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目的 探讨二甲双胍对破骨细胞体外分化的影响及其可能机制.方法 采用RANKL诱导鼠巨噬细胞系Raw264.7细胞破骨分化模型,给予不同浓度的二甲双胍(400 μmol/L、800 μmol/L和1000μmol/L)和雷帕霉素(100 hmol/L)处理后,通过抗酒石酸酸性磷酸酶(tartrate-resistant Acid Phosphatase,TRAP)染色和破骨细胞骨架结构荧光染色观察破骨细胞数量,骨吸收培养板观察骨陷窝面积,RT-PCR技术检测破骨细胞特异性基因TRAP、组织蛋白酶K、降钙素受体和金属基质蛋白酶-9的表达,ELISA法检测肿瘤坏死因子-α(tumor necrosis factor,TNF-α)表达水平,Western-b1ot检测c-Fos蛋白以及哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin complex l,mTORC1)信号通路下游底物S6K1 Thr389、S6 Ser235/236、4EBP1 Thr37/46的表达及磷酸化水平.结果 二甲双胍和雷帕霉素均可使RANKL诱导的破骨细胞数量减少,抑制破骨细胞特异性基因的表达、抑制TNF-α、c-Fos蛋白以及mTORC1信号通路下游底物S6K1 Thr389、S6 Ser235/236、4E-BP1 Thr37/46的磷酸化,且二甲双胍的抑制作用具有浓度依赖性.结论 二甲双胍可抑制RANKL诱导的破骨前体细胞分化,其机制可能与抑制TNF-α和c-Fos蛋白的生成,以及抑制mTORC1信号通路激活有关.Abstract: Objective To investigate the effects of mefformin on the differentiation of osteoclastas well as relative mechanism.Methods Raw264.7 cells from the murine macrophage cell line was used.Receptor activator of NF-κB ligand (RANKL) was used to stimulate osteoclast differentiation from Raw264.7 cells.Osteoclast differentiation was assessed by tartrate-resistant acid phosphatase (TRAP) and actin fluorescence staining and counting the TRAP-positive cells after exposure to different concentrations of mefformin (0 μmol/L,400 μmol/L,800 μmol/L and 1000 μmol/L) or rapamicin (100 nmol/L) in the presence of 50 ng/ml RANKL for 5 days.Bone-resorbing activity was evaluated by BD BioCoatTM OsteologicTM Bone Cell Culture System.The expression of osteoclast-specific genes like TRAP,capthesin K,calcitonin receptor (CTR) and matrix metalloproteinase (MMP-9) was evaluated by RT-PCR.The expression of tumor necrosis factor-α(TNF-ct) S6K1Thr389,S6 Ser235/236,4E-BP1Thr37/46 and c-Fos protein was evaluated by ELISA kit and Western blot analysis,respectively.Results Mefformin dose-dependently inhibited RANKL-stimulated osteoclasts differentiation in Raw264.7 cell culture,as manifested by decrease of TRAP-positive multinucleated cells and pit erosion area,down-regulation of TRAP,cathepsin K,CTR and MMP-9 mRNA and reduction of TNF-α and c-Fos protein expression.Further study revealed that RANKL activated mTOR complex 1(mTORC1) signaling,while mefformin impaired RANKL-stimulated mTORC1 signaling.Rapamycin,an mTORCl-specific inhibitor and immunosuppressive macrolides could also prevent RANKL-induced osteoclast differentiation and bone resorption in vitro.Conclusion Mefformin inhibits osteoclastogenesis in vitro,which may due to reduction of TNF-α and c-Fos protein expression,and mTORC1 signaling is involved in this process. 相似文献
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目的 探讨切开复位内固定治疗髋臼后柱骨折的临床疗效.方法 58例已经行切开复位内固定治疗的髋臼后柱不稳定性骨折患者,完全随访平均3年(2~9年),其中6例随访2年.采用Letournel和Judet评分标准进行临床功能评估.分析患者骨折情况、影像学改变与功能的联系.结果 手术后X线片见56例骨折达解剖复位,2例骨折复位不良.结果 优:29例;良:21例;可:6例;差:2例;影像学结果:优:43例;良:7例;可:6例;差:2例.临床功能与影像学分级有很密切的联系.引起临床功能不满意的因素包括:髋关节脱位复位前超过24 h,受伤年龄大于或等于60岁,涉及关节面的粉碎性骨折和骨质疏松.结论 及时的髋关节复位对髋关节功能恢复是必要的.老年人骨折、骨质疏松和粉碎性骨折是髋关节功能恢复不满意的重要影响因素.解剖复位和内固定治疗髋臼后柱骨折是达到长期的优良临床疗效的治疗方法. 相似文献
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目的:探讨X线监视下颈椎前路经椎弓根植入螺钉的安全性。方法取南昌大学基础医学院解剖学教研室经10%中性甲醛浸泡的新鲜尸体标本10具,以标本下颈椎C3-7为研究对象,CT扫描、重建并测量双侧椎弓根最大宽度及高度,并在X线透视定位下经前路植入椎弓根螺钉,CT 扫描、三维重建分析螺钉位置,同时根据Tomasino 法评价植入螺钉的安全性。结果10具尸体标本中共测量100个椎弓根的高度及宽度,其中有6个椎弓根宽度<4.0 mm和2个椎弓根高度<4.0 mm而不能植入螺钉。经前路椎弓根植入螺钉92枚,其中横断位:Grade 178枚,Grade 213枚,Grade31枚;矢状位:Grade 187枚,Grade 23枚,Grade 32枚。结论在术前个体化的CT扫描、重建分析颈椎椎弓根形态的前提下,X线监视下经前路植入下颈椎椎弓根螺钉是安全的。 相似文献
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MRI终板异常的下腰痛患者的临床表现探讨 总被引:1,自引:0,他引:1
目的:探讨MRI检查有终板异常的下腰痛患者的临床表现特征。方法:对10例MRI检查表现终板异常的下腰痛患者系统地询问下腰痛症状,并与10例MRI检查无终板异常者进行比较。结果:终板异常组的10例下腰痛患者中有8例表现为持续性疼痛;7例表现为站立、行走位加重。而对照组10例下腰痛患者中只有3例表现为持续性疼痛;2例表现为站立、行走位加重。结论:终板异常的患者更多地表现为持续性疼痛以及站立、行走位加重,终板及其下方骨组织在MRI上的改变应该引起重视。 相似文献