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Objective: The purpose of this study was to investigate the effect of TNF-α on osteoclast differentiation in primary murine bone marrow cell culture with and without RANKL.

目的:在诱导破骨细胞分化的体外骨髓细胞培养系统中,研究破骨细胞分化因子存在和不存在的情况下,肿瘤坏死因子α对破骨细胞分化的影响。

However, it is not known how orthodontic force influences the function of cementoblast and what is the relationship between the function change of cementoblast with root resorption. So we studied the effect of mechanical stimuli on root resoption by examining the expression OPG and RANKL and exploring the related signal transduction pathway in cementoblast. Methods: Cementoblasts OCCM30 were cultured in DMEM with 10% FBS for 48 hours and starved in DMEM with no FBS for 24 hours, then subjected to mechanical strain by four-point bending system with tension and compression stress at 2000μstrain at 0.5Hz frequency. The flow cytometry was used to examine the cell cycle and proliferation activity after 3h, 6h, 12h, 24h loading respectively. OPG and RANKL mRNA were analysed with real time quantitive RT-PCR after the same loading period as FCM.

本课题利用四点弯曲力学加载系统,采用流式细胞术、实时荧光定量PCR、Western免疫印迹等方法,研究体外培养的成牙骨质细胞株OCCM-30在不同力学环境中增殖活性的变化,以及其表达骨保护素和破骨细胞分化因子的mRNA的变化,探讨应力刺激对成牙骨质细胞表达破骨细胞分化调控蛋白的影响,并通过对应力刺激下成牙骨质细胞分裂原活化蛋白激酶家族中的ERK1/2、P38MAPK活性的变化及其与OPG、RANKL基因表达变化关系的研究,对力学刺激调节破骨细胞分化调控蛋白的相关上游信号转导通路进行初步探讨。

Result show that TRAP-positive mononucleated and multinucleated cells were markedly induced at the compression side at 3, 5, and 7 days. Mononucleated osteoclast reached peak level at 3 days after tooth movement, while multinucleated cells reached peak level at 5 days.

破骨细胞酶组化标记和计数统计显示,TRAP阳性细胞数量在牙移动第3、5和7天时与对照组比较有显著性差异;其中单核破骨细胞在第3天、多核破骨细胞在第5天时达到峰值。

Results: At the concentrations of 0.1, 1 and 10 μmol/L, berberine dose-dependently suppressed the formation of TRAP-positive multinucleated cells, the TRAP activity and the osteoclastic bone resorption. The strongest inhibitory effect was exhibited at the concentration of 10 μmol/L, with the inhibiting rate of 60.45%, 42.12% and 72.69% respectively.

结果:小檗碱在0.1~10μmol/L范围内,浓度依赖性地抑制TRAP阳性多核破骨细胞的形成和TRAP活性,减少破骨性骨吸收陷窝的面积;在10μmol/L浓度下,对破骨细胞的抑制作用最强,对TRAP阳性多核破骨细胞的形成和TRAP活性的抑制率分别达到了60.45%和42.12%,骨吸收陷窝面积减少72.69%。

Methods Applying 1,25-_2D_3 induced osteoclast-like multinucleated cells and isolated mature rabbit osteoclasts cocultured with bone slices, the effects of icariin on the differentiation and bone resorbing activity of osteoclasts were investigated.

方法采用1,25-2D3诱导兔骨髓单核细胞形成破骨细胞样细胞以及原代分离的乳兔成熟破骨细胞与骨片共培养的方法,考察淫羊藿苷对破骨细胞的分化及骨吸收功能的影响。

This study is to further investigate the mechanisms of osteoclastic bone resorption induced by IL-1 with established osteoclasts culture and osteoclasts, bone marrow stromal cells co-culture system.

本文利用建立的破骨细胞培养和破骨细胞、骨髓基质细胞共培养体系,在体外进一步研究IL-1诱导破骨细胞性骨吸收的机制,旨在探讨细胞因子在骨质疏松症骨丢失发生中的作用。

The effect of PTH on the osteoclastic bone resorption,and how the osteo-blasts mediat the reaction was studied in vitro by using the method of isolating and cul-turing rabbit osteoclasts and osteoblasts.

采用分离、培养兔破骨细胞和成骨细胞的方法,体外研究甲状旁腺激素对破骨细胞骨吸收功能的影响,以及成骨细胞和破骨细胞之间的相互作用。

The biochemistry and image analyzing were used to investigate the effects of the PDGF-AA at high concentration on the osteoclastic bone resorption in the osteoblast-osteoclast co-culture system.

在高浓度PDGF-AA的作用下,应用生化检查和图象分析技术,检测PDGF-AA对成骨细胞-破骨细胞共育体系中破骨细胞骨吸收功能的影响;应用免疫荧光检测破骨细胞合成抗酒石酸酸性磷酸酶的变化;应用免疫组化和RT-PCR技术检测骨保护素配体的表达。

Third passage osteoblasts were plated at the density of 5×104/ml in 75ml culture flask, twenty four hours later treatment factors were added. Total RAN was extracted from osteoblasts using guanidine, and mRNA expression of OPG and RANKL were examined using RT-PCR.

OPG作为RANKL的诱骗受体阻断其与表达于破骨细胞前体细胞和成熟的破骨细胞表面的功能性受体RANK的作用,从而抑制破骨细胞的分化和功能。

The third passage osteoblasts were plated at the density of 5×104/ml in 75ml culture flask, twenty four hours later treatment factors were added. Total RAN was extracted from osteoblasts using guanidine, and mRNA expression of OPG and RANKL were examined using RT-PCR.

OPG作为RANKL的诱骗受体阻断其与表达于破骨细胞前体细胞和成熟的破骨细胞表面的功能性受体RANK的作用,从而抑制破骨细胞的分化和功能。

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