姜黄素调控MicroRNA-1246及对膀胱癌T24细胞的放疗增敏机制探究

目的:研究在膀胱癌化疗中使用姜黄素的增敏功能原理。方法:在T24细胞经过姜黄素及放射处理后,探讨分析其细胞活力(细胞增殖试验试剂盒)、microRNA表达(miRNA芯片测序)、集落形成、凋亡(AnnexinV-F)分析、miR-1246及p53 mRNA及蛋白质(Westernblot)表达、ITC/7-AAD流式细胞计数(ITC/7-AAD)。结果:通过测试,发现17个异常microRNA表达,细胞在经过姜黄素处理后,T24细胞活力相较于常规组明显下降,并出现浓度依赖性的现象。在T24细胞中,miR-1246相较于人膀胱上皮永生化细胞(SV-HUC-1细胞)表达明显较高。姜黄素的浓度较高时,T24细胞miR-1246的表达会受其影响并明显下降。研究组使用浓度为20μg/mL的姜黄素,并结合放疗处理的方式,与常规组相比,研究组在控制miR-1246的表达、集落形成及细胞活力方面,效果更为显著,T24细胞比例会因转染antagomiR-1246而增加,细胞死亡现象一般发生于转染antagomiR-NC细胞时期。结论:膀胱癌细胞中,姜黄素和放射治疗都可促进microRNA-1246表达下调,在对肿瘤的治疗中可以使姜黄素及放疗结合,共同产生抗肿瘤作用。     

Health Care Utilization After Ventricular Tachycardia Ablation: A Propensity Score-Matched Cohort Study

Background

Catheter ablation of ventricular tachycardia (VT) can reduce the burden of ventricular arrhythmia (VA) but its effect on health care utilization and costs after such therapy is poorly known. We sought to compare the rates of cardiovascular (CV)-related hospitalizations, survival, and health care costs in patients with recurrent VT treated either with VT ablation or with medical therapy.

The Effect of Cardiac Rehabilitation on Health-Related Quality of Life in Patients With Coronary Artery Disease: A Meta-analysis

Background

The clinical effectiveness of cardiac rehabilitation (CR) on health-related quality of life (HRQOL) is an area that has not been consistently explored. The objective of this systematic review was to evaluate the effectiveness of providing any core component of CR on HRQOL domains.

基因间长链非编码RNA152靶向微小RNA?103a?3p对非小细胞肺癌细胞增殖和侵袭迁移的影响

目的探讨基因间长链非编码RNA 152(LINC00152)靶向调控微小RNA-103a-3p(miR-103a-3p)表达及对非小细胞肺癌(NSCLC)细胞增殖和侵袭迁移的影响。方法采用实时定量PCR(QPCR)检测正常肺上皮细胞BEAS-2B及NSCLC细胞(ANIP-973、NCI-H157、A549和NCI-H1975)的LINC00152水平。选取LINC00152水平最高的细胞分别转染LINC00152特异性小干扰RNA(si-LINC00152组)或无关序列(si-NC组),另设未转染细胞为对照组。QPCR检测LINC00152水平,活细胞计数CCK-8法、Transwell小室和划痕实验测定细胞增殖、侵袭和迁移能力,Western blotting检测基质金属蛋白酶(MMP)-2、MMP-9和第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)的水平;荧光素酶报告实验验证LINC00152靶向结合miR-103a-3p的能力。结果NSCLC细胞的LINC00152水平均高于BEAS-2B细胞(P<0.05),尤其是NCI-H1975细胞的最高。si-LINC00152组的LINC00152水平为0.352±0.087,低于对照组的1.058±0.219和si-NC组的1.126±0.139(P<0.05)。与si-NC组和对照组相比,si-LINC00152组NCI-H1975细胞转染48、72 h的增殖活力下降(P<0.05);si-LINC00152组的划痕愈合率和穿膜细胞数分别为(27.386±2.428)%和(78.840±5.031)个,低于si-NC组的(77.675±4.803)%和(179.208±13.264)个及对照组的(76.371±5.385)%和(174.003±15.678)个(P<0.05);与si-NC组和对照组相比,si-LINC00152组的MMP-2和MMP-9水平均降低,而PTEN水平升高(P<0.05)。对照组和si-NC组上述指标的差异无统计学意义(P>0.05)。双荧光素酶报告分析证实,miR-103a-3p模拟物降低了野生型LINC00152的荧光素酶活性(P<0.05),但对突变型无影响(P>0.05)。结论LINC00152在NSCLC细胞中高表达并发挥促癌作用,与NSCLC的迁移侵袭密切相关,LINC00152与miR-103a-3p间的相互作用在NSCLC靶向治疗中有一定潜能。     

Molecular mechanism of miR-382 in the pathogenesis of renal tubulointerstitial fibrosis

Objective To investigate the roles of microRNA-382 (miR-382) in the pathogenesis of renal tubulointerstitial fibrosis (TIF). Methods Human kidney epithelial cells (HK2)transfected with miR-382 inhibitor (antagomiR-382) were used to examine the effect of miR-382 abundance on cell polarity, as well as to test the complementary relationship between miR-382 and its predicted target gene heat shock protein 60 (HSPD1), which was further verified by 3′-untranslated region luciferase assay and site-directed mutagenesis. The role of miR-382 played in the development of renal interstitial fibrosis and redox regulation was examined in a mouse unilateral ureteral obstruction (UUO) model. Locked nucleic acid (LAN)-modified anti-miR-382 was intravenous delivered via tail vein 30 min prior to UUO, and repeated the dosage 24 h after the surgery. For clinical verification, renal biopsy specimens from 12 IgA nephropathy (IgAN) patients were collected, 6 patients with moderate to severe TIF and 6 patients without TIF. The relative abundance of miR-382 and HSPD1 protein was analyzed by using in situ hybridization and immunohistochemistry. Results HSPD1 was confirmed to be a new, direct target gene of miR-382 by in vitro 3′-untranslated region luciferase assay and site-directed mutagenesis. The development of epithelial transition in HK2 cells was accompanied with up-regulation of miR-382 [(6.54±0.96) vs (1.12±0.26), P＜0.05]. Blocking the expression of miR-382 could reversed the progression of epithelial transition partially. In UUO mice the abundance of miR-382 was up-regulated [(6.89±2.47) vs (1.00±0.42), P＜0.01] while HSPD1 and Trx were down-regulated compared with the sham group. Down-regulation of miR-382 was associated with significant decrease in TIF, but increase in HSPD1 and thioredoxin protein compared with UUO group [HSPD1: (0.34±0.10) vs (0.14±0.05); Trx: (0.79±0.18) vs (0.36±0.16); all P＜0.05]. The expression of miR-382 was up-regulated and HSPD1 was significantly down-regulated in IgAN patients with TIF. Conclusions miR-382 play an important role in renal tubulointerstitial fibrosis in human and mice. HSPD1 is one of the target genes of miR-382. The down-regulation of HSPD1 and the decrease ability of anti-oxidative stress may be the important mechanism of miR-382 involved in renal tubulointerstitial fibrosis.     

脑梗死患者血清miR-497,TNF-α表达水平变化及其临床意义

目的 探讨急性脑梗死(Acute cerebral infarction,ACI)患者血清微小RNA-497(MicroRNA-497,miR-497)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)的表达水平变化及其临床意义。方法 选取2016年1月-2019年11月本院收治的96例ACI患者,称ACI组,并选取本院同期98例体检健康者,称对照组; 采用实时荧光定量PCR(Real-time fluorescent quantitative PCR,qRT-PCR)法检测所有研究对象血清miR-497表达水平; 采用酶联免疫吸附法(Enzyme-linked immunosorbent assay,ELISA)检测所有研究对象血清肿瘤坏死因子-α水平; 评估ACI患者神经功能缺损程度、计算脑梗死体积,比较不同神经功能缺损程度/脑梗死体积的ACI患者血清miR-497、TNF-α水平; Pearson法分析ACI患者血清miR-497、TNF-α水平与神经功能缺损程度评分(National institutes of health stroke scale,NIHSS)、脑梗死体积的关系; 采用受试者工作特征曲线(Receiver operating characteristic curve,ROC)评价血清miR-497、TNF-α对ACI的诊断价值。结果 ACI组血清miR-497、TNF-α水平均明显高于对照组(P<0.05); ACI患者血清miR-497、TNF-α水平随神经功能缺损程度加重、脑梗死体积增加均呈递增趋势(P均<0.05); ACI患者血清miR-497、TNF-α水平与脑梗死体积、NIHSS评分均呈正相关(r=0.423,0.514,0.542,0.399,P均<0.05); 血清miR-497、TNF-α对ACI诊断的曲线下面积(Area under curve,AUC)为0.848、0.806,截断值分别为1.29、1.27,相应灵敏度分别为82.3%、81.3%,特异度分别为76.5%、77.6%; 两者联合诊断ACI的AUC为0.907,其灵敏度、特异度分别为81.3%、90.8%。结论 miR-497、TNF-α在ACI患者血清中表达均上调,且与神经功能缺损程度、脑梗死体积有关,均可能在ACI进展中起一定作用,两者联合可有效提高ACI的诊断效能,有助于诊断、评估ACI患者的病情。     

Investigation of the miRNA146a and miRNA155 gene expression levels in patients with multiple sclerosis

Multiple sclerosis (MS) is a chronic inflammatory disease and the most common neurodegenerative status. MicroRNAs play an important role in macrophage response to inflammatory processes, and alterations in miRNA levels trigger the inactivation of specific T lymphocytes. As a result, these factors can lead to autoimmune diseases such as MS. Therefore, to determine the role of MicroRNA-146a and MicroRNA-155 in MS patients, their expression levels in serum of MS patients were compared with healthy controls.In this study, the expression levels of MicroRNA-146a and MicroRNA-155 in 30 serum samples of MS and healthy patients as a control group. MicroRNA extraction and cDNA synthesis was performed according manufacture protocols. The expression levels of MicroRNAs were evaluated by Real Time-PCR.MicroRNA-146a and MicroRNA-155 levels were increased in patients with MS compared to controls. The results demonstrated that EDSS score are increased with increasing level of MicroRNA-146a and MicroRNA-155. ROC curve analysis showed that the area under curve (AUC) was significant for MicroRNA-146a and MicroRNA-155.Increased expression levels of MicroRNA-146a and MicroRNA-155 may be associated with the pathogenesis of MS disease. If this study is conducted in a larger sample population and the above results can be used to identify patients or control patients who are under medical care.     

miR-124a和miR-449a对非小细胞肺癌的诊断价值

目的探讨微小RNA-124a(miR-124a)和微小RNA-449a(miR-449a)对非小细胞肺癌(NSCLC)的诊断价值。方法选取行手术切除肿瘤的NSCLC患者90例(NSCLC组),出院随访36个月,根据术后复发情况分为复发组(56例)和未复发组(34例)。另选取肺部良性结节患者60例(良性结节组)、健康体检者80名(正常对照组)。收集所有对象的基本资料,同时检测血清神经元特异性烯醇化酶(NSE)、miR-124a和miR-449a水平。采用Spearman相关分析评估各项指标的相关性。采用受试者工作特征(ROC)曲线评估各项指标诊断NSCLC的效能。结果NSCLC组血清NSE水平高于正常对照组和良性结节组(P<0.05),血清miR-124a和miR-449a水平低于正常对照组和良性结节组(P<0.05)。正常对照组与良性结节组之间血清NSE、miR-124a和miR-449a水平差异均无统计学意义(P>0.05)。与未复发组比较,复发组血清NSE水平升高(P<0.05),血清miR-124a和miR-449a水平降低(P<0.05);2个组之间年龄、性别、体质量指数(BMI)、吸烟情况、肿瘤直径、病理类型和TNM分期之间差异均无统计学意义(P>0.05)。ROC曲线分析结果显示,NSE、miR-124a和miR-449a单项诊断NSCLC的曲线下面积(AUC)分别为0.660、0.703、0.759。NSE+miR-124a+miR-449a联合检测诊断NSCLC的AUC(0.895)和敏感性(96.25%)均最高,miR-124a+miR-449a联合检测诊断NSCLC的特异性最高(90.31%)。结论miR-124a和miR-449a或可作为NSCLC的辅助诊断指标。