细胞的

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。

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。

Light microscope and transmission electron microscopy showed that SMMC-7721 cells induced by SAHA had undergone the restorational alteration in morphology and ultrastructure, which were different from those of nontreated cells but were similar to those of normal cells, and the changes were as follows: the cells turned to be flat and spread; the nucleo-cytoplasmic ratio lessened and nuclear shape became rather regular; the number of nucleolus reduced and its volume lessened; euchromatin increased while heterochromatin decreased in nucleus; in the cytoplasm, mitochondria grew in number with relatively consistent structure and well-developed mitochondria cristae; Golgi complex turned to be well-developed and typical; rough endoplasmic reticulum increased. Immunocytochemistry assay showed that the expression of AFP and PCNA were declined significantly. FCM analysis showed SAHA could arrest SMMC-7721 cells in G0/G1 phase, with an accumulation of the cells in G0/G1 phase while a decrease of cells in S phase. Semi-quantitative RT-PCR detection revealed that the expression of p21WAFl mRNA was upregulated remarkably in the cells treated with SAHA.

结果：倒置显微镜和透射电镜观察显示，经SAHA处理的细胞增殖速度显著减慢，细胞体积增大，细胞核较小，形状较为规则，核仁数量减少、体积变小，核内常染色质增多而异染色质减少，核质比例减小，细胞质内线粒体数量增多、线粒体嵴发达，高尔基体较为典型，粗糙型内质网增多，呈现出与正常上皮细胞相似的形态变化；MTT比色法测定结果显示不同浓度(2.5、5.0、7.5、10.0uM)SAHA对SMMC-7721细胞的增殖均有抑制作用，并有明显的剂量依赖和时间依赖关系；免疫细胞化学检测显示SAHA能显著降低PCNA和AFP在SMMC-7721细胞中的表达；流式细胞仪检测结果显示，SMMC-7721细胞经SAHA处理后，G0/G1期细胞明显增加，S期细胞则明显减少，细胞被阻滞于G0/G1期；RT-PCR检测结果表明，SAHA作用12h后SMMC-7721细胞中p21WAF1 mRNA的表达即有增加，24h后更为明显。

Many factors may be involve in the course. To investigate the regulation activity of mesenchymal cells to differentiation of epithelial cells from hair follicle and to study its differentiation property, mesenchymal cells gel was made by nubby dermal papilla cells, free dermal papilla cells, skin fibroblasts. Skin keratinocytes and epithelial cells from hair follicle were inoculated on the gel surface and cultured in air-liquid interface. Three-dimensional model of DPC using to induce epithelial cells differentiation is built in vitro.

为了进一步研究毛囊细胞间的相互作用，探讨毛囊间质细胞对毛囊上皮细胞分化的调节作用，研究毛囊上皮细胞的分化特性，我们利用团块状的毛乳头细胞，游离分散的毛乳头细胞或皮肤成纤维细胞制成间质细胞胶原凝胶，表面接种皮肤角质形成细胞或毛囊上皮细胞，进行气-液界面培养，在体外建立了毛乳头细胞诱导毛囊上皮细胞分化的立体模型。

After the cell growth curves was recorded, RPE cells of the 3-5th passages were utilized. 2、Three different siRNA (siRNAl,siRNA2,siRNA3) targeting against human cx43 gene and one negative control siRNA were designed and transfected into cultured human RPE cells via liposome reagent. The most effective siRNA can be determined by semi-quantitative reverse transcription PCRRT-PCR. 3、To the most effective siRNA, after transfected into human RPEs with different concentration, the cellular proliferate activities were messured by MTT colorimetry ; the percentages of RPE in different cell circle phase was assayed by FCM; the changes of phenotypical properities were observed with SCM; the protein expression of cx43 was studied through immunocytochemistry stain and Weston blot; the communication intercellular was calculated with FRAP; and the ability of recovery was assessed by using an in vitro wound healing model.4、The total proteins of siRNA1 and RPE were seperated by two-dimensional gel electrophoresis and visualized by silver staining. Proteins with significant expression alterations were selected and their peptide mass fingerprints (PMFs were obtained by matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS).The PMFs were used to search NCBInr database by Auto MS-Fit software.

实验方法：1、培养原代的人RPE细胞，经过细胞角蛋白、S-100和神经胶质原纤维酸性蛋白免疫细胞化学鉴定后，通过AO/PI染色技术确定培养细胞的存活率，描记其生长曲线，第3-5代用于以下细胞实验2、生物合成针对人cx43基因的三条小干扰RNA和一条阴性RNA通过脂质体转染RPE细胞后，通过RT-PCR的方法确定抑制效率最高的干扰片断3、将该片段以不同浓度通过阳离子脂质体转染培养的人RPE细胞后，采用MTT法观察其对细胞的增殖力的作用；通过流式细胞仪观察其对细胞周期的影响；通过扫描电镜观察其对细胞形态的影响；通过免疫细胞化学和Weston blot观察其对cx43蛋白表达的作用；采用激光共聚焦和荧光淬灭恢复技术观察荧光恢复速率平均百分率，评价其对细胞间通讯功能的影响；通过制作RPE细胞损伤模型，观察其对损伤修复能力的作用4、分离纯化转染siRNA的RPE组和正常对照组RPE细胞的全部蛋白质，应用等电聚焦电泳和SDS-PAGE双向电泳技术，银染显示分离出的蛋白质斑点，经凝胶图像分析软件对两个样本进行胶图分析，寻找差异蛋白点。

The results indicated that the typical apoptosis was induced by PRRSV in lungs and uteri. The ultrastructural changes showed different characterizations at different stages of the infection. During the early stage from 8th h to day 3 postinfection,the apoptosis cells shrank,the cytoplasm concentrated,the chromatin condensed,the kernel disaggregated and the endoplasmic reticulum dilated. During the middle stage from day 5 to day 9 postinfection,the apoptosis cells evidently shrank,the cell membranes protu berated,the plastosome manifolded,and apoptosis bodies were found. During the last stage from day 10 upwards postinfection,the apoptosis bodies degenerated and disappeared,and no inflammation occurred. Few apoptosis cells were necrotic.

结果表明，PRRSV可诱导宿主肺和子宫发生典型的细胞凋亡，表现为细胞的超微形态结构随着病毒感染进程出现不同的特征性变化，早期（感染后第8 h至第3 d）凋亡细胞多表现为细胞体积缩小，细胞质密度增强，染色质浓缩，核仁解体，内质网扩张；中期（感染后第5 d至第9 d）凋亡细胞体积显著缩小，细胞膜突起，线粒体增多，有凋亡小体形成；晚期（感染后第10 d以后）凋亡细胞形态多表现为凋亡小体降解和消失，少数凋亡细胞在凋亡晚期表现为坏死细胞。

Cell cycle analysis was performed using flow cytometry and proliferous index was calculated.(9) Drug sensitivity assay was performed using MTT assay.

8借助流式细胞仪对基因转染细胞及其对照细胞进行细胞周期分析，并计算细胞的增殖指数(proliferous index，PI)。

Methods Pterygial samples were extracted and collected and the pterygial endothelial cells and pterygial fibroblasts were cultured alone, conditional and co-cultured to form different culture systems. The methods included that to select the suitable intensity of ultraviolet by MIT, to detect the changes of curves of growth about two kinds of cells by MIT and to explore the developments of protein and RNA of vascular endothelial growth factor and fibroblast growth factor-basic in three culture systems under ultraviolet whose intensity is 20 mJ/cm^2 by ELISA and RT-PCR.

收集翼状胬肉标本，采用血管内皮细胞和成纤维细胞单独培养、条件培养和共同培养的方法构建体系，用MTT法选择紫外线照射细胞的适宜强度；采用MTT法绘制强度20mJ/平方公分紫外线照射下细胞生长曲线；采用ELISA和RT-PCR检测紫外线照射下3种体系中细胞上清液和细胞中血管内皮细胞生长因子和成纤维细胞生长因子的蛋白和RNA含量变化。

These results indiacated that tranferring macrophages combinedwith cytokines could be a new adoptive immunotherapy protocol foradvanced cancer,and rTNF used in vivo locally could induce andactivate macrophages to kill tumor cells.Monocytes when activatedunderwent a series of phenotypic and functional changes includingthe expression of IL-2R which may provide an important immunoregu-latory pathway.The presence of lectin-like molecules on thesurface of monocytes and tumor cells may bring theeffector/target cells together,thus facilitating the inductionof apoptosis in target cells by triggering the production ofcytolytic factors and the modification of target of targer cell surface anti-gens.(such as HLA-DR).

综合以上结果可以得出以下结论：转输巨噬细胞并结合细胞因子的联合应用是肿瘤继承性免疫中的又一新的、有潜力的方法；抗肿瘤细胞因子rTNF可通过激活巨噬细胞在体内抑制对其不敏感的肿瘤细胞的生长；人单核细胞在一定条件下激活后，可表达IL-2R，进而一方面增强其自身对IL-2的敏感性，另一方面在体内也具有调节T细胞功能的作用；单核细胞和肿瘤细胞上存在的凝集素类受体可促使两种细胞之间在凝集素介导下相互接近，并诱发单核细胞产生细胞毒因子，以及通过调节靶细胞上表面抗原的表达，促进诱导靶细胞的细胞凋亡。

The pattern of aerenchyma formation could originate by the combination of cortical cell autolysis and enlarged intercellular space. The process occurred in some stages: cortical cells stopped expansion and the intercellular space became larger at the initial stage. Cortical cells to collapse and autolysis were found with the disintegration of cell contents. After the evacuation of cell contents, cortical cells became shrinking and invaginated.

形成过程大体分为：皮层细胞生长停止，细胞间隙增大；皮层细胞衰老，并发生自溶，内含物解体；解体物逐渐撤走，皮层细胞开始收缩、内陷，当细胞内含物全部撤走，相邻细胞的残留细胞壁叠合即在两侧形成通气空腔；径向上相邻的皮层细胞逐步发生凹陷和细胞壁叠合，结合方形的细胞间隙一起构成通气组织。

We have no common name for a mime of Sophron or Xenarchus and a Socratic Conversation; and we should still be without one even if the imitation in the two instances were in trimeters or elegiacs or some other kind of verse--though it is the way with people to tack on 'poet' to the name of a metre, and talk of elegiac-poets and epic-poets, thinking that they call them poets not by reason of the imitative nature of their work, but indiscriminately by reason of the metre they write in.

索夫农 、森那库斯和苏格拉底式的对话采用的模仿没有一个公共的名称；三音步诗、挽歌体或其他类型的诗的模仿也没有——人们把&诗人&这一名词和格律名称结合到一起，称之为挽歌体诗人或者史诗诗人，他们被称为诗人，似乎只是因为遵守格律写作，而非他们作品的模仿本质。

The relationship between communicative competence and grammar teaching should be that of the ends and the means.

交际能力和语法的关系应该是目标与途径的关系。