小鼠miR-214重组慢病毒载体的构建和鉴定

目的构建携带miR-214的重组慢病毒表达载体,为研究miR-214的功能和作用机制奠定基础。方法从小鼠基因组DNA扩增miR-214基因,测序鉴定后克隆至pCDH-CMV-MCS-EF1-copGFP慢病毒载体上,测序鉴定、病毒包装后转染HEK293细胞,测定病毒滴度。将重组慢病毒载体感染HEK293细胞,应用萤光定量PCR法检测miR-214表达。结果重组慢病毒载体pCDH-CMV-miR-214-EF1-copGFP测序完全正确,pCDH-CMV-miR-214-EF1-copGFP感染的HEK293细胞中miR-214表达显著增强。结论成功构建小鼠miR-214重组慢病毒载体,并可在HEK293细胞中稳定表达出miR-214,为进一步进行miR-214的功能和机制研究奠定了试验基础。     

miRNA-155与恶性血液病

微小RNA（miRNA）是一系列在植物、动物及病毒中都可发现的非编码RNA,在转录后水平调控基因表达。miRNA-155（miR-155）是一种表达在造血细胞的miRNA,通过转录后调控靶基因的表达参与恶性血液病的发病机制。近来研究表明,miR-155通过影响参与细胞增殖、分化、凋亡的细胞信号转导途径,作为癌基因在恶性血液病的发生发展中起到重要作用。miR-155对恶性淋巴瘤、急性髓系白血病及骨髓增生异常综合征等恶性血液病的诊断治疗、预后具有重要临床意义。把能够下调miR-155表达的药物如反义寡核苷酸与经典的细胞毒性治疗联合应用,可有效控制恶性血液病的发展。本文综述了近年来miR-155对恶性血液病发病机制作用的研究进展并对其潜在的治疗靶点作用进行了展望。     

自发性高血压大鼠血液miRNAs表达分析

目的 本试验检测自发性高血压大鼠（SHR）血液中miR-448 5p、miR-128、miR 146a和miR-24 2＊四种小分子RNA（miRNA）的表达,探讨高血压疾病与miRNA表达水平相关性,为人类高血压诊断及治疗提供新的靶标.方法 应用荧光定量PCR方法对SHR大鼠和正常大鼠血液中miR 448-5p、miR-128、miR-146a和miR 24-2＊表达量进行检测.结果 与正常对照组相比,SHR大鼠血液中miR 448-5p、miR-128、miR-146a和miR-24 2＊的表达量均显著上调2倍以上.结论 miR-448 5p等四种miRNA表达量与高血压疾病呈相关性,为人类高血压患者血液诊断标记物及为高血压治疗的研究提供新的思路.     

闭塞性细支气管炎模型大鼠微小RNA差异表达谱分析

背景：目前对于气管移植后并发闭塞性细支气管炎尚无有效的治疗方法。miRNA分子水平机制的治疗策略在防治器官移植后并发症方面具有重要意义。 目的：分析大鼠原位气管移植模拟肺移植后发生闭塞性细支气管炎的微小RNA 表达谱变化。 方法：通过近交系大鼠原位气管移植,模拟建立肺移植闭塞性细支气管炎的动物模型并经病理学证实;通过微小 RNA芯片筛选出供体移植气管闭塞性细支气管炎组织中显著差异表达的微小 RNA,并选取差异表达显著的miR-146a、miR-155和miR-451进行相对定量研究,应用实时定量RT-PCR（RT-qPCR）法验证芯片结果的可靠性。 结果与结论：原位气管移植后4周经病理检查证实大鼠闭塞性细支气管炎模型成功建立。microarray检测获得29个与闭塞性细支气管炎相关的微小RNA,包括15个微小RNA表达显著上调,14个微小RNA表达显著下调,其中显著上调miR-146a、miR-155和显著下调miR-451的功能涉及免疫炎症反应等。提示微小RNA在肺移植后闭塞性细支气管炎病理进程中发挥着重要的调控作用。     

微小RNA与冠心病的研究进展

微小RNA(microRNA,miRNA)是内源性的大小在20～23核苷酸的一类非编码的RNAs,广泛存在于真核生物体内,并在进化中高度保守.其中一些miRNA在心血管系统中高度表达和特异表达,它们在冠心病的发病机制及治疗中也起重要的作用,其中miR-126与miR-21参与内皮细胞功能的调节,miR-15和miR-320的下调可预防缺血诱导的细胞凋亡,miR-221/miR-222、miR92a参与调节血管的再生,miR-29的下调可抑制梗死后的纤维化.本文简要概述近年来有关miRNA与冠心病研究的部分进展并进行相应的展望.     

辛伐他汀干预对高血压治疗的影响分析

目的总结我院2 a来高血压患者在降血压治疗基础上加用辛伐他汀干预治疗,了解辛伐他汀在降压治疗中的作用。方法将唐山市丰润区人民医院120例原发性高血压患者随机分为常规治疗组和辛伐他汀干预组,常规治疗组施行常规降压治疗,辛伐他汀干预组在降压基础上加用辛伐他汀干预治疗,分别观察疗效。结果两组中3级高血压患者治疗后血压比较差异有统计学意义。结论他汀类药物有进一步降低原发性高血压3级患者血压的作用。     

大鼠miR-1慢病毒表达载体的构建与鉴定

目的构建并鉴定大鼠miR-1慢病毒过表达系统。方法采用EcoRⅠ酶双酶切将目的基因从合成载体上酶切后连接入EcoRⅠ线性化慢病毒载体,连接产物转化后进行阳性克隆鉴定。成功构建的miR-1重组质粒与慢病毒辅助包装质粒及Lipofectamine 2000共转染293T细胞,产生慢病毒浓缩液并标定病毒滴度。重组慢病毒载体转染大鼠骨髓间充质干细胞（MSCs）,观察转染效率及miR-1表达水平。结果测序结果显示目的基因与Genebank序列完全一致;孔稀释法检测病毒滴度为3×108 TU/μL;以感染复数（MOI）50感染大鼠MSCs,感染效率达90%以上,聚合酶链反应显示miR-1高水平表达。结论成功构建大鼠miR-1慢病毒表达载体并可高效转染大鼠MSCs,为进一步研究miR-1基因的相关功能提供了合适的载体。     

MiR-126在脓毒症所致心肌损伤的作用研究

目的 探讨miR-126在脓毒症心肌损伤中的作用并探讨其机制.方法 在整体动物水平,将C57BL/6小鼠随机分为两组,即假手术组与脓毒症组.采用盲肠结扎穿刺术构建脓毒症小鼠模型,收集小鼠心脏组织,采用qRT-PCR检测小鼠心肌组织中miR-126的表达水平.在离体细胞水平,采用LPS刺激H9C2细胞,转染miR-126...     

microRNA-181b影响U87胶质瘤细胞干细胞对替莫唑胺化疗耐受性的实验研究

目的 本研究旨在建立miR-181b慢病毒载体感染U87胶质瘤干细胞模型后,检测其对TMZ化疗耐受性的变化,并探讨miR-181b对于恶性脑胶质瘤（GBM）辅助化学治疗的意义.方法 通过miR-181b慢病毒表达载体转染U87胶质瘤干细胞,提高miR-181b在U87胶质瘤干细胞中的表达.进一步通过RT-PCR、二次神经球形成实验、琼脂糖集落形成实验、MTT实验来分析miR-181b对于U87胶质瘤干细胞在TMZ化疗耐受性方面的作用.结果 miR-181b慢病毒表达载体转染U87胶质瘤干细胞后,U87胶质瘤干细胞中的miR-181b明显上调,而且U87胶质瘤干细胞的生长受到明显抑制;miR-181b可以加强TMZ抑制U87胶质瘤干细胞的生长的作用.结论 miR-181b可以加强TMZ抑制U87胶质瘤干细胞的生长,miR-181b可能对于提高U87胶质瘤干细胞对TMZ的敏感性,降低化疗耐受性起到一定作用.     

microRNA-185下调AKT1信号通路对淋巴母细胞淋巴瘤细胞增殖和凋亡的影响及相关的分子机制的研究

目的探讨miR-185对淋巴母细胞淋巴瘤(T-LBL)细胞增殖和致肿瘤性的影响及相关的分子机制。方法 q PCR检测人T-LBL组织和正常的胸腺组织中miR-185表达水平。miR-185模拟物及NC-miRNA转染Jurkat细胞,MTT法测定细胞增殖情况,q PCR和MTT法分别检测miR-185表达水平和细胞增殖情况,细胞流失术检测细胞凋亡。生物信息学软件Target Scan预测miR-185的靶作用位点,并通过q PCR、Western blot和荧光素酶活性实验证实miR-185的靶作用位点。慢病毒介导的miR-185过表达Jurkat细胞分别通过皮下注射的方式注射入裸鼠左右侧,皮下注射后4 d、8 d、12 d、16 d、20 d、24 d、28 d、32 d分别测量肿瘤体积,以探讨miR-185对体内肿瘤生长的影响。结果与人正常胸腺组织相比,T-LBL组织中miR-185表达显著下调(P0.05);MTT检测细胞增殖结果表明,与NCmiRNA转染组相比,miR-185转染组Jurkat细胞增殖速率显著下降(P0.05),且转染miR-185后,Jurkat细胞凋亡比例显著升高(P0.05);Target Scan生物学软件预测表明AKT1为miR-185潜在的靶基因,实验结果进一步证实AKT1的3'非翻译序列(3'UTR)是miR-185的直接靶点;裸鼠移植瘤试验结果表明,miR-185能抑制体内肿瘤生长。结论miR-185通过靶向作用于AKT1基因抑制Jurkat细胞增殖,其可能为T-LBL的临床治疗提供新的靶标。     

Can nasal continuous positive airway pressure decrease clinic blood pressure in patients with obstructive sleep apnea?

Obstructive sleep apnea (OSA) is commonly associated with systemic hypertension and now recognized as an independent risk factor for daytime hypertension. We aimed to study the short- and long-term effect of nasal continuous positive airway pressure (CPAP) in hypertensive and normotensive patients with OSA. Forty-six patients with moderated to severe OSA were treated with nasal CPAP and followed after one year of treatment. Clinic blood pressure, heart rate, and body weight were taken before and followed up for one year after beginning nasal CPAP. In this study 25 patients with OSA were found to have hypertension (54.3%). The hypertensive group showed a significant reduction in clinic blood pressure after nasal CPAP, whereas the normotensive group showed no changes. The subgroup of hypertensive patients with OSA who had no anti-hypertensive medication revealed a decrease in clinic blood pressure comparable to those with anti-hypertensive drugs. The heart rate was not significantly changed in any patients. There was no significant correlation between the decrease in body weight and the reduction in blood pressure. These results suggest that nasal CPAP alone might have a substantial blood pressure lowering effect in hypertensive patients with OSA. This effect could decrease the morbidity and mortality related to cardiovascular complications in patients with OSA.     

原发性高血压合并颈椎病患者倾斜后血压和心率的变化

目的 探讨原发性高血压(EH)合并颈椎病患者头高位倾斜后血压和心率的变化及可能机制.方法 EH 49例,根据临床症状,运用X光机、颈椎CT或MRI和椎动脉彩超等检查方法,确诊合并颈椎病23例(合并颈椎病组),不合并颈椎病患者26例(单纯EH组),测定2组患者血压、血脂、血糖,并测定头高位倾斜后的血压、心率.结果 合并颈椎痛组和单纯EH组头高位倾斜后血压变化差异有统计学意义[4(-21～14)mm Hg与-9(-27～-3)mm Hg,H=25.44,P<0.01],心率变化差异有统计学意义[2(-1-14)次/min与5(-2～11)次/min,H=9.30,P<0.01].结论 合并颈椎病患者头高位倾斜后血压不下降与交感神经兴奋可能有关.     

厄贝沙坦及倍他乐克对高血压患者动态血压的影响

目的观察厄贝沙坦及倍他乐克对高血压患者动态血压的影响。方法选择80例原发性高血压患者,分成厄贝沙坦组及倍他乐克组,每组40例。治疗前及治疗后3个月行24h动态血压检测,观察血压控制情况。结果治疗前两组患者血压无显著差异,经治疗后两组患者血压都有显著性下降,两组在治疗后24h夜均舒张压没有显著差异性,其余24h收缩压、24h舒张压、日间收缩压和舒张压、夜间收缩压均有显著差异性,厄贝沙坦组血压下降更明显。结论厄贝沙坦和倍他乐克对原发性高血压降压疗效均明显,厄贝沙坦组血压下降效果更佳。     

恒定磁场对不同高血压模型大鼠的选择性作用

目的探讨不同高血压种类和程度用磁疗康复的疗效评价 . 方法在恒定弱磁场的作用下观察正常大鼠、去甲肾上腺素引起血压升高的大鼠及肾性高血压模型大鼠血压及心率的变化 . 结果该磁场对正常 Wistar大鼠的血压、心率均无影响 ; 对去甲肾上腺素引起的中、轻度高血压有较好的抑制作用 , 且降压程度与时间、剂量呈正相关 ; 对肾性高血压模型大鼠则没有影响 . 结论恒定弱磁场对大鼠血压的生物学效应取决于实验动物的功能状态 . 提示高血压患者应用磁疗进行的保健和康复效果应与高血压种类和程度有关 .     

The urinary excretion of catecholamines and their derivatives in primary hypertension in man

1. The 24 h urinary excretion of adrenaline, noradrenaline, normetadrenaline, metadrenaline and vanilloylmandelic acid has been compared in 17 male normotensive subjects and 25 age-matched male hypertensive patients studied under similar in-patient conditions. 2. 24 h urinary metadrenaline was significantly lower in the hypertensive patients. With this exception, no significant differences were found between the two groups when the total 24 h excretion of free catecholamines and their metabolites was analysed. 3. Diurnal variation in free catecholamine excretion was found in both normotensive and hypertensive subjects. There was no corresponding variation in metabolite excretion. 4. No correlation could be established between systolic or diastolic blood pressure and the amounts of the catecholamines or their metabolites in the urine of either group. 5. The results are considered in the light of recent work demonstrating high plasma catecholamine concentrations in hypertension. They lend no support to the concept that excessive circulating catecholamines are responsible for the elevated blood pressure in essential hypertension.     

超声心动图评价老年高血压患者伴瓣膜病变对心功能的影响

目的通过超声心动图探讨老年高血压病患者伴心脏瓣膜退行性改变对心功能和血流动力学影响。方法根据126例老年高血压病患者心脏各瓣膜变化情况分为瓣膜病变组(36例)、无瓣膜病变组(90例),32例健康老人作为正常对照组。应用超声心动图检测3组患者左心室舒张末期内径(LVDD)、室间隔厚度(IVST)和左心室壁厚度(LVPWT),左心房面积(LAA)、左心室射血分数(LVEF)和肺动脉收缩压(PSAP),再行心电图检查心房颤动情况,并将3组数据进行组间比较分析。结果高血压无瓣膜病变患者,瓣膜反流改变以二尖瓣最多(占86.1%),反流程度重度者以三尖瓣反流最多;瓣膜钙化以主动脉瓣多见。瓣膜病变组与无瓣膜病变组以及正常对照组LVDD、LAA、LVEF、PASP以及心房颤动发生率比较差异有统计学意义,而在瓣膜病变组与无瓣膜病变组之间心室壁厚度差异无统计学意义。结论老年高血压患者合并心脏瓣膜改变是加重心功能损害的重要因素。     

Hemodynamic and creatine kinase changes after a 12-week equipment-based Pilates training program in hypertensive women

Exercise therapy is a conservative strategy to manage hypertension. The aim of this study was to investigate the effects of a 12-week equipment-based Pilates training program on the hemodynamics of medication-controlled hypertensive women. Forty-five women were classified into two groups: medication-controlled hypertensive (n = 30) or control (n = 15). To be eligible for the hypertensive group, participants had to have a clinical diagnosis of hypertension, be taking medication to control their blood pressure and not to have done any exercise activity in the previous three months. In the control group, the participants needed to have normal blood pressure levels and not to have done any exercise activity in the previous three months. All women participated in a 12-week equipment-based Pilates training program (2 × 60min sessions per week). Systolic and diastolic blood pressure (SBP, DBP), heart rate (HR) and creatine kinase levels (CK) were assessed before and after the program. SBP, DBP and heart rate were also assessed before and after each session. There was no statistically significant difference after Pilates training for systolic (normotensive MD 4.1, 95%CI -9.2 to 17.5, hypertensive MD 3.8, 95%CI -5.3 to 13.1) and diastolic (normotensive MD 5.8, 95%CI -2.0 to 13.7, hypertensive MD 4.0, 95%CI -4.3 to 12.4) blood pressure. Heart rate was reduced after Pilates training in both normotensive (MD 4.5, 95%CI 1.1 to 7.8) and medication-controlled hypertensive (MD 7.9, 95%CI 4.4 to 11.4) women. Creatine kinase activity was reduced after Pilates training in medication-controlled hypertensive women (p = 0.019). Blood pressure was not altered, but heart rate and creatinine kinase activity were reduced following 12 weeks of Equipment-based Pilates training.     

New controversies in hypertension: questions answered, answers questioned.

Despite multiple, interdisciplinary group recommendations, we are still on uncertain ground when it comes to treatment of most aspects of hypertension. Seven major areas of controversy include mild hypertension, the relevance of hypertension and lipids, hypertensive agents and electrolyte imbalance, treatment and regression of left ventricular hypertrophy, isolated systolic hypertension, ambulatory blood pressure monitoring and overtreatment of hypertension--the "j shaped curve." Although our knowledge of these aspects has advanced tremendously, significant doubts exist as to our present approach. Key publications are reviewed to evaluate our present knowledge and recommendations are made. The 1988 recommendations of the Joint National Committee on Detection, Evaluation and Treatment of Hypertension both answered and raised some questions regarding treatment of high blood pressure. We lack information on the treatment outcomes and many of us remain unconvinced that our present approach is the best we can do. Many other questions abound. Should the treatment of mild hypertension be as aggressive as it is at present or should systolic hypertension in the elderly be treated at all? There are striking variations and recommendations of other groups outside the United States which reaffirm our lack of evidence. Ideally, we ought to be able to reduce or abolish the recognized poor outcomes of treated hypertension: heart attack, heart failure, stroke, renal failure and retinopathy. Adequate control of blood pressure has gone a long way towards preventing stroke, accelerated hypertension and hypertensive encephalopathy. Congestive heart failure has also been reduced. There is a singular lack of evidence of the influence on either total mortality or morbidity from coronary events.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between a genetic predisposition to hypertension, blood pressure levels and pain sensitivity.

INTRODUCTION: The aim of this study was to determine whether the degree of blood pressure elevation and/or a genetic predisposition to hypertension have a major role in determining a reduced pain perception in hypertensives. The reasons underlying the relationship between blood pressure elevation and pain perception mechanisms are not completely understood. METHODS: One hundred and four untreated hypertensive patients (65 subjects with and 39 without a positive parental history of hypertension) together with a control group of 42 subjects (20 normotensive offspring of normotensive parents, and 22 normotensive offspring of hypertensive parents) were submitted to standard blood pressure evaluation, 24-h blood pressure monitoring and dental pain perception evaluation. RESULTS: Both pain threshold and tolerance were found to be higher in hypertensive than normotensive subjects (P < 0.0001 and P < 0.015, respectively). Positive significant correlations were found between both 24-h systolic and diastolic pressure and the pain perception variables. When a 2 x 2 ANOVA test was performed, factoring for the effects of both blood pressure status and family history of hypertension on pain sensitivity, a significant effect was revealed only for blood pressure status. Moreover, after controlling for blood pressure by a covariate analysis, no significant difference was found between the subjects with or without hypertensive parents as regards pain perception variables. CONCLUSIONS: Pain sensitivity is correlated to blood pressure levels whereas the parental history of hypertension per se does not affect the pain perception pattern. Thus, the degree of blood pressure elevation, more than a genetic predisposition to hypertension may influence the mechanisms leading to hypalgesia in hypertension.     

养肝清肝平肝法干预原发性高血压患者血压及血管紧张素Ⅱ的变化

背景:前期临床观察血压平胶囊对原发性高血压患者有确切的降压效果。目的:观察具有养肝清肝平肝作用的血压平胶囊对阴虚阳亢或肝火旺盛型原发性高血压的疗效及其对血管紧张素Ⅱ的影响。设计:随机对照病例观察。单位:湖南中医药大学第一附属医院心内科。对象:以1999-04/2000-04湖南中医药大学第一附属医院住院和门诊患者为观察对象,纳入标准:符合原发性高血压诊断标准,辨证属于阴虚阳亢或肝火旺盛的患者,能耐受和进行本试验者,并经精神紧张性测试符合精神紧张性高血压的患者,均知情同意。排除标准:严重心衰、类风湿关节炎等,及不能耐受或进行本试验者。符合标准者60例,男29例,女31例,数字表法随机分为血压平组和维拉帕米组两组,每组30例。方法:所有患者惊醒精神紧张性测试结果阳性者进入实验,血压平组给予血压平胶囊(由天麻、酸枣仁、汉防已等组成,0.43g/粒,湖南中医药大学第一附属医院制,卫药剂字05-047,批号990503)5粒/次,3次/d。维拉帕米组给予维拉帕米(40mg/片,上海黄河利亚制药有限公司制,批号980301B)80mg/次,3次/d。两组均服药4周,为1个观察疗程。①血压疗效:治疗前连续3次非同日上午的血压均值为治疗前血压,治疗后3次血压均值为治疗后血压,以此作为评定标准。②症状疗效:采用计分法,于治疗前后按主要症状如眩晕、头痛、腰膝酸软等程度分4级计分。③冷加压试验前后分别测定其血管紧张素Ⅱ(放免法)。采用降压疗效、临床症状疗效、临床疗效评定标准,并观察其对血管紧张素Ⅱ影响。主要观察指标:临床症状、冷加压试验前后的血压、心率及血管紧张素Ⅱ水平等。结果:入选原发性高血压患者60例,均进入结果分析。①血压平组治疗原发性高血压患者症状的总有效率稍高,但两组降压疗效的差异无显著性意义(P>0.05)。②两组心率治疗前后比较差异均有非常显著性意义(P<0.001),但维拉帕米组下降幅度更大,两组比较差异有显著性意义(P<0.05)。③两组基础和冷加压试验后收缩压与舒张压均有降低(P<0.001),组间差异均无统计学意义(P>0.05)。④两组临床疗效,组间差异无显著统计学意义(P>0.05)。⑤治疗后两组基础状态的血管紧张素Ⅱ浓度明显下降,差异有非常显著性意义(P<0.001);两组治疗后冷加压试验后血管紧张素Ⅱ浓度与治疗前比较均有下降,但差别无显著性意义(P>0.05)。结论:血压平胶囊降压平稳,从降压疗效、症状疗效等方面,均达到甚至超过维拉帕米的疗效,在一定程度上降低血浆血管紧张素Ⅱ的水平。适用于阴虚阳亢或肝火旺盛证、精神紧张性原发性高血压患者的降压治疗。