MicroRNAs (miRNAs)are a class of small non-coding RNA that regulate gene expression at the posttranscrip-tional level and play important roles in cell proliferation,differentiation,apoptosis and metabolism.Moreover,specific miR-NAs can be secreted or released from tissues and cells in different physiological or pathological statusand enteredinto blood circulation,and secreted miRNAs can be delivered into recipient cells and emerged as powerful regulatorsof a wide range of biological processes.Many recent studieshave shown that miRNAswere involved in the development and progression of type 2 diabetes (T2DM).Due to the widely source,highly stability,and specific expression patternunder different physiological or pathological conditions,circulating miRNAs may serve as a novelbiomarker for T2DM and may also participate in the devel-opment of type 2 diabetes.

Objective To develop a high performance liquid chromatographic method (HPLC) for the analysis of of cholesterol in erythrocyte membranes.Methods The study included 167 consecutive chest pain patients who underwent coronary artery angiography in the Department of Cardiology,Nanjing General Hospital of Nanjing Command between September 2012 and February 2013.According to the clinical symptoms and t angiographic results,patients were divided into three groups:acute coronary syndrome (ACS) group (n =46),stable angina pectoris (SAP) group (n =76) and the control group (n =45).After the erythrocyte sample was hypotonically lysed and washed,saponification was carried out in a polassium hydroxide solution at 70 ℃.After extraction by Hexane/isopropanol mixture,the sample was separated on a Lichrospher column and detected by ultraviolet absorbance at 208 nm.A mobile phase composed of acetonitrile-isopropyl alcohol was found to be the most suitable for this separation.Concentrations of cholesterol in erythrocyte membranes were tested.Analysis of variance with covariates (ANOVA) was used to evaluate differences in CEM levels among groups.The relationship between continuous variables was evaluated by Spearman's correlation coefficient.Results Under the chromatographic conditions described,retention time of the cholesterol was approximately 6.1 min.Good separation and detectability of cholesterol in erythrocyte membranes were obtained.The method proved to be linear in the injection range of cholesterol from 0.05 g to 2.00 g.Cholesterol content in erythrocyte membranes were (87.0 μg/mg,75.4-98.9 μg/mg),(92.9 μg/mg,83.8-109.0 μg/mg) and (173.9 μg/mg,140.0-188.8 μ g/mg) in the control,SAP and ACS groups,respectively.Cholesterol content in erythrocyte membranes was significantly higher in ACS group than that in SAP and control groups (P ＜ 0.01).Conclusion We have successfully developed a method for the determination of cholesterol in erythrocyte membranes with good sensitivity,specificity and repeatability.

Objective To investigated the correlation between the effects of the compatibility of fried Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma Praeparata cum Melle on chemical composition and anti-inflammatory effects.Methods An HPLC-DAD method was employed for the simultaneous determination of various components of fried Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma Praeparata cum Melle,including paeoniflorin,albiflorin,benzoylpaeoniflorin,oxypaeoniflorin,glycyrrhizic acid and glycyrrhizin.An adjuvant-induced arthritis rat model was established.Administration groups were given fried Paeoniae Radix Alba,Glycyrrhizae Radix et Rhizoma Praeparata cum Melle,and fried Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma Praeparata cum Melle liquid by gavage for 2 weeks,respectively.Enzyme-linked immunosorbent assay was conducted to detect the levels of inflammatory factors IL-1β,TNF-α and PGE2 in serum of each group.Factorial design,bivariate correlation analysis,and multiple linear regression were employed to explore the effect of compatibility on chemical composition and its correlation with anti-inflammatory efficacy.Results The established HPLC-DAD analysis method revealed a significant increase in the content of oxypaeoniflorin,albiflorin and benzoylpaeoniflorin following the compatibility of fried Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma Praeparata cum Melle(P<0.05).Compared with the model group,the contents of IL-1β,TNF-α and PGE2 in serum of each administration group decreased(P<0.01),and the contents of IL-1β,TNF-α and PGE2 in serum of compatibility group were lower than fried Paeoniae Radix Alba group and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle group(P<0.05,P<0.01).The results from the factorial design indicated the presence of an interaction effect between fried Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma Praeparata cum Melle.Bivariate correlation analysis results showed negative correlations between oxypaeoniflorin,albiflorin,paeoniflorin,glycyrrhizin,benzoylpaeoniflorin and glycyrrhizic acid with the contents of IL-1β,TNF-α and PGE2,respectively.Multiple linear regression results revealed that glycyrrhizin and glycyrrhizic acid were negatively correlated with the contents of IL-1β,TNF-α and PGE2.Conclusion The established method for analyzing multiple constituents in the compatibility of fried Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma Praeparata cum Melle is straightforward,rapid,highly specific,accurate,and reliable,suitable for quality evaluation and control.The compatibility of fried Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma Praeparata cum Melle can inhibit the expressions of IIL-1β,TNF-α and PGE2,and the content of chemical composition is correlated with anti-inflammatory effects.