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To produce the scientific evidence for developing and manufacturing new antitumor drugs.Methods: 1 The inhibitory effect on cell growth of Hela was measured by MTT assay in treated or untreated groups (3.125, 6.25, 12.5, 25, 50μg/ml TAM and control) for three different treatment times (24h, 48h and 72h).2 Apoptosis and cell cycle were measured by FCM in four experimental groups (0, 4, 16, 40μg/ml TAM) for 48h.3 Adopting Wright and Giemse's staining to observe the morphology of Hela cells which treated with 40μg/ml TAM.4 Using invasion experiment to detect the Hela cells'invasive abilities which treated with 40μg/ml TAM.5 The protein expressional levels of P-ERK, ERK, C-myc and Cyclin D1 in Hela cells untreated or treated with 4, 16, 40μg/ml TAM for 24h were measured by Western blot.6 Expression of anti-apoptotic gene bcl-2, apoptotic gene bax and MMP-9 in Hela cells of four experimental groups (0, 4, 16, 40μg/ml TAM for 24h), were observed by revers transcription PCR.7 The protein expression of P-ERK, ERK, Bcl-2 and Bax in Hela cells treated with 40μg/ml TAM for 24h observed by laser scanning microscopes.

1采用四甲基偶氮唑蓝法检测不同浓度北豆根总碱(3.125、6.25、12.5、25、50μg/ml)处理不同时间(24、48和72小时)对Hela细胞增殖反应的抑制作用。2采用流式细胞技术(flow cytometry,FCM)检测不同浓度北豆根总碱(0、4、16、40μg/ml)作用48小时,对Hela细胞凋亡及周期变化的影响。3瑞氏-姬姆萨染色后显微镜观察北豆根总碱(0、40μg/ml)作用24小时后Hela细胞形态学变化。4采用Transwell小室法检测北豆根总碱(0、40μg/ml)作用24小时后对Hela细胞侵袭性的影响。5采用免疫印迹方法检测不同浓度北豆根总碱(0、4、16、40μg/ml)作用24小时后,Hela细胞中磷酸化ERK、ERK、C-myc、CyclinD1的表达变化。6采用逆转录-聚合酶链反应(revers transcription PCR,RT-PCR)半定量检测北豆根总碱(0、4、16、40μg/ml)作用24小时,Hela细胞抗凋亡基因bcl-2、促凋亡基因bax、基质金属蛋白酶-9(MMP-9)的表达变化。7应用激光共聚焦显微镜(laser scanning microscope,LSM)观察北豆根总碱(0、40μg/ml)作用24小时后,Hela细胞内P-ERK、ERK、Bcl-2、Bax蛋白的表达变化。

We detected that EGF mRNA was expressed sflungly lii the oocyte, and is also found hi gmnulosa cells, the cell fium smaller foflicular expressed stronger than fium bigger one. In the corpus hemonbaglcwn corpus luteurn, lean type and pseudocorpus-luteum, EGF rnRNA was detected,, no distinct difference can be seen in them. The EGF mRNA expressed strongly in fimbria end, ampulla and isthmus of oviduct, in the big follicular stage, ovulation stage, pregnancy stage and spurius pregnancy stage, we can not see any distinct change in them, but hi the medium follicuar stage,it is weaker.

结果发现:猪卵母细胞中EGF的mRNA强烈表达,且小卵泡卵母细胞→中卵泡卵母细胞→大卵泡卵母细胞中,EGF的mRNA表达量有逐渐减少的趋势;猪卵泡的颗粒细胞中有EGF的mRNA表达,小卵泡颗粒细胞→中卵泡颗粒细胞→大卵泡颗粒细胞中,EGF的mRNA表达也有逐渐减少的趋势;猪卵巢中的红体、黄体、白体和假黄体中都有EGF的mRNA表达,看不出几部分的表达量有明显的强弱变化;猪输卵管伞部、壶腹部和峡部,都有EGF的mRNA表达,在大卵泡期,排卵期,孕期和假孕期都强烈表达,各期间看不出明显的强弱变化,中卵泡期表达较弱;猪子宫中EGF的mRNA在大卵泡期,排卵期,孕期和假孕期都强烈表达,看不出表达量的明显变化,而小卵泡期表达量明显减弱。

First we first establish the apoptosis of related protein glossary database, then downloads the neurodegenerative disorders of apoptosis related abstract from the on-line biomedicine literature , and searchesfor the protein information database of the neurodegenerative disorders of apoptosis related pathway, then uses the data mining technology,with apoptosis of protein related glossary database use already establishment. Automatically searches for the neurodegenerative disorders of apoptosis related protein by the program in the biomedicine literature abstract separately in,(1) a sentence,(2) two sentences,(3) aliterature, finding simultaneously appears protein-protein interation. And comparing to the neurodegenerative disorders of apoptosis related protein database, looking for their repetition. This result comparing to the known neurodegenerative disorders of apoptosis related pathway, thus looking for unestablished neurodegenerative disorders of apoptosis pathway.

首先我们先建立细胞凋亡讯息蛋白质相关词汇字库,再从线上生物医学文献下载神经退化性疾病细胞凋亡相关文献摘要,并且搜寻线上蛋白质资料库中神经退化性疾病之细胞凋亡讯息蛋白质传递路径,接著利用文献探勘技术,使用已建立之细胞凋亡讯息蛋白质相关词汇字库,以程式自动在生物医学文献摘要中搜寻神经退化性疾病细胞凋亡相关蛋白质分别在,(1)一个句子、(2)二个句子、(3)一篇文献,中同时出现的两两蛋白质关系,再与已知线上神经退化性疾病细胞凋亡讯息传递路径资料库的蛋白质两两关系做比较,找出两者重复建立的部分,把这个结果与线上已知之细胞凋亡讯息传递路径资料库蛋白质关系作比较,由此找出尚未建立之细胞凋亡讯息传递路径图。

The results showed that the changes of the types were not caused by new cell mutation, but by translocation of the different cell types of variegation. The changes of division direction of white cells and green cells resulted that some of the green epidermal cells translocated into the second layer and it pushed furthermore the origin white cells of the second layer into the third layer, so that the variegation altered.

Pollock'突变型异化不是由于细胞突变形成,而是由于质体突变型白色细胞和原型绿色细胞在生长发育过程中相互竞争转轨所致,即白色细胞和原型绿色细胞改变了分裂方向,部分表皮绿色细胞进人第二层,挤压第二层白色细胞转轨进人第三层,导致细胞类型的移位,从而形成异型斑。

The results show that the normal process begins with archesporial cell and undergoes stages of primary and secondary sporogenous cell,microspore mother cell,dyad,tetrad,central nucleus microspore,vacuolated microspore,mature microspore,twocell pollen and threecell mature pollen.

对平流层辐射处理SP3谷子和对照CK3谷子雄性细胞发育的研究表明,雄性细胞正常发育过程从孢原细胞开始,经初生造孢细胞、次生造孢细胞、小孢子母细胞、二分体、四分体和单核小孢子中央期、单核小孢子液泡期、单核小孢子成熟期直到二细胞花粉、三细胞成熟花粉结束。

In the first section, the effect of sodium houttuyfonate on the level of lysozyme secreted by macrophages was first tested by bacteriolytic method; then the effect of sodium houttuyfonate on the level of tartaric acid resistant acid phosphatase produced in macrophages was investigated by analyzing the phenol generated in the reaction that was catalyzed by the enzyme; the third, the effect of sodium houttuyfonate on the phagocytic activity of macrophages was observed by microscopy; the forth, the effect of sodium houttuyfonate on the production of complement C4 from macrophages was tested by hemolytic method; the fifth, the effect of sodium houttuyfonate on the expression of C3b receptors on the surface of macrophages was analyzed by observing the formation percentage of zymosan wreath; the sixth, the effect of sodium houttuyfonate on the respiratory burst in macrophages was tested by flow cytometry; the seventh, the effect of sodium houttuyfonate on the production of IL-1α and IL-1β from macrophages was tested by ELISA method.

在这一章中,首先,采用溶菌法考察合成鱼腥草素对巨噬细胞分泌溶菌酶水平的影响;其次,采用游离酚法研究合成鱼腥草素对巨噬细胞产生的耐酒石酸酸性磷酸酶水平的影响;第三,采用显微镜观察法考察合成鱼腥草素对巨噬细胞吞噬功能的影响;第四,采用溶血法测定合成鱼腥草素对巨噬细胞产生的补体C4水平的影响;第五,采用酵母花环法检测合成鱼腥草素对巨噬细胞表面C3b受体水平的影响;第六,采用流式细胞术法观察合成鱼腥草素对巨噬细胞呼吸爆发的影响;第七,采用ELISA法探讨合成鱼腥草素对巨噬细胞产生IL-1α及IL-1β水平的影响。

This result indicates that WT1 gene plays an important role in differentiation and development of fetal kidney and may be the factor that promotes metanephric blastemal cell to differentiate into epithelial cell.

结果显示小胎龄肾组织中WT1蛋白在胚基细胞和幼稚肾小球细胞核表达而大胎龄组肾组织中WT1在肾小管细胞胞浆表达,阳性率分别为57.1%(8/14)和46.2%(6/13),提示WT1基因在胚胎肾分化发育的过程中起着重要作用,WT1蛋白可能是促进后肾胚基细胞向上皮细胞分化的调控因子,其表达在时间上和空间上都受到严格的调控,WT1的表达异常可能导致胚基细胞分化停滞。17例肾母细胞瘤WT1蛋白表达阳性率为41.2%(7/17),阳性部位在胚基型和上皮型肿瘤细胞核,表达部位和阳性率与早期胚胎肾相似,其中间质型肾母细胞瘤均为阴性,胚基型和上皮型肾母细胞瘤阳性率70%(7/10),两组间阳性率有显著差异。

When electric fusion method was used for nuclear transfer, the fusion rate (46. 0%), cleavage rate (53. 9%) and blastocyte development rate (10.9%) of adult ear fibroblasts were significantly lower than that of fetal fibroblasts (64. 5%, 70.1%, 21. 6% respectively), fetal skin cells (71. 5%, 70.8%, 22. 1% respectively) and ovary granulosa cells (88. 2%, 79. 1%, 25. 5% respectively). There was no significant difference among other donor cells in the cleavage and blastocyst development rate of resconstituted embryos.

当用电融合法进行核移植时,成体耳部成纤维细胞的融合率(46.0%),卵裂率(53.9%)和囊胚发育率(10.9%)均显著低于胎儿成纤维细胞(64.5%,70.1%和21.6%),胎儿皮肤细胞(71.5%,70.8%和22.1%),以及卵巢颗粒细胞(88.2%,79.1%和25.5%);另外三种细胞间的卵裂率,囊胚发育率无显著差异,但卵巢颗粒细胞的融合率显著高于胎儿成纤维细胞和胎儿皮肤细胞(88.2%vs 64.4%,71.5%,P<0.05)。

Results The number of cellular apoptosis was obviously increased in multiple organs from the six patients died from SARS. The cellular apoptosis occurred predominantly in cytokeratin-positive pneumoncytes, terminal bronchiolar epithelium, CD3 + and CD8 + lymphocytes, as well as a part of CD20 + lymphocytes and CD68 + macrophages.

结果 与正常组织比较,SARS患者全身多器官组织细胞凋亡数目明显增多,尤以肺脏和免疫器官为甚,主要的凋亡细胞类型为CK阳性肺泡上皮细胞、终末支气管黏膜上皮细胞、CD3+细胞、CD8+细胞,以及少部分CD2 0 +细胞和CD6 8+巨噬细胞

Methods: A 5ml bone marrow was extracted from the lilac of human volunteers. By Percoll fluid and density gradient centrifugation, the MSC was obtained; after the cells filled the bottom of vessel, subcultured them, when they subculture in third generation, redigested them, 500 R/min centrifugate, alter the completed medium to chemical definition medium, examined the form change and prolifration of cells by invert microscope, toluidine blue stain、immunocytochemical stain and RT-PCR to test the type Ⅱ collagen mRNA and proteoglycan.

取健康成人髂后上棘处骨髓5ml,经percoll液分离后密度梯度离心,〓/ml密度接种培养,观察原代细胞的贴壁、增殖状况,细胞长满瓶底后进行传代培养;传至第三代细胞,重新消化后以500转/分钟轻度离心5分钟,〓/ml接种,改用化学限定培养基代替完全培养基培养,倒置显微镜观察细胞生长情况及形态变化,甲苯胺蓝染色观察诱导细胞合成细胞外基质中的蛋白多糖,免疫细胞化学染色检测ECM中Ⅱ胶原的蛋白合成,RT-PCR鉴定诱导细胞Ⅱ胶原mRNA的表达。

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