高血压性脑出血患者血清miR-141-3p、 miR-29a-3p水平变化及临床意义

目的 观察高血压性脑出血(HICH)患者血清miR-141-3p、miR-29a-3p水平变化,并探讨其与病情以及预后的关系.方法 选择167例HICH患者(HICH组),根据斯堪纳维亚卒中量表(SSS)评分将患者分为轻型组(0～15分,42例)、中型组(16～30分,67例)和重型组(31～45分,58例),根据出院3个月后格拉斯哥预后(GOS)评分分为预后良好组(>4分,87例),预后不良组(≤4分,80例),另选择同期于我院体检的64例健康志愿者(对照组).比较组间血清miR-141-3p、miR-29a-3p和肿瘤坏死因子-α(TNF-α)、IL-1β和IL-6水平.采用Pearson相关分析miR-141-3p、miR-29a-3p与炎性因子的相关性,多因素Logistic回归分析HICH患者预后不良的危险因素,受试者工作特征(ROC)曲线分析miR-141-3p、miR-29a-3p预测HICH患者预后不良的价值.结果 HICH组血清miR-141-3p、miR-29a-3p水平低于对照组,血清TNF-α、IL-1β和IL-6水平高于对照组(P均<0.01).重型组血清miR-141-3p、miR-29a-3p水平低于中型组和轻型组,血清TNF-α、IL-1β、IL-6水平高于中型组和轻型组(P均<0.01);中型组血清miR-141-3p、miR-29a-3p水平低于轻型组,血清TNF-α、IL-1β、IL-6水平高于轻型组(P均<0.01).HICH组血清miR-141-3p、miR-29a-3p水平与TNF-α、IL-1β、IL-6水平均呈负相关(P均<0.01).高水平miR-141-3p(OR=0.656,95％CI:0.500～0.862,P<0.01)、miR-29a-3p(OR=0.733,95％CI:0.581～0.926,P<0.01)是HICH预后的保护因素.miR-141-3p、miR-29a-3p预测HICH预后不良的曲线下面积为0.675、0.688,联合预测曲线下面积为0.898,高于单独指标预测(Z分别为4.950、5.325,P均<0.05).结论 HICH患者血清miR-141-3p、miR-29a-3p水平均降低,且与神经损伤程度加重以及预后不良有关.     

阿尔茨海默病患者血清miR-98-5p和miR-142-5p水平变化及其意义

目的 探讨阿尔茨海默病（AD）患者血清微小RNA-98-5p(miR-98-5p)、微小RNA-142-5p(miR-142-5p)水平变化及其临床意义。方法 选择AD患者103例（观察组）、体检健康的志愿者50例（对照组）,采集两组清晨空腹外周静脉血,离心留取血清,采用RT-qPCR法检测血清miR-98-5p、miR-142-5p,采用ELISA法检测血清Aβ1-40、Aβ1-42,同期采用简易精神状态检查量表（MMSE）评分评估认知功能。采用Pearson/Spearman相关分析法分析AD患者血清miR-98-5p、miR-142-5p水平与血清Aβ1-40、Aβ1-42水平和MMSE评分的关系。采用受试者工作特征（ROC）曲线分析血清miR-98-5p、miR-142-5p水平对AD的诊断价值。结果 观察组血清miR-98-5p、miR-142-5p水平均显著高于对照组（t/Z分别为7.557、5.827,P均<0.01）。观察组血清Aβ1-40水平显著高于对照组（Z=7.519,P<0.01）,血清Aβ1-42水平和MMSE评分均显著低于对照组（t/Z分别为5....     

急性胰腺炎患者血清miR-148a-3p和miR-551b-3p的表达水平及其临床价值

目的:探讨AP患者血清miR-148a-3p和miR-551b-5p的表达水平及其临床价值。方法:选取2017年1月至2020年9月间儋州市人民医院收治的152例AP患者的临床资料,根据病情严重程度将患者分为MAP组(70例)、MSAP组(40例)和SAP组(42例),SAP组又根据患者预后情况分成生存组(25例)和死...     

帕金森病患者血清miR-103a-3p、miR-486-5p水平变化及临床意义

目的 探讨帕金森病(PD)患者血清miR-103a-3p、miR-486-5p水平变化及临床意义.方法 选取183例PD患者为PD组,根据Hoehn-Yahr分级分为重度组(n=53)、中度组(n=71)、轻度组(n=59),另选取75例体检健康者为对照组.采用qRT-PCR检测血清miR-103a-3p、miR-48...     

新生儿急性呼吸窘迫综合征患儿血清miR-877-5p表达变化及其与病情严重程度和预后的关系

目的 探讨新生儿急性呼吸窘迫综合征（NARDS）患儿血清微小RNA-877-5p(miR-877-5p)表达变化及其临床意义。方法 选择NARDS患儿95例（观察组），病情严重程度：轻度38例、中度35例、重度22例，28 d内临床结局：死亡21例、存活74例，同期选择健康新生儿40例作为对照组。采集所有研究对象外周静脉血，离心留取血清，采用实时荧光定量聚合酶链式反应检测血清miR-877-5p表达，采用酶联免疫吸附试验检测血清IL-1β、IL-6、TNF-α。采用Spearman相关分析法分析NARDS患儿血清miR-877-5p表达与氧指数和血清IL-1β、IL-6、TNF-α水平的关系。采用受试者工作特征（ROC）曲线分析血清miR-877-5p表达对NARDS诊断和预后的预测价值。结果 观察组血清miR-877-5p表达低于对照组，血清IL-1β、IL-6、TNF-α水平高于对照组（P均<0.01）。随着病情严重程度增加，NARDS患儿血清miR-877-5p表达逐渐降低，血清IL-1β、IL-6、TNF-α水平逐渐升高（P均<0.01）。Spearman相关分析显...     

血浆miR-1228-5p、miR-34a-5p、miR-192-5p和miR-30a-3p水平与早发冠心病的相关性及其初筛价值

目的]探讨血浆miR-1228-5p、miR-34a-5p、miR-192-5p和miR-30a-3p水平与早发冠心病(PCAD)的相关性及其对PCAD的初筛价值。[方法]根据纳入标准及排除标准,纳入6例明确诊断的PCAD患者作为PCAD组,纳入6例健康受试者作为对照组,收集PCAD组和对照组血液,提取血清样本并保存,使用DNBseq平台检测两组血清中miRNA水平,筛选差异水平显著的miRNA。根据纳入标准及排除标准,收集78例PCAD患者、75例晚发冠心病患者和69例健康对照者的血液并对筛选的miRNA进行实时荧光定量PCR验证。分析PCAD患者冠状动脉造影报告,采用Gensini评分评估冠状动脉病变的严重程度。Spearman相关性检验分析有关miRNA水平与冠状动脉狭窄程度的相关性。ROC曲线分析血浆miR-1228-5p、miR-34a-5p、miR-192-5p及miR-30a-3p水平对PCAD的诊断价值,多因素Logistic回归分析PCAD发生的影响因素。[结果]DNBseq平台分析显示,差异表达miRNA 33个,其中上调miRNA 17个,下调miRNA 16个,差异水平最为显著的5个miRNA分别为miR-1228-5p、miR-34a-5p、miR-192-5p、miR-424-3p和miR-30a-3p;实时荧光定量PCR结果显示,与对照组相比,PCAD患者血浆miR-1228-5p升高1.7倍,miR-34a-5p升高1.4倍,miR-192-5p升高0.7倍,miR-30a-3p升高2.5倍(P＜0.05),两组间血浆miR-424-3p水平差异无统计学意义(P＞0.05);血浆miR-1228-5p和miR-34a-5p水平与PCAD患者冠状动脉狭窄程度均呈正相关(r＝0.307,P＝0.004;r＝0.238,P＝0.036);ROC曲线分析显示,miR-1228-5p、miR-34a-5p、miR-192-5p和miR-30a-3p诊断PCAD的ROC曲线下面积分别为0.903、0.832、0.731及0.798,其联合诊断PCAD的ROC曲线下面积为0.990,95%CI为0.976～1.000。[结论]PCAD患者血浆miR-1228-5p、miR-34a-5p、miR-192-5p和miR-30a-3p水平显著升高,其联合检测诊断PCAD具有较高的准确性,有望成为初筛PCAD的新型生物标志物。     

血清miR-92a-1-5p及miR-92a-2-5p表达水平与老年卒中后抑郁的关系

目的 分析血清微小核糖核酸-92a-1-5p ( miR-92a-1-5p)、 miR-92a-2-5p 表达水平与老年卒中后抑郁的关系.方法 选取2018年2月至2020年10月胶州中心医院收治的129例老年卒中患者为研究组,另选取110名同期健康体检者为对照组,均检测血清miR-92a-1-5p、miR-92a-2...     

miR-181a-5p调控LIF的表达调节胰腺腺泡细胞凋亡的分子机制

背景急性胰腺炎(acute pancreatitis, AP)发病迅速,并发全身感染,因此早期的高效治疗对控制病情起到关键作用.微小RNA(microRNA, miRNA)在AP中的作用机制及临床应用的研究成为近几年的热点.其在AP发生发展及转归中的作用有助于为AP的诊断和治疗提供新思路.目的探讨miR-181a-5p对雨蛙素诱导的大鼠胰腺腺泡细胞AR42J凋亡的影响及机制.方法运用ELISA法检测雨蛙素诱导的大鼠胰腺腺泡细胞中AMY、肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)、白介素-6(interleukin-6,IL-6)的表达;将Cerulein+anti-miR-con组(转染anti-miR-con)、Cerulein+anti-miR-181a-5p组(转染anti-miR-181a-5p)、Cerulein+si-con组(转染si-con)、Cerulein+si-白血病抑制因子(leukemia inhibitory factor, LIF)组(转染si-LIF),均用脂质体法转染至AR42J细胞,再用15 nmol/L的雨蛙素处理8h;流式细胞术法检测各组细胞的凋亡; qRT-PCR检测各组细胞中miR-181a-5p mRNA和LIF mRNA的表达; Western blot检测各组细胞中LIF、caspase-3的蛋白表达;双荧光素酶报告基因检测实验检测各组细胞的荧光活性.结果与对照组相比,雨蛙素处理8 h是AMY、TNF-α和IL-6的含量均升高的时间点,且细胞凋亡率显著降低(P 0.05);与Control组相比,Cerulein组AR42J细胞中miR-181a-5p mRNA表达显著下调, LIF mRNA和蛋白表达显著上调(P0.05);过表达miR-181a-5p、敲减LIF均可抑制雨蛙素诱导的AR42J细胞凋亡的促进作用; miR-181a-5p可抑制野生型LIF细胞的荧光活性,且可负向调控LIF的蛋白表达.结论 miR-181a-5p可抑制雨蛙素诱导的AR42J细胞的凋亡,其机制可能与靶向LIF有关,将可为AP的治疗提供新方向.     

miR-186-5p基于JAK/STAT3通路对三叉神经痛模型大鼠胶质细胞活化的影响

目的 基于酪氨酸蛋白激酶/信号传导与转录激活因子(JAK/STAT)3通路探讨微小RNA-186-5p(miR-186-5p)对三叉神经痛模型大鼠胶质细胞活化的影响。方法 选取50只健康雄性大鼠，随机选取10只为正常组，剩余40只建立三叉神经痛模型，最终39只建模成功，随机分为三叉神经痛组、上调组、下调组各13只。建模完成后上调组注射10μl miR-186-5p过表达慢病毒悬液，下调组注射10μl miR-186-5p沉默慢病毒悬液，正常组、三叉神经痛组不做处理。对比各组体质量、机械痛阈，酶标分析仪检测白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α、IL-6、胶质细胞源性神经营养因子(GDNF)、胶质纤维酸性蛋白(GFAP)水平，Western印迹法检测非受体型JAK2、STAT3表达。结果 与正常组比较，其他3组GDNF、GFAP、TNF-α、IL-1β、IL-6表达水平显著较高(P<0.05)。与三叉神经痛组比较，上调组GDNF、GFAP表达水平较低，TNF-α、IL-1β、IL-6表达水平较高，下调组GDNF、GFAP表达水平较高，TNF-α、IL-1β、IL-6表...     

非酒精性脂肪性肝病患者血清miR-183-5p和miR-96-5p水平变化及其临床意义探讨

目的 探讨非酒精性脂肪肝病(NAFLD)患者血清miR-183-5p和miR-96-5p水平变化及其临床意义。方法 2019年11月～2022年1月我院诊治的NAFLD患者62例和同期性别和年龄相匹配且无过量饮酒史的健康体检者80例,采用实时荧光定量RT-PCR法检测血清miR-183-5p和miR-96-5p水平,应用多因素Logistic回归分析NAFLD发生的危险因素,应用受试者工作特征曲线(ROC)下面积(AUC)评估指标诊断严重脂肪性肝病的效能。结果 NAFLD患者BMI、胰岛素抵抗指数(HOMA-IR)、空腹血糖(FPG)和血脂水平显著高于健康人(P<0.05),血清miR-183-5p和miR-96-5p水平分别为(0.8±0.2)和(1.2±0.3),显著低于健康人【分别为(1.4±0.4)和(2.3±0.6),P<0.05】; 17例重度NAFLD患者血清miR-183-5p和miR-96-5p水平分别为(0.4±0.1)和(0.8±0.2),显著低于21例中度患者【分别为(0.8±0.2)和(1.2±0.3),P<0.05】或24例轻度患者【分别为...     

Diabetic Cardiomiopathy Progression is Triggered by miR122-5p and Involves Extracellular Matrix: A 5-Year Prospective Study

ObjectivesThe purpose of this study was to follow the long-term progression of diabetic cardiomyopathy by combining cardiac magnetic resonance (CMR) and molecular analysis.BackgroundThe evolution of diabetic cardiomyopathy to heart failure affects patients’morbidity and mortality. CMR is the gold standard to assess cardiac remodeling, but there is a lack of markers linked to the mechanism of diabetic cardiomyopathy progression.MethodsFive-year longitudinal study on patients with type 2 diabetes mellitus (T2DM) enrolled in the CECSID (Cardiovascular Effects of Chronic Sildenafil in Men With Type 2 Diabetes) trial compared with nondiabetic age-matched controls. CMR with tagging together with metabolic and molecular assessments were performed at baseline and 5-year follow-up.ResultsA total of 79 men (age 64 ± 8 years) enrolled, comprising 59 men with T2DM compared with 20 nondiabetic age-matched controls. Longitudinal CMR with tagging showed an increase in ventricular mass (ΔLVMi = 13.47 ± 29.66 g/m²; p = 0.014) and a borderline increase in end-diastolic volume (ΔEDVi = 5.16 ± 14.71 ml/m²; p = 0.056) in men with T2DM. Cardiac strain worsened (Δσ = 1.52 ± 3.85%; p = 0.033) whereas torsion was unchanged (Δθ = 0.24 ± 4.04°; p = 0.737), revealing a loss of the adaptive equilibrium between strain and torsion. Contraction dynamics showed a decrease in the systolic time-to-peak (ΔTtP = ?35.18 ± 28.81 ms; p < 0.001) and diastolic early recoil-rate (ΔRR = ?20.01 ± 19.07 s^-1; p < 0.001). The ejection fraction and metabolic parameters were unchanged. Circulating miR microarray revealed an up-regulation of miR122-5p. Network analysis predicted the matrix metalloproteinases (MMPs) MMP-16 and MMP-2 and their regulator (tissue inhibitors of metalloproteinases) as targets. In db/db mice we demonstrated that miR122-5p expression is associated with diabetic cardiomyopathy, that in the diabetic heart is overexpressed, and that, in vitro, it regulates MMP-2. Finally, we demonstrated that miR122-5p overexpression affects the extracellular matrix through MMP-2 modulation.ConclusionsWithin 5 years of diabetic cardiomyopathy onset, increasing cardiac hypertrophy is associated with progressive impairment in strain, depletion of the compensatory role of torsion, and changes in viscoelastic contraction dynamics. These changes are independent of glycemic control and paralleled by the up-regulation of specific microRNAs targeting the extracellular matrix. (Cardiovascular Effects of Chronic Sildenafil in Men With Type 2 Diabetes [CECSID]; NCT00692237)     

社区获得性肺炎外周血miR-182及miR-126的表达及临床意义

目的分析社区获得性肺炎(CAP)患者外周血微小RNA-182(miR-182)及微小RNA-126(miR-126)的表达及临床意义。 方法选取我院2018年4月至2021年5月收治的58例CAP患者为对象,于患者入院24 h内检测外周血miR-182及miR-126表达水平,于患者治疗4周后评估临床预后,根据预后将其分为预后良好者47例和预后不佳者11例。采用Logistic多因素回归分析影响CAP患者预后的相关因素;计算评估miR-182、miR-126水平对CAP的预后。 结果58例CAP患者中预后不佳者11例(18.96%),预后良好者47例(81.04%)。预后不佳者miR-182相对表达量均高于预后良好者,miR-126相对表达量低于预后良好组,差异有统计学意义(P<0.05)。Logistic多因素回归分析结果显示,PSI评分、WBC计数水平高均为CAP患者预后不佳的危险因素(P<0.05)。miR-182、miR-126水平单一及联合对CAP患者临床预后分别为0.742、0.724、0.833,且miR-182、miR-126水平对CAP患者临床预后的高于单一预测(P<0.05)。 结论预后不佳的CAP患者miR-182相对表达量升高,miR-126相对表达量降低,且miR-182、miR-126水平对预后具有临床意义。     

Cocktail of eternity: HDAC meets miR

In this issue of Blood, Sampath and colleagues provide an important missing link in how microRNAs (miRs) can be silenced in chronic lymphocytic leukemia (CLL):histone deacetylases (HDACs) that are overexpressed in CLL block critical miRs in the malignant B cell resulting in pro-survival signals. Thus,HDAC inhibition is an attractive new therapeutic strategy in CLL.     

miR495、miR551a靶向干扰SGC7901细胞PRL-3基因的表达

目的:构建靶向PRL-3基因的miRNA495和miRNA551a真核表达载体,观察其转染胃癌SGC7901细胞后对胃癌细胞PRL-3基因表达的影响.方法:设计并合成两条针对PRL-3基因的特异性miRNA495和miRNA551a干扰序列,分别与pcDNA6.2-GW/Em GFP-miR载体连接,转化大肠杆菌,纯化并鉴定后利用LipofectamineTM2000转染胃癌SGC7901细胞,实时定量PCR及Western blot技术鉴定重组体对PRL-3基因表达的干扰效果.结果:针对PRL-3基因的miRNA495和miRNA551a干扰质粒构建成功,胃癌SGC7901细胞分别转染两种质粒后miRNA495及miRNA551a的表达明显升高,并且明显抑制了PRL-3基因mRNA及蛋白的表达.结论:靶向PRL-3基因的miRNA495和miRNA551a真核表达载体构建成功,其可有效提高胃癌SGC7901细胞miRNA495和miRNA551a的表达,并抑制PRL-3mRNA及PRL-3蛋白的表达.     

miR-21-5p promotes NASH-related hepatocarcinogenesis

Background and Aims

The mechanisms governing the progression of non-alcoholic fatty liver disease (NAFLD) towards steatohepatitis (NASH) and hepatocellular carcinoma (HCC) remain elusive. Here, we evaluated the role of hsa-miRNA-21-5p in NASH-related hepatocarcinogenesis.

Methods

Hepatic hsa-miR-21-5p expression was evaluated in two cohorts of patients with biopsy-proven NAFLD (n = 199) or HCC (n = 366 HCC and n = 11 NAFLD-HCC). Serum/liver metabolomic profiles were correlated with hsa-miR-21-5p in NAFLD obese patients. Wild-type (WT) and Mir21 KO mice were fed a choline-deficient, amino acid-defined (CDAA) diet for 32 and 66 weeks to induce NASH and NASH-HCC, respectively.

Results

In obese individuals, hsa-miR-21-5p expression increased with NAFLD severity and associated with a hepatic lipotoxic profile. CDAA-fed WT mice displayed increased hepatic mmu-miR-21-5p levels and progressively developed NASH and fibrosis, with livers presenting macroscopically discernible pre-neoplastic nodules, hyperplastic foci and deregulated cancer-related pathways. Mir21 KO mice exhibited peroxisome-proliferator-activated receptor α (PPARα) activation, augmented mitochondrial activity, reduced liver injury and NAS below the threshold for NASH diagnosis, with the pro-inflammatory/fibrogenic milieu reversing to baseline levels. In parallel, Mir21 KO mice displayed reduced number of pre-neoplastic nodules, hepatocyte proliferation and activation of oncogenic signalling, being protected from NASH-associated carcinogenesis. The hsa-miRNA-21-5p/PPARα pathway was similarly deregulated in patients with HCC- or NASH-related HCC, correlating with HCC markers and worse prognosis.

Conclusions

Hsa-miR-21-5p is a key inducer of whole-spectrum NAFLD progression, from simple steatosis to NASH and NASH-associated carcinogenesis. The inhibition of hsa-miR-21-5p, leading to a pro-metabolic profile, might constitute an appealing therapeutic approach to ameliorate NASH and prevent progression towards HCC.     

Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia

Micro-RNAs (miR genes) are a large family of highly conserved noncoding genes thought to be involved in temporal and tissue-specific gene regulation. MiRs are transcribed as short hairpin precursors ( approximately 70 nt) and are processed into active 21- to 22-nt RNAs by Dicer, a ribonuclease that recognizes target mRNAs via base-pairing interactions. Here we show that miR15 and miR16 are located at chromosome 13q14, a region deleted in more than half of B cell chronic lymphocytic leukemias (B-CLL). Detailed deletion and expression analysis shows that miR15 and miR16 are located within a 30-kb region of loss in CLL, and that both genes are deleted or down-regulated in the majority ( approximately 68%) of CLL cases.     

胰腺癌患者血浆miR-155检测及其诊断价值

目的 检测胰腺癌患者血浆miR-155表达量,评价其对胰腺癌的诊断价值.方法 收集62例胰腺癌、61例慢性胰腺炎(CP)及36例正常对照者的血标本,抽提血浆RNA,应用实时PCR检测miR-155表达量,并分析其与胰腺癌临床参数的关系.应用接受者操作特征(ROC)曲线下面积(AUC)评价血浆miR-155表达量对胰腺癌的诊断价值.结果 胰腺癌、CP和正常对照组的血浆miR-155表达量分别为5.41±3.14、2.59±2.49和0.77±1.17,胰腺癌血浆miR-155表达量显著高于CP及正常对照组(P＜0.01).胰腺癌血浆miR-155表达量与患者年龄、性别、肿瘤大小等均无显著相关性,而与肿瘤TNM分期呈显著负相关(r=-0.323,P=0.01).经ROC分析,胰腺癌对正常对照组的AUC为0.943(95%CI0.902～0.985),敏感性和特异性分别为87.1%和83.3%;胰腺癌对CP的AUC为0.762(95%CI0.678～0.846),敏感性和特异性分别为64.5%和73.8%;胰腺癌对正常对照组+CP组的AUC为0.829(95%CI0.767～0.892),敏感性和特异性分别为62.9%和84.5%.结论 胰腺癌患者血浆miR-155表达量显著升高,对胰腺癌的诊断可能有一定的应用价值.     

No difference in kinetics of tau or histone phosphorylation by CDK5/p25 versus CDK5/p35 in vitro

CDK5/p35 is a cyclin-dependent kinase essential for normal neuron function. Proteolysis of the p35 subunit in vivo results in CDK5/p25 that causes neurotoxicity associated with a number of neurodegenerative diseases. Whereas the mechanism by which conversion of p35 to p25 leads to toxicity is unknown, there is common belief that CDK5/p25 is catalytically hyperactive compared to CDK5/p35. Here, we have compared the steady-state kinetic parameters of CDK5/p35 and CDK5/p25 towards both histone H1, the best known substrate for both enzymes, and the microtubule-associated protein, tau, a physiological substrate whose in vivo phosphorylation is relevant to Alzheimer’s disease. We show that the kinetics of both enzymes are the same towards either substrate in vitro. Furthermore, both enzymes display virtually identical kinetics towards individual phosphorylation sites in tau monitored by NMR. We conclude that conversion of p35 to p25 does not alter the catalytic efficiency of the CDK5 catalytic subunit by using histone H1 or tau as substrates, and that neurotoxicity associated with CDK5/p25 is unlikely attributable to CDK5 hyperactivation, as measured in vitro.