Objective To investigate the regulatory effect of miR-21 on ADAMTS-1 expression in pulmonary fibrosis model. Methods A mouse model of pulmonary fibrosis was established using bleomycin (BLM) anda model of pulmonary fibrosis was constructed in vitro , the expression level of miR-21 was measured by quantitative real-time polymerase chain reaction (qRT-PCR), while the protein expression of ADAMTS-1 was measured by Western blot. NIH3T3 were transfected with miR-21 mimics and inhibitor in vitro and the cellular expression of ADAMTS-1 was measured by Western blot. Results Compared with the blank group , in mouse models of pulmonary fibrosis , the miR-21 expressions in lung tissues at three time points after BLM-treatment were significantly up-regulated while an evident decrease in ADAMTS-1 expressions were observed (P < 0.01). In vitro pulmonary fibrosis model , NIH3T3 cells after TGF-β1 in concentration 5 μg/L stimulation down-regulated ADAMTS-1 expression and up-regulated miR-21 expression (P < 0.01). NIH3T3 transfected with miR-21 mimics and inhibitor, up-regulated miR-21 expression, while down-regulated ADAMTS-1 protein expression. Conclusions Up-regulation of miR-21 and Down-regulation of ADAMTS-1 might be involved in the progression of pulmonary fibrosis model; miR-21 could negatively regulate ADAMTS-1 expression.

Objective To investigate the effect of resveratrol on autophagic flux of nasopharyngeal carcinoma CNE-2 cells, and to explore the underlying mechanism. Methods Nasopharyngeal carcinoma CNE-2 cells were divided into control group and resveratrol group. Cells in control group were normally cultured at 37℃and received no further treatment. Resveratrol group was added 40 μmol/L resveratrol 2 h before cells were culture at 37 ℃. Western blot analysis was performed to detect protein expressions of LC3B, p62, Beclin-1, phospho-mTOR (p-mTOR) and phospho-S6 (p-S6). The autophagic flux was detected under the confocal laser scanning microscopy through different color spots, after cells were transfected with adenovirus encoding GFP-mRFP-LC3. Results (1) The protein expression of LC3B was significantly increased and the protein expression of p62 was significantly decreased in resveratrol group compared with those of control group (P<0.05). There was no significant difference in Beclin-1 expression between two groups. (2) Compared to control group, expressions of p-mTOR and p-S6 were significantly decreased in resveratrol group (P<0.05). (3) Compared to control group, the red mRFP puncta were significantly increased, and the yellow GFP puncta were significantly decreased in resveratrol group (P<0.05). Conclusion Resveratrol promotes the autophagic flux of nasopharyngeal carcinoma CNE-2 cells, and the effects are possibly dependent on the activation of mTOR pathway-related proteins.

Objective To investigate the adjustment of miRNA-155 on CD4+ CD25+ Treg regulative T cell in peripheral blood in patients with acute cerebral infarction (ACI) and its pathogenesis. Methods Sixty patients with ACI were divided into three groups according to clinical neurological deficit score. Twenty healthy volunteers were enrolled into the control group. The expression levels of plasma miR-155 mRNA and Foxp3 mRNA were detected by real-time quantitative PCR(qRT-PCR). IL-10 levels in plasma were detected by ELISA. Results Expression of miR-155, Treg, Foxp3 mRNA and levels of IL-10 were significantly increased in patients with ACI compared with normal control group, with statistical differences; Expression of miR-155, Treg, Foxp3 mRNA and levels of IL-10 were gradually increased. The values showed significant statistical difference among the mild, moderate and severe ACI groups (P < 0.01). Among the patients,the levels of miR-155, Treg, Foxp3 mRNA and levels of IL-10 in the survival group were obviously lower than those in the non (P<0.05 or P<0.01). There was a positive correlation between miR-155 and Treg, Foxp3 mRNA (P < 0.01). Conclusion This study suggests that miR-155 is involved in the cell proliferation regulation of CD4+ CD25+ Treg cells,and plays some role in the immunological dissonance with ACI.

Objective To investigate the effect of microRNA(miR)-21 on proliferation and differentiation of murine pulmonary fibroblasts. Methods C57BL/6 mice of SPF grade (n=24) were randomly divided into Sham group and Pulmo?nary fibrosis model group with 12 mice in each group. Pulmonary fibrosis model was established by trans-tracheal jet ventila?tion of bleomycin into mice. The transcription levels of miR-21 were examined by quantitative real-time PCR in various pul?monary fibrosis tissues. Primary fibroblast were isolated and digested by Trypsin then inoculated into 6 well plate to reach confluence of 30%-50%. PBS (2.5μL), negative control stock solution and miR-21 mimic stock solution (20μmol/L) were added into Opti-MEM (50μL) as control group, blank group and miR-21 mimic group respectively.The cell viability was as?sessed by CCK-8. Expressions of ADAMTS-1 and TGF-β1 in the pulmonary fibroblasts were tested using Western blot. Re?sults The expression of miR-21 was significantly increased in lungs of mice in pulmonary fibrosis model group than that in sham group. Expression of miR-21 was higher in miR-21 mimic group than that in control group and blank group. Expres?sion of miR-21 was significantly higher with better cell viability in miR-21 mimic group than that in control group and blank group. The expression of ADAMTS-1 was significantly decreased in miR-21mimic group, while the expression of TGF-β1, a target gene of miR-21, was significantly increased in miR-21 mimic group compared with the other two groups. There is no significant different in expressions of ADAMTS-1 and TGF-β1 between control group and blank group. Conclusion Over?expression of miR-21 in pulmonary fibroblasts disrupts TGF-β1 signaling pathway by reducing expression of ADAMTS-1, which promotes the proliferation and differentiation of pulmonary fibroblast.

Objective : To analyze the effects of transglutaminase 2(TGM2) gene interference on the malignant proliferation and movement of lung cancer cells . Methods : A549 cells in logarithmic growth phase were divided into control group , shRNA⁃NC group and TGM2⁃shRNA1 group . After transfection , expression levels of TGM2 mRNA A549 cells in logarithmic growth phase were divided into control group , shRNA⁃NC group and TGM2⁃shRNA1 group . After transfection , expression levels of TGM2 mRNA clone formation assay . The proliferation activity of cells was detected by CCK⁃8 . The apoptosis rate was detected by flow cytometer. The invasion of cells was detected by Transwell . The number of tubule formation in each group was observed under microscope . The expression of E ⁃cadherin (E⁃cad) , N ⁃cadherin (N⁃cad) , fibronectin (FN) and vascular endothelial growth factor (VEGF) in each group was detected by Western blot . Results : Compared with shRNA⁃NC group , the mRNA level and protein expression of TGM2 in TGM2⁃shRNA1 group were decreased (P <0. 05) , and the rate of clone formation , proliferation activity , number of invasive cells and number of tubules formation were decreased ( P < 0. 05) . The apoptosis rate and the expression of E ⁃cad protein were increased ( P <0. 05) , while the expression of N ⁃cad , FN and VEGF protein were decreased (P < 0. 05) . Conclusion TGM2 gene silencing can inhibit the expression of VEGF protein , inhibit the malignant proliferation and invasion of A549 cells , and promote apoptosis .

Objective : To analyze the effects of miR⁃346 on alleviating cardiac function damage and oxidative stress in rats undergoing myocardial ischemia/reperfusion (I/R) . Methods : A total of 48 rats were randomly divided into control group , I/R group , I/R + agomiR⁃NC group and I/R + agomiR⁃346 group. I/R model was replicated by the ligation of left anterior descending coronary artery. The recombinant adeno⁃associated virus miR⁃346 was injected through the tail vein for overexpression intervention. After 120 minutes of reperfusion , heart rate (HR) , left ventricular ejection fraction (LVEF) , fractional shortening (FS) and left ventricular wall thickness (LVWT) , levels of serum creatine kinase MB (CK⁃MB) , myoglobin (Mb) , lactate dehydrogenase (LDH) and cardiac troponin I (cTnI) , and levels of superoxide dismutase ( SOD) , glutathion peroxidase ( GSH⁃Px ) and malondialdehyde (MDA) in myocardial tissues were detected. The apoptosis of myocardial tissue cells was detected by TUNEL staining. The expression of B⁃cell lymphoma/leukemia⁃2 (Bcl⁃2) gene , Bcl⁃associated x protein (Bax) , cysteine protease 3 ( Cas⁃3 ) and Cas⁃9 in myocardial tissues was detected by Western blot. Results : HR , LVEF , FS and LVWT in I/R group were lower than those in control group , the levels of serum CK⁃MB , LDH , Mb and cTnI were higher than those in control group , the expression of MDA , Bax/Bcl⁃2 , cleaved Cas⁃3/Cas⁃3 and cleaved Cas⁃9/Cas⁃9 proteins in myocardial tissues was higher than that in control group , the levels of SOD and GSH⁃px were lower than those in control group , and the apoptosis rate of myocardial tissue cells was higher than that in control group (P < 0. 05) . HR , LVEF , FS and LVWT in I/R + agomiR⁃346 group were higher than those in I/R + agomiR⁃NC group , the levels of serum CK⁃MB , LDH , Mb and cTnI were lower than those in I/R + agomiR⁃NC group , the expression of MDA , Bax/Bcl⁃2 , cleaved Cas⁃3/Cas⁃3 and cleaved Cas⁃9/Cas⁃9 proteins in myocardial tissues was lower than that in I/R + agomiR⁃NC group , the levels of SOD and GSH⁃px were higher than those in I/R + agomiR⁃NC group , and the apoptosis rate of myocardial tissue cells was lower than that in I/R + agomiR⁃NC group ( P <0. 05) . Conclusion The miR⁃346 overexpression can reduce oxidative stress level in I/R rats , alleviate myocardial tissue damage , and improve cardiac function.