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.

Proteasome activator PA28γ,also known as Ki antigen,REGγ or PSME3,is first found as Ki antigen in the serum of a patient with systemic lupus erythematosus,which belongs to 11S proteasome activator family together with PA28α and PA28β.In the amino acid sequence,PA28γ has 25%homology with PA28α and PA28β and its seven-subunit homomer is a 20S proteasome acti-vator,which mainly exists in the nucleus and participates in ubiquitin and ATP-dependent or independent protein degradation.More and more studies have reported the role of PA28γ in human immune-related diseases.This article reviews the function of PA28γ as a proteasome activator and its role in immune-related diseases including cancer,inflammation and virus infection-related diseases and other diseases in recent years,so as to reveal the role of PA28γ in the occurrence and development of immune-related diseases and its potential as a target for immunotherapy.

A new definition of metabolic dysfunction-associated fatty liver disease (MAFLD) has recently been proposed. We aim to examine the associations of MAFLD, particularly its discordance from non-alcoholic fatty liver disease (NAFLD), with the progression of elevated brachial-ankle pulse wave velocity (baPWV) and albuminuria in a community-based study sample in Shanghai, China. After 4.3 years of follow-up, 778 participants developed elevated baPWV and 499 developed albuminuria. In comparison with the non-MAFLD group, the multivariable adjusted odds ratio (OR) of MAFLD group for new-onset elevated baPWV was 1.25 (95% confidence interval (CI) 1.01-1.55) and 1.35 (95% CI 1.07-1.70) for albuminuria. Participants without NAFLD but diagnosed according to MAFLD definition were associated with higher risk of incident albuminuria (OR 1.77; 95% CI 1.07-2.94). Patients with MAFLD with high value of hepamet fibrosis score or poor-controlled diabetes had higher risk of elevated baPWV or albuminuria. In conclusion, MAFLD was associated with new-onset elevated baPWV and albuminuria independently of body mass index, waist circumference, and hip circumference. Individuals without NAFLD but diagnosed as MAFLD had high risk of albuminuria, supporting that MAFLD criteria would be practical for the evaluation of long-term risk of subclinical atherosclerosis among fatty liver patients.
Humans ; Pulse Wave Analysis ; Albuminuria ; Ankle Brachial Index ; Non-alcoholic Fatty Liver Disease/diagnosis* ; Vascular Stiffness ; Prospective Studies ; Risk Factors ; China/epidemiology*