Objective To choose one protocol that can quickly ,safely and effectively provide amount enough of bone marrow derived mesenchymal stem cells(MSCs) for use of clinical or experimental test through comparison of their growth characteristics and growth factors levels in culture solution .Methods Cells extracted from bone marrow of C57BL/C mice respectively underwent two different isolation protocols :whole bone marrow adherent culture(WBMAC) or gradient density separation(GDS);characteris‐tic surface antigens of MSCs were identified by flow cytometry on cells isolated in different ways ;the distinct growth curve of pri‐mary stem cells cultured in vitro described their different proliferation rate;levels of vascular endothelial growth factor(VEGF) and stromal cell‐derived factor‐1α(SDF‐1α) in culture medium were detected by ELISA .Results Primary MSCs obtained by WBMAC proliferated at higher speed and exhibited shorter growth cycle than those separated by GDS ;on MSCs from both groups ,surface antigens CD29 were detected positively ,and antigens including CD31 ,CD34 and CD45 were assayed negatively ;concentration of VEGF and SDF‐1αin both two nutrient solution primarily keep at low levels ,comparatively ,level of VEGF and SDF‐1αin the di‐shes which contain MSCs by WBMAC was higher than the one in the dishes which contain MSCs by GDS .Conclusion MSCs ex‐tracted by WBMAC shows unimpaired cell function ,can build automatically more suitable microenviroment for their growth;this classic method was qualified for clinical and experimental use in a safe ,rapid ,effective way .

Objective To explore the inhibitory effect of curcumin on LPS-induced inflammation and the activation of Toll-like receptor 4 (TLR4 )/NADPH oxidase/reactive oxygen species (ROS)signaling pathway in vascular smooth muscle cells (VSMCs).Methods Primary VSMCs were cultured and divided into control group, LPS group,LPS + curcumin 5 μmol/L group,LPS + curcumin 10 μmol/L group and LPS + curcumin 30 μmol/L group.Cell activity was observed by MTT assay.The secretion of tumor necrosis factor-α(TNF-α)and interleukin-1 (IL-1)was measured by enzyme linked immunosorbent assay (ELISA)kits.The mRNA expressions of TLR4 and p22phox were detected by real-time PCR.Expression of intracellular ROS was measured by flow cytometry. Results The activities of VSMCs were not significantly affected by curcumin at the concentration between 0 and 80 μmol/L.Curcumin (5,10 and 30 μmol/L)significantly inhibited LPS-induced oversecretion of TNF-αand IL-1, as well as overexpression of TLR4 and p22phox at the mRNA and protein levels,and ROS production in VSMCs in a concentration-dependent manner.Conclusion Curcumin has a concentration-dependent inhibitory effect on the secretion of inflammatory cytokine,overexpressions of TLR4 and p22phox,and production of ROS in VSMCs stimulated by LPS.Furthermore,curcumin may partly depend on TLR4/NADPH oxidase/ROS signaling pathways to inhibit inflammation in LPS-induced VSMCs.

Objective To investigate the effect of curcumin (Cur)on AngⅡ-induced proliferation and oxidative stress of vascular smooth muscle cells (VSMCs).Methods Primary rat VSMCs were cultured and divided into control group,AngⅡ group,AngⅡ+Cur 5μmol/L group,AngⅡ+Cur 10μmol/L group,AngⅡ+Cur 20μmol/L group,and Cur 20μmol/L group.The proliferation of AngⅡ-induced VSMCs was measured by MTT assay.The mRNA and protein expressions of inducible nitric oxide synthase (iNOS)and p47phox were detected by real-time PCR and Western blot.Nitric oxide (NO)production was measured by Griess reaction.Production of intracellular reactive oxygen species (ROS)was measured by DCFH-DA staining,and the activities of superoxide dismutase (SOD)and glutathione peroxidase (Gpx)were detected by xanthine oxidase assay and visible spectrophotometer. small interfering RNA (siRNA)was used to silence the expression of p47phox to further explore the mechanism for Cur inhibiting the proliferation of AngⅡ-induced VSMCs and oxidative stress.Results VSMCs activities were not significantly affected by Cur at the concentration between 0 and 80μmol/L.Cur (5,10 and 20μmol/L)significantly inhibited AngⅡ-induced proliferation of VSMCs.Cur had an inhibitory effect on the overexpression of NO,iNOS, p47phox and ROS in VSMCs and upregulated the activities of SOD and Gpx in a concentration-dependent manner. AngⅡ-induced ROS production in VSMCs was significantly attenuated by pretreatment with p47phox specific siRNA.Conclusion Cur can inhibit the proliferation and oxidative stress of AngⅡ-induced VSMCs.

Objective To investigate the relationship between the volume of left atrial appendage and recurrence of atrial fibrillation(AF)after radiofrequency ablation. Methods In this retrospective cohort study, 66 cases of first atrial fibrillation radiofrequency catheter ablation in the Department of cardiovascular medicine of the First Affiliated Hospital of Zhengzhou University were enrolled from June 2014 to June 2016 and divided into the recurrence group(n=18)and the non recurrent group(n=48)based on the 1 year follow-up results ,Collecting the patient's clinical data and following up.64 layers of spiral CT scans were performed for all patients before operation,and the volume of left atrium( LAV)and left atrial appendage volume(LAAV)were measured. The general data ,laboratory examinationresults ,echocardiographic parameters and left atrial CT parameters of two groups were compared. The relationship between patient parameters and recurrence of atrial fibrillation after radio-frequency ablation were analyzed by multivariate logistic regression analysis. Results There was no significant difference in blood lipid and left ventricular ejection fraction(LVEF%)between the two groups in terms of sex, age ,hypertension ,coronary heart disease and other common diseases (P > 0.05).The volume of left atrial appendage and left atrial volume in the recurrent group were larger than those in the non recurrence group (P <0.05). The left atrial appendage volume(OR=1.518,95%CI:1.151-2.000,P = 0.003)can be used as an independent risk factor for postoperative recurrence of atrial fibrillation. The area under the ROC curve of left atrial appendage volume in predicting the recurrence of atrial fibrillation after radiofrequency ablation is 0.806(95%CI:0.689-0.922 ,P < 0.001). Conclusion Greater left ventricular volume is an independent risk factor for recurrence of atrial fibrillation after radiofrequency catheter ablation ,whether in paroxysmal atrial fibrillation or persistent atrial fibrillation.