ABSTRACT
Background and Objectives@#Acute myocardial infarction (AMI) often occurs suddenly and leads to fatal consequences. Ferroptosis is closely related to the progression of AMI.However, the specific mechanism of ferroptosis in AMI remains unclear. @*Methods@#We constructed a cell model of AMI using AC16 cells under oxygen and glucose deprivation (OGD) conditions and a mice model of AMI using the left anterior descending (LAD) ligation. The 3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide was employed to determine cell viability. The levels of lactate dehydrogenase, creatine kinase, reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), and iron were measured using corresponding kits. Dual luciferase reporter gene assay, RNAbinding protein immunoprecipitation, and RNA pull-down were performed to validate the correlations among AC005332.7, miR-331-3p, and cyclin D2 (CCND2). Hematoxylin and eosin staining was employed to evaluate myocardial damage. @*Results@#AC005332.7 and CCND2 were lowly expressed, while miR-331-3p was highly expressed in vivo and in vitro models of AMI. AC005332.7 sufficiency reduced ROS, MDA, iron, and ACSL4 while boosting the GSH and GPX4, indicating that AC005332.7 sufficiency impeded ferroptosis to improve cardiomyocyte injury in AMI. Mechanistically, AC005332.7 interacted with miR-331-3p, and miR-331-3p targeted CCND2. Additionally, miR-331-3p overexpression or CCND2 depletion abolished the suppressive impact of AC005332.7 on ferroptosis in OGD-induced AC16 cells. Moreover, AC005332.7 overexpression suppressed ferroptosis in mice models of AMI. @*Conclusions@#AC005332.7 suppressed ferroptosis in OGD-induced AC16 cells and LAD ligation-operated mice through modulating miR-331-3p/CCND2 axis, thereby mitigating the cardiomyocyte injury in AMI, which proposed novel targets for AMI treatment.
ABSTRACT
Objective To improve the purifying method of Jo-1 antigen from rabbit thymus used for detection of anti-Jo-1 antibody by dot-blotting immunoassay(DB).Methods The rabbit thymus glands were cut into pieces,homogenized and extracted by PBS.Total protein was precipitated by acetone to get acetone powder(RTAP).The RTAP was solved in PBS and separated by an by anti-Jo-1 IgG affinity column.Results 5～7 g RTAP was obtained from 100g rabbit thymus glands.There was 19%～24% of protein in RTAP.Jo-1 antigen was enriched around 1900 folds through affinity chromatography,with 2.5% recovery of antigenic activity.In this preparation,there were several bands on SDS-PAGE,but only one band about 50 ku,reacted with anti-Jo-1 antisera on immunoblotting.Dot-blotting also showed that the antigen only reacted with Jo-1 antisera.The purified Jo-1 antigen was not stable for long time,but the antigenic activity could maintain for a long time when there was MgCl2 in the solution.Conclusion Affinity chromatography was a simple and easy method for purifying Jo-1 antigen from rabbit thymus.The antigen purified by affinity chromatography could meet the requirement for detecting Jo-1 antibody bydot-blotting.
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Objective To make an investigation for autoantibodies detection in China. Method Forty-eight hospitals were included by mail or Email. The items of QC include ANA, anti-ENA antibody, anti-dsDNA antibody, anti-mitochondria antibody and anti-smooth muscle antibody. The distribution of samples and analysis of the testing results was double-blinded. Result The correct rate of ANA, anti-ENA antibody, anti-dsDNA antibody, anti-mitochondria antibody and anti-smooth muscle antibody were 47%, 70%, 46%, 95%and 40% respectively. Conclusion The overall results are not optimistic. The quality of detecting kits is not all good, neither level of technicians. This condition should be changed by selecting better methods and improving understanding.
ABSTRACT
Objective To prepare Mi 2 antigen,detect anti Mi 2 antibody and investigate its significance in autoimmune connective tissue diseases (CTD).Methods Mi 2 antigen was prepared from rabbit thymus acetone powder and purified by DE52 chromatography.Anti Mi 2 antibodies were tested in 40 normal controls and 315 patients with CTD by double immunodiffusion.The distribution of anti Mi 2 antibodies in CTD was analysed.The clinical characteristics of dermatomyositis between anti Mi 2 positive group and negative group were compared.Results The precipitation line developed between the antigen and the standard anti Mi 2 serum.In DE52 chromatography,Mi 2 antigen was found in elute with 0 1 mmol/L and 0 2 mmol/L NaCl.The positive rate of anti Mi 2 antibodies was 26 1%(12/46) in dermatomyositis and 4 5%(2/44) in polymyositis respectively.There were no positive cases in 50 patients with rheumatoid arthritis,30 patients with primary Sjgren syndrome (SS),60 patients with systemic lupus erythematosus,50 patients with systemic sclerosis,35 patients with other CTD and 40 normal controls.The specificity of anti Mi 2 antibody in dermatomyositis was 99 4%.In dermatomyositis,the patients with positive anti Mi 2 antibody usually presented skin lesions at beginning,and most patients had V sign and/or shawl sign during the course of disease.On the other hand,the antibody negative ones manifested muscle involvements initially and easily got fever during the course of disease.There were statistic differences between the two groups in above features ( P