T3 对 MG63 细胞株 TRAIL，QPG，QPGL 表达的影响

目的观察在不同浓度T3环境下人成骨肉瘤MG63细胞株肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其护骨素(OPG)、护骨素配体(OPGL)的表达,探讨甲亢性骨质疏松症的发病机制。方法用不同浓度T3(对照组,10-12、10-10、10-8mol/L组)分别刺激培养的MG63细胞24h,RT-PCR法检测TRAIL,OPG,OPGLmRNA的表达。结果T3对MG63细胞中TRAIL,OPGLmRNA的表达,均按照对照组、10-12mol/L组、10-10mol/L组、10-8mol/L组顺序递增(P<0.05),OPGmRNA的表达按照此顺序递减(P<0.05),10-8mol/L组TRAILmRNA的水平明显高于对照组(P<0.05)。同时,OPG/OPGL比率按照对照组、10-12mol/L组、10-10mol/L组、10-8mol/L组顺序递减。结论T3可能导致成骨细胞中TRAIL和OPGL表达增多,OPG的表达减少。这可能是甲亢性骨质疏松症的重要发病机制之一。     

葡萄糖、雌二醇对MG63细胞株TRAIL，OPG，OPGL mRNA表达的影响

目的 观察在不同浓度葡萄糖、雌二醇环境下人成骨肉瘤MG63细胞株肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其护骨素(OPG)、护骨素配体(OPGL)的表达,探讨绝经后糖尿病女性骨质疏松症的发病机制.方法 用不同浓度葡萄糖(5.5、16.7、33.3 mmol/L)和17β-E2分别刺激培养的MG63细胞24 h,RT-PCR法检测TRAIL、OPG、OPGL mRNA的表达.结果 葡萄糖对MG63细胞中TRAIL、OPGLmRNA的表达,均按照对照组、5.5 mmol/L组、16.7 mmol/L组、33.3 mmol/L组顺序递增(P＜0.05),OPGmRNA的表达按照此顺序递减(P＜0.05).33.3 mmol/L组TRAIL mRNA的水平明显高于对照组[(1.004±0.070)vs(0.740±0.023),P＜0.05],而17β-E2可增加OPG mRNA的表达,减少TRAIL、OPGLmRNA的表达.结论 高糖环境可能导致成骨细胞中TRAIL和OPGL表达增多,OPG的表达减少,从而导致骨质疏松.而雌二醇对葡萄糖环境下的MG63细胞中上述因子的表达可产生一定的对抗效应.这可能是绝经后糖尿病女性骨质疏松症患者雌激素替代治疗的依据之一.     

甲状旁腺激素对MG63细胞OPG、OPGL及其相关因子的调节

目的探讨甲状旁腺激素（PTH）调节骨吸收作用的途径和机理。方法用0．2-20μg PTH干预人成骨肉瘤MG63细胞2d，观察PTH对MG63细胞表达护骨素（OPG）、护骨素配体（OPGL）及其相关因子，如肿瘤坏死因子相关性凋亡诱导配体（TRAIL）、巨噬细胞集落刺激因子（M—CSF）的影响，用RT-PCR法检测OPG、OPGL、M—CSF和TRAIL mRNA表达情况，用Western blot分析法检测OPG和OPGL蛋白表达情况。结果PTH下调MG63细胞中OPG的表达，上调OPGL、M—CSF及TRAIL的表达。结论删能够减低成骨样细胞MG63中OPG／OPGL表达比率，增加M—CSF、TRAIL的表达。这可能有利于促进破骨细胞生成及活化，进而促进骨吸收。     

TiO2喷砂酸蚀处理对人成骨细胞骨保护素和骨保护素配体mRNA表达影响的研究

[目的]探讨纯钛钛片经过喷砂及喷砂酸蚀处理后对人成骨细胞系MG63细胞骨保护素(osteoprotegerin,OPG)及骨保护素配体(osteoprotegerin ligand,OPGL)mRNA表达水平的影响.[方法]纯钛钛片表面分别进行机械打磨、喷砂及喷砂酸蚀处理,将人成骨细胞系MG63细胞接种于钛片表面,采用荧光实时定量PCR法检测OPG、OPGL mRNA表达水平.[结果]MG63细胞在经过喷砂及喷砂酸蚀处理后的钛片上培养后其OPG mRNA水平增高,与机械打磨组相比有统计学意义(P＜0.05),而OPGL mRNA表达水平在各组之间没有明显差异(P＞0.05).[结论]经过喷砂及喷砂酸蚀处理的钛片均可促进人成骨细胞表达OPG,从而调节成骨细胞与破骨细胞之间的平衡,促进骨质重建.     

不同浓度钛微粒对护骨素/护骨素配体基因表达影响的体外研究

目的: 探讨磨损微粒引起假体周围骨溶解的机理, 为人工关节假体松动的预防研究打下基础。方法: 采用不同浓度钛微粒 (粒径 <3μm) 对成骨细胞MG 63细胞进行 24h干预, 半定量RT PCR观察其对护骨素 (OPG) 及护骨素配体 (OPG- L) 基因表达的影响。结果:钛微粒能上调MG 63细胞OPG及OPG- L基因表达, 且对后者作用强于前者。结论: OPG- L/OPG比率随着钛微粒浓度的加大而增高, 引起骨溶解大于骨形成, 这可能是磨损微粒引起人工关节假体松动的重要原因, 这表明降低OPG- L/OPG比率有可能预防或逆转此病理过程。     

雄激素对体外成骨细胞QPG及其配体基因表达的影响

目的检测小鼠胎鼠成骨细胞体外培养并将不同浓度雄激素干预后,OPG和OPGLmRNA表达的变化,探讨雄激素在成骨细胞介导破骨细胞分化、活化过程中的调控机制。方法分离得到小鼠胎鼠颅盖骨成骨细胞,培养传代并选择第二代细胞用于实验;对第二代成骨细胞实施含10-10mol/L、10-9mol/L、10-8mol/L3种浓度雄激素的培养液干预;抽提细胞RNA,采用RT-PCR方法半定量观察成骨细胞中OPG和OPGL基因mRNA表达的变化。结果实验选用的各浓度组均未出现细胞毒性反应,雄激素干预使成骨细胞中OPG基因表达上调,而OPGL与OPG比率随时间呈递减趋势。结论雄激素可以特异性地在转录水平调节成骨细胞中OPG和OPGL基因的表达。     

hsa-miR-655靶向CLCF1基因对人成骨肉瘤MG63 细胞OPG/RANKL系统表达的影响

目的探讨hsa-miR-655通过靶向调控心肌营养素样细胞因子1(cardiotrophin-like cytokine factor 1,CLCF1)基因表达,对人成骨肉MG63细胞系增殖、护骨素(osteoprotegerin,OPG)和核因子κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)表达的影响。方法通过hsa-miR-655慢病毒转染MG63细胞进行功能获得实验。实验分阴性对照组(NC组)和hsa-miR-655过表达组(OE组),荧光显微镜和流式细胞术(flow cytometry analysis,FCM)评估转染效率,Cell Counting Kit-8(CCK-8)法检测细胞增殖能力,FCM检测细胞周期分布,实时荧光定量PCR(qRT-PCR)检测hsa-miR-655、CLCF1、OPG和RANKL mRNA的水平,蛋白质印迹法(Western blot)检测CLCF1、OPG及RANKL蛋白的变化。结果荧光显微镜和FCM显示对照组和miR-655过表达组感染效率为80%以上;qRT-PCR结果显示,相对于对照组,过表达组MG63细胞hsa-miR-655表达上调,差异有统计学意义(P=0.0004);CLCF1 mRNA水平未见明显变化(P=0.0986);过表达组OPG(P=0.0384)、RANKL(P=0.0007)mRNA均有所上调,差异具有统计学意义(P0.05);Western blot结果显示,过表达组CLCF1蛋白水平表达下调(P=0.0053);与对照组比较,过表达组OPG(P=0.0021)、RANKL(P=0.0002)蛋白均上调,而OPG/RANKL比值却降低(P=0.0078),差异具有统计学意义(P0.05);CCK-8细胞增殖实验结果显示,miR-655过表达后抑制MG63细胞增殖;FCM检测细胞周期结果显示,miR-655过表达后MG63细胞G0/G1期细胞比例增加,S期减少,差异具有统计学意义(P0.05)。结论hsa-miR-655通过负调控CLCF1基因的表达,抑制MG63细胞的增殖,下调OPG/RANKL的比值。     

近β钛合金对人成骨细胞骨保护素/核因子-κB受体活化因子配体mRNA表达影响的研究

[目的]探讨近B钛合金Ti-5Zr-3Sn-5Mo-15Nb(TLM)对人成骨样MG63细胞骨保护素(osteoprotegerin,OPG)及核因子-κB受体活化因子配体(receptor activator of nuclear factor kappa B ligand,RANKL)mRNA表达水平的影响.[方法]将人成骨样MG63细胞分别接种于纯钛钛片、TLM及微弧氧化处理的TLM表面,采用荧光实时定量PCR法检测OPG/RANKL mRNA表达水平.[结果]TLM组OPG mRNA表达水平略高于纯钛钛片组,但没有统计学意义(P>0.05),而微弧氧化处理组OPG mRNA水平明显升高,与纯钛钛片组及TLM组相比有显著性差异(P<0.05),三组之间的RANKL mRNA水平没有明显差异(P>0.05).[结论]经过微弧氧化处理的TLM可以上调成骨细胞OPG mRNA水平,从而影响OPG/RANKL mRNA的比值,调节成骨细胞与破骨细胞之间的平衡,进一步促进骨质重建.     

利塞膦酸钠对人成骨样细胞MG63 OPG及TNFα表达的影响

目的研究利塞膦酸钠对人成骨样细胞MG 63护骨素（OPG）及肿瘤坏死因子α（TNFα）表达的影响，以阐明利塞膦酸钠发挥骨保护作用的机制。方法以不同浓度（10^-10mol／L；10^-9mol／L；10^-8mol／L；10^-7mol／L）的利塞膦酸钠干预MG 63细胞72h；ELISA法测定细胞条件培养液中OPG及TNFα的含量。结果利塞膦酸钠可下词MG 63细胞TNFα的表达；上调OPG的表达；并提高碱性磷酸酶（ALP）活性。结论利塞膦酸钠可通过调节成骨样细胞MG 63 OPG、TNFα的表达，从而发挥其抗骨吸收的作用．     

Distribution, elimination, and action of d-tubocurarine in neonates, infants, children, and adults

The relation of plasma concentration of d-tubocurarine (dTc) to neuromuscular blockade, and the distribution and urinary excretion of dTc was determined in neonates (n = 4), infants (n = 6), children (n = 8), and adults (n = 8). The plasma concentration-time course curves to 24 hr are best described for all groups by three-compartment models. Both neonates and infants exhibit decreased plasma clearance (CLP), 1.1 +/- 0.08 and 1.0 +/- 0.06 ml X kg-1 X min-1, and in addition a prolonged t1/2 terminal phase, 311 +/- 44 and 306 +/- 35 (mean +/- SEM, min). The neonates' 24-hr urinary excretion, 27 +/- 2 (mean +/- SEM, % total dose) is significantly less than the adult value, 45 +/- 4% total dose. There was no significant difference seen in the log plasma concentration-evoked compound electromyogram (ECEMG) response between 20-80% paralysis for adults, children, infants, and five of the seven neonates studied. Two of the neonates had a significant shift of their log concentration-response curve to the right. There was also no significant difference between any of the groups in the time for 50% return of ECEMG stimulus height or the time required for recovery of the ECEMG from 25 to 75% of control value. for recovery of the ECEMG from 25 to 75% of control value.     

Methotrexate, Azathioprine, Cyclosporine, and Risk of Fracture

We studied the fracture risk associated with use of methotrexate, azathioprine, and cyclosporine. The study was designed as a case-control study. All patients with a fracture (n = 124,655) in the year 2000 in Denmark served as cases. Information on fractures and confounders was retrieved from the National Hospital Discharge Register and a number of other national registers. For each case, three age- and gender-matched controls were randomly drawn from the general population (n = 373,962). Exposure was use of the drugs and a number of covariates including other immunosuppressive drugs, corticosteroids, any cancer, Crohn’s disease, ulcerative colitis, rheumatoid arthritis, psoriasis, liver and kidney disease, prior fracture, and alcoholism. Azathioprine was associated with an increase in overall fracture risk, but besides this, none of the drugs was significantly associated with overall fracture risk or risk of hip, spine, or forearm fracture. Liver [odds ratio (OR) = 1.55, 95% confidence interval (CI) 1.42–1.69] and kidney (OR = 1.26, 95% CI 1.16–1.37) diseases were significantly associated with increased risk of fractures. Azathioprine was associated with an increase in overall fracture risk but not in the risk of spine, hip, or forearm fractures. Methotrexate and cyclosporine were not associated with fracture risk. It thus seems that the underlying disease for which the treatment is administered may be responsible for much of the increase in fracture risk rather than the drugs used to treat the disorder in question.