高危型HPV阳性宫颈癌患者miRNA-34a-5p的表达及临床意义

目的　探讨高危型人乳头状瘤病毒（high risk-human papilloma virus, HR-HPV）阳性宫颈癌患者组织及血清中微小RNA（microRNA, miRNA）-34a-5p的表达及其对宫颈癌的诊断价值。方法　将100例HR-HPV阳性宫颈癌患者（宫颈癌组）、70例宫颈上皮内瘤变（cervical intraepithelial neoplasia, CIN）患者（CIN组）、70例HR-HPV阳性子宫良性病变或者HPV单纯阳性的健康体检者（对照组）作为研究对象。采集上述研究对象的血清标本及70例行手术治疗的宫颈癌组患者癌组织及癌旁组织。使用实时荧光定量PCR技术测定组织及血清中miRNA-34a-5p水平,并分析miRNA-34a-5p与宫颈癌临床、病理指标的关联及血清miRNA-34a-5p对宫颈癌的诊断价值。结果　①对照组血清中miRNA-34a-5p的相对表达量高于CIN组和宫颈癌组：（0.933±0.097）vs.（0.554±0.099）vs.（0.369±0.099）,差异有统计学意义（F=741.401,P=0.000）;宫颈癌组患者术前血清中miRNA-34a-5p相对表达量低于术后1周,差异有统计学意义（t=8.305,P=0.000）。有无淋巴结转移和有无远处转移的宫颈癌患者血清中miRNA-34a-5p相对表达量的差异有统计学意义（P均＜0.05）。②宫颈癌组患者癌旁组织中miRNA-34a-5p的相对表达量高于癌组织,差异具有统计学意义（t=40.313,P=0.000）。③宫颈癌组患者癌组织中的miRNA-34a-5p表达水平与血清中的表达水平呈正相关（r=0.908,P=0.000）。④以0.508为miRNA-34a-5p相对表达量的最佳临界值诊断宫颈癌,其灵敏度为83.6%,特异度为79.4%,AUC为0.860,95%置信区间为0.803～0.917。结论　HR-HPV阳性宫颈癌患者癌组织和血清中miRNA-34a-5p表达下调,对宫颈癌诊断及预后监测具有一定的参考价值。     

The Urinary Concentration of Sialic Acid is Increased in Non-insulin-dependent Diabetic Patients with Microangiopathy: a Possible Useful Marker for Diabetic Microangiopathy

In order to investigate the relationship between urinary excretion of sialic acid and the severity of diabetic microangiopathy, urinary levels of sialic acid were determined in patients with non-insulin-dependent diabetes mellitus. The urinary molar ratio of sialic acid to creatinine in the diabetic patients was significantly higher than in the healthy controls (p < 0.01). Moreover, the urinary ratio was found to be gradually increased with the degree of diabetic microangiopathy. Urine molar ratio of sialic acid to creatinine in patients with proliferative diabetic retinopathy was significantly higher than in patients without retinopathy (p < 0.01). Urinary excretion in patients with macroproteinuria was also significantly higher than in patients without nephropathy (p < 0.01). Since urinary levels of sialic acid are proportionally increased with the severity of diabetic microangiopathy, the measurement of urinary sialic acid could become a useful biochemical means to monitor the degree of diabetic microangiopathy.     

2型糖尿病患者HbA1c水平与糖尿病视网膜病变DR分期的相关性

目的 探讨2型糖尿病患者HbA1c水平与糖尿病视网膜病变DR分期的相关性.方法 选取2019年11月—2020年8月就诊于承德市中心医院内分泌科,并诊断为2型糖尿病的患者265例,根据眼底检查结果分为非糖尿病视网膜病变(NDR)组(n=156)及糖尿病视网膜病变(DR)组(n=109).收集两组患者的临床资料包括性别、...     

miRNA-1236通过靶向FOXO3a调控结直肠癌细胞增殖的作用及机制研究

目的探讨miRNA-1236对结直肠癌细胞增殖的作用及其相关机制。 方法通过实时定量PCR检测转染miR-1236后miRNA-1236的表达,使用CCK-8以及克隆形成方法检测转染miRNA-1236后HCT116细胞的活性,ki67方法检测转染miRNA-1236后HCT116细胞增殖能力的变化,生物信息方法筛选miR-1236的潜在靶基因FOXO3a。Luciferase报告检测miRNA-1236与FOXO3a的结合位点,通过实时定量PCR方法检测转染miRNA-1236后FOXO3a基因的变化;通过免疫组化的方法检测结直肠癌患者组织中FOXO3a的表达,通过western blot检测低表达miRNA-1236后HCT116细胞中FOXO3a、PCNA以及Bax蛋白的变化。通过TCGA数据库检测FOXO3a基因在结直肠癌患者癌-癌旁,不同肿瘤阶段与不同性别中的表达。 结果本研究发现高表达miRNA-1236后HCT116细胞活性相对于阴性对照组明显增加,而低表达miRNA-1236后活性降低。低表达miRNA-1236后HCT116细胞增殖能力相对于阴性对照组明显降低。生物信息学预测miR-1236与FOXO3a有潜在的结合位点。采用Western blot以及逆转录实时定量PCR验证,过表达miRNA-1236后FOXO3a表达相对于阴性对照组明显降低,而低表达miRNA-1236后,FOXO3a表达明显升高,同时luciferase结果显示,FOXO3a是miRNA-1236的靶基因。并且,FOXO3a在结直肠癌症组织中明显降低。 结论miR-1236通过靶向FOXO3a的表达调节结直肠癌细胞HCT116增殖,同时为miR-1236成为诊断、预防以及治疗结直肠癌的生物学标记物提供理论基础。     

胰岛素、干细胞因子和Cajal间质细胞在糖尿病胃轻瘫中的作用

糖尿病胃轻瘫是糖尿病的常见并发症，严重影响患者的生活质量和血糖控制。胃肠道Cajal间质细胞在维持胃肠功能中发挥重要作用，糖尿病智组织中存在Cajal间质细胞数量和结构的异常改变，从而影响胃动力。胰岛素、干细胞因子对糖尿病胃肠道Cajal间质细胞的病变具有重要调控作用。本文就胰岛素、干细胞因子和Cajal间质细胞在糖尿病胃轻瘫中的作用作一综述。     

阿司匹林对人胃黏膜上皮细胞生长和occludin蛋白表达的影响及其机制研究

背景：研究显示ERK信号通路激活可促进细胞生长,上调紧密连接蛋白oceludin表达,而阿司匹林可抑制ERK的磷酸化激活。目的：探讨阿司匹林对人胃黏膜上皮细胞生长和occludin蛋白表达的影响及其可能机制。方法：建立人胃黏膜上皮细胞株GES-1单层细胞模型．M1Tr法检测阿司匹林（5-20mmol／L）和MEK抑制剂U0126（10～40μmol／L）对GES-1细胞的生长抑制作用。将GES-1细胞分为阿司匹林（8．78mmol／L）组、U0126（14．91μmol／L）组、联合组（U0126＋阿司匹林）和阴性对照组,倒置相差显微镜下观察细胞形态学变化,流式细胞术检测细胞凋亡情况,蛋白质印迹法检测P-ERK1／2、occludin蛋白表达。结果：阿司匹林和U0126均能剂量依赖性地抑制GES．1细胞生长,联合组细胞凋亡较两药单用更为显著,贴壁细胞数量减少更为明显．细胞变圆、悬浮。阿司匹林组、U0126组和联合组P-ERK1／2、occludin蛋白表达量均显著低于阴性对照组,U0126组和联合组降低更为明显。结论：阿司匹林可抑制人胃黏膜上皮细胞生长．下调occludin蛋白表达,其机制至少部分与抑制ERK信号通路激活有关。     

糖尿病大鼠钙化血管中Msx2和Wnt3a的表达

目的 研究糖尿病对血管钙化的影响及Msx2和wnt3a基因在钙化血管中的表达变化.方法 将48只雄性wistar大鼠随机分为四组:维生素D3和尼古丁诱导的单纯血管钙化组(n=12)、链脲佐菌素诱导的单纯糖尿病组(n=12)、链脲佐菌素联合维生素D3和尼古丁诱导的糖尿病合并血管钙化组(n=12)和正常雄性Wistar大鼠为正常对照组(n=12).测定大鼠血糖、血清胰岛素、总胆固醇和甘油三酯水平,以血管von Kossa染色、血管钙含量和碱性磷酸酶活性作为判断血管钙化程度的指标,测定大鼠血管中Msx2和wnt3a mRNA 的表达.结果 与正常对照组相比,单纯血管钙化组大鼠血管中Msx2和wnt3a mRNA 相对表达量有所升高(P<0.05),但血管钙含量和碱性磷酸酶活性无明显变化.与正常对照组及单纯血管钙化组相比,糖尿病合并血管钙化组大鼠的血管中,可见沿中膜弹力层内广泛分布的钙盐沉积,大鼠血管中钙含量和碱性磷酸酶活性以及血管内Msx2和wnt3amRNA 相对表达量明显增高(P<0.05).结论 糖尿病可以明显加速血管钙化的发生和发展.在糖尿病大鼠钙化血管中,骨形成过程中的转录因子Msx2和Wnt3a表达增高,提示血管钙化是一个类似于骨形成的过程,Msx2和Wnt3a 参与血管钙化病变的发生.     

糖尿病患者糖化血红蛋白检测结果及意义探析

目的 探讨糖尿病患者糖化血红蛋白检测结果及意义.方法 选择该院2019年1—12月收治的79例糖尿病患者作研究组,选择同期79名健康人群作对照组,对比两组糖化血红蛋白检查结果,了解研究组当中糖化血红蛋白和空腹血糖的比例、糖化血红蛋白和并发症发生的关系.结果 ①研究组空腹血糖、餐后2h血糖、糖化血红蛋白均高于对照组,差异...     

糖尿病酮症酸中毒患者心肌酶谱与心电图变化分析

目的通过观察糖尿病酮症酸中毒患者的心肌酶谱以及心电图改变,了解糖尿病酮症酸中毒对心肌的损害,进一步明确心肌酶谱以及心电图改变的意义。方法检测并比较44例糖尿病酮症酸中毒患者急性期以及缓解期的心肌酶谱,包括肌酸磷酸激酶、肌酸磷酸激酶同功酶、天门冬氨基转移酶以及肌钙蛋白Ⅰ;同时观察并比较患者的心电图变化。结果44例患者急性期的肌酸磷酸激酶、肌酸磷酸激酶同功酶以及天门冬氨基转移酶与缓解期比较明显升高,有统计学意义(P<0.05);急性期患者的肌酸磷酸激酶同功酶与肌酸磷酸激酶无相关性(r=0.12,P>0.05)。44例急性期患者心电图异常者28例(63.6%),主要异常是窦性心动过速18例(40.9%)以及ST-T改变14例(31.8%)。急性期心电图异常组患者的酸磷酸激酶、肌酸磷酸激酶同功酶以及天门冬氨基转移酶明显高于心电图正常组(P<0.05)。结论糖尿病酮症酸中毒患者心肌酶谱可以出现异常升高,糖尿病酮症酸中毒伴有心肌细胞的一过性非特异性损伤;糖尿病酮症酸中毒越重,患者越容易发生心律失常。     

年龄相关性白内障晶状体上皮细胞中miR-181a和SIRT1的表达关系及意义

目的探讨白内障晶状体上皮细胞微小RNA-181a(miR-181a)与沉默信息调节因子(SIRT)1的表达关系,研究其在年龄相关性白内障患者的诊治及病情评估方面的临床意义。方法选取诊断为年龄相关性白内障患者62例为白内障组,另选取同期近视矫正手术患者42例为对照组,利用RT q-PCR法,检测两组晶状体上皮细胞miR-181a与SIRT1表达水平;Pearson法测定miR-181a与SIRT1相关性;利用Lipofectamine 2000转染miR-181a模拟物和抑制剂,调节人晶状体上皮细胞miR-181a表达水平,RT q-PCR法检测SIRT1表达水平;酶联免疫吸附试验(ELISA)检测晶状体上皮细胞中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱氨酸(GSH)、谷胱氨酸过氧化物酶(GSH-Px)水平。结果与对照组相比,白内障组miR-181a表达水平显著升高,SIRT1表达水平显著降低(P<0.05);Pearson相关分析显示,miR-181a和SIRT1呈负相关关系(r=-0.719,P<0.05);miR-181a模拟物组SIRT1表达水平显著低于模拟物阳性对照组(P<0.05),miR-181a抑制剂组,SIRT1表达水平显著高于抑制剂阴性对照组(P<0.05);白内障组SOD、GSH、GSH-Px水平明显低于正常组(P<0.05),MDA水平明显高于正常组(P<0.05);miR-181a表达水平与SOD、GSH、GSH-Px因子呈负相关关系(r=-0.702,-0.739,-0.776),与MDA水平呈正相关关系(r=0.679);SIRT1表达水平与SOD、GSH、GSH-Px呈正相关关系(r=0.699,0.743,0.763),与MDA水平呈负相关关系(r=-0.684)。结论白内障晶状体上皮细胞中miR-181a高表达,SIRT1低表达,miR-181a可调控人晶状体上皮细胞中SIRT1表达;SIRT1低表达可促使晶状体上皮细胞凋亡,从而影响或导致了年龄相关性白内障的发病。     

Prevalence of Diabetic Retinopathy and Cataract in Adult Patients With Type 1 And Type 2 Diabetes in Russia

AIM: The aim of the study was to identify the prevalence of diabetic retinopathy (DR) and diabetic cataract (DC) in type 1 and type 2 diabetic patients within the Russian Federation. Also, the stage of DR at the time of its identification and the proportion of new cases diagnosed with DR or DC were to be determined. METHODS: A random sample of 7,186 adult patients with diabetes was screened for DR and DC using fundoscopy and fundus photography. Levels of HbA1c, total cholesterol, triglycerides, creatinine and urinary albumin excretion rate were assessed. RESULTS: In diabetic patients, the prevalence of DR and DC was 45.9% and 30.6%, respectively. These complications appeared significantly more frequently in patients with type 1 diabetes than in type 2 diabetes. The prevalence of background, preproliferative and proliferative DR among diabetic patients was 28.1%, 8.1%, and 6.7%, respectively. Patients with DR were older, had a longer duration of diabetes, higher HbA1c, elevated plasma total cholesterol, increased triglicerides, and higher systolic BP, compared with patients without DR. Microalbuminuria and proteinuria were more prevalent among patients with DR compared with non-DR patients. CONCLUSIONS: The results showed that diabetic retinopathy and cataract are wide-spread complications among diabetic patients in Russia. However, the disease course is more aggressive and accelerated in patients with type 1 diabetes than in those having type 2 diabetes. Therefore, it is important to prevent DR by identifying diabetes and signs of retinopathy at the earliest possible stage of progression for timely and adequate retina laser coagulation or surgical treatment, compensation of carbohydrate and lipid metabolism, and normalization of blood glucose and pressure.     

胶质瘤中miR-34a靶向作用于MAPK13 mRNA并抑制其表达

目的预测并验证miR-34a与促分裂原活化蛋白激酶13（MAPK13）的靶向作用关系,寻找胶质瘤基因治疗的新方法。方法将miR-34a相似物转染八胶质瘤细胞A172中,实时定量PCR检测miR－34a在胶质瘤中的表达。目的基因的mRNA水平用实时定量PCR来评估,蛋白水平分别用荧光素酶试验和蛋白印迹来评估。miR－34a与目的基因3′非翻译区（3′UTR）的结合用MAPK13报告实验确认。结果miR－34a相似物转染入胶质瘤细胞A172后在A172细胞中大量表达,实时定量PCR、蛋白印迹和免疫荧光证明miR－34a降低MAPK13的表达量,MAPK13荧光素酶报告实验显示,MAPK13荧光素酶活性被miR－34a降低。结论miR－34a通过与MAPK133′UTR区的特异结合作用发挥对胶质瘤责任基因MAPK13的靶向沉默作用,可能成为未来胶质瘤基因治疗的新选择。     

Association of Exosomal miR-34a with Markers of Dyslipidemia and Endothelial Dysfunction in Children and Adolescents with T1DM

Objective:Dyslipidemia and endothelial dysfunction are common disorders and major causative factors for atherosclerosis in patients with type 1 diabetes mellitus (T1DM). However, their pathophysiology in young patients with T1DM is still under evaluated. We aimed, for the first time, to assess the expression of exosomal micro-RNA 34a (miR-34a) in serum of children and adolescents with T1DM and correlate this expression with markers of dyslipidemia and endothelial dysfunction.Methods:The study included 120 T1DM patients and 100 control subjects. Assessment of miR-34a was performed using quantitative real-time polymerase chain reaction. Lipid profile was assessed on an automated analyzer and serum endoglin and intracellular adhesion molecule (ICAM) concentrations were measured using immunometric methods.Results:Relative expression of miR-34a and serum endoglin and ICAM concentrations were higher in patients than controls (p=0.001) and in patients with dyslipidemia (42 patients) compared to patients without dyslipidemia (78 patients) (p=0.01). Linear regression analysis revealed a strong independent association between exosomal miR-34a expression and total cholesterol, low-density lipoprotein, serum endoglin and serum ICAM after adjustment for other cofactors. The utility of miR-34a as an indicator for associated dyslipidemia was tested using receiver operator characteristic curve analysis which revealed area under the curve: 0.73 with confidence interval: 0.63-0.83 and p=0.001.Conclusion:This was the first study to show the altered expression of exosomal miR-34a among children and adolescents with T1DM. Moreover, association of miR-34a with markers of dyslipidemia and endothelial dysfunction was identified, suggesting that it could play a role in regulation of lipid metabolism and endothelial function in T1DM.     

胰腺癌组织c-MET表达与循环miR-34a、miR-449水平的相关性

目的 探讨胰腺癌组织织间质表皮转化因子(c-MET)的表达与循环miR-34a、miR-449水平的相关性及其临床意义。方法 收集2015年3月至2017年3月间嘉兴医学院附属第二医院行手术治疗并经病理证实的41例胰腺癌患者的临床资料。通过免疫组织化学染色检测胰腺癌组织及其匹配的癌旁正常胰腺组c-MET表达,根据测定结果将患者分为c-MET阳性组与c-MET阴性组。采集患者手术前和手术后3个月外周血,应用荧光定量PCR法检测循环miR-34a和miR-449水平。分析癌组织c-MET表达与患者临床病理参数、预后及循环miR-34a、miR-449水平的关系。分析循环miR-34a和miR-449水平对胰腺癌TNM分期、淋巴结转移和预后的评估价值。结果 胰腺癌组织c-MET阳性率明显高于癌旁正常组织(63.4%比24.4%),差异有统计学意义(P<0.05)。与c-MET阴性患者比较,c-MET阳性患者TNMⅢ/Ⅳ期者居多(73.1%比33.4%),淋巴结转移率高(76.9%比46.7%),生存时间短(29.5个月比35.0个月),生存率低(38.5%比53.3%),差异均有统计学意义(P值均<0.05)。术前c-MET阳性患者循环miR-34a、miR-449水平明显低于c-MET阴性患者(0.228±0.068比0.524±0.106、0.252±0.063比0.432±0.094,P<0.05),术后c-MET阳性患者循环miR-449水平仍明显低于c-MET阴性患者(0.414±0.088比0.512±0.114,P<0.05),但两组间miR-34a水平的差异无统计学意义。术前循环miR-34a、miR-449水平对胰腺癌TNM分期、淋巴结转移及预后有预测价值(P<0.05)。结论 miR-34a、miR-449靶向结合胰腺癌组织c-MET,有可能成为潜在的胰腺癌标志物。     

Genetic Variation of a Collagen IV α1-Chain Gene Polymorphism in Danish Insulin-dependent Diabetes Mellitus (IDDM) Patients: Lack of Association to Nephropathy and Proliferative Retinopathy

In insulin-dependent (Type 1) diabetes mellitus (IDDM) the development of nephropathy is partly due to genetic susceptibility. Previously one study has demonstrated a relationship between a HindIII restriction polymorphism of the collagen IV α1-chain gene and diabetic nephropathy. The aim of the present study was to evaluate such as association in a case–control study including 207 Danish IDDM patients: 116 with nephropathy (urinary albumin excretion rate (AER) > 300 mg 24 h⁻¹) and 91 without nephropathy (AER < 30 mg 24 h⁻¹). Using genomic DNA, HindIII restriction fragment length analysis revealed a bi allele polymorphism visualized by Southern hybridization with a cDNA probe recognizing the collagen IV α1-chain gene. No differences in genotype frequencies or allele frequencies were demonstrated comparing patients with and without nephropathy: p = 0.39 and p = 0.96, respectively. Neither were there any difference in genotype frequencies or allele frequencies when the patients were stratified according to the presence of proliferative retinopathy: p = 0.44 and p = 0.84, respectively. Pooling the diabetic groups revealed genotype frequencies and allele frequencies comparable to those found in 57 healthy unrelated Danish individuals. We conclude that in a Danish IDDM population a HindIII restriction polymorphism of the collagen IV α1-chain gene is not associated with diabetic nephropathy, diabetic retinopathy or with diabetes per se. © 1997 by John Wiley & Sons, Ltd.