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用于分析瘦素和脂联素的唾液处理的有效技术。

An effective technique for the processing of saliva for the analysis of leptin and adiponectin.

机构信息

Department of Oral Medicine and Periodontology, Faculty of Dentistry, Mahidol University, Bangkok, Thailand.

出版信息

Peptides. 2013 Sep;47:60-5. doi: 10.1016/j.peptides.2013.06.010. Epub 2013 Jul 12.

Abstract

The recovery of protein from saliva has been extensively investigated as a method to monitor health. The aim of this study was to compare filtration and centrifugation as two methods of saliva processing necessary for determining the levels of salivary leptin and adiponectin. Thirty-seven healthy patients (median age of 45 years; range 35-73) participated in the study. Unstimulated whole saliva was collected by a drooling technique. An aliquot was filtered using a Millex-Millipore(®) (0.45μm PVDF Dura Pore membrane) syringe and a second aliquot was centrifuged at 15000×g for 15min at 4°C. Leptin and adiponectin levels were analyzed using an ELISA kit for serum (RayBio(®), GA, USA) with minor modifications. Leptin and adiponectin levels following the filtration technique yielded comparable results with those after centrifugation. Correlation was observed between filtered and centrifuged salivary leptin levels ((r=0.9155; 95% CI 0.8362-0.9573; p<0.0001) with concordance correlation coefficient k 0.9114 (95% CI 0.8332-0.9539)). Less correlation was observed for adiponectin ((r=0.5718; 95% CI 0.3041-0.7558; p=0.0002) with concordance correlation coefficient k 0.5586 (95% CI 0.2977-0.7419)). Using a Bland-Altman plot, similar measurements for both adipocytokines were observed with mean difference within a 95% CI, and interpreted as no systematic differences between the two processing techniques. This study showed that filtration is an alternative saliva processing technique to retrieve supernatant for protein analysis. Filtered saliva yielded leptin and adiponectin concentrations comparable with those obtained from centrifuged saliva.

摘要

从唾液中回收蛋白质已被广泛研究作为监测健康的一种方法。本研究旨在比较过滤和离心两种唾液处理方法,这两种方法都是确定唾液瘦素和脂联素水平所必需的。37 名健康患者(中位年龄 45 岁;范围 35-73 岁)参与了这项研究。采用流涎技术收集未刺激的全唾液。用 Millex-Millipore®(0.45μm PVDF Dura Pore 膜)注射器过滤一份唾液样本,另一份唾液样本在 4°C 下以 15000×g 离心 15min。使用 RayBio(®),GA,USA 的用于血清的 ELISA 试剂盒(略有修改)分析瘦素和脂联素水平。过滤技术后获得的瘦素和脂联素水平与离心技术后获得的结果相当。过滤和离心唾液瘦素水平之间存在相关性(r=0.9155;95%置信区间 0.8362-0.9573;p<0.0001),一致性相关系数 k 为 0.9114(95%置信区间 0.8332-0.9539))。脂联素的相关性较低(r=0.5718;95%置信区间 0.3041-0.7558;p=0.0002),一致性相关系数 k 为 0.5586(95%置信区间 0.2977-0.7419))。使用 Bland-Altman 图,两种处理技术之间的两种细胞因子的测量值相似,在 95%置信区间内的平均差异被解释为没有系统差异。本研究表明,过滤是一种替代的唾液处理技术,可以回收上清液进行蛋白质分析。过滤后的唾液产生的瘦素和脂联素浓度与离心后的唾液相当。

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