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The progress of the cell cycle of Spodoptera frgiperda IPLB-Sf21-AE clonal isolate 9 (Sf9) cells was directed by Flow cytometry analysis.The results showed that the whole time of the cell cycle was about 18 hours and the gapes between every two phases were about 6 hours.At 12-18 hours post AcNPV infection,Sf9 cells were arrested in G\-2/M phase.When the cells in G\-1/S phase synchronized by druge were infected by AcNPV,2/3 of the cells was in G\-2/M phase and 1/3 of the cells in S phase.

应用流式细胞仪FACS的荧光检测,测出Sf9细胞完成整个周期循环大约需要 18h ,G1、S、G2 /M各时相的时间间隔约为 6h ;AcNPV感染Sf9细胞 12 18h ,细胞被抑制于G2 /M期;Sf9细胞同步于G1/S期后释放细胞并用AcNPV感染,12h后,2 / 3的细胞处于G2 /M期,1/ 3的细胞处于S期

Objective The origin and the nature of the Hodgkin and Reed-Sternberg cell of Hodgkins lymphoma has been attracting a lot of medical researchers engaged in studying it,for its character by scattered large atypical cells residing in a complex admixture of inflammatory cells.Great improvement has been made since a new method of isolation of single H/RS cells from a frozen section had been set up by KUppers in 1994,and many studies have approved of the B-cell derivation of H/RS cell.lt has been reported that H/R-S cells might partly be originated from B-cell in our research before,but at the same time,we also found that only 18.8% of H/RS cells express CD20,31.3% of immunoglobulin heavy chain rearrangement have been revealed.

目的 霍奇金淋巴瘤(Hodgkins lymphoma,HL)由于它的恶性肿瘤细胞—H/RS细胞一般只占肿瘤组织的极少部分(不到1%),且散在分布在背景细胞间,因此对于H/RS细胞的来源和性质研究一直是人们探索的目标。1994年德国科隆大学Kūppers等发展了一种从冰冻组织切片上用显微操作仪挑取单个H/RS细胞的显微切割方法进行H/RS细胞基因分析后,人们对H/RS细胞来源的研究有了突破性的进展,多数支持B细胞来源。

The results of character evolution history reconstruction shows the SC in PT and AD clade might be a homoplasy character; also, the SC in PT clade might have multiple origins. SC in Pteridaceae has some special characteristics:(1) the cell long axes usually run parallel to the lateral veins;(2) Most of Pteris and Adianum species have VSC only in lower epidermis;(3) Species have ISC also have VSC.(4) Presence of spicular cells is highly correlated with the long veinal epidermal cells. Species have type III silica deposition type usually have longer and narrower veinal epidermal cell compared with ordinary epidermal cells; Species have type IV silica deposition type also have long epidermal cells, but not significantly different from ordinary epidermal cells.

本实验中亦发现凤尾蕨科之矽异形细胞具有几项特色:(1)矽异形细胞之长轴多与叶片侧脉方向平行(2)发现多数铁线蕨属以及凤尾蕨属植物仅下表皮具有脉上矽异形细胞;(3)具有脉间矽异形细胞的种类会同时具有脉上矽异形细胞;(4)长条形的侧脉上表皮细胞与矽异形细胞具有相关性,仅具脉上矽异形细胞的分类群之脉上表皮细胞均较一般表皮细胞细长;具有脉间矽异形细胞之物种之脉上表皮细胞亦为长条形但形态与一般表皮细胞常无明显形态差异。

The results indicated that the content of cytosolic CaM in cells treated with exogenous Ca2+ has increased indistinctively, while fluorescence intensity in cells treated with TFP decreased. So we believed that exogenous Ca2+ has little effect on the expression of CaM. High concentration of TFP can enter yeast cells and combine to CaM to make it inactive, which is the reason that TFP restrain the growth of yeast cells. 5mmol/L EGTA could completely arrest the cell proliferation of MFP7 after 28h, when the fluorescence intensity in cells wasobviously increased with flow cytometry and LSCM.

被较高浓度TFP处理过的MFP7胞内荧光强度则相对较弱,推测是因为TFP进入胞内与CaM结合从而使其失活,这也是高浓度TFP抑制酵母细胞增殖的原因。5mmol/L EGTA处理28h左右后,酵母细胞的生长被抑制在G1晚期,此时通过细胞流式法和在激光扫描共聚焦显微镜下观察到胞内荧光强度显著高于对照,说明CaM的表达水平在G1后期开始上升;回加10mmol/L Ca~(2+)处理24h后,细胞开始恢复生长,细胞流式法测定胞内荧光强度有所下降,表明多数细胞完成G1/S转换,进入生长对数期。

Rat glioma cell C6 was infected by the deficiency recombinant retrovirus (named C6TK). The sensitivity of C6TK cells to GCV or ACV increased 450 or 10 times than that of C6 cells respectively. Rat C6 glioma model was successfully built, and SD rats inoculated intracerebrally C6TK cells had normal gliomas. The average survival periods of the rats were about 15 days. However, after treated with ACV, the growth of C6TK tumors obviously regressed, and the survival periods extended to 57.8±8.1 days, especially in rats injected with mixed PA317TK and C6 cells or in situ PA317TK cells, which the tumors nearly disappeared after ACV administration and the survival periods extended to over 120 or 71.4±36.1 days respectively, P<0.001.

结果:构建了带有HSV-tk基因的反转录病毒载体GINaTK,应用脂质体转移方法将GINaTK导入反转录病毒包装细胞PA317,成为产病毒细胞PA317TK,用带有HSV-tk基因的复制缺陷型反转录病毒感染C6细胞,命名为C6TK细胞,对GCV和ACV的敏感性分别高于亲本450倍和10倍;成功地建立了大鼠脑胶质瘤模型,并证实转染细胞C6TK的成瘤效应未改变,存活期约为15天;而经ACV治疗后,含有C6TK细胞的肿瘤生长明显被抑制,大鼠生存期延长为57.8±8.07天,尤其是采用PA317TK细胞混和治疗组和原位治疗组,肿瘤基本消失,大鼠生存期显著延长,混和治疗组存活120天以上,原位治疗组存活至71.4±36.1天。t检验,P值均小于0.001。

Results: GINaTK retroviral vector containing HSV-tk was constructed and transduced into PA317 retrovirus packaging cell by lipofectin (named PA317TK). Rat glioma cell C6 was infected by the deficiency recombinant retrovirus (named C6TK). The sensitivity of C6TK cells to GCV or ACV increased 450 or 10 times than that of C6 cells respectively. Rat C6 glioma model was successfully built, and SD rats inoculated intracerebrally C6TK cells had normal gliomas. The average survival periods of the rats were about 15 days. However, after treated with ACV, the growth of C6TK tumors obviously regressed, and the survival periods extended to 57.8±8.1 days, especially in rats injected with mixed PA317TK and C6 cells or in situ PA317TK cells, which the tumors nearly disappeared after ACV administration and the survival periods extended to over 120 or 71.4±36.1 days respectively, P <0.001.

结果:构建了带有HSV-tk基因的反转录病毒载体GINaTK,应用脂质体转移方法将GINaTK导入反转录病毒包装细胞PA317,成为产病毒细胞PA317TK,用带有HSV-tk基因的复制缺陷型反转录病毒感染C6细胞,命名为C6TK细胞,对GCV和ACV的敏感性分别高于亲本450倍和10倍;成功地建立了大鼠脑胶质瘤模型,并证实转染细胞C6TK的成瘤效应未改变,存活期约为15天;而经ACV治疗后,含有C6TK细胞的肿瘤生长明显被抑制,大鼠生存期延长为57.8±8.07天,尤其是采用PA317TK细胞混和治疗组和原位治疗组,肿瘤基本消失,大鼠生存期显著延长,混和治疗组存活120天以上,原位治疗组存活至71.4±36.1天。t检验,P值均小于0.001。

Results An adequately cellular cervical cytology smear,the reduce of cells overlay, the integrality of cells morphology, the clear structure of cells and unconspicuous phenomenon of dissolving and cell shrinkage were observed , the cells were stained well and flamboyantly, the contrast ration was outstanding between cytoplasm and nucleu, the cytoplasm, nucleus,karyosome,and other components were easy to be resolved, the cells boundary,transparence showed clearly.

结果:涂片的细胞量充足,重叠减少、形态结构清晰,无皱缩自溶现象,着色良好,色彩艳丽,浆核对比突出,胞质、核染色质、核仁和其他细胞成分易于识别,细胞境界清楚,透明度良好。

Fig 1 Negative control Female umbilical cord sample has no Y-specific signal after in situ hybridization with the biotinylated Y-repeated sequence DNA probe PY3.4 Fig 2 Positive control Flow-sorted male umbilical cord cells hybridized to Y-specific DNA probe PY3.4,Every cell contains a Y-specific signal Fig 3 Fetal cells sorted from maternal blood Flow-sorted cells from a pregnant woman at 20 weeks of gestation hybridized to Y-specific DNA probe PY3.4,containing a Y-specific signal Fig 4 The result of polymerase chain reaction Lane 1:male umbilical cord NRBCs sorted by FITC-conjugated anti-monoclonal glycophorin A;Lane 2:sample of 2;Lane 3:male umbilical cord NRBCs sorted by FITC-conjugated anti-CD36;Lanes 4-5:samples of 1 and 3;Lane 6:50 cells of male;Lane 7:5 cells of male;Lane 8:200ng male DNA(positive+control);Lane 9:nonpregnant female DNA(negative+control);Lane 10:ΦX174 HaeⅢ Maker,271bp:amplified band of Y-specific gene SRY;383bp:amplified band of human β-globin gene

图1 阴性对照女性脐带血标本,经与生物素标记的Y-染色体重复序列DNA探针原位杂交后未见Y-染色体特异信号图2 阳性对照分选的男性脐血细胞经与Y-特异DNA探针杂交后,每一细胞都含有Y-染色体特异信号图3 母体外周血中分选的胎儿细胞从一位妊娠20周的孕妇外周血中分选出的细胞经与Y-特异DNA探针杂交细胞中含有Y-染色体特异信号图4 聚合酶链反应结果 1:GPA-FITC单抗分选男胎脐血NRBCs;2:2号标本;3:CD36-FITC单抗分选男胎脐血NRBCs;4、5:1、3号标本;6:50个男性细胞标本;7:5个男性细胞标本;8:200ng男性DNA;9:未孕女性DNA;10:ΦX174 HaeⅢ标准,271bp:为SRY基因扩增带;383bp:为人β-珠蛋白基因扩增带内参照

RESULTS: The microscopic observation showed chondrocytes in the transducted group proliferated stably, and triangle, and spindle cells were predominant in the seventh passage, accompanied by few long-spindle and root cells. In addition, the cells were of clear appearance and large lucency. However, in the non-transducted group, long-spindle cells were observed in the fourth passage, and almost all cells exhibited long-spindle shape in the fifth passage.

结果:①倒置显微镜观察,转导组兔关节软骨细胞生长增殖稳定,至第7代仍旧以三角形、梭形及纺锤形细胞为主,偶见长梭形和树根形细胞,且轮廓清晰,透亮度大,折光性强;而未转导组细胞培养至第4代时即己出现长梭形细胞,自第5代细胞形态几乎全部变成长梭形。

Results一. Culturing and identification of human conjunctiva cells1. morphous of conjunctiva cellsCells were harvested by tissue cultivation after digestion. Ponderosus cells adhered after 48 hours. Cells were shown in shape of round, ellipse and polygon. Cell body was loose and lucency with nucleus in center, cell membrane was also clearly seen. Cells were arranged in inlay and fuse in film after 12-14 days.2. Immunochemistry testNomogeneous immunochernical positive granules against CK13 were observed in endochylema of human conjunctiva cells.3. growth curve of conjunctival cellsSubcultured cells were aged and feeble after 5th passage.

结果一、正常人结膜细胞的培养与鉴定1、活体细胞的形态消化后组织块培养法:48小时后见大量上皮细胞贴壁,细胞不规则呈圆形、椭圆形、多边形,细胞体肥大透亮,细胞核位于中央,胞膜清楚,约12-14天后细胞呈镶嵌状排列,融合成膜状。2、免疫组化鉴定培养的人结膜上皮细胞CK13免疫组化染色见阳性颗粒均匀分布于胞浆中。3、培养结膜细胞生长曲线传代培养多数样本于P5代以后开始出现衰老征象。

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