A double-crosslinked antibacterial hydrogel (CS-Zn) was developed using small intestinal submucosa-derived collagen (SIS-Col), sodium alginate (SA), and zinc ions (Zn2+), and its potential for soft-tissue repair was systematically investigated. Through physical blending and ionic crosslinking, the hydrogel formed a dual-network structure with a tunable three-dimensional porous architecture, favorable mechanical properties, and good biocompatibility. In vitro studies demonstrated low cytotoxicity toward L929 cells and effective antibacterial activity against E. coli and S. aureus. In animal models, the hydrogel significantly promoted wound closure, angiogenesis, and organized collagen deposition in full-thickness skin wounds. In addition, preliminary evaluation of the myocardial ischemic injury model suggested that it had potential to improve tissue remodeling. In general, these findings indicate that CS Zn hydrogel is a promising multifunctional biomaterial and can be used for soft tissue repair.

ABSTRACT:Very elderly people (over 80 years) with cardiac implantable electronic devices (CIEDs) indications often have a higher prevalence of aging comorbidity, among which cognitive impairment is not uncommon. This study aimed to investigate periprocedural complications of CIED implantation among very elderly patients with and without cognitive impairment. One hundred eighty patients ≥80 years of age indicated for CIED implantation were included in our study. During hospitalization, the cognitive evaluation was performed according to the Diagnostic and Statistical Manual of Mental Disorders (fifth edition). According to the cognitive test results, patients were divided into 2 groups (90 patients with normal cognitive function and 90 patients with cognitive impairment). Meanwhile, their physical parameters and laboratory measurements were completed. The procedural data and periprocedural complications were collected from both groups. The association between cognitive impairment and periprocedural complications was analyzed using univariate and multiple logistic regression analyses. During a one-month follow-up, the most frequent periprocedural complications in very elderly patients were pocket hematoma and thrombosis events. Cognitively impaired patients had a higher incidence of complications than normal cognitive patients. Multivariate regression analysis showed that cognitive impairment was positively correlated with periprocedural complications in very elderly patients. Cognitive impairment is associated with increased periprocedural complications of CIED implantation in very elderly patients.

Background: Long noncoding RNAs (lncRNAs) represent a novel class of noncoding RNAs that are involved in a variety of biological processes and human diseases. Recent evidence suggested that lncRNAs were associated with cardiac disorders. However, the roles of lncRNAs in paroxysmal atrial fibrillation (PAF) remain elusive. The purpose of the present study was to identify differentially expressed lncRNAs in PAF and predict their potential functions. Methods: Between May 2014 and December 2015, a total of 67 patients, including 34 patients with PAF and 33 patients without PAF were recruited in this study. Of these participants, 3 PAF patients and 3 controls were used for the microarray analysis and a separate cohort (31 PAF patients and 30 controls) were used for further validation. LncRNA profiles in the leukocytes were detected by microarray. Results: A total of 2095 and 1584 differentially expressed lncRNAs and mRNAs, respectively, were identified between the PAF patients and controls. Four lncRNAs (uc002nvy.3, ENST00000561094, uc004aef.3, ENST00000559960) were randomly selected for quantitative real-time PCR (qRT-PCR) in a separate cohort, validating that ENST00000559960 was upregulated and uc004aef.3 was downregulated in the PAF patients. uc002nvy.3 and ENST00000561094 showed no significant difference between PAF and the controls. Multiple logistic analyses showed that ENST00000559960 (OR 1.47; 95% CI 1.09 to 2.00; P = 0.01) and uc004aef.3 (OR 0.63; 95% CI 0.41 to 0.96; P = 0.03) were independently associated with PAF. Receiver operating characteristic (ROC) curves analyses revealed that ENST00000559960 and uc004aef.3 were modest predictors of PAF. The area under the curve (AUC) was 0.67 +/- 0.07 (95% CI 0.54-0.81; P = 0.02) for uc004aef.3 and 0.70 +/- 0.07 (95% CI 0.56-0.83; P < 0.01) for ENST00000559960. Bioinformatic analyses (lncRNAs classification and subgroup, gene ontology analysis, pathway analysis and gene co-expression network construction) were performed for predicting the role of IncRNAs. Conclusions: Our results demonstrated that lncRNA profiles were differentially expressed in the PAF leukocytes, and two lncRNAs (ENST00000559960 and uc004aef.3) may help in prediction of PAF. This motivates further investigation of the role of lncRNAs for PAF.

Objective 　To investigate the effect of radiofrequency ablation (RFA) on miRNA expression in coronary sinus blood (CSB) of patients with atrial fibrillation (AF), especially after the termination of AF by RFA. And try to identify the possible miRNAs which can play real regulatory role in AF, so that to find the pathogenesis of AF and possible miRNA targets for intervention. Methods 　Thirty AF patients (paroxysmal, persistent and permanent AF, 10 each) were enrolled, alongside of 10 healthy subjects as controls. Peripheral blood (PB) samples were obtained before and 3 months after RFA, respectively. The total RNA was extracted and hybridized with the microRNA chips (microRNA v18.0), and the differential expressions of miRNA and clustering analysis in whole genome were performed with Volcano Plot and tMEV software respectively, and validated by Real-time PCR. The target gene of miRNAs was analyzed by retrieving the mirbase, miranda and targetscan databases. The important miRNAs were undergone with dual luciferase binding experiment to analyze and validate target genes. Results 　Comparing the CSB with PB of patient-self before RFA, 142 miRNAs expressed differently, of which 6 up-regulated and 8 down-regulated significantly (P<0.05). When comparing the PB of patients-self after and before RFA, 3 miRNAs up-regulated and another 3 down-regulated in above mentioned up-regulated 6 miRNAs, and miR-1266 was down-regulated 204.17-fold. Of 8 down-regulated miRNAs mentioned above, 7 were down-regulated again after RFA, and miR-3664-5p down-regulated 44.66-fold, while miR-574-3p up-regulated 5.25fold. SCN5A was the target gene of miR-1266, and CACNA1C was the target gene of miR-4279 confirmed by luciferase binding experiments. Conclusions 　The differential expression of miRNAs in CSB may directly reflect the regulatory status of miRNAs in cardiomyocytes in AF attack. RFA can reverse or change the abnormal expressions of miRNAs in CSB of AF patients in pro-RFA. The miRNAs which altered significantly before and after RFA, or regulate multi-ion channel proteins simultaneously, like miR-1266, may become a future target for AF intervention. DOI: 10.11855/j.issn.0577-7402.2016.10.06

The aim of the present study was to examine the effect of small interfering RNA (siRNA) methods on the expression of N-ethylmaleimide sensitive factor (NSF) and Weibel-Palade body (WPB) release in endothelial cells. A small hairpin RNA (shRNA), mediated with an adenovirus vector, was designed to target the N-terminal functional area of NSF. Subsequently, viruses were transfected into human aortic endothelial cells. The mRNA and protein expression levels of NSF were detected using reverse transcription-quantitative polymerase chain reaction and Western blot analyses, respectively, and the release of WPBs in the endothelial cells was examined using immunofluorescence. The mRNA expression of NSF in the endothelial cells, which were transfected with the adenoviruses carrying the NSF-shRNA was significantly decreased, compared with the negative control group (P=0.035) and blank control group (P=0.02). In addition, the mRNA expression of NSF was gradually decreased as duration increased; there were marked differences between the 24, 48 and 72 h groups (P<0.05). The protein expression of NSF was significantly decreased in the experimental group, compared with the negative control group (P=0.004) and blank control group (P=0.031), however, no difference was observed between the negative control and blank control groups (P=0.249). The immunofluorescence staining showed that the release of WPBs in the endothelial cells induced with thrombin was inhibited markedly following transfection with the virus carrying the NSF-shRNA. Therefore NSF-siRNA inhibited the mRNA and protein expression levels of NSF, and inhibited the release of WPBs in endothelial cells induced with thrombin. These results suggested that NSF-siRNA may be valuable for preventing and treating atherosclerosis and acute coronary syndrome.

The mechanism of miRNA regulation in atrial fibrillation (AF) occurrence and development is still unclear, especially, the regulating values of coronary circulating miRNAs has not been reported. Based on our AF radiofrequency ablation clinical practice and previous miRNA study, we proposed a hypothesis that the coronary circulating miRNA might much better reflect the regulating state and metabolic level of myocardial miRNA in AF patient. To investigate the regulating values of coronary circulation miRNA, 90 AF patients were selected and compared with 90 healthy subjects, the changes of coronary circulating miRNA differential expression profile in the whole genome were observed in this study. We found out that compared with autologous peripheral blood (PB), 6 miRNAs were upregulated and 8 miRNA downregulated in AF patients' coronary sinus blood (CSB) significantly, especially, the expression of miR-1266, miR-4279 and miR-4666a-3p were obviously increased. Compared with normal donors' peripheral blood, 16 miRNAs were upregulated and 24 miRNAs downregulated dramatically in patients' peripheral blood, among them, the miR-3171 decreased, but miR-892a and miR-3149 increased significantly from the early to end stages of AF. Our results indicated that the coronary circulating miRNA can really reflect the regulating values of miRNA in AF patient; the level of miRNA change in 3 types of AF may reflect the severity of AF clinical and pathophysiological advance; The miR-892a, miR-3171 and miR-3149 may be used as biomarkers for earlier diagnosis, while miR-1266, miR-4279 and miR-4666a-3p may serve as potential intervening targets for AF patient in future.

OBJECTIVES:To access the biocompatibility, effectiveness, and safety of biodegradable magnesium (Mg) alloy stent (BMAS) in the coronary artery and femoral artery.BACKGROUND:Atherosclerosis is a lesion of cardiovascular system, including the diseases in heart and blood vessels.METHODS:The aluminum (Al) and zinc (Zn)-based BMAS was designed by cold drawing methods. Forty healthy immunized mongrel dogs were randomly divided into 8 groups. Five dogs who have not been treated with stent were included in control group. The other dogs were implanted with an absorbable magnesium (Mg) alloy in the coronary and/or femoral artery, and their artery angiography were observed at 7 time points (1, 3, 5, 7, 14, 21, and 28 days; n = 5) follow-up. Dogs from each cohort were sacrificed following angiography for pathology assessment. The histological response including inflammatory response, thrombosis, and intimal hyperplasia were analyzed by hematoxylin-eosin staining. Lumen area (La), intimal hyperplasia area (IHa), and the ratio of IHa were calculated by image analysis software.RESULTS:The thin-walled BMAS were designed and produced by cold-drawing technology. Fifty-one devices were implanted into coronary artery of 35 dogs successfully. During the follow-up days, the angiography of coronary artery and femoral artery had confirmed that the lumen was clear and there were no elastic recoil and thrombosis. The stents were completely disappeared at 7 days after implantation. Moderate intimal hyperplasia was found at 14 days after implantation.CONCLUSION:The BMAS stent proved to be of good biocompatibility, safety, and effectiveness. (J Interven Cardiol 2015;XXXX:XX-XX).

OBJECTIVETo observe the change of circulating microRNAs(miRNAs), regulatory mechanism in patients with atrial fibrillation (AF) before and after radiofrequency ablation (RFA).METHODSFrom January 2011 to December 2013, peripheral blood samples were taken from 30 AF patients (10 paroxysmal, 10 persistent and 10 permanent AF) before and 3 months after RFA. The total RNA was extracted and hybridized with the miRNA chips, and the differential expression of miRNA and clustering analysis in whole genome were made with Volcano Plot and tMEV software respectively, and validated by real-time PCR. The target gene analysis of miRNAs was predicted through the Mirbase, Miranda and Targetscan databases. Results were compared with those from 10 healthy subjects (control group).RESULTSCompared with control group, the expressions of 25 miRNAs were down-regulated before RFA and up-regulated after RFA in AF group, while other 40 miRNAs expression changed in the opposite way; among them, the expressions of 7 miRNAs including miR-199a-3p/miR-199b-3p were down- regulated >1.5-fold before RFA and up-regulated>100-fold after RFA; oppositely, 6 miRNAs including miR-BART8-3p were up-regulated>1.5-fold before RFA and down-regulated>10-fold after RFA. Interestingly, 6 miRNAs including miR-30b-5p, which were involved in AF-related electrical and structural remodeling, were down-regulated>5-fold before RFA, but up-regulated>50-fold after RFA. Four miRNAs including miR-377-5p, which were involved in the regulation of CACNA1C ICaL channel protein, were different before and after RFA.CONCLUSIONmiRNAs regulate the occurrence and development of AF. RFA can change the expression of miRNAs in AF patients, which may be important for reversing the electrical and structural remodeling and maintaining sinus rhythm after RFA. miRNAs, such as miR-30b-5p, miR-377-5p and miR-199a-3p/miR-199b-3p etc., might become the target markers for early diagnosis and intervention of AF in future.

目的 探讨心房颤动(房颤)射频消融术是否通过对调控离子通道蛋白的微小RNA(microRNA,miRNA)的影响,实现心房离子流再平衡和逆重构,并试图发现有价值的调控miRNA.方法 选择行房颤射频消融术患者(阵发性、持续性和永久性房颤各10例)30例作为房颤组,健康体检者10例作为正常对照组.正常对照组体检时,房颤组射频消融术前和术后3个月分别取外周血,使用miRNA芯片(miRNA v18.0)进行全基因组miRNA表达谱微阵列分析,2组miRNA表达比值≥1.5倍为显著上调,实时定量PCR验证miRNA表达差异结果,并通过mirbase、miranda、targetscan数据库进行靶基因分析.结果 与正常对照组比较,房颤组射频消融术前主要参与离子通道蛋白调控的21个miRNA差异表达均有显著意义(P＜0.01);房颤组术后3个月与自身术前比较,上述21个miRNA表达亦有明显差异(P＜0.01).其中miR-1266等5个miRNA术前表达上调≥1.5倍,术后明显下调≥10倍;仅miR-3664-5p术前下调8.88倍,术后进一步下降46.06倍;其余15个miRNA均术前表达下调,术后显著上调.结论 房颤射频消融术通过影响调控离子通道蛋白的主要miRNA,实现了心房离子流逆重构.调控多个离子流的miR-1266,miR-377-5p,miR-101-5p和miR-151-3p有望成为房颤治疗新靶点.

Objective To study the relation between miR-1266 and atrial fibrillation(AF)-associated target gene of sodium ion channel protein by dual feuciferase reporter gene assay.MethodsFour target genes(SCN5A,KCNH2,KCNE1 and KCNJ5)of miR-1266 were retrived from TargetScan,miRanda and miRDB databases.A 3'UTR recombinant luciferase reporter plasmid system was constructed for the 4target genes.The 4target genes were divided into miR-1266 transfection group,miR-1266 negative control group and miR-1266 blank control group.Forty-eight hours after the recombinant luciferase reporter plasmids,miR-1266 plasmds,and control plasmids of the 4target genes in the first two groups were transfected into the HEK293 cells,the activity of luciferase in different groups was assayed.Results For SCN5 A,the activities of luciferase in miR-1266 transfection group were markedly lower than those in miR-1266 negative control group(0.100±0.007 vs 0.209±0.011,P=0.002).For KCNH2,KCNE1 and KCNJ5,no significant difference of the activities of luciferase were found between miR-1266 transfection group and miR-1266 negative control group(0.819±0.021 vs 0.886±0.012,P=0.1269;0.467±0.018 vs0.551±0.016,P=0.0652;0.727±0.055 vs 0.668±0.010,P=0.2326).Conclusion SCN5 Ais the direct target gene of miR-1266 while KCNH2,KCNE1 and KCNJ5are not the direct target genes of miR-1266,indicating that miR-1266 may become a new AF treatment target in future by inhibiting SCN5 A expression and participating in AF remolding.

目的 探讨房颤射频消融手术对房颤离子通道蛋白和离子流重构的影响和调控作用,寻找可能的miRNAs调控干预靶点,为房颤的预警干预奠定基础.方法 选择30例行房颤射频消融术患者(阵发性、持续性和永久性房颤各10例),健康体检者10名作为正常对照组.射频消融术前和术后3个月分别取外周血,使用miRNA芯片进行全基因组miRNA表达谱微阵列分析,Real-time PCR对调控离子通道蛋白的主要miRNAs表达差异结果进行验证,通过mirbase、miranda、targetscan数据库进行靶基因分析.结果 主要参与调控离子通道蛋白SCN5A、CACNA1C、KCNA5、KCNH2、KCNE1、KCNQ1、KCNJ2、KCNC4、KCND3、KC-NN3、HCN1、HCN3和HCN4的miRNAs共21个.射频消融术前房颤组与正常对照组比较,其表达有显著差异(P＜0.05).术后3个月房颤组与自身术前比较,这些miRNAs表达亦有明显差异(P＜0.05),其中miR-1266等5个miRNAs术前表达上调,术后明显下调,仅miR-3664-5p术后较术前进一步下调,其余15种miRNAs均术前表达下调,术后显著上调.调控外向型钾离子通道(如KCNA5)的miRNA手术前后调控趋势一致,调控幅度较大.结论 外向性K+离子流增多在房颤电重构中可能起主要作用.房颤射频消融手术对房颤的电重构起到了离子流再平衡和逆重构的作用.调控多个离子流的miR-1266等miRNAs,有可能成为未来房颤干预的靶点.

目的 探讨非瓣膜病心房颤动(房颤)患者血清小分子RNA(miRNA)全基因组表达差异及其可能的调控作用和早期预警价值.方法 15例房颤患者,分为阵发性、持续性和永久性房颤组,每组5例,对照组5例健康人.射频消融术前和术中分别取外周血和冠状窦血,提取血浆总RNA,使用microRNA芯片(microRNA v 18.0)进行全基因组miRNA表达谱微阵列分析,Volcano Plot法获得差异表达niRNAs,并用tMEV软件进行聚类分析,以及通过mirbase、miranda、targetscan数据库进行靶基因分析,并进行RT-PCR的差异表达验证.结果 房颤组冠状窦血与外周血比较有14个miRNAs表达差异显著,其中6个表达上调:即miR-1266、miR-4279、miR-4787-5p、miR-4666a-3p、kshv-miR-K12-6-3p和miR-3150a-5p,8个表达下调:即miR-892a、miR-3149 、miR-3171、miR-3664-5p、miR-3591-3p、miR-4423-5p、miR-4473和miR-574-3p.其中,miR-1266在阵发性、持续性和永久性房颤组均明显升高,而miR-3171则显著降低.房颤组与对照组外周血及冠状窦血比较miRNAs表达也有明显差异.结论 房颤患者冠状窦血与外周血比较miRNAs表达均有显著差异,而冠状窦血miRNAs更能反映心脏的代谢与调控状况;血清miR-3171、miR-892a、miR-3149在房颤发生发展早期出现且持续表达差异,有可能成为早期预警诊断的标志物;miR-1266、miR-4279、miR-4666a-3p有可能成为未来治疗房颤的干预靶点.

Objective To study the value of circulating miRNA expression in regulation,early diagonosis and treatment of atrial fibrillation(AF).Methods Fifteen AF patients(5with paroxysmal AF,5with persistent AF and 5with permanent AF)served as an AF group and 5subjects undergoing physical examination served as a control group.Peripheral blood(PB)and coronary sinus blood(CSB)samples were taken from the patients before and during radiofrequency ablation.Circulating miRNA expression in whole genome was detected by Volcano Plot and target genes in Mirbase,Miranda and Targetscan were analyzed.Results Of the 14 differently expressed miRNAs in CSB group and PB group,the expressions of 6miRNAs(miR 1266,miR 4279,miR4787-5p,miR 4666a-3p,kshv miR K126-3p and miR3150a-5p)were up-regulated and those of 8miRNAs(miR 892a,miR 3149,miR 3171,miR 3664-5p,miR 3591-3p,miR 4423-5p,miR 4473and miR 574-3p)were down-regulated.The expression level of miR 1266and miRNA was significantly higher and that of miR 3171was significantly lower in AF group than in control group(P0.05).Conclusion CSB can effectively predict the metabolism and regulation of miRNA in AF patients.The miR 3171,miR 892a,miR 3149are persistently expressed in early AF patients and can thus used as a diagnostic marker of AF.The miR 1266,miR 4279,miR 4666a-3p may serve as AF treatment targets.

<正>心力衰竭(简称心衰)是一种多种致病因素导致的临床综合征,可出现一系列细胞表型及重构改变,这些改变可能与心肌细胞内钙离子转运异常有关,并引起电机械功能障碍,如:心肌收缩力降低,QT间期延长[1],室性期前收缩频率增多及心源性猝死发生率增加。心室肌细胞内钙离子浓度的变化调控兴奋-收缩耦联(E-C coupling)。在衰竭的心肌中,细胞内钙离子的异常转运可导致机械及电生理功能异常。本文将综述正常心肌兴奋-收缩耦联以及心衰后的相关改变,重点表述心室肌细胞重构的变化

<正>流行病学及相关实验发现舒张性心力衰竭(DHF)发病机制与收缩性心力衰竭(SHF)不同[1-3]。虽然对定义为DHF或左心室射血分数(LVEF)尚保留的心力衰竭(HFPEF)存在争议,但有辅助影像学数据支持的临床诊断标准,认为DHF是一种单独的