高血压病血瘀证患者血清对内皮细胞功能及AMPK激活的影响

目的:探索高血压病血瘀证患者血清对内皮细胞功能及AMPK活性的影响.方法:高血压病血瘀证患者血清刺激人脐静脉内皮细胞(HUVECs)24 h,以高血压病非血瘀证患者血清作为对照.采用一氧化氮(NO)、内皮素(ET-1)检测试剂盒检测NO和ET-1的含量.Western blot法检测磷酸化一氧化氮合酶(e NOS)、总e NOS、细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)、磷酸化AMPK及总AMPK的表达.定量PCR法检测肿瘤坏死因子(TNF-α)、白细胞介素-1β(IL-1β)的表达.结果:高血压病血瘀证患者血清处理24 h后,显著降低NO含量、抑制e NOS的活性、升高ET-1的含量,升高ICAM-1、VCAM-1的表达,升高单核细胞黏附率,升高内皮细胞TNF-α、IL-1β的表达水平,与高血压病非血瘀证患者血清相比,有统计学差异(P0.05).此外,高血压病血瘀证患者血清显著降低蛋白激酶AMPKα的磷酸化水平,与高血压病非血瘀证患者血清相比,有统计学差异(P0.05).结论:高血压病血瘀证患者血清能诱导内皮功能障碍和炎症反应,其机制可能与抑制AMPK的活性有关.     

鱼腥草素钠对人脐静脉内皮细胞NO合成及NOS活力的影响

目的:研究鱼腥草素钠对脂多糖(LPS)诱导的人脐静脉内皮细胞(HUVECs)NO合成及一氧化氮合酶(NOS)活力的影响.方法:采用15μg/ml LPS作用于HUVECs,共同孵育12h建立炎性损伤细胞模型.考察不同剂量鱼腥草素钠对各组HUVECs细胞存活率、培养上清液中LDH活力、NO含量、内皮型NOS(eNOS)及诱导型NOS(iNOS)表达的影响.结果:鱼腥草素钠可明显提高HUVECs细胞存活率,增加NO的分泌与释放,提高eNOS与降低iNOS活力,从而保护脂多糖诱导的HUVECs炎性损伤.结论:鱼腥草素钠对脂多糖诱导损伤的内皮细胞具有保护作用,其机制与其促进内皮细胞NO分泌与释放,增强eNOS活性和表达,改善血管内皮功能状态相关.     

北五味子多糖通过ERK1/2通路对AngⅡ诱导的心肌成纤维细胞增殖的作用机制

目的观察北五味子多糖(polysaccharide of Schisandra chinensis,SCP)对血管紧张素Ⅱ(AngiotensinⅡ,AngⅡ)诱导的心肌成纤维细胞(CFb)增殖作用,并探讨其作用机制.方法以培养的新生Wistar大鼠乳鼠CFb为实验模型,实验分为对照组、模型组、3个药物剂量组.采用胰酶消化、差速贴壁法培养CFb;四氮唑盐(MTT)比色法检测CFb增殖;羟脯氨酸(Hyp)法测定胶原含量;分光光度计测定CFb中超氧化物歧化酶(SOD)活力、丙二醛(MDA)水平;硝酸还原酶法和分光光度法分别测定不同干预条件下CFb培养液中一氧化氮合酶(NOS)活性及一氧化氮(NO)水平;免疫组化技术检测CFb中细胞外调节蛋白激酶(ERK1/2)蛋白表达.结果 SCP能够显著抑制AngⅡ诱导的CFb增殖(P0.05或P0.01),降低Hyp含量,提高SOD活力,降低MDA水平(P0.05,P0.01),提高NOS活性及NO水平(P0.05,P0.01),减低ERK1/2蛋白表达(P0.05,P0.01).结论 SCP可通过抑制ERK1/2信号通路提高NOS活性,并升高NO水平,增强抗AngⅡ诱导的CFb增殖能力.     

艳山姜挥发油对TGF-β_1诱导内皮细胞氧化应激损伤的保护作用研究

通过体外离体培养人脐静脉内皮细胞(HUVECs),建立转化生长因子-β_1(TGF-β_1)诱导的血管内皮细胞氧化应激损伤模型,观察艳山姜挥发油(EOFAZ)对TGF-β_1诱导的HUVECs氧化应激损伤的保护作用,并初步分析其机制.实验结果显示TGF-β_1显著引起内皮细胞氧化应激损伤,艳山姜挥发油能够明显抑制内皮细胞存活率降低、细胞内ROS水平升高及细胞迁移能力增强,并上调Nrf2蛋白的表达.上述结果表明,艳山姜挥发油对TGF-β_1诱导的HUVECs氧化应激损伤具有保护作用,其机制可能与其调控Nrf2蛋白的表达相关.     

新鲜冰冻血浆（FFP）通过AMPK/Akt/eNOS通路促肺内皮细胞一氧化氮释放及机制研究

探讨新鲜冰冻血浆(FFP)对人肺正常微血管内皮细胞(HPMECs)一氧化氮(NO)释放的影响及其机制.采用NO/Nitrite/Nitrate分析法检测了体外培养内皮细胞HPMECs经FFP处理后不同时间点细胞上清NO含量.结果显示:与培养基空白对照组相比,FFP显著增加内皮细胞NO生成;采用磷酸化蛋白激酶抗体芯片和免疫印迹方法筛选和鉴定FFP处理后内皮细胞相关蛋白激酶磷酸化,结果为FFP显著增加内皮细胞AMPKα1,Akt1和eNOS蛋白的磷酸化.进一步分别采用Compound C(AMPK抑制剂),LY294002(PI3K/Akt抑制剂)或L-NAME(NOS抑制剂)预处理细胞,阻挡AMPKα1/Akt1/eNOS信号转导通路.结果显示上述三种抑制剂均能抑制FFP诱导eNOS激活和NO生成,表明AMPKα1/Akt1/eNOS信号转导通路介导FFP诱导NO分泌参与血管保护.     

H2S对ox-HDL致内皮细胞损伤的保护作用

探讨硫化氢对氧化高密度脂蛋白（ox-HDL）致人脐静脉内皮细胞（HUVECs）损伤的保护作用.方法 体外培养HUVECs细胞,应用形态学观察法和噻唑兰（MTT）比色法检测细胞活性,试剂盒测定培养液中乳酸脱氢酶(LDH) 及丙二醛(MDA)含量.结果 ox-HDL组细胞损伤明显,经硫化氢低、中、高剂量组的干预,受损情况均有较明显改善,细胞存活率增加,且培养液中LDH及MDA含量降低,与对照组比较差异有显著性(P<0.05).结论 硫化氢对ox-HDL所致内皮细胞损伤具有一定的保护作用,其机制可能与降低氧自由基抗氧化作用有关.     

红景天苷抑制Hcy诱导的血管内皮细胞氧化应激损伤的实验研究

研究红景天苷(SAL)对同型半胱氨酸(Hcy)诱导体外培养的人脐静脉内皮细胞(HUVECs)氧化应激损伤的保护作用及可能机制.选择3-8代生长状态良好的HUVECs用于实验,采用SAL预孵育HUVECs 2h后,再加入Hcy(1 mmol/L)共孵育12 h诱导内皮细胞氧化应激损伤.采用MTT和LDH法分析细胞损伤,荧光探针法DCFH-DA分析细胞内活性氧(ROS)水平,酶联免疫吸附法分析细胞内丙二醛(MDA),超氧化物歧化酶(SOD)的含量,qRT-PCR技术检测Nrf2、HO-1、NQO1的基因表达.结果显示,Hcy显著引起血管内皮细胞损伤,而10、50μmol/L的红景天苷干预给药后,能够提高Hcy诱导的细胞存活率,明显降低LDH、MDA的水平,增加胞内SOD活性.与Hcy处理组相比,SAL干预给药能够降低Hcy所致细胞内ROS的增高,上调Nrf2、抗氧化酶基因HO-1、醌氧化还原酶NQO1的mRNA表达.本研究表明,红景天苷有效抑制Hcy诱导的HUVECs氧化应激损伤,其作用与红景天苷增强细胞清除自由基能力,调控Nrf2信号有关.     

递增负荷运动通过NO信号通路降低高血压机制的探讨

为探讨大负荷运动通过NO信号通路降低高血压的可能机制,作者利用大鼠高血压模型进行大负荷运动后,再以酶化学方法检测大鼠血液中一氧化氮合酶(NOS)活性变化以及用反转录多聚酶链式反应检测大鼠大动脉组织中NO和NOS的mRNA水平;还以免疫印迹实验检测大鼠大动脉组织中VEGF蛋白水平的变化及动脉血管组织Akt,PI3K水平.结果显示,模型组心脏中NOS活性及NO的mRNA水平显著高于对照组(P<0.05),模型组心脏中NOS及NO的蛋白表达水平也显著高于对照组(P<0.05),模型组心脏组织中激活型Akt,PI3K水平高于对照组,而总Akt,PI3K蛋白水平没有明显增高.以上结果表明,大负荷运动可增强组织中的Akt,PI3K活性,从而调节血液中NOS活性,调节血液中NO水平,并最终降低高血压.     

瓜蒌皮水提物通过PI3K/Akt/eNOS信号通路抑制缺血缺氧大鼠原代心肌细胞的凋亡

以激活磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/内皮型一氧化氮合酶(eNOS)信号通路、抑制心肌细胞凋亡为切入点，探究瓜蒌皮水提物(TP-W)保护心肌缺血的机制.分离大鼠心肌细胞，通过缺氧气体+无血清培养液培养制造缺血缺氧损伤模型，以模拟急性心肌缺血时的体内心肌细胞环境；同时添加PI3K特异性抑制剂(LY294002)和TP-W处理细胞，利用倒置相差显微镜观察细胞形态，噻唑兰法、末端标记法分别检测细胞存活率及凋亡率，免疫荧光联合免疫印迹方法定位、定量地检测细胞内PI3K、Akt及eNOS蛋白表达及磷酸化水平.结果显示：对上述缺血缺氧损伤的心肌细胞，TP-W可显著改善其生长状态、提高存活率、降低凋亡率；同时显著上调上述3种蛋白表达水平及磷酸化水平，但磷酸化水平均可被LY294002处理显著削弱.可见，激活PI3K/Akt/eNOS信号通路、抑制心肌细胞凋亡可能是TP-W抗心肌缺血的重要机制，也是中药“开胸除痹”的关键生物学内涵.     

蛋白激酶D1对心肌梗死大鼠心肌组织eNOS及Ang1表达的调控作用

目的:探讨蛋白激酶D1(PKD1)对大鼠心肌梗死后心肌组织内皮型一氧化氮合酶(e NOS)及促血管生成素1(Ang1)表达调控的影响.方法:采用冠状动脉左前降支结扎造模方法复制大鼠心肌梗死模型,术后存活达2d以上的心肌梗死大鼠随机被分为模型组、PKD1处理组与CID755673处理组,每组8只.应用逆转录聚合酶链式反应(RT-PCR)分析PKD1及其阻断剂CID755673对大鼠心肌组织中e NOS及Ang1 mRNA表达的影响,应用免疫组化法和免疫印迹法分析e NOS及Ang1的蛋白表达.结果:和心肌梗死模型组相比较,PKD1处理组大鼠心肌组织中e NOS及Ang1的mRNA和蛋白表达水平显著升高(P0.01);和PKD1处理组比较,CID755673处理组大鼠心肌组织中e NOS及Ang1的mRNA和蛋白表达水平显著下降(P0.01),接近于模型组水平.结论:PKD1显著上调大鼠心肌梗死后受损心肌组织中e NOS及Ang1的表达.     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that