Effect of blood purification on serum miR-126 and VEGF levels in the process of atherosclerosis in uremic patients under maintenance hemodialysis

This study aims to study the influence of different blood purification approaches on the expression of miR-126 and VEGF serum levels in the development of atherosclerosis in uremic patients under maintenance hemodialysis (MHD). A total of 207 MHD patients with uremia were divided into HD (hemodialysis, n = 60), HDF (hemodiafiltration, n = 72), and HD + HP (hemodialysis + hemoperfusion, n = 75) groups based on different purification approaches. Eighty individuals who underwent physical examinations during the same period constituted the healthy controls. The serum levels of miR-126 and VEGF were examined by qRT-PCR and ELISA both before and after treatment, and the intima media thickness (IMT) value and plaque area were evaluated by color Doppler ultrasound. The serum miR-126 level was down-regulated in MHD patients compared with healthy controls, and this was negatively linked to VEGF. The post-treatment expression level of serum miR-126 in the HDF and HD + HP groups was remarkably increased, but VEGF was decreased in MHD patients, and especially significantly in the HDF group. In addition, IMT and plaque area were obviously improved in the HD group after treatment. Pearson correlation analysis showed a negative correlation of miR-126 with IMT and plaque area, but a positive association between VEGF and IMT and plaque area. miR-126 and VEGF are expected to become a valuable biomarker for monitoring the progression of atherosclerosis in uremic patients undergoing MHD.     

急性高原低氧对于大鼠血红蛋白和血浆miR－144表达影响的研究

目的探讨急性高原低氧对于大鼠血红蛋白和血浆miR－144表达的影响。 方法应用低压氧舱构建高原低氧大鼠模型,分为高原低氧暴露3、7、14、28 d实验组以及常压常氧对照组。检测各组大鼠血常规和血浆miR－144表达情况。 结果与对照组比较,低压低氧组大鼠血浆miR－144、红细胞计数、血红蛋白含量均随高原低氧时间延长而显著升高(P<0.05)。Spearman相关性分析提示血浆miR－144与红细胞计数、血红蛋白含量呈正相关。 结论急性高原低氧可使大鼠血红蛋白和血浆miR－144表达升高。血浆miR－144可能参与调控高原低氧相关的红系变化。     

肝癌患者血清外泌体内miR-665的表达及临床意义

[摘 要] 目的 探讨肝癌患者血清来源的外泌体内miR-665与肝癌临床病理指标及预后的关系。方法 采用试剂盒方法提取患者血清内的外泌体并用透射电镜观察验证。然后利用RT-PCR方法检测30例肝癌患者和10例体检者血清来源的外泌体内miR-665的表达水平,进行对照研究,探讨临床病理指标和预后的关系。结果 血清提取的外泌体呈囊泡状,直径约30～150 nm。肝癌患者血清来源的外泌体miR-665较正常体检者表达水平明显升高,中位表达水平是对照组的8.3倍。外泌体miR-665高表达与患者肿瘤大小（>5 cm） （P=0.0042）、有包膜浸润（P=0.0197）以及临床TNM分期（III～IV期） （P=0.0276）成明显相关,与患者年龄、性别、HBsAg以及AFP水平未见明显相关性（P > 0.05）,外泌体miR-665高表达组患者生存期较低表达组更短（P=0.036）。结论 肝癌患者血清中外泌体miR-665水平升高与肝癌进展相关,测定血清外泌体内miR-665的水平可能有助于肝癌的临床诊断和预后判断。     

不同miR-146a表达条件下Treg细胞体内回输对小鼠心脏移植免疫排斥反应的影响

目的:本研究探讨调控miR-146a Treg细胞体内回输对小鼠心脏移植免疫排斥反应的影响。方法:流式分选Treg细胞并进行体外扩增。转染试剂分别上调和下调Treg miR-146a表达。建立小鼠心脏移植模型,设立对照组,空白Treg组,miR-146a上调组,miR-146a下调组。移植后回输Treg,观察生存曲线,病理分级,测定受体脾T细胞亚群,RT-PCR检测供心IFN-γ、IL-4、IL-17表达。结果:细胞扩增10倍后miR-146a表达无明显变化,并能有效调控miR-146a表达。回输后空白Treg组（中位生存期11 d）及上调组（中位生存期13 d）生存时间延长,病理分级降低（P<0.05）;上调组更为明显（P<0.05）;下调组（中位生存期7 d）生存时间缩短,病理分级升高（P<0.05）,Th1细胞数量（28.6%±2.7%）及供心IFN-γ表达（1.12±0.11）升高（P<0.05）。结论:体外能有效地分选和扩增Treg细胞,并保证miR-146a的表达。回输Treg明显抑制小鼠心脏移植急性排斥反应,上调组抑制功能增强,下调组抑制功能减弱。     

肺鳞癌中miR-144靶基因预测及其生物信息学分析

目的:分析miR-144靶基因在肺鳞癌中参与的信号通路生物过程。方法:分析The Cancer Genome Atlas网站中肺鳞癌患者的miR-144的表达量及生存曲线;预测miR-144靶基因并通过表达数据筛选影响肺鳞癌进展基因;BinGo软件对候选靶基因进行GO注释分析,利用DAVID网站预测其信号通路。结果: miR-144表达量高有利于延长肺鳞癌患者生存时间。综合4个靶基因数据库发现miR-144有72个预测结果,其中45个在肺鳞癌患者癌与癌旁组织间表达存在差异。其中部分候选靶基因参与胞核及星形微管的组成,参与上皮细胞增生、RNA合成。通路分析显示部分候选靶基因参与Ras、Wnt、Hippo等信号通路。结论:45个miR-144候选靶基因通过多条信号通路及生物过程调节细胞生命活动,影响肿瘤患者生存预期。     

miR-34a-5p对HaCaT细胞增殖及其靶基因Notch1表达的影响

目的: 检测miR-34a-5p对人角质形成细胞株HaCaT细胞体外增殖及其靶基因Notch1 mRNA表达的影响。方法: HaCaT细胞体外培养,优化转染浓度,分为4组进行转染：miR-34a-5p 模拟物组（minic 组）,阴性对照组（NC组）,miR-34a-5p 抑制物组（inhibitor组）,抑制物阴性对照组（inhibitor NC组）。MTT检测细胞增殖情况; RT-qPCR检测Notch1的mRNA表达情况。结果: 转染后48小时minic 组、inhibitor组细胞增殖活力分别为39.7%和15.7%,minic 组、inhibitor组与NC组（21.3%）相比,差异有统计学意义(Ps<0.05)。HaCaT细胞转染后,minic 组和inhibitor组Notch1表达量分别为2.78±1.30和0.37±0.46,与NC组(1.0±0.06)比较,差异均有统计学意义(P<0.05)。结论: miR-34a-5p促进HaCaT细胞体外增殖,机制可能与Notch1 mRNA表达上调有关。     

Plasma Levels of miR-125b-5p and miR-206 in Acute Ischemic Stroke Patients After Recanalization Treatment: A Prospective Observational Study

Introduction: Multiple microRNAs (miRNAs) participate in the response to hypoxic/ischemic and ischemia-reperfusion events. However, the expression of these miRNAs in circulation from patients with acute ischemic stroke (AIS) receiving recanalization treatment has not been examined, and whether they are associated with the severity and outcome of stroke is still unknown. Materials and methods: In this prospective cohort study, plasma levels of miR-125b-5p, miR-15a-3p, miR-15a-5p, and miR-206 were measured at 24 hours after thrombolysis with or without endovascular treatment in 94 patients with AIS, as determined by qRT-PCR. Stroke severity was assessed based on National Institutes of Health Stroke Scale (NIHSS) score and infarct lesion. Intracranial haemorrhage (ICH) was recorded. An unfavorable outcome was defined as a modified Rankin Scale score greater than 2 at day 90 after stroke. Results: miR-125b-5p and miR-206 levels were correlated with NIHSS scores (P = .014 and P = .002) and cerebral infarction volumes (P = .025 and P = .030). miR-125b-5p levels were significantly higher in patients with an unfavorable outcome than in patients with a favorable outcome (P = .002) and showed good diagnostic accuracy in discriminating the presence of an unfavorable outcome (area under the curve .735, 95% confidence interval .623-.829, P < .001). No association was found between different miRNAs and ICH. Conclusions: In AIS patients after thrombolysis with or without endovascular treatment, miR-125b-5p is a novel prognostic biomarker highly associated with an unfavorable outcome. miR-125b-5p and miR-206 levels are associated with stroke severity.     

Heart Transplant and Hepato-Renal Dysfunction: The Model of End-Stage Liver Disease Excluding International Normalized Ratio as a Predictor of Postoperative Outcomes

IntroductionPreoperative liver and renal dysfunction remain surgical risk factors for both postoperative morbidity and mortality. The Model of End-Stage Liver Disease Excluding INR (international normalized ratio), or MELD-XI, score calculation may help as a predictor in patients with advanced heart failure. We analyzed the impact of progressive elevated MELD-XI values among recipients of heart transplant at our institution.MethodsThe data of a total of 425 consecutive adult patients who underwent heart transplantation, between January 2000 and August 2018, have been reviewed and divided into 3 cohorts according to preoperative MELD-XI calculations (MELD-XI < 11; MELD-XI 11-18; and MELD-XI > 18). Early and late outcomes have been analyzed.ResultsPatients with a MELD-XI score > 18 had a more critical clinical condition preoperatively and had a higher risk of early mortality (hazard ratio [HR] 1.45 [1.11-1.67], P < .001). They showed high risk for postoperative dialysis (HR 2.8 [1.5-5.3], P < .001), rethoracothomy for bleeding (HR 2.1 [1.2-4.1], P = .001), prolonged time of mechanical ventilation, time of intensive care unit stay (HR 2.2 [1.3-3.8], P = .005), and graft failure requiring mechanical circulatory support (HR 1.9 [1.1-3.3], P = .003). After risk adjustment per MELD-XI cohort, ischemic dilated cardiomyopathy, redo operation, and cold ischemic time > 240 minutes resulted in being the strongest predictors of early mortality (P < .001). The 5-year and 10-year survival for MELD-XI > 18 cohort was 63% and 47% vs 72% and 59% in the control group (MELD-XI < 18) (log-rank, P < .001).ConclusionsPatients with an elevated preoperative MELD-XI profile presented more comorbidities and significantly lower survival. This suggests the MELD-XI score may provide further insight into appropriate recipient and eventual donor selection. Renal insufficiency and congestive hepatopathy should be properly optimized before heart transplantation.     

MicroRNA-139 is a predictor of prostate cancer recurrence and inhibits growth and migration of prostate cancer cells through cell cycle arrest and targeting IGF1R and AXL

Background: We previously identified a panel of five microRNAs (miRNAs) associated with biochemical recurrence and metastasis following prostatectomy from prostate cancer patients using next-generation sequencing-based whole miRNome sequencing and quantitative polymerase chain reaction-based validation analysis. In this study, we examined the mechanism of action of miR-139-5p, one of the downregulated miRNAs identified in the panel. Methods: Using a cohort of 585 patients treated with radical prostatectomy, we examined the prognostic significance of miR-139 (dichotomized around the median) using the Kaplan-Meier method and Cox proportional hazard models. We validated these results using The Cancer Genome Atlas (TCGA) data. We created cell lines that overexpressed miR-139 to confirm its targets as well as examine pathways through which miR-139 may function using cell-based assays. Results: Low miR-139 expression was significantly associated with a variety of prognostic factors in prostate cancer, including Gleason score, pathologic stage, margin positivity, and lymph node status. MiR-139 expression was associated with prognosis: the cumulative incidence of biochemical recurrence and metastasis were significantly lower among patients with high miR-139 expression (P = .0004 and .038, respectively). Validation in the TCGA data set showed a significant association between dichotomized miR-139 expression and biochemical recurrence (odds ratio, 0.52; 95% confidence interval, 0.33-0.82). Overexpression of miR-139 in prostate cancer cells led to a significant reduction in cell proliferation and migration compared with control cells, with cells arrested in G2 of cell cycle. IGF1R and AXL were identified as potential targets of miR-139 based on multiple miRNA-binding sites in 3′-untranslated regions of both the genes and their association with prostate cancer growth pathways. Luciferase assays verified AXL and IGF1R as direct targets of miR-139. Furthermore, immunoblotting of prostate cancer cells demonstrated IGF1R and AXL protein expression were inhibited by miR-139 treatment, which was reversed by the addition of miR-139 antagomir. Examination of the molecular mechanism of growth inhibition by miR-139 revealed the downregulation of activated AKT and cyclin D1, with upregulation of the CDK inhibitor p21. Conclusions: miR-139 is associated with improved prognosis in patients with localized prostate cancer, which may be mediated through downregulation of IGF1R and/or AXL and associated signaling pathway components.