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To investigate the effect of p38MAPK signal conducting pathway on livercancinoma cell's malignant phenotype induced by VEGF,we take theexperiment with cell growth test,scanning microscope and laser scanningconfocal microscope so that observe effect of p38MAPK signal conductingpathway on liver cancinoma cell growth,pseudopodium formation andframework of cytoskeleton induced by VEGF.results indicate that the cellbecame ellipse and there were more and thick pseudopodium in the cell'ssurface after being treated by VEGF,and destroyed framework ofcytoskeleton,which can be blocked by pretreated with a special inhibitor ofp38MAPK SB203580 so that VEGF promote metastasis by enhancing livercell migration and movement via p38MAPK signal conducting pathway,butVEGF promote cell growth without p38MAPK signal conducting pathway.

为进一步探讨VEGF通过p38信号传导通路诱导肝癌细胞转移时，对肝癌细胞恶性表型的影响，采用细胞增殖实验、扫描电子显微镜、激光扫描共聚焦显微镜观察VEGF对肝癌细胞增殖作用、细胞伪足形成、细胞骨架微丝结构的影响以及p38MAPK信号通路调控作用。结果显示，VEGF能够不通过p38信号通路促进肝癌细胞增殖；VEGF诱导肝癌细胞丝状伪足增多、增长，使细胞骨架微丝结构破坏、甚至消失。阻断p38MAPK信号通路，可以抑制VEGF诱导的肝癌细胞伪足形成，细胞骨架丝状结构呈束状，排列较规整。上述结果表明，VEGF可以通过p38信号传导通路诱导肝癌细胞伪足增多、增长，促使细胞骨架微丝结构破坏，使肝癌细胞迁移、运动能力增加，促进肿瘤转移。VEGF并不通过p38信号通路诱导肝癌细胞增殖作用。

Some key techniques related to the close and continuous process were investigated by the application of H9N2 avian influenza virus with Vero cells, such as the susceptibility of cell to influenza virus, virus production with cell microcarrier culture method, cell bead-to-bead transfer, virus production through bead-to-bead transfer, cell culture and virus production with serum free medium, metabolism analysis, and repetitiously intermittent bead-to-bead transfer of cell for virus production to simulate the close and continuous process.

通过使用Vero细胞增殖禽流感病毒H9N2，本文针对封闭连续工艺过程的一些关键技术开展研究，包括细胞对流感病毒敏感性分析、细胞微载体培养生产病毒工艺、细胞珠到珠转移、转移后细胞对病毒增殖敏感性验证、细胞无血清培养生产流感病毒、代谢分析、可模拟连续操作的多次间歇式珠到珠转移培养细胞生产流感病毒等方面。

Cell lines was established in vitro cell culture system. Only one of it grows well during gene targeting operation. The sex of these cell lines was identified by Sry-PCR.Using lipotransfection method to transfer pZJ into pig fetal fibroblast cell. After 7 days G418 selection, 30 neo-positive cell clones were obtained, among them, 12 of which is non-green ones.

用Sry-PCR法和染色体核型分析法鉴定细胞系的性别，采用优化的脂质体转染法将pZJ转染猪胎儿成纤维细胞，经7天G418(600μg／ml)药物筛选，共获得30个neo阳性细胞克隆，其中12个为非绿色细胞克隆，仅有7个可以扩大培养。

Inaddition,we extract total RNA from cultural U251 cell of humanglioma,make coding gene extron amplification of TfR through ReverseTranscription-Polymerase Chain Reaction,construct expressionplasmid of pCDNA3.1-TfR after digestion of enzyme andconjunction,do transfection to U251 cell by liposome afteridentification,cause excess expression,detect the efficiency of transientexpression of TfR protein by pEGFPN1-TfR, sieve positive cell ofpCDNA3.1-TfR through G418, choose positive cell clone, detect theexpression of TfR in anteroposterior period through Western blot,FCM,cell immunochemistry and mRNA through RT-PCR, identify the effect of transfection.

另外，培养人脑胶质瘤U251细胞，进行细胞总RNA抽提，运用RT-PCR方法进行TfR基因的完整编码区外显子的扩增，经过酶切、连接后构建pCDNA3.1-TfR的表达质粒，鉴定正确后按脂质体转染方法转染人胶质瘤U251细胞，使其过量表达，通过pEGFPN1-TfR检测其瞬时转染的效率；用G418对稳定表达pCDNA3.1-TfR的细胞进行筛选，挑选阳性细胞克隆，运用Western blot、流式细胞检测技术和细胞免疫化学以及mRNA半定量的方法检测TfR在转染前后的表达水平，鉴定转染的效果。

ResultsMorphological features with diagnostic value were established as follows:①Irregular or round-mesh basket type nucleolus;②Malformed nucleus and lobate nucleus with fine bridge;③Perinuclear microfilaments;④Nuclear bodies;⑤Nuclear inclusion bodies;⑥Marked cell pleomorphism arranged irregularly,like a disordered stone pavement;⑦Strange cell and/or giant cancer cell were found;⑧Peculiar papillae or cell clusters were formed;⑨Specific function structure on the surface of cell differentiated poorly.

结果 确定了具有诊断意义的形态学特征：①不规则奇形核仁或网孔状核仁；②畸形核，分叶核伴细桥－核间桥形成；③核旁微丝；④核体；⑤核包涵体；⑥细胞重度多形性，排列紊乱，似乱石铺堆；⑦出现奇形细胞和/或癌巨细胞；⑧形成奇形乳头和奇形细胞簇；⑨细胞表面特异功能结构分化不良。

The cancer cells are in different differentiation periods: the chromatin of the young cancer cell's nucleus is rich in color and its cytoplasm is basophilous. The young cancer cells don't form into typical glandular cavity; The mature cancer cell is columnar or cubical and its nucleus is located at the base of the cell in gland tube-like arrangement; The decrepit cancer cell stain thin while its nucleus stain dense. The severer's nucleus disintegrate into small fragments. The decrepit cancer cell's arrangement is disorganized, only keeping its glandular shape.

显微镜下癌细胞呈现不同的分化程度：幼稚型癌细胞胞核染色质丰富，胞质嗜碱性，不形成典型腺腔；成熟型癌细胞呈柱状或立方状，细胞核位于细胞基部，呈腺管样排列；衰老型癌细胞胞桨染色变淡，胞核浓染，严重者胞核碎裂成细小碎片，衰老型癌细胞排列紊乱，仅保留腺体样结构轮廓。

The ultrastructure characters of pollen grain in Brassica napus are:1There are vacuoles in cytoplasm but no starch grain in uniceelluar pollengrain.2The veget- ative nucleus of bicellular pollen grain is generally spheroidal;while the generative cell is spinde-shaped,with a big nucleus,thin layer of cytoplasmand few cell orgens,no cell wall.3In three-celled pollen grain the sperm cell is separated from the cytoplasm of vegetative cell by two layers of plasmic membrances.

Brassica napus的花粉粒，在不同发育时期超微结构的特征如下：1单胞花粉粒有一个球形的细胞核和明显的核仁，细胞质内出现液泡，缺少典型的淀粉粒。2双胞花粉粒的营养核多为球形。生殖细胞纺锤形，无细胞壁，细胞核比例大，细胞质呈薄层，细胞器少。3三胞花粉粒有一个裂片状的营养核和两个纺锤形的精细胞，精细胞无壁，以两层质膜与营养细胞的细胞质相隔。

Results The IC50 of SPG-Rg3 was (155.7±0.71) mg·L-1,when the concentration of SPG-Rg3 was arranged from 37.5 to 600.0 mg·L-1,the growth inhibitory rate of MCF-7 cells was higher following the increase of its concentration,the cell growth was greatly inhibited compared with control group(P.05); cell cycle was changed,the cell number of S period increased compared with control group(P.01),the cell number of G2/M period decreased compared with control group(P.01),in experiment group there was an apoptotic apex before G1 apex,and the number of apoptotic cells increased; most of cell nuleus in SPG-Rg3 (150 mg·L-1)experiment group appeared yellow or chrysoidine fluorescence,contracted,beaded and crescent; the result of immunocellularchemical staining of caspase-8 indicated that caspase-8 protein had no expression in control group,but stronger expression in SPG-Rg3 group.

结果：SPG-Rg3 IC50为（155.70±0.71） mg·L-1，当SPG-Rg3浓度在37.5~600.0mg·L-1时，MCF-7细胞的生长抑制率随SPG-Rg3浓度的增加而增大，与对照组比较，细胞增殖受到明显抑制（P.05）；MCF-7细胞的生长周期也发生变化，S期细胞数增加，明显高于对照组（P.01，G2/M期细胞数则明显少于对照组（P.01；并且在G1峰前出现明显的凋亡峰，凋亡细胞数增加。

The relative cell groth rate of L929 cell represented the proliferation of cells on materials with respect to control over time. The result showed that cytotoxicity of thermo-sensitive cell culcure dish was none or little,which can be used for lab use.Additional cell culture was performed with two types of celland cell sheets detachment were observed by reducing the culture tempareture after the confluently growth of the two types of cells.

进一步利用两种不同的细胞永生化的成牙本质细胞系和人牙周膜细胞在接枝有PNIAPm的培养皿上进行培养，观察到细胞可以正常贴壁，生长并增殖，并且在形成连续的细胞层之后，通过降低温度，获得了完整的细胞层结构，结合前两部分实验结果，证实了利用等离子体预处理和紫外线照射接枝制备温敏性培养皿的可行性，为后续研究提供了实验基础。

Results Of 62 cases of PMLGI, 6 cases were low grade malignant B-cell lymphoma of MALT type, 14 cases high grade malignant B-cell lymphoma of MALT type with a low grade malignant component, 1 case lymphomatous polyposes, 2 cases Butkitt-like lymphoma, 38 cases B-cell large cell lymphoma and 1 case intestinal T-cell lymphoma.

结果：62例PMLGI中，低度恶性B细胞粘膜相关组织型淋巴瘤6例，高度恶性B细胞粘膜相关组织型淋巴瘤伴低恶成分14例，淋巴瘤性息肉病1例，Burkitt样淋巴瘤2例，B大细胞淋巴瘤38例，肠道T细胞淋巴瘤1例。

Do you know, i need you to come back

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Yang yinshu、Wang xiangsheng、Li decang,The first discovery of haemaphysalis conicinna.

1〕 杨银书，王祥生，李德昌。安徽省首次发现嗜群血蜱。