细胞

In the model of tumor cells metastases with cell line A673, 10〓 tumor cells can be detected after only one ampliation with outer primers, which means we can find 1 tumor cells in 10〓 normal PBMC, and enlarged again with inner primers, 10〓 tumor cells can be detected, which means we can find 1 tumor cell in 10〓 normal PBMC. In the simulative model of tumor cells metastases with RNA, 10〓 tumor cell can be detected that is to say we can find 1 tumor cell in about 10〓 normal PBMC. In all 15 samples of 11 patients, 2 patients'EWS/FLI1 expressions were negative, no evidence of metastases was found followed up by 8 months and 30 months respectively. Semi-quantitative study was used to detect the expression level of EWS/FLI1 mRNA in the other 9 positive patients, and metastases occurred in 6 high-level expression patients in 2-24 months after the operation, no evidence of metastases in the other 3 low-level expression patients up to now.

应用A673细胞系建立的肿瘤转移模型，单独应用外引物进行一次扩增，可以检测到10〓级肿瘤细胞，即可以从10〓个正常细胞中检测到1个肿瘤细胞；而在此基础上再次应用内引物扩增，则可以检测到10〓级肿瘤细胞，即可以从10〓个正常细胞中检测到1个肿瘤细胞；在用RNA模拟的肿瘤细胞转移模型中，两次扩增可以检测到10〓级肿瘤细胞，即可以从10〓个正常细胞中检测到1个肿瘤细胞。11例患者的15份标本中，2例没有检测到EWS/FLI1 mRNA的表达，分别随访8个月和30个月，没有发现临床转移的证据；对其余9例阳性表达的病例进行半定量的研究，6例EWS/FLI1 mRNA高表达的病例，分别在术后2-24个月发生转移，而3例EWS/FLI1 mRNA低表达的病例，则至今没有发现转移的证据。

Beijingensis under three droughttreatments, normal, moderate and severe stresses. The results showed:(1) the canker disease ofboth cultivars was serious gradually with increased drought;(2) the bark tissue cells sufferedplasmolysis, more evident with the severity of drought stress;(3) under the drought andinoculation with B. dothidea, cells of two cultivars damaged at different degree, mailyrepresented in the changes of organelles, such as chloroplast swollen and distorted, number ofmitochondria increased and membrane system indistinct; then organelles suffered furtherdamagement with inoculation time, thinned mitochondrias stroma, decreased cristae, crumpledand partly broken membrane of chloroplasts with stroma exosmosis. At last, the chloroplastspartly disorganized;(4) the hyphae growed mainly intercellular in resistant cultivar and notonly intercellular but also intracellular in susceptible cultivar, which directly caused thenecrosis of cells;(5) under the severe drought, the damage of cells enhanced the infection ofpathogen and drought and pathogen stressed together and promoted the disease development;the damage from pathogen on cells was more serious than that from drought.

结果显示：(1)随着干旱胁迫程度的增加，2种杨树溃疡病害发生渐趋严重；(2)干旱胁迫下，杨树树皮组织细胞发生质壁分离，并随胁迫程度的增加而严重；(3)干旱胁迫下接种病原菌，2种杨树细胞发生不同程度的损伤，主要表现为细胞器发生较大变化，如出现叶绿体肿胀变形、线粒体数量增多，质膜模糊不清等现象；随接种时间的延长，细胞器受到进一步损伤，叶绿体被膜折皱，严重时局部破裂，基质外渗，并部分最终解体；(4)毛白杨中的菌丝主要在细胞间隙中穿行，而北京杨的菌丝除在细胞间隙中生长之外，侵入细胞内部也较多，直接导致细胞的解体；(5)干旱胁迫下细胞的损伤促进了病原菌的侵染，干旱和病原菌的双重胁迫加剧了病害的发生程度，并且病原菌侵染对细胞的破坏程度大于水分胁迫。

Results:Whith the time and doses of ART incubation extended, ART significantly inhibited the proliferation of SGC-7901 cells and the inhibited effect shows dose-and -time-dependent. Obvious changes of apoptotic morphology were observed by invert microscope, fluorescence microscope, scanning electron microscope and transmission electron microscope, they showed that cells had marked nuclear condensation or the fragmentation of chromation as well as apoptotic , mitochondrial edema and vesicle , with the time of incubation extended , the proliferation rate become slow and the volume became small and transformed. FCM assay indicated that most of the cells were arrested in Go/Gi and the apoptotic peak appeared. During the prolong of incubation time, the apoptosis rate was increased. At the same time, the number of S and G2/M phase cells were decreased. The result of TUNEL indicant that there are apoptosis and necrosis.

结果：随着药物浓度的增加和作用时间的延长，蒿甲醚对胃癌SGC-7901细胞的抑制作用呈时间和浓度依赖关系：在倒置显微镜、荧光显微镜、扫描电镜和透射电镜下可观察到典型的凋亡细胞的形态改变，表现为：核固缩或染色质边聚或凝聚成大块状，可见凋亡小体，线粒体肿胀增殖，严重的空泡化，随作用时间的延长，细胞增殖速度减慢，细胞体积缩小变形；流式细胞术显示胃癌SGC-7901细胞出现明显的凋亡峰，随着作用时间的延长，其凋亡率逐渐升高，细胞周期阻滞在G_0/G_1期，S及G_2/M期细胞数大量减少；流式细胞术TUNEL检测结果显示细胞凋亡和坏死同时存在。

The SMMC-7721 hepatoma cell line of experimental groups and control groups were cultured with the addition of sodium selenite, potassium iodide or triiodothyronine (T3). The cell quantity was checked by the MTT assay , and the cell cycle was observed by flow cytometry and levels of AFP and ALB in the culture solution were determined by RIA.The SD rats were divided into positive control group, negative control group, T3 group and mixture group (the mixture of sodium selenite, KI and T3). After two weeks of the administration of sodium selenite、 KI and T3,Walker256 cells were transplanted into the abdominal cavity. The administration was kept on for another two weeks, then observed the morbility and mortality of the rats with ascites carcinoma. At the end, the ascites were for the cells counting and the cell cycles analysising.

方 法：在体外，应用细胞培养的方法，培养SMMC－7721肝癌细胞系，分为试验组和对照组，试验组加入不同浓度Se、I或T3干预，培养一段时间后，通过噻唑蓝试验、流式细胞仪，观察细胞数量及细胞周期的变化，测定细胞培养上清液中甲胎蛋白、白蛋白的水平，观察细胞分化情况；在体内，将试验用SD大鼠分成阴性对照组、阳性对照组、T3组和混合组（为Se、I和T3混合物），首先预防性用药2周，然后给试验组大鼠和阳性对照组大鼠腹腔注射Walker256细胞，制造腹水癌大鼠模型；继续用药2周，观察各组大鼠发病率及死亡率，测量不同时期大鼠血清FT3含量，通过流式细胞仪测量大鼠腹水细胞数及细胞周期变化。

The comparison was made of amplitudes of the postsynaptic potential evoked in AP cells by the stimulation of T, P and N cells. The results showed that for each type of T, P and N cell the contralateral member of the pair evoked larger synaptic potentials in the AP cell that did its ipsilateral homologue and the T, P and N cell with a dorsally situated receptive field produced a larger synaptic input on the AP cell that its counterpart with a ventrally situated receptive field.

分别比较T、P和N细胞在AP细胞上引起的突触后电位的幅度，可见对侧T、P和N细胞在AP细胞上引起的突触后电位幅度比同侧的T、P和N细胞在AP细胞上引起的突触后电位幅度大；具有背部感受野的T、P和N细胞在AP细胞上引起的突触后电位的幅度比具有腹部感受野的T、P和N细胞在AP细胞上引起的突触后电位的幅度大。

It showed that for each type of P and N cell the contralateral member of the pair evoked larger synaptic potentials in the AP cell that did its ipsilateral homologue and the P and N cell with a ventrally situated receptive field produced a larger synaptic input on the AP cell that its counterpart with a dorsally situated receptive field.

比较P和N细胞在AP细胞上的突触后电位幅度，可见对侧P和N细胞在邻节的AP细胞的突触后效应强于同侧的P和N细胞在AP细胞上的突触后效应；具有腹部感受野的P和N细胞在邻节AP细胞上的突触后效应强于具有背部感受野的P和N细胞在AP细胞上的效应。

The differentiation states of costae indicates the ways they absorb and transport water, and their ability adapting to the dry conditions, which was elucidated by the following examples: Brachythecium plumosum, because of its thin cell-walls in the costae and the absence of hydrome, assistant and steroid cells, can absorb water and nutrition under shady and moist conditions; Plagiomnium rostratum, though often grows in shady and wet condition, has hydrome and steroid cells, a feature similar to those of xeric mosses, which endows it to grow in the conditions with periodical drought stress during its life cycle; Hygrohypnum luridum is characterized by its slender leaves with only one layer of cells, the thin cell-walls, the absence of filaments on the leaf surface, and the fewer layer cells in the costae, the absence of hydrome and steroid cells. These features make it adaptable to aquatic environments. For Pogonatum inflexum and Atrichum undulatum, their ventral surfaces covered with lamella, while for Racomitrium japonicum, Thuidium cymbifolium, Macromitrium ferriei, Diphyscium fulvifolium, Barbula unguiculata and Ceratodon purpureus, their leaves are strongly mammillose or papillos. Such appendiculate structures made them adaptable to thy conditions.

例如，荫湿生环境下的羽枝青藓Brachythecium plumosum，其中肋细胞胞壁较薄，无导水主细胞和副细胞的分化，也没有厚壁细胞分化，能够在阴湿环境下吸收水分和养分；钝叶匍灯藓Plagiomnium rostratum具有与旱生藓类植物相似的中肋结构，叶片较厚，中肋具导水主细胞，中肋背面具厚壁细胞，这些特点使该种藓类植物能够分布于间隙性干旱胁迫的环境中；水灰藓Hygrohypnum luridum叶片纤细柔弱，仅1层细胞，细胞胞壁薄，叶表无附属结构，中肋细胞层数少，无导水主细胞分化，也没有厚壁细胞，这些特点使得水灰藓'能够生长在水生环境中；东亚小金发藓Pogonatum inflerum和波叶仙鹤藓Atrichum undulatum的叶腹面覆盖着栉片，东亚砂藓Racomitrium japonicum、大羽藓Thuidium cymbifolium、福氏蓑藓Macromitrium ferriei、东亚短颈藓Diphyscium fuhifolium、扭口藓Barbula unguiculata和角齿藓Ceratodon purpureus的叶片表面有乳头状突起或疣状物，这些附属结构使它们能够适应于旱生的环境中。

The study showed the recombinant wt/mCREG protein depressed the VSMC proliferation depending on dose and the optimal concentration was 400nM;2biologic function of CREG protein and the membrance receptor mechanism:①effect on VSMC migration: the wound healing experiment showed the OB2 cells migration was slower significantly after added wt/mCREG(400Nm) in supernatant. The HITASY cells migration were very slowly and no remarkable change. The gelatinase digestion and Western blot analysis showed the matrix metalloproteinase was decreased and TIMPs was increased;②effect on differentiation: after added wt/mCREG(400nM), the expression of myocardin, SMα-actin, MHC and caldesmin were increased and that of LM-1 and FN were decreased in OB2 cells. These effects were more significant when adding wtCREG.;③effect on VSMC proliferation: Cell cycle assay and BrDU stain showed: after added the wtCREG and mCREG protein, the ratio of cell in G0/G1 phase increased to 0.5773 and 0.5572 from 0.5308 respectively in OB2 group, which increased to 0.7369 and 0.7034 respectively from 0.6297 in HITASY group;3Role of M6P/IGF2R in CREG biologic function:①ELISA and co-immunoprecipitation showed the wt/mCREG binding to M6P/IGF2R directly.②antibody blocking test: when the anti-IGF2R was added to medium at the same time with wt/mCREG at different concentration(2μg/mL、4μg/mL、8μg/mL),the effects of CREG protein which depressing proliferation, migration, secretion and promoting differentiation were blocked, which had the positive correlation to the concentration of added anti body. The studies showed two combinant CREG promoted VSMC switch to differentiation phaenotype, at the same time, depress VSMC proliferation, migration and secreting extracellular matrix.

上述实验结果证实：两种重组CREG蛋白对VSMC增殖均有剂量依赖性的抑制作用，并且相同浓度的糖基化的CREG蛋白对细胞增殖的抑制效应更为显著，最佳效应浓度为400nM；2两种重组CREG蛋白添加后对HITASY和OB2细胞生物学行为的影响：①CREG蛋白对VSMC迁移的影响：刮伤实验发现，加入最佳效应浓度的wtCREG和mCREG蛋白24h后，OB2组迁移能力下降，HITASY组无明显变化；细胞外基质金属蛋白酶-2，9(Matrix metallo-proteinase 2，9，MMP2 ，9)明胶酶电泳检测和Western blot检测结果证实，两种CREG蛋白均可以使OB2细胞合成细胞外基质MMP2，9减少，而组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases，TIMPs)增加；②CREG蛋白对VSMC分化的影响：加入400nM的wtCREG和mCREG蛋白12h后，OB2细胞myocardin、SMα-actin、MHC、caldesmin表达增加，LM-1、FN表达减少；③流式细胞仪分析细胞周期和BrDU染色分析证实，加入400nM的wtCREG和mCREG蛋白后，OB2组G0/G1期细胞由0.5308分别增加至0.5773和0.5572，HITASY组G0/G1期细胞由0.6297分别增加至0.7369和0.7034；3M6P/IGF2R在重组CREG蛋白的生物学功能中的调控作用：①免疫共沉淀和免疫荧光双染色分析结果显示，CREG蛋白与M6P/IGF2R存在直接结合；②应用抗体阻断实验：将不同浓度的anti- M6P/IGF2R(2、4、8μg /mL)与两种CREG蛋白同时加入培养液中，CREG蛋白抑制VSMC增值、迁移和合成细胞外基质、促进分化的效应减弱，而且与加入anti- M6P/IGF2R浓度正相关。

Both the HepG2 cells and L-O2 cells have the expression of RhoA protein. However, the expression level of RhoA in HepG2 is much higher than that in L-O2. we also discover that the number of HepG2 cells mults more greatly with the detection of Millicell cabin than that of L-O2 cells through the hole, and that the number of HepG2 cells mults obviously and predominantly if added to RhoA excitomotor and decreases, obviously if added to RhoA antagon.

RhoA蛋白在HepG2及L-O2两株细胞中均有表达，但肝癌细胞株HepG2的表达量远远高于正常细胞株L-O2；通过Millicell小室检测，发现HepG2细胞通过细胞小室虑孔的细胞比L-O2细胞明显增多；HepG2细胞在加入RhoA激动剂后通过细胞小室虑孔的细胞显著增多；HepG2细胞在加入RhoA拮抗剂后剂通过细胞小室虑孔的细胞显著减少。

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 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。