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Results AQP1 is expressed at the apical and basolateral membrane of the microvascular endothelium; AQP3 was detected at basal cells of both the bronchiole epithelium and submucosal gland acinus; AQP4 is present in the basolateral membrane of columnar cells in bronchiole; while AQP5 is expressed in the apical membrane of type Ⅰ pneumocytes, and also at the apical of columnar cells of superficial epithelium and submucosal gland acinar cells.

结果本研究发现AQPs基因在羊肺中的表达分布与人相似,AQP1在肺内的毛细血管内皮细胞表达;AQP3在小支气管黏膜上皮的基底细胞的基侧膜表达,AQP4存在于小支气管黏膜上皮的柱状纤毛细胞的基侧膜;AQP5存在于Ⅰ型肺泡上皮细胞的顶质膜,存在于小支气管黏膜上皮柱状纤毛细胞,以及在气道黏膜下腺的腺细胞的顶质膜表达。

We got the result: Bufo Raddei Strauch Tadpole (1) Bufo Raddei Strauch Tadpole was capable of lens regeneration, which originated from the epithelial cells at iris dorsal margin by dipigmentation.(2) Dipigmentation of Bufo Raddei Strauch by two methods: one was licked up by amoebocyte, the other one was flowed from the pigment cells.(3) In the lens regeneration cells of Bufo Raddei Strauch profiles of the mitochondria became complicated ribosome manifold, and the endoplasmic reticulum was cisternal dilation. During the process of deferentiation of lens fiber the cell nucleolus became little and chromatin shrinked.(4)The crystalline electrophoresis of Bufo Raddei Strauch Tadpole lens regeneration showed that during lens regeneration P-crystalline was expressed earliest then was γ-crystalline and α-crystallin, this result is consistent with newt.

结果表明:花背蟾蜍蝌蚪(1)具有晶状体再生的能力,其晶状体的再生来源于虹膜背缘的上皮细胞;(2)虹膜背缘去色素是转分化的前提,主要有两种方式:一种也是最主要的方式是在晶状体被摘除后虹膜外侧产生巨噬细胞,巨噬细胞吞噬虹膜色素上皮细胞中的色素颗粒;另一种方式是虹膜色素上皮细胞自身释放色素颗粒;(3)电镜观察发现虹膜色素上皮细胞转分化过程中线粒体有明显变化,核糖体增多,粗面内质网增多,晶状体纤维分化过程中细胞核逐渐凝聚;(4)SDS-PAGE电泳发现,花背蟾蜍蝌蚪晶状体再生中相关蛋白表达顺序与有尾类蝾螈相似,蛋白表达顺序依次是β-晶状体蛋白、γ-晶状体蛋白和α-晶状体蛋白,并且后两种蛋白几乎同时表达。

RESULT: 1.Ouabain act on lens sodium-pump,under the LM,lens anterior capsular membrane discontinuous,epithelium cells clustered,occluding zonule seperated,lens fiber layers fractured.Under the EM,cells totally hollowed,mitochondria swelling,myelin figure appeared.RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1、α2 and α3-isoform are all decreased.2.Digoxin act on lens sodium-pump,under the LM,lens cell oedema,linkage distructed,extensive exfoliation.Under the EM,plasma appeared little half-transparant hollow region,mitochondria swelling and ridge disappeared. RT-PCR examine,α1、α2 and α3-isoform are all decreased.3.Amphotericin B act on lens sodium-pump,under the LM,lens epithelium cells linked tightly,arranged in-line,lens fiber layers arranged tightly and regularily.Under the EM,abbundant cellular organes,exuberant cells function indicated. RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1 and α3-isoform are increased significantly,demonstrated isoform-specific action.4D-thyroxine act on lens sodium-pump,under the LM,lens plasmalemma integrated,cells arranged tightly and regularily.Under the EM,nucleus fission appeared,desmosome half-desmosome and tensile microfilaments linked the cells. RT-PCR examine,α2 and α3-isoform are increased, also demonstrated isoform-specific action.5.Vitamin E act on lens sodium-pump,under the LM,lens anterior capsular membrane continuous and smooth,epithelium cells tightly linked,lens fiber layers appearede hollow region occasionally.Under the EM,lateral membrane high density belt appeared,abundant nucleolus. RT-PCR examine,onlyα1-isoform are increased, demonstrated significantly isoform-specific action.6.DMSO act on lens sodium-pump,under the LM,lens anterior capsular membrane slightly thicker,cells linkage partly distructed.Under the EM,plasmalemma denaturation,mitochondria swelling.RT-PCR examine,α1、α2 and α3-isoform are all altered slightly and haven't significant meanning.

结果:1、哇巴因作用于晶状体钠泵后,光镜下晶状体前囊膜断裂、上皮细胞聚积、闭合连接分离、纤维板层裂隙,电镜下全层细胞空泡化、线粒体肿胀出现髓样结构,RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达均减弱。2、地高辛作用于晶状体钠泵后,光镜下晶状体细胞水肿、细胞连接破坏、广泛剥离,电镜下胞质见少许半透明空化区、线粒体肿胀嵴消失,RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达均减弱。3、两性霉素B作用于晶状体钠泵后,光镜下晶状体上皮细胞紧密连接、线状排列、纤维板层紧密规整,电镜下细胞器丰富、细胞功能旺盛,RT-PCR法检测α1及α3表达显著增强、具有一定的重整异构作用特异性。4、D甲状腺素作用于晶状体钠泵后,光镜下晶状体质膜完整、细胞排列紧密规整,电镜下胞核见分裂像、细胞间有桥粒、半桥粒及张力微丝,RT-PCR法检测α2及α3表达均增强、亦有一定的重整异构作用特异性。5、维生素E作用于晶状体钠泵后,光镜下晶状体前囊膜连续光滑、上皮细胞紧密连接、纤维板层偶见空化,电镜下囊侧膜内有高电子密度带、细胞核仁丰富,RT-PCR法检测仅有α1的表达显著增强、具有极强的重整异构作用特异性。6、二甲基亚砜作用于晶状体钠泵后,光镜下晶状体前囊膜轻度增厚、细胞连接部分破坏,电镜下质膜变性、线粒体肿胀,RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达无显著改变。

The results shown that:(1) Bufo raddei Strauch tadpole was capable of lens regeneration, which originated from the epithelial cells at dorsal iris margin by depigmentation.(2) Depigmentation of Bufo raddei Strauch during lens regeneration was by two methods: First, pigment granules initially dispersed all over the cytoplasm were moved towards the periphery of the cell, and then were directly taken up by the amoeboid cells. Second, pigment granules in a cell were first crowded in a mass, surrounded by a membranous structure in the cytoplasm, which were eventually discharged as a mass from the cell.

结果表明:花背蟾蜍蝌蚪(1)具有晶状体再生的能力,其晶状体的再生来源于虹膜背缘的上皮细胞;(2)虹膜背缘去色素是转分化的前提,主要有两种方式:一种也是最主要的方式是在晶状体被摘除后虹膜外侧产生巨噬细胞,巨噬细胞吞噬虹膜色素上皮细胞中的色素颗粒;另一种方式是虹膜色素上皮细胞自身释放色素颗粒;(3)电镜观察发现虹膜色素上皮细胞转分化过程中线粒体有明显变化,核糖体增多,粗面内质网增多,晶状体纤维分化过程中细胞核逐渐凝聚。

In the normal uterus, Cytokeratins immunolabelling were detected in glandular cell, luminal epithelial cell, Vimentin immunolabelling were detected in stromal cell and endoblastic cell; CK7 immunolabelling were not detected in any tissue of the yak utenus.

研究结果显示:未妊娠时,泛角蛋白在子宫内膜腺上皮细胞、腔上皮细胞内表达,波形蛋白在子宫内膜基质细胞内表达,平滑肌肌动蛋白在子宫平滑肌和血管平滑肌内表达,牦牛子宫任何部位均不表达角蛋白7;妊娠30天左右时,泛角蛋白在子宫内膜腺上皮细胞、子宫内膜腔上皮细胞、滋养层细胞、内胚层细胞和尿囊细胞内表达,波形蛋白在子宫内膜基质细胞和内胚层细胞内表达,平滑肌肌动蛋白在子宫平滑肌和血管平滑肌内表达,角蛋白7在尿囊细胞内表达,偶尔在腔上皮细胞的细胞核边缘表达;消化法进行原代培养时,组织经胶原酶消化并通过100目和400目筛网组合可以有效地分离原代子宫内膜基质细胞和子宫内膜腺上皮细胞;分离得到的子宫内膜基质细胞活率达90%以上,并可在体外传代7次以上;分离得到的子宫内膜腺上皮细胞活率可达85%以上,并可在体外传代5次以上;RPMI1640培养基最适合子宫内膜基质细胞和子宫内膜腺上皮细胞的生长,维持子宫内膜基质细胞正常生长的FBS添加量为20%,维持子宫内膜腺上皮细胞正常生长的胎牛血清添加量为30%。

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。

Result: 1. The situation of HIV infection in gastro-mucosal tissues:(1) Our study found HIV gag sequence in not only CD4~+ T cells but also mucosalepithelial cells, gland epithelial cells in lamina propria and spindle stromal cells;(2) HIV p24 was expressed in T cells, plasmocyte, and a few mucosa epithelial cells, gland epithelial cellls and dell epithelial cellls.

结果:1.HIV在胃黏膜中的感染状况:(1)HIV感染者胃黏膜内HIVgag区基因不仅见于CD4~+T细胞等免疫细胞中,少数胃黏膜上皮、腺上皮、小凹内上皮细胞和间质梭形细胞中亦有阳性杂交信号;(2)HIV感染者尸检胃黏膜内HIVp24蛋白于T细胞、浆细胞及少部分胃黏膜黏膜上皮、腺上皮及小凹上皮细胞中呈阳性表达。2。

MTT assay FAK signaling pathway inhibitor genistein on human corneal epithelial cell cytotoxicity;RT-PCR detection of human corneal epithelial cells adhesion to fungus at different times,extracellular matrix protein including laminin,fibronectin,FN glass,Ⅳcollagen,transmembrane protein integrinαⅤ,integrinβ1(ITGβ1),as well as the FAK signaling pathway FAK1,FAK2 and Paxillin gene expression;Western blot detection of the signal transduction pathway adhesion-associated protein ITGβ1,FAK and PAX expression and the inhibition of genistein. Immunocytochemical method was used to observe the LN,FN and FAK expression in human corneal epithelial cells during interaction with the fungues;Laser scanning confocal microscope had a cell positioning on FN,FAK and PAX,observed the changing of the human corneal epithelial cytoskeleton after stimulated by fungues;Quantitatived by flow cytometry to detect of human corneal epithelial cells with PAX at ITGβ1 fungal expression after adhesion;Optical microscopy quantitied the fungues and human corneal epithelial cell adhesion and recorded to determination the integral optical density afrer adhesion;Scanning and transmitted electron microscope observed the changing of cell ultrastructure after fungues and human corneal epithelial cell adhesion.

第一部分真菌激活人角膜上皮细胞FAK信号转导通路的体外实验研究将三种常见致病真菌(白色念珠菌、烟曲霉菌和茄病镰刀菌)分别与人角膜上皮细胞共孵育,MTT法检测FAK信号通路抑制剂染料木黄酮的对人角膜上皮细胞的细胞毒性作用;RT-PCR检测真菌黏附人角膜上皮细胞后不同时间细胞外基质层连蛋白、纤连蛋白、玻连蛋白、Ⅳ型胶原、跨膜蛋白整合素αV、整合素β1(ITGβ1),以及FAK信号通路中FAK1、FAK2和桩蛋白基因的表达情况;Western blot的方法检测黏附信号转导途径相关蛋白ITGβ1、FAK和PAX的表达,以及染料木黄酮对真菌刺激人角膜上皮细胞FAK信息通路活化的抑制作用;免疫细胞化学方法观察人角膜上皮细胞与真菌相互作用过程中LN、FN和FAK的表达;激光共聚焦显微镜对FN、FAK和PAX进行了细胞定位,并观察真菌刺激后人角膜上皮细胞骨架的变化;流式细胞仪定量检测人角膜上皮细胞ITGβ1与PAX在真菌黏附后表达的改变;光学显微镜观察真菌与人角膜上皮细胞黏附数量,记录并测定了黏附后积分光密度值OD扫描及投射电镜观察了真菌与人角膜上皮细胞黏附后,细胞超微结构的改变。

Results: in contrast with model group, SSM can (1) increase the survival rate of ARF in rats, cut down Scr and BUN at 24hr, improve renal function, slightly surpass Vp in its therapeutic effect;(2) mitigate the extent of renal pathologicchanges, decrease the numbers of renal tubule necroses and casts;(3) protect renal ultrastructure such as microvilli, mitochondria, endoplasmic reticulum of TEC, podocytic process of glomerulus epithelial cell,endothelial cell etc, better than Vp;(4) enhance SOD activity, lower MDA contents in renal cortex;(5) enhance NO contents in serum, reduce ET levels in plasma;(6) evidently cut down TNF- a contents in serum, superior to Vp;(7) antagonize calcium overload in TEC;(8) alleviate renal pathological changes in ARF metaphase (at 3d), advance regeneration and recovery of TEC, restore renal structure in ARF convalescence (at 5d), and better than Vp;(9) increase the expression of TEC PCNA and prepro EGFmRNA in ARF metaphase.

结果:与模型组相比,SSM能(1)提高ARF大鼠的存活率,降低24hr时Scr及BUN,改善肾功能,其疗效略优于维拉帕米;(2)减轻ARF肾脏病变程度,减少24hr肾小管坏死数及管腔内管型数;(3)对肾脏的超微结构如肾小管上皮细胞微绒毛、线粒体、内质网、肾小球上皮细胞足突、内皮细胞等形态结构均有一定的保护作用,其疗效略优于维拉帕米;(4)提高肾皮质SOD活性,降低MDA含量;(5)提高血清NO水平,同时降低血浆ET含量;(6)显著降低血清TNF-α的水平,疗效明显优于维拉帕米;(7)拮抗小管上皮细胞[Ca~(2+)]_i的升高;(8)使ARF中期(第3d时)肾组织病变较同期其它两组明显减轻,并使肾小管细胞再生修复提前,至ARF恢复期(第5d时)肾组织结构恢复重建良好;作用明显优于维拉帕米;(9)促进ARF中期肾小管上皮细胞PCNA的表达及EGF前体mRNA的合成,从而增加EGF的合成释放,而维拉帕米无此作用。

The proliferative capacity of LEC from human had directly relevance to the age of donor(r=-0.996).Conclusions The formation of lens-like spherules is characteristics of LEC lines in the cultured cells.Under identical conditions,the proliferative rate of LEC from bovine and rabbit is fast than that of LEC from human,but dedifferentiation of LEC from bovine and rabbit is easier than that of LEC from human;LEC from the three species exhibit similar limited growth potential.The proliferative rate of LEC from human has a inversely proportion with age.

&晶状体小体&的形成可作为确定晶状体上皮细胞株的一项特征性依据,而体外培养的人、牛、兔晶状体上皮细胞具有相同的有限生长潜能,在相同的条件下,牛、兔晶状体上皮细胞的生长增殖速度比人晶状体上皮细胞快,但易于发生去分化;此外,人晶状体上皮细胞的生长增殖率与年龄密切相关,年龄越小,晶状体上皮细胞的生长增殖速度越快。

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从鲍尔点的形成原理出发,分析对称连杆曲线上鲍尔点的产生条件,提出等边机构的对称连杆曲线上有单鲍尔点和双鲍尔点。

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