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BMSCs were isolated, depurated, cultivanted, and identified,then incubated with the concentration of 25μg Fe per milliliter at 37℃in 5% CO2. The labeled cells were stained by Prussian blue/trypan blue,and observed under fluorescent microscope.2. The labeled cells of different density (1×104/ml,5×104/ml,1×105/ml,5×105/ml,1×106 /ml,5×106/ml)were imaged by MRI with T1WI, T2WI and T2*WI sequences;and the same density (5×104/ml,1×105/ml)labeled cells were imaged by T2*WI sequences at different time.Then the signal intensities were measured and statistically analyzed.3. The model of rabbit renal ischemia-reperfusion injury was set up and treated. Then BMSCs(5×105)were injected into 16 recipient rabbits(1abeled cells in 10,unlabeled cells in 6)from ear vein.MR images of kidneys were obtained respectively at the time points of 0,1,3,5, 8 days after transplantation and before transplantation. MR imaging findings were analyzed,which were correlated with histological findings.

实验方法1分离、纯化、培养、鉴定兔BMSCs并以SPIO以25μg Fe/ml培养液浓度标记,对标记后不同时间的细胞行普鲁士蓝染色和台盼蓝拒染后显微镜观察。2将不同细胞浓度标记细胞管(1×104/ml、5×104/ml、1×105/ml、5×105/ml、1×106/ml、5×106/ml),以不同扫描序列T1WI,T2WI,T2*WI(GRE进行MR成像,再选择相同细胞浓度组(5×104/ml、1×105/ml)进行不同时相MR成像,并测量信号强度,进行统计学分析。3缺血再灌注肾损伤模型建立和处理,然后将标记和未标记细胞(5×105个)经耳缘静脉移植入家兔体内(共16只:注入标记细胞者10只,注入未标记细胞者6只),两组均于注射前、注射后第0、1、3、5、8天应用MRI对移植细胞进行活体示踪并与肾脏组织切片对照,然后对收集的信号强度进行统计学分析。

Results The morphology of endocrine cells in digestive tract was of multiplicity.The density of 5-HT cells was the highest in the colon,but not found in the esophagus,cardia gastric and cecum.The results showed that SS cells were distributed in the pylorus mostly,but not detected in the cardia,colon,cecum and rectum.Gas positive cells were the most in the duodenums,while in the esophagus and cardia was not discovered.In the cardia VIP cells were maximum,but in the esophagus,duodenums and ileum did not appear.

结果 消化道中4种免疫阳性细胞形态多样,大多呈椭圆形和锥体形。5-HT阳性细胞数量以结肠最多,空肠和回肠次之,幽门腺区、十二指肠和直肠较少,食管、贲门腺区、胃底腺区和盲肠中未见;SS阳性细胞在幽门腺区数量最多,贲门腺区、结肠、盲肠和直肠中未见;Gas阳性细胞大量出现于十二指肠,在食管、贲门腺区和盲肠未见;除个别器官未见VIP阳性细胞外,在消化道的其余各段均有分布。

The results showed that, after induced for 4~5 d by dexamethasone, beta-glycerophosphate and ascorbate-2 phosphate, the volume of some cells of the goat MSCs gradually largened and changed their form from spindle/polygon to cuboid/ovoid; With the induce time continuing, the numbers of the cuboid/ovoid cells continuous increased; At 15th day, there were many cuboid cells, and the modality of the cells changed from cuboid to ovoid; At 25th day, the oval and cuboid cells accumulation were observed.

结果表明,山羊MSCs在地塞米松、β-磷酸甘油和抗坏血酸的作用下,从诱导的第4~5天开始,即可见有些MSCs体积逐渐增大,由梭形或多角形转变为立方形或卵圆形;随着诱导时间延长,立方形或卵圆形细胞不断增多,到第15天时,有较多的立方形细胞,且这种变化经历了从立方形向卵圆形转变的过程;第25天,可见卵圆形和方形细胞堆积,诱导分化率为90.2%± 2.97%。

Methods: By the adherence , to isolate the bone marrow mesenchymal stem cells and adipose mesenchymal stem cells , to compare the characteristics of the two cells , including obtain the quantity of the cells 、 the morphos of the cells 、 the rate of adherence 、 cell kinetics 、 cell surface marker .

利用细胞贴壁性对骨髓间充质干细胞和脂肪间充质干细胞进行分离,比较两种细胞的情况,包括(1)单位质量组织获得两种细胞的量、(2)细胞形态、(3)细胞贴壁率、(4)细胞动力学、(5)细胞表面标记。

In order to study the effects of pre-heating on the sensitivity of cells to tumor necrosis factor-α, the K562 cells were pre-heated at 40℃ for different times to induce heat shock protein 70(HSP70). To observe the effects of TNF-α concentration on the sensitivity of pre-heated cells, cells were pre-heated for 60 min and treated with TNF-α(50,100,200 and 400U/ml) for 16 hours, and the cells vitalities were detected by MTT.

为探讨预热对肿瘤细胞肿瘤坏死因子α敏感性的影响及其机理,将白血病患者胸腔渗出细胞系(K562细胞)在40℃预热不同时间(0、30、60、90、120、150和180min),使细胞热应激蛋白70(HSP70)高表达;然后将预热60min细胞和未预热细胞分别暴露于50、100、200和400U/ml的TNF-α16小时,用MTT比色法检测细胞活力。

Mechanism of oncolysis of Newcastle disease virus CN strain is studied in vitro in five carcinoma cells including HEP3B, T24, A549 and Hela cells, The pseudopods of the infected cells were retracted after 16h infected with 16HU/mL CN strain, the cells changed round and lost adhesion, the survival rates of the infected cells were lower than 10% by 120h.

本文研究了NDV-CN株对5株不同的人肿瘤细胞的体外杀伤作用。

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。

The total proteins from TGIF transfectant and vector control cells were separated by 2D electrophorosis. 760±41 and 680±38 spots were detected in TGIF transfectant and vector control cells respectively. Locus repeat analysis of protein spots showed that the mean deviation in IEF direction in TGIF transfectant and vector control cells were 0.864±0.123 mm and 0.843±0.115 mm respectively whereas the mean deviation in SDS-PAGE direction in these two cells were 0.812±0.109 mm and 1. 125±0.123 mm respectively. So the well-resolved and reproducible 2DE patterns from TGIF transfectant and vector control cells were established.

通过双向凝胶电泳分别分离TGIF转染细胞和PcDNA3.1空白质粒转染细胞的总蛋白质,测得两组细胞的蛋白质斑点数分别为760±41和680±38个;位置重复性分析表明TGIF转染细胞在等电聚焦方向上的平均偏差为0.864±0.123mm,在垂直板SDS-PAGE电泳方向上的平均位置偏差为0.812±0.109mm;PcDNA3.1空白质粒转染细胞在IEF方向上的平均偏差为0.843±0.115mm,在垂直板SDS-PAGE电泳方向上的平均位置偏差为1.125±0.123mm。

METHODS: The subcutaneous adipose tissue was obtained from adult New Zealand rabbits under aseptic condition, and cultured in vitro with collagenase digestion. All cells were divided into 3 groups: in the BMP-2 group, cells were cultured with medium containing 0.1 g/L vitamin C, 10 mmol/Lβ-sodium glycerophosphate and 10μg/L BMP-2 for 10 minutes, followed by 4-14 days inoculation with density of 18×104 cells per pore. In the BMP-7 group, cells were cultured with BMP-7 with the same methods as BMP-2 group. The cells were cultured with simple culture medium in the control group.

无菌切取成年新西兰大白兔皮下脂肪,胶原酶消化法体外分离培养脂肪干细胞,分为3组:骨形成蛋白2组加入含0.1 g/L维生素C、10 mmol/L β-甘油磷酸钠、10 μg/L骨形成蛋白2的诱导培养基培育15 min,然后按18×104个细胞/孔接种,再培育4~14 d;骨形成蛋白7组培养方法基本相同,仅将骨形成蛋白2更换为骨形成蛋白7;对照组同法加入单纯培养基进行培育。

Nowadays, n stem-like cells have been found, which exist in the tumors of some tissues, such as haematogenesis system, brain, lung, mammary gland and so on. We defined them as tumor stem cells for their similar characteristics to the normal stem cells, but they are a minor population of tumor cells and the origin of them may be tumor cells.

随着对干细胞的研究不断深入,使人们对肿瘤的发生机制重新进行了审视,并在造血系统、脑、肺、乳腺等部位肿瘤中发现极少量的具有与干细胞非常类似生物学特性的细胞,称之为肿瘤干细胞,它们很可能是肿瘤细胞的起源。

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