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Results showed endothelialcells inoculated by shimen virulent strain of CSFV have no obvious change after 12h,24h;At36h,cells become be elongate and appearanced defluvium;At 48h,a few of cells started tobecome round and intercellular space become auxesis;At 72h, intercellular space becomemore auxesis, defluvium of cells was acute.

结果表明,血管内皮细胞于接种 CSFV 石门株后 12h,24h 无明显变化,至 36h 时,细胞开始拉长,个别细胞开始脱落;至 48h 时,部分细胞变圆,脱落,细胞间隙增大;至 72h,细胞拉长呈长梭状,细胞间隙很大,细胞脱落严重。3 个对照组的细胞均未出现明显变化。

The expression level of metabolitic enzyme (CYP1A1, CYP1B1) and its correlation with cell differentiation /apoptosis were studied as well. The influence of resveratrol in cell cycle was determined with FCM. Results 1 Resveratrol not only suppresses the growth of medulloblastoma cells in a dose and time related fashion but also induces cell differentiation and apoptosis. Resveratrol promotes differention of UW228-1,-2 cells to forward glia, UW228-3 and Med-3 cells to neuron. A treatment of resveratrol in the concentration of 100μM for 48hrs comit most of cells die of apoptosis. 2 Among the four cell lines Fas and Caspase-3 were constantly expessed, whereas FasL was absent irrespective to the drug treatment. 3 ICC, RT-PCR, Western-blot hybridization and EROD enzymatic activity assay demonstrated that expression of CYP1A1 is different after treatment with resveratrol in four cell lines, up-regulated in three UW228 cell lines in a dose dependent manner but down-regulated in Med-3 cells. Suppressive effect of resveratrol on CYP1B1 expression is the same among four medulloblatoma cell lines. 4 Cell cycle distribution of UW228-3 was greatly changed after treatment.

结果:1、白藜芦醇以时间和剂量依赖性方式抑制髓母细胞细胞生长,同时促进细胞的分化和凋亡:白藜芦醇促使UW228-1、-2靶细胞向神经胶质细胞方向成熟分化,UW228-3、Med-3细胞向神经元细胞方向成熟分化,以100μM作用48小时效果显著;2、四个细胞系均有Fas和Caspase-3表达,但无Fas-L基因表达和蛋白活性产生,因此难以形成Fas相关性死亡通路;3、白藜芦醇处理前后CYP1A1基因的表达在各细胞系略有不同:UW228-1,-2,-3三个细胞系白藜芦醇以剂量相关性方式上调CYP1A1的表达,而Med-3细胞系,白藜芦醇却抑制CYP1A1的表达;四个细胞系在白藜芦醇处理后CYP1B1的表达均受到抑制;4、UW228-3细胞系经白藜芦醇作用后细胞周期有较大改变,表现在G0/G1期在整个细胞周期中所占的比例增加,而其它时期则相应减少。

The present studies showed that two cell populations were found in haemocytes: large cell with high granularity and small cell with low granularity by flow cytometry FCM on light scanttering pattern. Two distinct cell types were identified based on phase contrast microscope: one type of cell was dark and dioptric aberration, while the other was bright and dioptric strong. By Giemsa and H.E staining, cytoplasmic staining were heterogeneous and internal particles were obvious in one type of cell, while cytoplastic staining were homogeneous and internal particles were inexistent in the other type of cell. By transmission electron microscope, we found that the mitochondria, Golgi apparatus organelles were rich and internal particles were obvious in one type of cells, and contrary to the another cells.

流式细胞术光散射图谱显示血细胞被分两类,一类为颗粒度高的大细胞,另外一类为颗粒度低的小细胞;相差显微镜观察显示,血细胞可分为胞体暗、折光性差和胞体明亮、折光性强的两类; Giemsa和H.E染色显示细胞分为胞质染色不均一、胞内颗粒明显和胞质染色均一、胞内颗粒不明显的两类;透射电镜超薄切片观察显示,颗粒明显的细胞胞质内线粒体、高尔基体等细胞器较丰富,颗粒不明显的细胞胞质内细胞器较少;负染结果表明血细胞主要分为表面不光滑、突起明显和细胞表面光滑、突起较不明显的两类。

The living cell of L428 cell has experienced from monoclonal to the multi-cell forming process,and presents a big cell around many small clone cell encystation phenomenon.finally,gigantic cells is dead in the life time.3.The cell counting result showed that the proportion of big cell or the H/RS type cell(diameter≥25um)is for(11.6±1.5)%in the L428 group,for(4.6±0.7)%in L428-MVC group and for(13.1±1.3)%in L428-EVC group,respectively.

经持续稀释成单个大细胞、单个小细胞的L428细胞,小细胞可分裂转化成大细胞,大细胞亦可生成小细胞;常见单个大细胞周围出现多个小细胞围绕的现象,在此过程中NF-kB一直持续活化。3。

This paper reviews the pathways of ES cells in vitro being committed and differentiated to hematopoietic cells, cardiomyocytes, neural cells, adipocytes, pancreatic islets, endothelial cells, epithelia, osteoblasts, chondroblast and melanocytes.

文章综述了体外定向诱导ES细胞分化为造血细胞、心肌细胞、神经细胞、脂肪细胞、胰岛细胞、内皮细胞、上皮细胞、成骨细胞、软骨细胞和黑素细胞等的途径和方法。

As the DDP group A549DDP cells were treated with IC10 of cisplatin for 48 hours. As the DDP+Rh2 group A549DDP cells were treated with IC10 of cisplatin and IC5 of gensenoside Rh2 for 48 hours. As the controlling group A549DDP cells were not treated with any kinds of drugs. Changes in cellular morphology were observed by fluorescence microscopy. Peak value apoptosis of A549DDP cells were inspected by flow cytometry. State of PTP was evaluated by ultraviolate spectrofluorometer. Changs of calcium in cells and membrane potential of mitochondrion were determined by flow cytometry. Release of cyt-c from mitochondrion with westton blot Radiative activity of〓Tc-MIBI ingested by cells was measured withγcounter. Results: IC50 of cisplatine to A549 cells was 24uM.

以无毒浓度的Rh2单独作用于A549DDP细胞作为Rh2组,以低效浓度的顺铂单独作用于A549DDP细胞作为DDP组,以无毒浓度的Rh2和低效浓度的顺铂联合作用于A549DDP细胞作为Rh2+DDP组,作用时间为48小时,以不加药物干预作为对照组,Hoechst33258染色荧光显微镜下观察细胞形态的变化,流式细胞仪检测细胞凋亡峰;紫外光分光光度计检测线粒体PTP开放情况,流式细胞仪检测细胞内钙离子浓度及线粒体跨膜电位的变化,Westton blot检测线粒体内细胞色素C释放情况及Caspase-3的活性,γ计数仪检测细胞摄取〓Tc-MIBI放射性活性。

Result 1 Magnetic nanoparticles, magnetic nanoparticles modified with antisense oligodeoxynucleotide of human telomerase reverse transcriptase induced HL-60 tumor cells to apoptosis, we could see typical morphologic change of apoptosis cells: karyopyknosis, chromation"s condensing and aggregation in nuclear, forming crescent-shaped or annulus structures to lean on edge of cell nucleus"s membrane and posing apoptosis body by Atomic Force Microscope, Fluorescence microscope, transmission electron Microscope 2 There was a significant difference compared with control group(p.01), inhibition ratio had significant positive correlation with medication dosage and time ;during 0.8-8μM dosage amplitude, inhibition ratio accrescenced by dosages increasing. However, the inhibition ratio would decrease when dosage over 8-80μM.

结果 1 磁性纳米粒子、修饰有端粒酶反义寡核苷酸的磁性纳米粒子诱导HL-60细胞发生凋亡,原子力显微镜、光学显微镜、荧光显微镜和透射电镜下均观察到HL-60细胞呈现典型的凋亡细胞的形态变化:细胞核固缩,核内染色质浓缩、凝聚、形成新月形或环状结构紧靠在细胞核膜边缘,并形成凋亡小体。2 磁性纳米粒子、修饰有端粒酶反义寡聚脱氧核苷酸的磁性纳米粒子对HL-60肿瘤细胞的生长和增殖有明显的抑制作用,与对照组相比有显著性差异(p<0.01),在剂量为0.8-8μmol/L范围内,抑制率随剂量的增加而增加,当剂量超过8μmol/L时,抑制率反而下降;3 磁性纳米粒子、修饰有端粒酶反义寡聚脱氧核苷酸的磁性纳米粒子可增强p53基因的表达活性,引起DNA降解损伤,反向调节细胞周期活动,促使细胞从G0期进入G1期,抑制肿瘤细胞的生长。4 修饰有端粒酶反义寡聚脱氧核苷酸的量子点能通过内吞作用进入HL-60肿瘤细胞细胞核,可以在细胞内进行定位和促进HL-60肿瘤细胞的凋亡。

Results: The results showed that:(1) The three types of NOS immunoreactivity were not found in rat testis until 30 days after birth;(2) NOS1 immunoreactivity was found in a few spermatocytes in rat testes on postnatal 30 days, and in spermatozoa present at the lumen surface of seminiferous tubules and some Leydig cells on postnatal 60 days;(3) NOS2 immunoreactivity appeared in a few spermatocytes, Sertoli cells and peritubular myoid cells on postnatal 30 days. On postnatal 60 days, NOS2 immunoreactivity appeared in some Leydig cells, peritubular myoid cells, Sertoli cells, very few spermatocytes and the head of immature spermatozoa in some seminiferous tubules;(4) NOS3 immunoreactivity was detected in a few spermatocytes on postnatal 30 days, as well as in the smooth muscle cells of blood vessels on postnatal 30 days and 60 days.

结果:(1)生后4、7、14 d大鼠睾丸未见3种NOS免疫阳性反应;(2)生后30 d少数精母细胞及生后60 d生精小管腔面精子和间质细胞呈NOS1阳性;(3)生后30 d少数精母细胞、支持细胞和管周类肌细胞呈NOS2阳性,而生后60 d NOS2阳性反应见于睾丸间质细胞、管周类肌细胞、支持细胞、极少数精母细胞和不成熟精子头部;(4)生后30 d睾丸内少数精母细胞和血管壁呈NOS3阳性,生后60 d NOS3阳性反应仅见于血管壁。

We found that 1 mutant EG4 cells showed typical characteristics of pluripotent stem cells which had no obvious difference with wild cells; 2 When induced by 10〓 M retinoic acid , mutant EG4 cells differentiated into adipocytes with high frequency compared with mutant cells, suggested that EGFR plays a role in adipocyte differentiation; 3 when injected into nude mice, mutant teratocarcinomas contained a large amount of connective tissues as well as skeletal muscle, while wild EG4 cells produced frequently cartilage, keratinocyte and neuroepithelium.

我们建立了稳定表达胞内区功能缺失的外源EGFR cDNA片段的EG4细胞,分析其生长分化特性,发现 1)突变型细胞可在未分化状态下维持长时间的增殖,表明EGFR对EG多能干细胞表型无明显影响;2)〓 M的维甲酸A(retinoid acid A,RA)诱导后,大部分对照组细胞分化为脂肪细胞,而突变型细胞分化为脂肪细胞的比例明显较少,表明EGFR在脂肪细胞的发育分化中具有一定的调节作用;3)畸胎瘤切片分析显示,突变型瘤组织分布有大量的未分化细胞和结缔组织,分化细胞以肌肉细胞为主;对照组瘤组织含丰富的角质上皮、软骨、神经管等依赖EGFR的分化组织。

To understand the infectivity by porcine endogenous retrovirus with porcine skin fibroblast cell in vitro and in vivo, porcine skin fibroblast cell established by our laboratory were co-cultured with neo/HEK293 cell for the infection of RERV in vitro, and were subcutaneously transplantated to SCID (severe combined immuno-deficiency) mice for the infection of PERV in vivo, laying the foundation for valuation of biologic safety of xenotrans-plantation. The event of neo/HEK293 cells infected by PERV occurred during co-culture of porcine skin fibroblast cells with neo/HEK293 cells, expanding the rang of the infection of porcine endogenous retrovirus. Afterpig cells transplantated subcutaneously in SCID mice, the microchimerism (78.57%) of pig cells occurred widel, and there was phenomena of integration of PERV provirus (85.71%) in several organs or tissues remote from the injected sites, indicating infection of PERV in SCID mice in vivo. yet, there is no evidence of active viral replication in analysis of PERV env RNA of these tissues or organs.

为了解猪皮肤成纤维细胞PERV在体外和体内的感染性,通过建立猪皮肤成纤维细胞系,将所建细胞系与人胚胎肾293细胞体外共培养,并移植于严重联合免疫缺陷鼠皮下进行猪皮肤成纤维细胞PERV的体外和体内感染性实验,结果表明,猪皮肤成纤维细胞与人胚胎肾细胞共培养过程中,猪内源性逆转录病霉感染人胚胎肾细胞,进一步证实和拓宽了猪细胞PERV感染人细胞的范畴;猪皮肤成纤维细胞移植SCID鼠皮下后,导致SCID鼠发生猪细胞微嵌合(78.57%)和PERV在体内感染(85.71%)并且波及远离移植部位的多种组织或器官,但是并未检测出SCID鼠组织中表达PERV env RNA。

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