人血浆及粪便中微小RNA-92a-1表达水平在结直肠肿瘤筛查中的意义

目的 探讨联合检测人血浆及粪便中微小RNA-92a-1(miRNA-92a-1)表达水平作为结直肠肿瘤筛查标志物的可行性及临床意义.方法 收集2011年8月至10月间60例结直肠癌患者、23例结直肠腺瘤患者及30名健康对照者的粪便及血浆标本,采用实时定量RT-PCR的方法检测miRNA-92a-1的表达水平.组间采用Mann-Whitney U检验进行差异性检验,然后根据受试者工作特征曲线(ROC曲线)确定截断点,对实验结果的敏感度及特异度进行分析.结果 结直肠癌和结直肠腺瘤患者血浆中miRNA-92a-1的表达水平均高于健康对照者(U=288.5和151.0,P均＜0.01).结直肠癌患者粪便中miRNA-92a-1的表达水平高于健康对照者(U=627.5,P=0.0199).根据ROC曲线分析,当截断点值＞1.22时,miRNA-92a-1在结直肠癌和结直肠腺瘤患者血浆中的敏感度分别为85.0％(51/60)和73.9％(17/23),在健康对照者血浆中的特异度为76.7％(23/30);当截断点值＞1.14时,miRNA-92a-1在结直肠癌和结直肠腺瘤患者粪便中的敏感度分别为31.7％(19/60)和26.1％(6/23),在健康对照者粪便中的特异度为90.0％(27/30).联合血浆及粪便中的检测结果,miRNA-92a-1在结直肠癌和结直肠腺瘤患者中的敏感度分别为88.3％(53/60)和82.6％(19/23),在健康对照者中的特异度为73.3％(22/30),敏感度高于单样本检测.结论 联合检测结直肠癌及结直肠腺瘤患者血浆及粪便中miRNA-92a-1的表达水平,具有较高的敏感度和特异度.miRNA-92a-1可作为结直肠癌诊断及筛查的一个潜在指标.     

阿尔茨海默病患者血浆microRNA-101a-3p的表达

目的探讨microRNA在阿尔茨海默病(AD)患者血浆中的差异变化,以寻求一个可能作为辅助诊断AD的早期生物标志物。方法实验分为患者组和健康对照组,分别取其血浆并提取microRNA,利用microRNA基因芯片和荧光定量PCR的方法寻找有表达差异的microRNA。结果基因芯片筛选出68个AD相关的差异表达microRNAs基因(P0.05),其中miR-101a的表达水平在AD患者中显著低于对照组(log=-5.53,P=1.19×10-5),选定为感兴趣miRNA;荧光定量PCR示:患者组中miR-101a的相对表达量为8.834×10-4±2.842×10-4,对照组为6.305×10-3±1.161×10-3。患者组表达量仅为对照组14%,两组间差异有统计学意义(F=21.76,P0.001),结果与miRNA芯片检测结果一致。ROC示:如果以0.002 163为诊断临界值,即miR-101a相对表达量低于0.002 163提示诊断AD。结论 miR-101a可能通过多种途径影响AD的发生发展,可能作为一种辅助诊断AD的早期生物标志物。     

hARD1在人大肠腺癌中的表达及与肿瘤分化的相关性

目的:检测人大肠腺癌标本中hARD1的表达及其与肿瘤分化的相关性.方法:收集昆明医学院第一附属医院手术切除的大肠癌患者大肠腺癌组织、癌旁组织及大肠正常切缘组织标本100例,排除术前化疗的2例,共98例,其中高分化腺癌30例,中分化腺癌27例,低分化腺癌31例,黏液腺癌10例.细胞免疫化学法检测hARD1在大肠腺癌组织、...     

生物钟基因hClock蛋白在结直肠肿瘤中的表达研究

目的探讨人类生物钟基因hClock蛋白在结直肠肿瘤中的表达及其意义。方法应用免疫组织化学法检测不同Dukes分期结直肠肿瘤及相应癌旁组织中hClock基因蛋白产物（Clock蛋白）的表达,并比较其差异。结果Clock蛋白红结直肠肿瘤中呈现中或强阳性表达,与肿瘤分化程度及Dukes分期无显著相关性（P〉0.05）。癌旁组织呈现弱阳性表达。结论Clock蛋白与结直肠肿瘤的发生有相关性,与侵袭、转移的关系有待更深入地研究。     

Increased tumor necrosis factor receptor 1 expression in human colorectal adenomas

AIM: To determine the expression statuses of tumor necrosis factor (TNF)-α, its receptors (TNF-R) and downstream effector molecules in human colorectal adenomas. METHODS: We measured the serum concentrations of TNF-α and its receptors in 62 colorectal adenoma patients and 34 healthy controls. The protein expression of TNF-α, TNF-R1, TNF-R2 and downstream signals of the TNF receptors, such as c-Jun N-terminal kinase (JNK), nuclear factor-κ B and caspase-3, were also investigated in human colorectal adenomas and in normal colorectal mucosal tissues by immunohistochemistry. Immunofluorescence confocal microscopy was used to investigate the consistency of expression of TNF-R1 and phospho-JNK (p-JNK). RESULTS: The serum levels of soluble TNF-R1 (sTNF-R1) in adenoma patients were significantly higher than in the control group (3.67 ± 0.86 ng/mL vs 1.57 ± 0.72 ng/mL, P 0.001). Receiver operating characteristic analysis revealed the high diagnostic sensitivity of TNF-R1 measurements (AUC was 0.928) for the diagnosis of adenoma, and the best cut-off level of TNF-R1 was 2.08 ng/mL, with a sensitivity of 93.4% and a specificity of 82.4%. There were no significant differences in the serum levels of TNF-α or sTNF-R2 between the two groups. Immunohistochemistry showed high levels of TNF-R1 and p-JNK expression in the epithelial cells of adenomas. Furthermore, a high incidence of co-localization of TNF-R1 and p-JNK was identified in adenoma tissue. CONCLUSION: TNF-R1 may be a promising biomarker of colorectal adenoma, and it may also play an important role in the very early stages of colorectal carcinogenesis.     

稳定性心绞痛患者血糖水平与循环微小RNA-92a表达关系的研究

目的探讨稳定性心绞痛(SAP)患者循环微小RNA-92a(microRNA-92a,miR-92a)表达与血糖的关系。方法将93例SAP患者分为A组:降糖治疗+空腹血糖(FPG)<7.0 mmol/L 18例,B组:降糖治疗+FPG≥7.0 mmol/L8例,C组:非降糖治疗+FPG<7.0 mmol/L 61例,D组:非降糖治疗+FPG≥7.0 mmol/L 6例。比较4组患者血糖水平与循环miR-92a表达的关系。结果 SAP患者合并2型糖尿病发病率为34.4%,降糖治疗后,仍有30.8%FPG≥7.0 mmol/L。A组与B组miR-92a是否>-0.1或>0.5水平的OR值分别为2.67或0.17,C组与D组OR值分别为0.36、0.48。A组miR-92a>-0.1的患者为88.9%,显著高于C组的41.0%(P<0.01)。B组miR-92a>0.5的患者为75.0%,高于A组的33.3%(P>0.05)。结论单纯测定FPG诊断2型糖尿病漏诊率高。循环miR-92a有助于FPG<7.0 mmol/L的SAP合并2型糖尿病的诊断及疗效评价。     

High expression levels of microRNA-629, microRNA-525-5p and microRNA-516a-3p in paediatric systemic lupus erythematosus

Paediatric systemic lupus erythematosus (pSLE) refers to childhood-onset systemic lupus erythematosus. pSLE has its own unique characteristics, and its pathogenesis is unclear. To study the relationship between microRNAs (miRNAs) and pSLE, we selected three pSLE patients who were newly diagnosed and had not yet been treated, and two controls were also included. We collected their peripheral blood mononuclear cells to perform Agilent human miRNA (8?×?15 k) 12.0 analysis. To verify the results, we next selected 12 other pSLE patients who had different disease activities and 3 healthy controls and conducted real-time PCR. The results showed high expression of miRNA-516a-3p, miRNA-629 and miRNA-525-5p in pSLE patients with active disease; these levels were normal in patients without active disease. Increased expression levels of these three miRNAs were positively correlated with the score obtained from the systemic lupus erythematosus disease activity index scoring system (SLEDAI) 2000 and C-reactive protein (CRP) levels. Furthermore, the target genes of these three miRNAs were important to the pathogenesis of pSLE. Therefore, these three miRNAs might be specific to pSLE and may be used as novel biomarkers of pSLE to diagnose and monitor the disease.     

Nuciferine downregulates microRNA-33a-5p expression and upregulates ABCA1/G1 expressionin THP-1 derived macrophages

正Objective To explore the effects of nuciferine on the expression of microRNA-33a-5p(miR-33a-5p)and ATPbinding cassette transporter A1/G1(ABCA1/G1)in THP-1 derived macrophages.Methods Macrophages were derived from THP-1 monocytes by PMA.Macrophages were randomly divided into four groups:normal     

血浆中miR-144*直接扩增在大肠癌非侵入性诊断中的应用

目的:研究miR-144*在大肠癌(CRC)、大肠腺瘤、炎症性肠病(IBD)及健康对照组的血浆标本中直接扩增后的表达量,初步探索其在大肠癌的发生、发展中的作用.方法:分别收集CRC、大肠腺瘤、IBD及全结肠镜检查阴性的健康人的血浆标本55例、30例、30例、30例,上述标本均取自患者未治疗前;分别收集CRC、大肠腺瘤患者行肿瘤切除术后7d的血浆标本43例、30例.利用TRIzol试剂进行上述标本中RNA的提取,得到纯化后的RNA样本,逆转录及实时荧光定量PCR反应检测miR-144*的表达,进行统计学分析.结果:在人大肠癌组血浆标本中,miR-144*较非大肠癌组中表达增高,miR-144*的表达量与肿瘤的大小及浸润深度相关,肿瘤越大,浸润深度越深,miR-144*表达量越高.在43例大肠癌患者术后7d的血浆标本中,30例行大肠癌根治术患者中27例miR-144*的表达量较术前降低,13例行大肠癌姑息性手术患者的血浆miR-144*的表达量较术前无明显差异.大肠腺瘤组中,进展期腺瘤和非进展期腺瘤在行腺瘤切除术前后miR-144*的表达量无明显变化.结论:血浆中miRNA-144*表达水平的检测,可作为大肠癌非侵入性诊断方法并可以预测大肠癌的复发.     

MiR-92a-3p_R+1和miR-92a-1-5p在ox-LDL诱导大鼠VSMC表型转化和增殖中的作用

目的探讨miR-92a-3p_R+1和miR-92a-1-5p在氧化型低密度脂蛋白(ox-LDL)诱导大鼠血管平滑肌细胞(VSMC)表型转化和增殖中的作用及其与ERK1/2信号通路的相关性。方法分离、培养大鼠VSMC,以ox-LDL(50 mg/L)诱导VSMC,采用ERK1/2特异性抑制剂U0126(10μmol/L)阻断ox-LDL(50 mg/L)诱导VSMC的ERK1/2信号激活,MicroRNA微阵列分析VSMC的miR-92a-3p_R+1和miR-92a-1-5p表达,CCK-8法和Brdu流式细胞术检测细胞增殖;免疫荧光法检测VSMC收缩表型标志蛋白SM22α的变化;Western blot检测VSMC的ERK1/2通路信号激活情况(ERK、p-ERK)、表型标志蛋白SM22α、细胞周期相关蛋白(PCNA、cyclin D1、p21、p27)的表达情况。结果 ox-LDL诱导下,VSMC的miR-92a-3p_R+1和miR-92a-1-5p表达明显上调,VSMC的ERK1/2磷酸化水平明显增加,SM22α的表达降低,同时细胞周期相关蛋白PCNA、cyclin D1高表达,p21、p27低表达;ERK1/2通路特异性抑制剂U0126干预后,ERK1/2磷酸化水平受抑制,相应的VSMC miR-92a-3p_R+1和miR-92a-1-5p表达明显下调(P0.05),VSMC增殖显著下降,SM22α的表达上调(P0.05),提示VSMC由合成表型转化为收缩表型,并下调PCNA、cyclin D1的表达,上调p21、p27蛋白的表达(P0.05),说明其表型转化和增殖明显受抑制。结论 miR-92a-3p_R+1和miR-92a-1-5p在ox-LDL诱导VSMC表型转化和增殖中起重要作用,并与ERK1/2信号通路密切相关。     

Effect of miR-92a-3p_R+1 and miR-92a-1-5 in ERK signaling pathway of ox-LDL induced phenotypic transformation and proliferation of VSMC in rats

<正>Objective To investigate the effect of miR-92a-3p_R+1 and miR-92a-l-5p on phenotypic transformation and proliferation of rat vascular smooth muscle cells (VSMC)induced by oxidized low-density lipoprotein (ox-LDL) in the ERK 1/2 signaling pathway.Methods VSMC was isolated from rats,and ox-LDL (50 mg/L) was used to induce VSMC.ERK1/2 inhibitors U0126 (10μmol/L)was used for intervention.The miRNA microarray profi-     

Bmi-1和Mel-18基因在大肠癌组织中的表达及意义

目的:探讨Bmi-1和Mel-18基因在大肠癌中的表达及其临床意义.方法:收集我院68例大肠癌组织标本,68例癌旁正常组织作为对照.利用免疫组织化学的方法检测大肠癌及正常组织中的Bmi-1基因和Mel-18基因的表达,并结合临床资料,对该两种基因的表达与大肠癌患者临床表现的相关性进行分析.结果:(1)Bmi-l在大肠癌组织中的表达明显高于正常组织(73.5%vs23.5%,P<0.05),且与大肠癌的侵袭深度、淋巴结转移及临床分期有关(P<0.05);(2)Mel-18基因在大肠癌组织中的表达明显低于正常组织(41.2%vs66.2%,P<0.05),且与大肠癌的淋巴结转移及临床分期呈负相关(P<0.05);(3)相关性分析发现,Bmi-1和Mel-18基因在大肠癌组织中的表达呈负相关(r=-0.545,P<0.05).结论:Bmi-1和Mel-18基因与大肠癌的发展、转移关系密切,检测Bmi-1和Mel-18对大肠癌的诊断及判断预后可能有重要意义.     

Changes of microRNA-1 and microRNA-21 in human atrial fibrillation

Objectives To investigate changes of microRNA- 1(miR-1) and microRNA-21(miR-21) expressions in human atrial fibrillation(AF).Methods Right atrial appendages were obtained from 12 rheumatic valvular heart disease patients with sinus rhythm(SR)and 14 patients with chronic AF.Quantitative real time polymerase chain reaction(qRT-PCR) was applied to assess expressions of miR-1 and miR- 21.Immunohistochemistry was used to evaluate Kir2.1 expression and masson staining was used to estimate collagen deposition.Results Expressions of miR-1 was decreased (relative expression:0.68±0.09 vs.1.76±0.15,PmiR-21 was increased(relative expression:2.23±0.31 vs.0.94±0.24 vs. 0.043±0.005 vs.8.3±0.01;PmiR-l was negatively correlated to that of left atrium diameters(r=-0.47,P=0.02).Expression of miR-21 was positively correlated to collagen content (r=0.45,P<0.01).Conclusions Expression of miR-1 was down-regulated in AF patients,which may be associated with the increase in Kir2.1.miR-21 level was significantly up-regulated, which may contribute to the increase in collagen content in the atrium.     

microRNA-92a expression in the sera and dermal fibroblasts increases in patients with scleroderma

Objectives. microRNAs (miRNAs) play a part in various cellular activities. However, the role of miRNA in SSc is not fully understood. This study investigated the expression and role of miR-92a in SSc patients and evaluated the possibility that miR-92a is involved in the pathogenesis of this disease. Methods. Serum samples were obtained from 61 SSc patients. mRNAs were purified from serum and levels of miR-92a and miR-135 were measured with quantitative real-time PCR. miR-92a expression in dermal fibroblasts was also determined by quantitative real-time PCR. Immunoblotting was performed to detect MMP-1 protein. Results. The median serum levels of miR-92a, not miR-135, were significantly higher in SSc patients than normal subjects. The constitutive up-regulated miR-92a expression was also found in cultured dermal fibroblasts from SSc skin, which was decreased by the transfection with siRNA of TGF-β. Furthermore, the forced overexpression of miR-92a in normal dermal fibroblasts using miR-92a mimic resulted in the down-regulation of MMP-1 expression. Conclusion. The increase of miR-92a in SSc may be due to the stimulation of intrinsic TGF-β activation seen in this disease. There is also a possibility that MMP-1 is the target of miR-92a and that increased miR-92a expression therefore plays a role in excessive collagen accumulation in SSc via the down-regulation of MMP-1. Clarifying the role of miRNAs in SSc may result in a better understanding of this disease and the development of new therapeutic approaches.     

钙卫蛋白在大肠癌中的表达及意义

目的研究钙卫蛋白(calprotectin)在大肠癌、大肠腺瘤、正常大肠黏膜中的表达情况。方法采取免疫组织化学法对大肠癌、大肠腺瘤、正常大肠黏膜标本中钙卫蛋白进行检测。结果钙卫蛋白在大肠癌细胞、大肠腺瘤细胞、正常大肠黏膜细胞中无表达,表达在以上组织间质的中性粒细胞中,癌组织中阳性率为75.61%,显著高于大肠腺瘤组织、正常结肠黏膜(4.87%、3.57%,P0.05);钙卫蛋白表达与大肠癌部位、病理类型、分化程度无关(P0.05)。结论大肠癌组织中钙卫蛋白含量增高,表达在大肠癌组织间质中。     

节律基因hBmal1在人结肠癌中的表达及临床意义

目的检测hBmal1在人结肠癌中的表达,探讨其临床意义。 方法应用免疫组织化学方法检测湖北省肿瘤医院30例人结肠癌患者中hBmal1的表达,以图像分析软件进行相关测定,并分析其表达与临床及病理常用指标的关系。 结果人结肠癌组织和癌旁正常对照组织中hBmal1表达阳性率为76.67%（23/30） vs 26.67%（8/30）（χ²=15.016,P<0.01）;hBmal1在人结肠癌Dukes分期B、C期中强阳性表达率为46.15%（6/13） vs 88.24%（15/17）（χ²=6.212,P=0.013）。 结论hBmal1在人结肠癌组织中有高表达,可能与癌症的发生发展侵袭转移有关。     

Kinetics of plasma microRNA-499 expression in acute myocardial infarction

Background

MicroRNA (miRNA) is reported to be present in human plasma and has been increasingly suggested as a biomarker for diseases. Our study aimed to investigate the kinetics of cardiac-specific microR-499 (miR-499) in acute myocardial infarction (AMI).

Methods

Circulating concentrations of cardiac enriched miR-499 were measured by quantitative PCR in 73 patients with acute coronary syndrome (ACS), including 53 with AMI and 20 with unstable angina (UA). Thirty healthy subjects were used as controls. Plasma samples in AMI group were obtained immediately after admission and at 12 h, 24 h, 3 d and 7 d after onset of symptoms. Plasma samples in UA and healthy control groups were collected immediately after admission. The severity and extent of coronary stenotic lesions were evaluated on the basis of coronary angiography using Gensini score.

Results

miR-499 expression levels were significantly higher in the 53 AMI patients than in the 20 UA patients and 30 healthy controls immediately after admission (P<0.01). A measurable increase in miR-499 levels was observed in AMI patients within 24 h of the last onset of chest pain and the levels returned to the baseline after 7 d. Plasma miR-499 levels in the patients with AMI were positively-correlated with cTnI (r=0.384, P<0.01) and CK-MB (r=0.402, P<0.01). In addition, miR-499 levels in AMI patients with two- and three-vessel coronary artery disease (CAD) were significantly higher than those in patients with single-vessel CAD (P<0.05). Gensini scores were used to evaluate the severity of coronary stenosis. miR-499 were positively correlated with Gensini scores (r=0.52, P<0.01). miR-499 levels at admission were significantly higher than that those 24 h after percutaneous coronary intervention (PCI) in AMI patients (P<0.01) and were negatively correlated with LVEF (r=0.36, P=0.008).

Conclusions

Cardiac-specific miRNA-499 levels were found to be linearly proportional to myocardial damage. MiRNA-499 might prove to be a new biomarker for AMI and a predictor of the risk of myocardial ischemia.