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Objective To analyze the relationship between the level of microRNA-29b in circulation and left ventricular hypertrophy in hypertensive patients.Methods A total of 240 subjects from Henan Province People's Hospital from June 2015 to June 2018 were included in the present study.Among them,160 were hospitalized patients,and were divided into two groups.Patients with simple hypertension and had no left ventricular hypertrophy (80 cases) were in the simple hypertension group (HBP-NLVH),and patients with hypertension combined with left ventricular hypertrophy (80 cases) were in the high blood pressure with left ventricular hypertrophy group (HBP-LVH).Normal control subjects (80 cases) were those with no hypertension and randomly selected from the medical center of Henan Province People's Hospital.Serum microRNA-29b expressions were detected by real time fluorescence quantitative PCR.The thickness of interventricular septum (IVSD) and left ventricular posterior wall thickness (LVPWD) were measured by echocardiography.Results Compared with the normal control group (1.95±0.79),the relative expression of microRNA-29b in the patients both in the HBP-NLVH group (2.67±0.92) and the HBP-LVH group (5.12 ± 1.23) was up-regulated,and the difference between normal control and patients was statistically significant (P＜0.05).In patients,the microRNA-29b level in the HBP-LVH group was significantly higher than that in the HBP-NLVH group (P＜0.05).The expression level of microRNA-29b was positively correlated with IVSD (r=0.71,P＜0.05) and LVPWD (r=0.74,P＜0.05),respectively.The sensitivity and specificity of serum microRNA-29b levels in the diagnosis of left ventricular hypertrophy in hypertension patients were 96.8％ and 91.3％,respectively.Conclusion Serum microRNA-29b level is elevated in hypertensive patients with left ventricular hypertrophy,and is positively correlated with left ventricular hypertrophy.The circulation microRNA-29b might be a useful biomarker with prognostic value in left ventricular hypertrophy in hypertension patients.
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Objective To investigate the relationship between p53 and miR-29b in myofibroblasts (MFs)induced from human Tenon fibroblasts (HTFs).Methods Human Tenon tissues were obtained from strabismus surgery patients in the First Affiliated Hospital of Anhui Medical University from March to July 2016.The primary HTFs were cultured by tissue culture method and passaged to 2-4 generations.The cells were divided into 4 groups:HTFs group cells were normally cultured,the TGF-β1 activation group cells was activated with 10 ng/ml tumor necrosis factorβ1 (TGF-β1),the siRNA transfection group and the negative control group were transfected with p53 siRNA and negative siRNA,respectively.The expression of p53 protein and cell migration ability of HTFs and MFs were detected by immunohistochemical and cell scratch assay.The relative expression level of miR-29b and p53 mRNA in each group was detected by real-time PCR.This study followed the Helsinki declaration and the patients or their guardians signed informed consent.Results The primary HTFs had long fusiform shape,and were arranged in a spiral shape.After activated by TGF-β1,the cells were transformed into MFs,and the cell morphology was irregular and the arrangement was disordered.The staining intensity of p53 protein in TGF-β1 activation group was significantly stronger than that in HTFs group (t =-10.384,P＜0.05).The migration distance of TGF-β1 activation group cells was significantly larger than that of HTFs group (P＜0.05).The relative expression level of p53 mRNA was 1.00±0.00,3.95±2.61,0.06±0.06 and 0.98±0.16 in HTFs group,TGF-β1 activation group,siRNA transfection group and negative control group,respectively;the relative expression levels of miR-29b was 1.00±0.00,0.54±0.09,5.10±2.31 and 1.03 ±0.09 in HTFs group,TGF-β31 activation group,siRNA transfection group and negative control group,respectively;the overall differences were statistically significant (p53 mRNA:F =6.688,P =0.006;miR-29b:F=13.640,P=0.000).The relative expression of p53 mRNA was significantly higher and the relative expression of miR-29b was significantly lower in the TGFβ1 activation groups than those in the HTFs group and siRNA transfection group,the differences were statistically significant (all at P＜0.05).Conclusions During the process of HTFs transforming to MFs,the expression of p53 is upregulated and the expression of miR-29b is downregulated,Supressing the expression of p53 can upregulate the expression of miR-29b.There may exist interactions between p53 and miR-29b,which regulate the formation of fibrotic scars.
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Objective To identify the expression differences of mircoRNA-29 b (miR-29b),microRNA? 24 (miR-24) and microRNA-200 c (miR-200c) in plasma of infants with primary congenital glaucoma (PCG) and normal,and analyze its clinical significance.Methods The expression quantity of microRNAs (miR-29b,miR-24,miR-200c) in plasma of PCG group (16 cases) and normal control group (49 cases) were detected by RT-PCR,and the relationship between their expression differences and severity of disease were analyzed.The diagnostic value of miRNAs for PCG was evaluated by receiver-operating characteristic curve (ROC).Results The expression quantity of miR-29b,miR-24,miR-200c in PCG group (0.31 ±0.19,0.17 ±0.16,0.55 ±0.18,respectively) were significantly lower than those in normal control group (1.18-±0.52,2.86 ±2.65,1.62 ± 0.76,respectively) (all P ＜ 0.05);The expression quantity of miR-24 and miR-29b in plasma was related to the severity of PCG,the more severe the disease,the lower the expression;ROC curve indicated that miR-24 and miR-29b had a higher diagnostic value for PCG disease than miR-200c.Conclusion Free miRNAs in plasma may be used as a new plasma markers for auxiliary diagnosis of PCG.
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Objective:To construct a lentiviral vector overexpression of micrRNA-29b and investigate the biological characteristics in mouse neuronal cell lines GT1-7.Methods:We chemically synthesized two oligonucleotide single-stranded,complete the comple-mentary by bridging extension into DNA double-stranded to form miR-29b precursor structure.The restriction enzyme digested vector plasmid FUGW was ligated to the precursor structure ofmiR-29b by homologous recombination to construct the corresponding lentiviral vector of microRNA-29b overexpression,and the stable cells were obtained in the mouse neuronal cell line GT1-7 by bleomycin drag screening.RT-PCR was used to detect the expression level of related genes at mRNA transcription level,Results:The recombinant lentiviral expression plasmid f-F-miR-29b was successfully constructed,and the expression level was about 30 times higher than that of the control group.The expressions of DCX,Vdac1 and pten were inhibited,have no changes in sex developmental related genes LH-β,kiss-l,Inshulin,IGF-I,GPR54,GnRH and leptin-R.Conclusion:Using the method of lentivirus screening,the microRNA-29b overexpressing stably transformed cells was successfully obtained in mouse neuron GT1-7 cells,which laid a foundation for the study of biological characteristics ofmicroRNA-29b.
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Objective:To investigate microRNA-29b ( miR-29b) expression in cerebral cortex , spinal cord, fore limb muscle, and serum of SOD1-G93A amyotrophic lateral sclerosis ( ALS) mice, and to identify the biomarker and to assess diagnostic values for ALS .Methods:Cerebral cortex , spinal cord , fore limb muscle and serum from 16 SOD1-G93 A ALS mice and 16 wild-type mice were taken and then microRNA extracted , detecting the expression of miR-29 b by real-time quantitative polymerase chain re-action ( RT-qPCR ) .The diagnostic performance of miR-29b for ALS was estimated by the receiver operating characteristic ( ROC ) curve . Results: The results from the validation indicated that the differences in miR-29b between the cerebral cortex of SOD1-G93A ALS and the healthy control subjects were statistically significant (P=0.001).Meanwhile, the expressions 8, 12, and 16 weeks later were higher than those of the controls ( ALS vs.Control: 8 weeks, P=0.044; 12 weeks, P=0.018; 16 weeks, P=0.045).When the relative expression level of miR-29b was used to diagnose ALS in SOD1-G93A ALS mice, the area under the ROC (area under the curve, AUC) was 0.885, if the diagnostic threshold was set at 0.185 6, the sensitivity and specificity were 92.9%and 71.4%.Conclusion:MiR-29 b may act as medical monitoring indices of ALS in early time .