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These cancer cells might histogenetically be related to the transitional or metaplastic epithelium of prostate according to morphological analysis,(2) Mucinous adenocarcinoma, Xanthomatous carcinoma, ductal carcinoma, medullary carcinoma, endometrioid carcinoma, papillary carcinoma and signet-ring cell carcinoma were positive for PSA and 35βH11, these carcinomas might histogenetically be related to prostatic secretory epithelium,(3) Prostatic carcinoid showed positive to PSA, 35βH11, NSE and CgA, corresponded with endocrine cell originator,(4) Small cell carcinoma were negative for PSA, 35βH11, NSE and CgA, whether or not it originates from endocrine cells, storage cells or basal cell of prostate had yet to be proved,(5) 34βE12 marking was negative in cancerous areas of 27 cases, and the basal cells were absent in PPTC.

从形态分析,这两种癌可能同源于移行上皮或化生上皮:(2)粘液腺癌、黄色瘤样癌、导管癌、髓样癌、宫内膜样癌、乳头状癌及印戒细胞癌均显PSA及35βH11阳性,提示这几种癌可能来源于分泌上皮,(3)类癌对PSA、35βH11、NSE及CgA均显阳性,符合内分泌细胞来源,(4)小细胞癌无PSA、NSE及CgA表达,对c-erbB-2及35βH11显阳性,是否来源于前列腺内分泌细胞、储备细胞或基细胞有待证实,(5)27例癌区均无34βE12表达,提示PPTC中基细胞缺失。

Methods:MTT assay was used to detect PGPG homotypic adhesion and PGmatrix adhesion,and PGHUVEC adhesion was determined by Rose Bengal stain.

小檗碱(5、10 μg/ml)处理PG细胞24小时,采用MTT法观察PG细胞与PG细胞的黏附,PG细胞细胞外基质(FN和Martrigel)的黏附;用虎红染色法观察PG细胞与人脐静脉内皮细胞的黏附。

Methods:The subcutaneous adipose tissue was obtained from the fold inguen of four SD rats, then digested with collagenase type Ⅰ and cultured in endothelial medium for seven days.

获取4只雄性SD大鼠鼠蹊部皮下脂肪,通过机械分割和组织消化法获取脂肪基质细胞,在内皮细胞培养液中培养7d后,经免疫细胞化学及透射电镜鉴定证实95%以上细胞已具内皮细胞表型特征,再将这些细胞进行成脂(DMEM/F-12培养基内加入0.5mmol/L1-甲基-3-异丁基-黄嘌呤,1μmol/L地塞米松,10μmol/L胰岛素,200μmol/L吲哚美星)定向诱导7d,形态学观察和油红O特殊染色鉴定诱导后的细胞

The cilia of retinal pig merit epithelium disappeared completely by TEM, the granola of retinal pigment epithelium decreased, the rough endoplasmic retinal, the mitochondrial crista breaks, the outer nuclear layer arrangemen disorderin, disc broad, the vacuole timer and outer plexi-form layer shaped, the ganglion cells and membranous of eye cell outer side got think, intermembranous space gotorgancelle disappeared mostly, the crista of cytoplasm ganglion cells layer swelled.

透射电说可见:视网膜色素上皮细胞表面纤毛完全消失,视网膜色素上皮细胞内颗粒减少,粗而内质网、线粒体嵴断裂,外颗粒层细胞排列紊乱,视细胞外段盘膜粗大,盘膜间隙增宽,内外丛状层空泡形成,神经节细胞细胞器大部分消失,神经节细胞层可见细胞质有嵴性肿胀。

We introduced improved primary mixed glial culture and different-attachment method to isolate and purify the OPCs, the cells were proliferated in serum-free medium, flow cytometry and immunohischemistry methods were employed to estimate the purity of cultured OPCs. Their abilities of differentiation and expression of trophic factors were identified by RT-PCR and immunostaining. Several methods including TUNEL and MTT were adopted to estimate the protective effects of conditioned culture medium from oligodendrocyte lineage cells on the primary cultured cerebellar granule neurons. Intravitreal transplant of OPCs, combined with retrograde fluorescent labeling the superior colliculus and intraorbital optic nerve transection, were used to investigate the protective effects of OPCs on the axotomized RGCs in vivo. Intravitreal transplantof OPCs or NSCs on the newborn rats, and retinal transplant of OPCs on the young rats were performed, to observe the myelin formation in the retina at different stages after cellular transplantation. Optic nerve transection was carried out on some rats with myelinated retinae, to study the influence of myelination on the injuried RGCs.

为此,本研究采用改良的胶质细胞混合培养与差速贴壁方法获得大鼠OPCs,使用无血清培养基进行扩增、培养,用免疫组织化学和流式细胞技术对培养细胞的纯度进行鉴定,对少突胶质系细胞表达部分营养因子的情况进行检测;采用TUNEL、MTT等方法对少突胶质系细胞条件培养基对原代培养小脑颗粒神经元的保护作用进行检测;将OPCs移植入成年SD大鼠玻璃体内,利用上丘逆行荧光标记技术,观察眼内移植的OPCs对眶内视神经切断时的视网膜神经节细胞的保护作用及其持续时间;将OPCs或NSCs移植入新生和幼年SD大鼠玻璃体或视网膜内,观察不同时期视网膜内髓鞘形成与分布特点,分析髓鞘的超微结构,并观察眼内髓鞘形成对损伤神经节细胞的保护作用。

The expression changes of relating-apoptosis gene proteins (bcl-2, bax) were detected by immunohistochemistry before and after treating with the Agaricus Blazei Murill extract to explore the possible mechanisms of inducing apoptosis for MGC-803 cells in vitro. Results: The Agaricus Blazei Murill extract significantly inhibited the proliferation of gastric cancer cells in vitro and the inhibitive effects were depended on the medicine concentration and treating times. After treating 24 hours on the gastric cancer cells with the morphologic changes of apoptosis with chromatin margination, karyopyknosis, karyorrhexis were found by the light.

结果:(1)通过细胞培养和MTT法,表明长白山姬松茸在体外对MGC-803细胞株有明显的抑制作用,加药组与对照组相比其生长抑制率有显著性差异(P<0.01),而且这种抑制作用呈现浓度和时间的依赖性;(2)通过集落形成率的测定结果表明,加药组与对照组相比其集落形成率和集落形成抑制率有明显差异(P<0.01),说明长白山姬松茸对MGC-803胃癌细胞株有明显抗增殖作用;(3)光镜观察结果表明,加药组可见细胞脱水浓缩伊红染色增强,胞体缩小,内含高度浓缩的胞核呈深蓝色等典型细胞凋亡形态;经AO/EB荧光染色观察结果表明,当终浓度为1.0mg/ml的姬松茸提取物作用于MGC-803胃癌细胞24h后,其凋亡率和破膜率与自然凋亡率与破膜率相比,均有显著性差异,其凋亡率86.3%(P<0.001),破膜率为41.6%(P<0.01),说明姬松茸确实有诱导MGC-803胃癌细胞凋亡的作用,同时也有细胞毒作用,但以诱导细胞凋亡为主;(4)免疫组化结果表明,用药前后凋亡相关基因的BCl-2、Bax蛋白均有显著性的改变(P<0.001)。

Leucocytic differentiation antigen 40 sign-al access participates in regulation of inflammatory reaction accommodation of major cell component (vascular endothelial cell、vascular smooth muscle cell and macrophage), it is playing critical role in AS process, participates ather-osclerotic occurrence and development ,reside in atherosclerotic plaque cell thro-ugh CD40-CD40L interaction, induce activate by itself , express and secrete profit to happen immune response、inflammatory reaction、hemagglutination t-hrombogenic proteo-cytokine.

细胞分化抗原40 (CD40)信号通路参与了动脉粥样硬化斑块内主要细胞成分(血管内皮细胞、血管平滑肌细胞以及巨噬细胞)炎症反应的调节,其在AS进程中扮演着关键性的角色,参与了动脉粥样硬化的发生和发展;存在于AS斑块内的细胞通过CD40-CD40L相互作用,导致自身活化,表达和分泌了有利于免疫应答发生、炎症反应、血凝和血栓形成的蛋白质细胞因子,如激活在AS斑块中的关键性细胞成分如黏附分子、炎症因子、基质金属蛋白酶等的产生,导致斑块的不稳定和破裂,而最终导致粥样硬化斑块病变的恶化。

The effects of several components in the culture medium on Lithospermum erythrorhizon cell growth and secondary metabolite synthesis were studied, as well as structural dynamic model. The two-liquid-phase culture of Lithospermum erythrorhizon was carried out by choosing the proper organic solvent as the second phase. The bioactive carrier for adsorption was prepared and the condition of cell immobilization was determined. We combined the technique of two-liquid-phase culture and immobilization to carry out the culture. We chose the suitable type of reactor, studied its characteristics and results of cell culture using this reactor. The fed-batch operation was also studied on the basis of twoliquid-phase culture and immobilization used in culture in the reactor.

本文研究了紫草细胞悬浮培养中培养基中多种成分对细胞生长与次生代谢产物合成的影响,进行了结构化的动力学模型研究;通过选择合适的有机溶剂对紫草细胞进行了双液相培养研究;通过确定以吸附为细胞的固定化方法,进行了生物活性吸附载体的制备与固定化细胞的制备研究;并结合双液相培养技术,对紫草细胞进行了固定化培养及其动力学模型的研究;对反应器进行选型,并进行冷模与热模研究;在反应器中进行了固定化紫草细胞的双液相培养条件下的流加操作研究。

That GER cells which are likely hair cell progenitors could be cultured in vitro and generate stereociliary bundle-like structure when forced to express Hath1 suggests that the misexpression of Hath1 probably can induce the predifferentiation of pure hair cell progenitors into hair cells in vitro.

GER细胞作为毛细胞前体细胞可以实现体外原代培养,在Hath1基因重表达情况下,部分细胞的表面可见不成熟的纤毛样结构,提示Hath1基因也许可以诱导离体培养下的纯的毛细胞前体细胞向毛细胞的初步分化。

When reached logarithm growth phase, it was poured or the medium was removed. 5 mL medium containing 10 mg/L mitocin-C was added , kept at 37 ℃, and then cultured in saturated humidity warm box with the CO2 of 0.05 fraction volume for 2 or 3 hours. It was coated with 0.1% gelatin in 6-hole plate, laid for over 30 minutes, and then threw away or the medium with mitocin-C was removed, washed with phosptat buffer for 5 times so as to remove the mitocin-C. 2 mL 0.05% trypsin digestive cells were added, and it was observed under microscope. When crevice appeared, cells became round (about 2-4 minutes), digestion was stopped by adding medium of the same volume, and blew up with straws repetitively to make it into monoplast suspension. Special-used cover glass was put in the center of cell counting chamber. Cells were sucked in by glass siphon, and cell suspension flew out at bucket of up or down-sides of counting chamber, until the cover glass was filled with fluid. Living cell were inoculated at concentrations of 3×108, 5×108, 1×109 L-1 after their number counted.

选取2~5代的小鼠胚胎成纤维细胞,待其至对数生长期倒掉或吸掉培养基,加入含10 mg/L丝裂霉素C的细胞全培养液5 mL,置37 ℃,体积分数0.05的CO2饱和湿度温箱中培养二三小时,在6孔板中加入0.1%明胶包被,放置30 min以上,倒掉或吸掉含丝裂霉素C的培养基,磷酸盐缓冲液清洗5遍,尽量除去丝裂霉素C,加入2 mL 0.05%的胰蛋白酶消化细胞,镜下观察,当细胞间出现裂隙,细胞变圆时(约2~4 min),立即加入等量的全培养液终止消化,并用吸管反复吹打,使之成为单细胞悬液,在细胞计数板中央放置专用的盖玻片,用玻璃虹吸管吸取细胞,让虹吸管在盖玻片上或下侧的计数板凹槽处流出悬液,至盖玻片下被液体充满为止。

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