ABSTRACT
Objective: To detect the expression levels of miR-194-5p and magnesium transporter protein 1 (MagTl) in peripheral blood mononuclear cells (PBMCs) of the patients with chronic hepatitis B virus (HBV) infection, and to investigate their possible clinical significances. Methods: The clinical materials of 40 healthy subjects (healthy control group). 40 cases of asymptomatic hepatitis B carriers (HBV carrier group). 40 patients with mild-moderate chronic hepatitis B (mild-moderate CHB group) and 40 patients with severe chronic hepatitis B (severe CHB group) were collected. The peripheral blood of the subjects in various groups was collected and the PBMCs were isolated by Ficoll-Hypaque density gradient centrifugation. The expression levels of miR-194-5p and MagTl mRNA in PBMCs of the subjects in various groups were detected by RT-PCR assay, the expression levels of MagTl protein in PBMCs of the subjects in various groups were detected by Western blotting method, the levels of Mg in peripheral blood and PBMCs of the subjects in various groups were detected, and the expression levels of CD6+ T lymphocyte surface molecules programmed death receptor 1 (PD-1) and natural killer cell receptor 2 group D molecule (NKG2D) in PBMCs of the subjects in various groups were analyzed by flow cytometry. The miR-194-5p mimic (miR-194-5p mimic group) and miR-jNC (miR-NC group) were transfected into the 293T cells, and then the dual luciferase reporter gene assay was used to detect the luciferase activities in 293T cells in two groups. Results: Compared with healthy control group, the expression levels of MagTl mRNA and protein, the expression levels of CD8 + T lymphocyte surface molecule NKG2D and the levels of Mg in PBMCs of the patients in HBV carrier group, mild-moderate CHB group and severe CHB group were decreased ( P<-0. 05) . the expression levels of miR-194-5p mRNA in PBMCs and CD8 + T lymphocyte surface molecule PD-1 of the patients were increased (P<.0. 05). Compared with HBV carrier group and mild-moderate CHB group, the expression levels of MagTl mRNA and protein, the expression level of CD8+ T lymphocyte surface molecule NKG2D and the level of Mg in PBMCs of the patients in severe CHB group were decreased ( P<0. 05). and the expression levels of miR-194-5p mRNA in PBMCs and CD8 + T lymphocyte surface molecule PD-1 of the patients were increased (P<0. 05). Compared with HBV carrier group, the expression levels of MagTl mRNA and protein, the expression level of CD8+ T lymphocyte surface molecule NKG2D and the level of Mg in PBMCs of the patients in mild-moderate CHB group were decreased ( P
ABSTRACT
Objective@#To investigate the levels of urinary exosomal miR-194-5p in children with idiopathic nephrotic syndrome (INS) and its clinical value as a non-invasive auxiliary diagnostic marker. @*Methods@#Urine samples were collected from 101 INS children and 98 sex- and age-matched healthy children, and the levels of urinary exosomal miR-194-5p were determined by quantitative real-time polymerase chain reaction (qRT-PCR). The clinical value of urinary exosomal miR-194-5p in the diagnosis of children′s INS was evaluated by the ROC curve and correlation analysis. @*Results@#The levels of urinary exosomal miR-194-5p in INS children (2.420 [0.650,9.515] fmol/L) were significantly higher than that in healthy controls (0.360 [0.220,0.653] fmol/L, U=1 552, P＜0.01). Compared with the INS children before treatment, the levels of urinary exosomal miR-194-5p in clinical remission period decreased significantly (0.320 [0.145,0.523] fmol/L vs 0.975 [0.375,4.358] fmol/L, W=708, P＜0.01). Moreover, the levels of miR-194-5p in INS children with heavy urine protein (8.430 [7.225,13.070] fmol/L) were significantly higher than that with light urine protein (2.130 [1.180,3.090] fmol/L, U=0, P＜0.01). The area under the ROC curve (AUC ROC ) of miR-194-5p was 0.843 (95%CI: 0.789-0.897) for the diagnosis of INS children. Spearman correlation analysis showed that the levels of urinary exosomal miR-194-5p in INS children were negatively correlated with serum albumin levels (r=-0.300), and were positively correlated with serum total cholesterol levels (r=0.278) and 24-hour urine protein content (r=0.296, all P＜0.01). @*Conclusion@#The levels of urinary exosomal miR-194-5p in INS children increase obviously, and are closely associated with 24-hour urine protein, serum albumin and total cholesterol levels, indicating that urinary exosomal miR-194-5p may be serve as a new non-invasive fluid biopsy molecular marker for the auxiliary diagnosis of INS in children.