Objective To investigate the expression and the binding characteristics of leptin receptor (Ob-R) on human osteoblast. Methods The human osteoblasts were cultured and identified by cell biological criteria. 125I-lepti...Objective To investigate the expression and the binding characteristics of leptin receptor (Ob-R) on human osteoblast. Methods The human osteoblasts were cultured and identified by cell biological criteria. 125I-leptin was prepared by chloramine-T method and purified by PAGE. Radioligand binding assay of receptor and Scatchard plot were performed to identify the binding properties of the osteoblasts. Results Morphologically, the cultured cells showed analogical phenotype to osteoblast, and were positive in histochemical and immuno-fluorence staining for alkaline phosphatase and osteocalcin, respectively. The Alizarin Reds staining showed that mineralization was also developed by supplementation with ascorbate and β-glycerophosphate. The percentages of 125I-leptin binding to osteoblast were 16.7%±2.3% and 6.1%±1.4% without and with unlabelled leptin, respectively. From the saturation assay and by Scatchard plotting, a single class of high-affinity binding sites of Ob-R for leptin was identified with an apparent kd of 0.11±0.06 nmol/L and a Bmax of 1.7±0 3 nmol/10 5 cells. Conclusion The human osteoblasts are target of leptin, with expression of Ob-R. The leptin could possibly regulate the growth, prolifiration, function of osteoblasts and even bone remoldeling.展开更多
目的探讨胰岛素抗体（IA）对电化学发光免疫分析法（ECLIA）检测胰岛素的影响。方法放射免疫法检测糖尿病患者血清IA,IA结合率〈5%组10例,IA结合率≥5%组53例。聚乙二醇（PEG）沉淀法去除IA,Roche E lecsys 2010测定胰岛素浓度。结果IA...目的探讨胰岛素抗体（IA）对电化学发光免疫分析法（ECLIA）检测胰岛素的影响。方法放射免疫法检测糖尿病患者血清IA,IA结合率〈5%组10例,IA结合率≥5%组53例。聚乙二醇（PEG）沉淀法去除IA,Roche E lecsys 2010测定胰岛素浓度。结果IA结合率〈5%组和IA结合率5%-10%组PEG处理前后的胰岛素浓度、游离胰岛素/直接胰岛素比值、胰岛素浓度改变值差异均无统计学意义（P〉0.05）。IA结合率10%-20%、20%-30%、〉30%组PEG处理前后的胰岛素浓度、游离胰岛素/直接胰岛素比值、胰岛素浓度改变值差异有统计学意义（P〈0.05）。IA结合率〉10%时,IA结合率与游离胰岛素/直接胰岛素比值呈负相关（r=-0.729,P=0.000）,与胰岛素浓度的改变值呈正相关（r=0.821,P=0.000）。结论IA结合率≤10%时,IA对ECLIA测定血清胰岛素浓度没有影响。IA结合率〉10%时,IA使ECLIA测定胰岛素结果假性升高。展开更多
文摘Objective To investigate the expression and the binding characteristics of leptin receptor (Ob-R) on human osteoblast. Methods The human osteoblasts were cultured and identified by cell biological criteria. 125I-leptin was prepared by chloramine-T method and purified by PAGE. Radioligand binding assay of receptor and Scatchard plot were performed to identify the binding properties of the osteoblasts. Results Morphologically, the cultured cells showed analogical phenotype to osteoblast, and were positive in histochemical and immuno-fluorence staining for alkaline phosphatase and osteocalcin, respectively. The Alizarin Reds staining showed that mineralization was also developed by supplementation with ascorbate and β-glycerophosphate. The percentages of 125I-leptin binding to osteoblast were 16.7%±2.3% and 6.1%±1.4% without and with unlabelled leptin, respectively. From the saturation assay and by Scatchard plotting, a single class of high-affinity binding sites of Ob-R for leptin was identified with an apparent kd of 0.11±0.06 nmol/L and a Bmax of 1.7±0 3 nmol/10 5 cells. Conclusion The human osteoblasts are target of leptin, with expression of Ob-R. The leptin could possibly regulate the growth, prolifiration, function of osteoblasts and even bone remoldeling.
文摘目的探讨胰岛素抗体（IA）对电化学发光免疫分析法（ECLIA）检测胰岛素的影响。方法放射免疫法检测糖尿病患者血清IA,IA结合率〈5%组10例,IA结合率≥5%组53例。聚乙二醇（PEG）沉淀法去除IA,Roche E lecsys 2010测定胰岛素浓度。结果IA结合率〈5%组和IA结合率5%-10%组PEG处理前后的胰岛素浓度、游离胰岛素/直接胰岛素比值、胰岛素浓度改变值差异均无统计学意义（P〉0.05）。IA结合率10%-20%、20%-30%、〉30%组PEG处理前后的胰岛素浓度、游离胰岛素/直接胰岛素比值、胰岛素浓度改变值差异有统计学意义（P〈0.05）。IA结合率〉10%时,IA结合率与游离胰岛素/直接胰岛素比值呈负相关（r=-0.729,P=0.000）,与胰岛素浓度的改变值呈正相关（r=0.821,P=0.000）。结论IA结合率≤10%时,IA对ECLIA测定血清胰岛素浓度没有影响。IA结合率〉10%时,IA使ECLIA测定胰岛素结果假性升高。
