endothelial, myogenic, cardiac and smoothes muscle cells, but the committed differentiation into epidermal stem cells have not been reported.

ES细胞为研究基因功能、胚胎发生和个体发育，及组织细胞分化机理提供了理想的模型。近年来的研究表明，适当条件下，ES细胞可定向分化为三个胚层的各种类型的细胞，其中包括向神经元、造血细胞、成骨细胞、心肌细胞、血管内皮、软骨细胞、平滑肌特方向的定向分化。

The cardiomyocyte viability were detected by MTT assay for calculating the survival rate and the inhibitory rate of the virus. Myocardial F-actin and VEGF were analyzed by immunofluorescent staining with confocal microscopy. Mean fluorescent intensity of F-actin and VEGF were determined by flow cytometry.Results There is no toxicity on cardiomyocytes when PD is at concentration of 0.02 mmol/L and 0.2 mmol/L while there is toxicity at concentration of 2 mmol/L. CVB3 could reduce the viability of cardiomyocytes, depolymerize F-actin cytoskeleton and reorganize VEGF protein.

体外培养心肌细胞，用100 TCID50柯萨奇B3病毒（CVB3）感染心肌细胞，建立病毒性心肌炎实验模型，然后分别用0.02mmol/L、0.2mmol/L、2mmol/L三种不同浓度的PD处理感染CVB3病毒的心肌细胞，观察心肌细胞的形态和搏动，用细胞免疫荧光技术标记纤维状肌动蛋白和VEGF蛋白，四唑蓝比色法测定心肌细胞活性和病毒抑制率，流式细胞仪检测各组心肌细胞VEGF蛋白和F-actin平均荧光强度。

Human umbilical venous endothelial cells were cultured, and were exposed to different concerntrations of HCY with/or not with CuSO4; Cell counting of detached cells were performed with a hematocytometer; the cell survival was monitored by MTT assay and the cellular injury was evaluated by LDH release; Hoechst 33258 staining was used to observe nuclear morphological changes and the apoptosis was measured by DNA agarose gel electrophoresis ;the percentage of apoptosis was quantified by flow cytometry.

体外培养人脐静脉血管内皮细胞，用不同浓度的HCY伴／不伴生理浓度Cu~(2+)与内皮细胞作用；细胞计数板检测脱落细胞率；MTT法及乳酸脱氢酶释放率的测定来衡量细胞的存活与损伤；Hoechst 33258荧光染色观察细胞核形态变化和DNA琼脂糖凝胶电泳检测细胞凋亡，并采用流式细胞术定量测定细胞凋亡率。

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。

Results are as followed:1 Exposure of HELF cells to BP caused c-Jun activation,and increased the activity of MAPK,PI-3K,p53 and cyclin D1 pathway.2 BP-induced c-Jun activation was inhibited by dominant negative mutants of extracellular signal-regulated protein kinase or c-Jun NH_2-terminal kinase,but not by p38,impling that JNK and ERK pathways medicate c-Jun activation induced by BP.3 Overexpression of dominant-negative mutants PI-3K and Akt potently blocked phosphorylations of c-Jun and ERK,but not JNK in response to BP,suggesting that PI-3K/Akt pathway positively regulates BP-induced c-Jun activation through ERK.4 Inhibition of p53 by its chemical or molecular inhibitor markedly increased the phosphorylation levels of c-Jun,Akt and ERK upon BP stimulation,indicating that p53 negatively medicates BP-induced c-Jun activation through PI-3K/Akt/ERK pathway.5 The cell lines expressed TAM67 exhibits no significant affecting normal cell growth properties.6 TAM67 was able to significantly block G_1-S transition and subsequent cell proliferation,suggesting that c-Jun is essential for cell cycle alternations elicited by BP.7 Overexpression of TAM67 impaired BP-induced cyclin D1 activation,decreasing expression of E2F1 and pRb,indicating that c-Jun participates in the modulation of BP-induced activation of cyclin D1/pRb/E2F1 pathway.8 Stably expression of TAM67 led to the increases in the expression levels of p53 and p21,elevating phosphorylation level of p53,clearly indicating that c-Jun regulates p53/p21 pathway activation induced by BRCollectively,PI3K/Akt/ERK pathway mediated BP-induced c-Jun activation through p53-dependent mechanism.

结果显示：1BP刺激细胞可促进c-Jun活化，并伴随着MAPK、PI-3K、p53和cyclinD1通路各组成成分的活性增强。2利用MAPK通路的显性失活突变体分别阻断细胞外信号调节激酶和c-Jun氨基末端激酶活性，均可明显抑制BP诱导的c-Jun活化，但阻断p38活性对BP引起的c-Jun活化无明显影响，提示JNK和ERK通路参与调控BP诱导的c-Jun活化。3过表达PI-3K和Akt的显性失活突变体也可显著抑制BP诱导的c-Jun活化，并降低磷酸化ERK的表达水平，但对磷酸化JNK的表达水平无明显影响，说明PI-3K/Akt通路通过ERK正性调控了BP诱导的c-Jun活化。4p53的化学/分子抑制剂能使BP作用的细胞内c-Jun活性明显增加，并同时诱导Akt和ERK的磷酸化水平的升高，表明p53可通过PI-3K/Akt/ERK通路对BP诱导的c-Jun活化进行负性调控。5随后观察转染细胞的生长情况，发现TAM67对细胞正常生长和形态无明显影响。6稳定表达TAM67可有效抑制BP诱导的S期细胞数的增加，提示c-Jun在BP致细胞周期改变的过程中发挥了重要作用。7TAM67过表达能够抑制BP诱导的cyclin D1活化，降低磷酸化Rb以及E2F1蛋白表达水平，表明c-Jun参与调控BP诱导的cyclin D1/Rb/E2F1通路的活化。8过表达TAM67可使BP刺激的细胞中p53、p21总蛋白以及p53磷酸化的表达水平明显升高，可见c-Jun也参与调控BP诱导的p53/p21通路活化。

HBV transplacental transmission route may be realized by cell to cell transfer in placenta, which means that HBVs in maternal blood and/or intervillous space infect DCEC and/or DC and/or TC, then transmit to VMC, then to VCEC, and finally to fetus.

HBV经胎盘感染胎儿的路径可能是通过胎盘细胞至细胞的传递而实现的。即母血和/或绒毛间隙HBV→蜕膜毛细胞管内皮细胞和/或蜕膜细胞和/或滋养层细胞→绒毛间质细胞→绒毛毛细血管内皮细胞→胎儿。

In the whole Lauraceae or in some large genera, the evolutionary trend of the distribution types of oil cells and mucilage cells might be as follows: typeⅠ→ type Ⅱ→type Ⅲ→ type Ⅳ.

在本科内甚至在某些属内，油细胞和粘液细胞分布的4种类型的演化趋势为：只有油细胞分布→油细胞与粘液细胞共存→只有粘液细胞分布→油细胞与粘液细胞皆无。

The contributions of photon radiations are ignored because they are negligible compared to the contributions of the particle radiations, or can be calculated and added to the S-values using MIRD formulae.

如果细胞群较小，则光子的剂量贡献忽略不记。细胞群中每个细胞内的放射性活度相同。计算得到细胞群中各个周围细胞中心到靶细胞中心的距离和周围细胞中的核素对靶细胞的剂量贡献。

However, in 2 cases with intestinal tuberculosis, another chronic inflammatory disease of the large bowel, reticulum cells and countless neutrophilic leucocytes were seen in both cases , and plasma cells , atypical epithelial cells, and multinucleated giant cells in one case, In 7cases with amebic colitis countless neutrophilic leucocytes were seen in 2 cases (28.6%), plasma cells, reticulum cells, atypical epithelial cells, and multinucleated giant cells in one case (14.3%) respectively.

然而，在肠结核（另一种大肠慢性感染疾病）2例中，网状细胞和无数嗜中性白细胞在2例中均可见，浆细胞、网状细胞何多核巨噬细胞仅在1例中可见。在7例变形虫结肠炎中，2例无数嗜中性白细胞可见，浆细胞、网状细胞、不规则上皮细胞和多核巨噬细胞分别在1例中可见。

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。