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Objective To determine the activities of matrix metalloproteinase-2 and -9 in odontoblast of healthy and carious teeth and explore the roles of MMP-2 and MMP-9 in pathogenic mechanism of dental caries.

目的 比较健康牙齿和龋齿成牙本质细胞中基质金属蛋白酶-2MMP-2)和基质金属蛋白酶-9(MMP-9)的活性,以探讨MMP-2和MMP-9在龋病发病机制中的作用。

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。

Positive staining of Stro-1 was found in 7.6% of SCAP. Morphologically, PDLSCs were fusiform-shaped, with many short and long ramifications under a scanning electron microscope, and the secreted extracellular matrix were around them.

透射电镜下可见,第1~3代牙周膜干细胞及根尖牙乳头干细胞生长良好,胞浆内线粒体,高尔基体和粗面内质网丰富,随着培养时间的延长,分泌的细胞外基质增多。

Objective To explore an experimental method of transfecting the marrow stromal stem cells with the reconstructed PGL3-transforming growth factor-β1 (TGF-β1) gene and to evaluate the feasibility of self-induction of MSCs to the chondrocytes in vitro so as to provide a scientific and experimental basis for a further "gene enhanced tissue engineering" research.

目的 探讨将编码细胞转化生长因子β1(transforming growth factorβ1,TGF—β1)的重组PGL3-TGF-β1质粒转染兔骨髓基质干细胞(marrow stromal stemceils,MSCs),体外通过自分泌、诱导作用向软骨细胞方向分化的可行性,为基因加强组织工程学修复软骨损伤提供体外实验依据。

Then the concentration of NO in supernatant of culture and the changes in histomorphometry were examined.

体内NO主要来源于成骨细胞及骨髓细胞[3] ,而骨髓基质细胞是成骨细胞的主要来源。

Intimal areas were measured using morphometric analysis of perfusion-fixed vein graft specimens, and intimal thickness was calculated using circumferential measurements. The SMC proliferation was studied by the immunohistochemical detection of proliferating cell nuclear antigen. Expression of MMP-2、MMP-9 mRNA in vein grafts and unoperated control Vein grafts was detected by reverse transcription polymerase chain reaction. Substrate gel zymography was used to determine the proteolytic activity.

分别行HE染色、Verhoeff弹性纤维染色观察组织病理变化,计算机病理图象分析系统测量新生内膜厚度及面积,免疫组织化学方法检测静脉壁增殖细胞核抗原表达以观察细胞增殖情况,半定量逆转录-聚合酶链反应检测静脉壁基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)mRNA的表达,明胶酶谱法检测MMP-2、MMP-9活性,比较各组之间的差别。

In this study, astroglia infected with TS-4 strain T. gondii tachyzoite and to investigate the changes of matrix metalloproteinase-12 (MMP-12) and its substrate elastin in the pathogenesis.

本研究我们利用弓虫速殖子(TS-4 strain)感染星状胶细胞的细胞模式进行研究,探讨基质金属蛋白-12(matrix metalloproteinase-12, MMP-12)和其受质弹性蛋白的变化之致病机转。

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。

Now MSC become the chief seed-cell in cartilage tissue engineer, but the technology to stimulate MSC differentiate into chondroblast is not very successful,,the standard technology is culture the cell-ball in centrifuge tube,the reconstructured cartilage-like tissue is not achieve the mechanics of hyaline cartilage.

软骨组织工程的材料学研究主要体现在两类基质材料方面的进展,一类为天然基质材料,如胶原、壳聚糖、纤维素等;一类为人工合成可降解材料,如PGA、PLA等,但两者在来源、塑形能力及细胞识别信号等方面存在不足。

The aim are:lTo examine the proliferation ability and potential chondroblast differentiation ;2To find an ideal condition stimulated BMSC differentiate into chondroblast;3To examine the chondroblast proliferation in porous scaffolds and to explore the interaction between cells and materials;4To examine the release of cytokine in vitro.

取健康成人略后上棘处骨髓 5ml,经 percoll液分离后密度梯度离心,10'砌l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,10V加 l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QCM)中的蛋白多糖,免疫细胞化学染色检测ECM中*胶原的蛋白合成,RT干CR鉴定诱导细胞*胶原mRNA的表达。

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