ObjectiveTo investigate whether lysyl oxidase(LOX) has significant relation to persistent atrial fibrillation(AF) with mitral valvular diseases. MethodsWe included 184 consecutive lone mitral valvular disease patients who needed surgery in our hospital between March 2012 and February 2014. Patients who had persistent AF formed the AF group, and those who still kept sinus rhythm(SR) comprised the SR group. In the AF group, patients were separated into two groups by the subgroup of mitral valvular disease(mitral stenosis and mitral regurgitation), then formed a MS+AF group and a MR+AF group. There were 97 patients with 44 males and 53 females at age of 52.76±11.35 years in the AF group and 90 patients with 48 males and 42 females at age of 47.95±14.22 years in the SR group. Blood specimens were obtained from patients for the first time peripheral venous blood after admitted to hospital. LOX levels were measured by ELISA test kits of LOX. ResultsAF was diagnosed in 51.87%(97/187) of lone mitral valvular disease patients. Mitral stenosis patients were easy to have AF(60.31% vs. 34.43%, P<0.05). The plasma level of LOX was significantly higher in the AF group than that in the SR group(73.78±25.42 IU/L vs. 51.05±18.96 IU/L，P<0.05). In the AF group, the LOX level in the mitral stenosis group was higher than that in the mitral regurgitation group(84.21±32.15 IU/L vs. 59.74±35.21 IU/L， P<0.05). Mitral stenosis patients more frequently had a history of stroke than mitral regurgitation patients did. AF correlated significantly with the level of LOX(r=0.124， P=0.036) and left atrial dimension(r=0.531，P=0.042). ConclusionWe validate and extend the hypothesis that increasing LOX level predicts an increasing risk of AF in mitral valvular diseases. Lysine oxidase is a potential diagnostic biomarker for AF. It is expressed significantly in mitral stenosis patients with AF especially.
ObjectiveTo investigate whether miR-93-5p suppresses osteogenic differentiation of mouse mesenchymal stem cells (C3H10T1/2) by targeting Smad5, a predicted target in silicon. MethodsSmad5 3'-UTRluciferase vector (pmiR-RB-REPORTTM) was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-93-5p on Smad5 3'-UTR-luciferase activity to identify whether Smad5 was the target gene of miR-93-5p. miR-93-5p mimics (group M), miR-93-5p inhibitor (group In), miR-93-5p mimics negative control (group MC), and miR-93-5p inhibitor negative control (group InC) were transfected into the C3H10T1/2 cells, respectively, and followed by induction of osteogenic differentiation. After 48 hours, the real-time fluorescent quantitative PCR (qRTPCR) and Western blot assays were performed to detect the relative expressions of Smad5 mRNA and protein. At 14 days, to realize the regulation role of miR-93-5p in osteogenic differentiation, the extracellular calcium deposition during the osteogenesis of C3H10T1/2 cells was tested by Alizarin red staining. ResultsDual-luciferase reporter gene assay showed that miR-93-5p could combine with Smad5 mRNA 3'-UTR specificity, and inhibited its luciferase activity (P<0.05). After 48 hours, no significant difference was shown in the relative expression of Smad5 mRNA between group M and group MC as well as between group In and group InC by qRT-PCR assay (P>0.05); however, the results of Western blot assay showed that the relative expression of Smad5 protein was significantly decreased in group M and increased in group In when compared with groups MC and InC (P<0.05). At 14 days after osteogenic induction, Alizarin red staining showed that the extracellular calcium deposition of group M was obviously less than that of group MC, and it was obviously more in group In than in group InC. ConclusionSmad5 may be the target gene of miR-93-5p. And miR-93-5p can suppress osteogenic differentiation of C3H10T1/2 cells by directly targeting Smad5.
ObjectiveTo explore if Smad7 protein can inhibit growth of keloids by observing the gene and protein expressions of Smad7, collagen type Ⅰ, and collagen type Ⅲ and cell proliferation after over-expression vectors of Smad7 transfecting keloid fibroblasts (KFb). MethodsFibroblasts were acquired from 10 male patient with keloids at the age of 20 to 25 years. After in vitro culture, KFb were divided into 3 groups: untransfected group (group A), pcDNA3.1 (-) transfected group (group B), and pcDNA3.1 (-)-smad7 transfected group (group C). The mRNA and protein expression levels of Smad7, collagen type Ⅰ, and collagen type Ⅲ were detected by real-time fluorescence quantitative PCR and Western blot at 48 hours after transfection. The cell proliferation ability was detected by MTT assay at 24 hours after transfection. ResultsThe relative expression levels of mRNA and protein of Smad7 in group C were significantly higher than those in group A and group B (P < 0.01). The relative expression levels of mRNA and protein of collagen type Ⅰ and collagen type Ⅲ in group C were significantly lower than those in group A and group B (P < 0.01). The relative expression levels of mRNA of collagen type Ⅰ and collagen type Ⅲ in group B were significantly higher than those in group A (P < 0.01); and the relative expression levels of proteins of Smad7, collagen type Ⅰ, and collagen type Ⅲ were significantly lower than those in group A (P < 0.01). The cell proliferation ability in group C was significantly lower than that in group A and group B at each time point by MTT assay (P < 0.05), but no difference was found between group A and group B (P>0.05). ConclusionGene expressions of collagen type Ⅰ, and collagen type Ⅲ and cell proliferation will be inhibited after KFb are transfected by over-expression vector of Smad7.