阿托伐他汀对HMGB1诱导血管内皮细胞激活的影响

目的探讨阿托伐他汀能否抑制高迁移率族蛋白1(HMGB1)诱导的血管内皮细胞激活,阐明其潜在的分子机制。方法体外培养大鼠胸主动脉内皮细胞,分别用不同浓度的阿托伐他汀、HMGB1、TLR4特异性抑制剂CLI-095预处理内皮细胞,荧光定量分析中性粒细胞与内皮细胞的黏附活力;实时定量RT-PCR与Western blot分别检测TLR4、细胞间黏附分子1(ICAM-1)和E-选择素mRNA和蛋白的表达水平;电泳迁移率实验(EMSA)测定核因子κB(NF-κB)p65的DNA结合活性。结果阿托伐他汀(0.1～10μmol/L)呈剂量依赖性地抑制HMGB1诱导的血管内皮细胞活化。阿托伐他汀预处理能明显下调HMGB1诱导的TLR4 mRNA和蛋白表达水平(P均0.05);阿托伐他汀、CLI-095均能有效抑制HMGB1诱导的NF-κB p65的DNA结合活性以及ICAM-1和E选择素的表达水平(P均0.05)。结论阿托伐他汀可通过调节黏附分子(ICAM-1和E-选择素)的表达而显著抑制HMGB1诱导的血管内皮细胞活化效应,其机制可能与它抑制TLR4的表达及NF-κB激活有关。     

促血管生成素1和血管内皮生长因子基因合适剂量配比有效促进血管生成

目的在大鼠后肢缺血模型中,探讨联合转染促血管生成素1和血管内皮生长因子基因治疗对新生血管数量和侧支循环形成的影响,并观察不同基因剂量配比对新生血管形态和功能的影响。方法制备大鼠后肢血管闭塞病变模型,采用电脉冲法介导转基因,按不同剂量配比进行肌肉组织转染携带促血管生成素1基因的质粒和/或携带血管内皮生长因子基因的质粒。采用逆转录聚合酶链反应检测外源基因的表达、免疫组织化学染色检测缺血局部毛细血管和小动脉的数量和分布、通过后肢血管造影检测侧支循环建立的情况。结果单独转促血管生成素1基因或血管内皮生长因子基因治疗组血管生成数量略有增加,联合基因治疗组中,血管内皮生长因子质粒剂量为100μg时,随着促血管生成素1质粒剂量的增加,小动脉计数较同期空载质粒对照组分别增加0.90倍、1.40倍和1.45倍;当促血管生成素1质粒剂量为100μg时,随着血管内皮生长因子质粒剂量的增加,小动脉计数较同期空载质粒对照组分别增加1.90倍、1.07倍和1.39倍。血管通透性检测表明,随着血管内皮生长因子质粒剂量的增加,血管通透性渐增加,促血管生成素1质粒能降低新生血管的通透性;其中促血管生成素1质粒200μg联合血管内皮生长因子质粒100μg组在有效促进小动脉形成的同时血管通透性与正常对照组最为接近。结论联合血管生成素1和血管内皮细胞生长因子基因治疗能更有效促进小动脉形成,其中血管内皮生长因子质粒与促血管生成素1质粒比例为1:2时血管通透性最接近生理状态,是比较理想的联合基因治疗剂量配比。     

糖耐量减低患者血小板活化和血管内皮功能的变化

为观察糖耐量减低对血小板活化和血管内皮功能的影响,选择40例糖耐量减低患者,测定血小板α－颗粒膜蛋白浓度,并应用高分辨超声测量右肱动脉在静息时（基础内径）,反应性充血（流量介导血管舒张）,舌下含服硝酸甘油（硝酸甘油介导血管舒张）时的舒张期内径。取30例查体健康者作对照组,结果发现,糖耐量减低组空腹血糖与对照组无显著差异（P＞0．05）,而空腹时血小板α－颗粒膜蛋白较对照组显著升高（P＜0．01）,流量介导血管舒张时内径较对照组显著降低（P＜0．01）,而两组间硝酸甘油介导血管舒张内径和基础内径无显著差异（P＞0．05）。口服75g葡萄糖负荷后2h,糖耐量减低组血糖浓度显著高于对照组,血小板α－糖粒膜蛋白较空腹状态时显著升高（P＜0．01）,同时2h流量介导血管舒张时内径较空腹时进一步降低,差异显著IP＜0．01）,相关分析表明,糖耐量减低组空腹血糖与血小板α－糖粒膜蛋白和流量介导血管舒张时内径无显著相关,而糖负荷后,血糖变化程度与血小板α－颗粒膜蛋白变化程度显著正相关（P＜0．01）,与流量介导血管舒张时内径的变化程度显著负相关（P＜0．01）,血小板活化增强和血管内皮功能受损,并且在糖负荷后病程程度进一步加重。     

内皮抑素和血管内皮生长因子与2型糖尿病大血管病变的关系

目的探讨内皮抑素和血管内皮生长因子与2型糖尿病大血管病变发生的关系。方法用酶联免疫吸附测定法检测100例2型糖尿病患者(34例2型糖尿病无合并症、40例伴发一种大血管病变、26例伴发多种大血管病变)和30例正常对照者血清内皮抑素和血管内皮生长因子表达水平的变化。结果内皮抑素和血管内皮生长因子在2型糖尿病大血管病变各组(包括合并一种大血管病变组、合并两种以上大血管病变组)血清含量分别显著高于2型糖尿病无合并症和正常对照组(内皮抑素为32.4±15.6μg/L和35.1±20.2μg/L比11.2±8.6μg/L和9.9±6.7μg/L;血管内皮生长因子为133.5±36.8ng/L和302.1±52.4ng/L比90.2±42.4ng/L和81.3±33.5ng/L,P<0.01),两者在正常对照和2型糖尿病无合并症组间比较差异不显著;血管内皮生长因子在伴发一种大血管病变、多种大血管病变组间比较表达水平显著上调(133.5±36.8ng/L比302.1±52.4ng/L,P<0.01),而内皮抑素的表达差异不显著;在2型糖尿病伴发一种大血管病变组,内皮抑素和血管内皮生长因子成显著正相关(r=0.540,P<0.01)。结论血管内皮生长因子的表达水平与2型糖尿病大血管病变程度密切相关,内皮抑素和血管内皮生长因子可能以自稳态调节机制参与动脉粥样硬化的发生和发展。     

重型肝炎患者血清血管内皮细胞生长因子的含量及其临床意义

目的 :研究重型肝炎患者血清血管内皮细胞生长因子 (VEGF)的含量与重型肝炎预后的关系。方法 :采用ELISA定量检测55例重型肝炎患者 (其中急性重型肝炎 2例 ,亚急性重型肝炎 9例 ,慢性重型肝炎 44例 )及健康体检者血清VEGF含量。结果 :正常对照组、重型肝炎患者存活组、死亡组血清VEGF含量分别为 :44.73± 1 4 .71pg/ml,80 .48± 1 0 .47pg/ml,37.99± 1 2 .70pg/ml。提示重型肝炎患者存活组血清VEGF水平与正常对照组相比显著升高 (P <0 .0 5) ,死亡组略降低但无统计学意义 (P >0 .0 5)。结论 :(1 )VEGF与肝细胞再生密切相关 ;(2 )动态检测血清VEGF水平变化可以预测重型肝炎患者的预后     

葡萄糖对血管内皮细胞小凹蛋白1和血管内皮生长因子表达的影响

为了探讨高浓度葡萄糖损伤血管内皮细胞及其对小凹蛋白-1和血管内皮生长因子表达的影响。将人脐静脉内皮细胞株ECV304．分别培养在对照组和含5.5mmol／L、11.1mmol／L、22.0mmol／L、33.0mmol／L葡萄糖的培养基中。经葡萄糖培养24h后，噻唑蓝法测定细胞增殖活性，硝酸还原酶法测定培养上清液中一氧化氮浓度，免疫组织化学和免疫印迹方法检测细胞中小凹蛋白-1和血管内皮生长因子的表达。结果发现，随着葡萄糖浓度的增加，内皮细胞增殖活性呈浓度依赖性抑制(r=-0.776，P=0.000)；一氧化氮浓度呈浓度依赖性增加(r=0.698，P=0.000)；小凹蛋白-1和血管内皮生长因子为棕黄色颗粒，主要分布于胞浆中；血管内皮生长因子的表达呈浓度依赖性增加(r=0.645，P=0.009)；小凹蛋白-1的表达也呈浓度依赖性增加(r=0.808，P=0.000)。提示高糖可诱导血管内皮细胞的血管内皮生长因子和小凹蛋白-1的表达，此变化可能与糖尿病患者高糖致血管病变有关。     

Utility of Platelet Parameters in Serologically Proven Dengue Cases with Thrombocytopenia

Worldwide, there is a rise in the incidence of dengue infection associated with multiple serotypes. Epidemiological studies have reported a higher incidence of severe dengue in secondary dengue infections. A rapid fall in platelet count associated with an increase in hematocrit above the baseline is one of the warning signs of plasma leakage. This study was undertaken to determine the utility of platelet indices such as mean platelet volume (MPV), platelet distribution width (PDW) and plateletcrit in dengue fever cases with thrombocytopenia. A hospital-based cross-sectional study was carried out between April and September 2014, among dengue patients with thrombocytopenia using platelet histograms. The study population included all the laboratory confirmed cases of dengue infection with thrombocytopenia admitted at Kasturba Medical College, Manipal, Karnataka during the study period. The blood samples collected from serologically confirmed dengue patients with thrombocytopenia were analyzed using automated analyzer within 2 h of venipuncture. The platelet histograms (MPV, PDW, Plateletcrit) generated by the Beckman Coulter counter LH755? and LH780? series were assessed in dengue fever cases with thrombocytopenia. The mean platelet volume (MPV) was observed to be 9.01 fL (SD = 0.09). The mean platelet distribution width and median plateletcrit were 17.2% (SD = 0.98) and 0.47 (IQR 0.2–0.8) respectively. None of the study participants presented with bleeding manifestations. The present study revealed no significant changes of platelet parameters in dengue cases with thrombocytopenia.     

Vascular and Endothelial Function in Youth with Type 2 Diabetes Mellitus

Purpose of Review

This review aims to discuss the burden of type 2 diabetes in youth and summarize the studies that have utilized noninvasive techniques to assess early vascular disease in youth with type 2 diabetes.

Recent Findings

Noninvasive imaging modalities provide researchers with tools to investigate the vasculature in adolescents with type 2 diabetes. The data published to date consistently show adolescents with type 2 diabetes have greater vascular thickness and stiffness and worse endothelial function compared to their obese and lean peers.

Summary

As the prevalence of type 2 diabetes continues to increase adolescent youth, there is concern adolescents with type 2 diabetes are at risk to develop early onset cardiovascular disease and complications. Future studies need to address treatments that have the potential to improve or reverse vascular dysfunction and decrease the rate of cardiovascular disease and complications.

     

内皮祖细胞与糖尿病血管并发症的研究进展

糖尿病的血管并发症是糖尿病致残致死的主要原因,其病理基础为血管内皮功能失调,而内皮祖细胞是血管内皮修复的重要物质,且与糖尿病以及血管疾病密切相关,被认为是心血管疾病的预测因子。现就内皮祖细胞与糖尿病及其血管并发症之间的关系以及研究进展进行综述。     

高血压前期体检人群血管内皮功能及其与脉搏波传导速度的关系

目的 探讨高血压前期体检人群血管内皮功能情况及与高血压和正常人群的差别,并分析其与动脉硬化的相关性。方法 随机选取810例年龄43.06±8.54岁的体检人群作为研究对象,分为三组：高血压组、高血压前期组、正常组。以臂踝脉搏波传导速度（baPWV）和肱动脉血流介导的舒张功能（FMD）作为评估动脉硬化和内皮功能的指标。结果 高血压前期组baPWV比高血压组低（1419.98±209.18 cm/s比1487.92±316.99 cm/s,P＜0.001）,FMD比高血压组高（6.84%±2.64%比6.45%±2.46%,P＜0.001）,与正常组相比差异有统计学意义（分别为1243.33±170.52 cm/s和8.01%±2.78%）。在全人群、高血压组和高血压前期组中FMD与baPWV存在负相关（r分别为－0.316、－0.354、－0.266,P＜0.001）,但在正常组中两者无明显相关性（r＝－0.104,P＝0.097）。在高血压前期组中对影响baPWV的因素进行多元线性逐步回归分析,FMD进入回归方程。结论 在高血压前期人群中,血管内皮功能障碍已经开始,因此在高血压前期人群中开展血管内皮功能的检测和干预可以避免动脉硬化的进展和恶化。     

2型糖尿病合并糖尿病肾病脂联素水平变化及与血管内皮功能的关系

目的探讨糖尿病肾病患者血清脂联素水平的变化,及与血管内皮功能的关系。方法 50例无明显临床大血管并发症的2型糖尿病患者,按24 h尿白蛋白排出量分为正常白蛋白尿组、微量白蛋白尿组及大量白蛋白尿组。检测血清脂联素、可溶性血管细胞黏附分子1、生物化学指标、肱动脉内皮依赖性舒张功能(FMD)、含服硝酸甘油后肱动脉内皮依赖性舒张功能(NID)、心脏结构参数及颈动脉内膜-中膜厚度。结果大量白蛋白尿组脂联素水平是正常白蛋白尿组的4倍,是微量白蛋白尿组的2倍(P<0.01和P<0.05)。微量白蛋白尿组脂联素水平比正常白蛋白尿组升高(P<0.05)。脂联素/血肌酐在三组中有同样的变化(均P<0.05)。大量白蛋白尿组和微量白蛋白尿组可溶性血管细胞黏附分子1均高于正常白蛋白尿组(P<0.01和P<0.05)。大量白蛋白尿组FMD、NID随脂联素水平的增加而下降(P<0.05)。颈动脉内膜-中膜厚度在三组间无统计学差异。脂联素与可溶性血管细胞黏附分子1、24 h尿白蛋白排出量、血肌酐、左心室后壁厚度、内膜-中膜厚度呈正相关(r值分别为0.338、0.704、0.470、0.331、0.324,P<0.05),与FMD、NID呈显著负相关(r值为-0.397、-0.413,P<0.01)。结论糖尿病肾病伴随高脂联素血症,且与血管内皮功能损害密切相关。脂联素可作为糖尿病肾病患者早期血管内皮功能障碍的预测指标。     

血管内皮生长因子与自发性高血压大鼠血管重构的关系

目的探讨血管内皮生长因子与血管重构的关系。方法12只13周龄雄性自发性高血压大鼠(SHR组)作为观察组,12只同周龄雄性WKY大鼠作为正常血压对照组(WKY组)。分别于实验的第4、8周末每组处死大鼠各6只。采用酶联免疫吸附法测定血浆血管内皮生长因子浓度;放射免疫法测定颈动脉血管紧张素Ⅱ浓度;病理图象管理系统测定颈动脉管腔横截面积、内弹力层围绕面积、外弹力层围绕面积,评价内膜和中膜增生程度;免疫组织化学法检测颈动脉血管内皮生长因子蛋白表达。结果与WKY组比较,SHR组血浆血管内皮生长因子浓度明显下降,颈动脉血管内皮生长因子蛋白表达明显减弱,颈动脉血管紧张素Ⅱ浓度却显著升高(P<0.01),8周末这一作用更加显著(P<0.01);SHR组内膜增生较WKY组明显,中膜面积显著增大(P<0.01)。结论在血管重构过程中,血管内皮生长因子水平下降,提示血管内皮生长因子可能具有改善血管重构的作用。     

ACE2 Activation Confers Endothelial Protection and Attenuates Neointimal Lesions in Prevention of Severe Pulmonary Arterial Hypertension in Rats

Background

Angiotensin-converting enzyme 2 (ACE2), an ACE homolog, hydrolyzes angiotensin II and opposes its actions, and plays a protective role in the pathogenesis of pulmonary arterial hypertension (PAH). However, the underlying mechanisms involved in the effect of ACE2 on PAH are still uncertain. In this study, we observed the effects of ACE2 activation on endothelial dysfunction and vascular remodeling in the development of severe PAH in rats.

Methods

Severe PAH was induced by monocrotaline injection 1 week following left pneumonectomy, and ACE2 was activated by continuous injection of resorcinolnaphthalein. The PAH-related hemodynamics, pathological changes, and endothelium-dependent vasorelaxation were examined to assess the effects of ACE2 activation. In addition, the changes of the main components of the renin-angiotensin system were identified by ELISA or Western blotting.

Results

Severe PAH was established at 3 weeks and was characterized by high pulmonary arterial pressure (45 mmHg), significant right ventricular hypertrophy, neointimal occlusive lesions, and impaired endothelium-dependent relaxation in pulmonary arteries. Coadministration of resorcinolnaphthalein reduced pulmonary arterial pressure, right ventricular hypertrophy, and neointimal formation and shifted the endothelial-dependent responses toward values measured in normal rats. Theses changes were associated with an increase in ACE2 and angiotensin-(1–7) levels and a decrease in ACE and angiotensin II levels, in addition to a decrease in the ACE/ACE2 ratio and the angiotensin II/angiotensin-(1–7) ratio. The beneficial effects of resorcinolnaphthalein were abolished by A-779.

Conclusions

These findings suggested that ACE2 activation by resorcinolnaphthalein improved endothelial function and suppressed neointimal formation in the prevention of severe PAH by the mechanism of mediating the levels of the components of the renin-angiotensin system.     

The Association of Cytokines with Severe Dengue in Children

Background: Dengue virus infection is a major public health problem. A hypothesis put forward for severe dengue is the cytokine storm, a sudden increase in cytokines that induces vascular permeability. Previous studies and our recent meta-analysis showed that IL-6, IL-8, IFNγ, TNFα, VEGF-A and VCAM-1 are associated with dengue shock syndrome. Therefore, in this study we aim to validate the association of these cytokines with severe dengue. Methods & Findings: In a hospital based-case control study in Vietnam, children with dengue fever, other febrile illness and healthy controls were recruited. Dengue virus infection was confirmed by several diagnostic tests. Multiplex immunoassay using Luminex technology was used to measure cytokines simultaneously. A positive association with dengue shock syndrome was found for VCAM-1, whereas a negative association was found for IFNγ. Furthermore, multivariate logistic analysis also showed that VCAM-1 and IFNγ were independently correlated with dengue shock syndrome. Conclusion: IFNγ and VCAM-1 were associated with dengue shock syndrome, although their role in the severe dengue pathogenesis remains unclear. Additional studies are required to shed further light on the function of these cytokines in severe dengue.     

葡萄糖对人血管内皮细胞多元醇通路的激活作用及其机理

观察葡萄糖对人血管内皮细胞多元醇通路的影响 ,并探讨其作用机理。体外培养人脐静脉内皮细胞 ,加不同浓度葡萄糖或作用不同时间 ,采用高效液相色谱仪、硝酸还原酶法、生物化学检测及逆转录聚合酶链反应等方法测定内皮细胞山梨醇、一氧化氮、醛糖还原酶活性及醛糖还原酶基因mRNA。结果发现 ,经高浓度葡萄糖处理的人脐静脉内皮细胞山梨醇浓度明显高于对照组 (P <0 .0 5) ,一氧化氮浓度明显低于对照组 (P <0 .0 5)。醛糖还原酶基因mRNA水平及其活性均呈浓度及时间依赖性 ,但是内皮细胞经 2 2mmol L葡萄糖作用 48h或 44mmol L葡萄糖作用 2 4h后 ,醛糖还原酶基因mRNA水平及其活性均不再升高 ,而呈下降趋势 (P <0 .0 5)。结果提示 ,高浓度葡萄糖能引起内皮细胞功能改变 ,其机制可能是高浓度葡萄糖能增强醛糖还原酶基因的转录并提高其活性 ,从而活化多元醇通路。     

Liver Function Derangement in Patients with Severe Fever and Thrombocytopenia Syndrome

Background and AimsPatients with severe fever with thrombocytopenia syndrome (SFTS) commonly show liver function impairment. This study aimed to characterize the liver function indices in SFTS patients and investigate their association with mortality.MethodsClinical information and laboratory results of 459 laboratory-confirmed SFTS patients, including 78 deceased and 381 surviving patients, were retrospectively analyzed. To explore the infectivity of SFTS caused by novel Bunyavirus (SFTSV) in hepatocytes, Huh7 human hepatoma cells were infected with various concentrations of SFTSV in vitro.ResultsThe proportion of SFTS patients developing liver injury during hospitalization was 73.2% (336/459); the hepatocellular injury was the predominant type. The median time to occurrence of liver injury from disease onset was 8 d. Liver injury in the deceased group occurred earlier than that in the surviving group. Alanine aminotransferase (ALT) level between 2–5 times upper limit of normal (ULN) at 4–6 d and between 5–15 ULN at 7–12 d of disease course were independent predictors of mortality. Alkaline phosphatase (ALP) >2 ULN at 7–9 d and elevated ALP at 10–12 days after disease onset were risk factors for death. ALT and aspartate transaminase (AST) levels were correlated with lymphocyte count and platelet-to-lymphocyte ratio (PLR). Total bilirubin (TB), ALT, AST levels showed positive correlation with viral load. In the in vitro experiment, SFTSV infected and replicated inside Huh7 cells.ConclusionsLiver injury is common in SFTS patients. ALT and ALP were independent predictors of SFTS-related mortality. Frequent monitoring and evaluation of liver function indices are needed for SFTS patients.