NADPH氧化酶NtrbohD参与烟草悬浮细胞ABA诱导的H2O2快速产生过程

研究了烟草BY-2悬浮细胞在脱落酸（ABA）诱导下产生过氧化氢（H2O2）的机制,发现ABA能够显著诱导H2O2快速产生,且H2O2主要是由超氧化物歧化酶催化超氧阴离子产生的．ABA处理可导致烟草悬浮细胞质膜NADPH氧化酶活性以时间依赖的方式快速增加,其增加的幅度和时间与ABA诱导H2O2积累一致,而且,这些增加的效应能够被NADPH氧化酶抑制剂碘二苯和咪唑显著抑制,说明质膜NADPH氧化酶参与烟草悬浮细胞H2O2快速积累过程．另外,用RT-PCR和蛋白印迹技术分析了烟草质膜NADPH氧化酶基因NtrbohD的表达水平,发现该基因在mRNA和蛋白水平的表达受ABA上调,表明NtrbohD参与烟草悬浮细胞ABA诱导H2O2快速产生过程．结果说明,ABA能够诱导悬浮细胞通过NADPH氧化酶快速产生活性氧,提示NAD-PH氧化酶和H2O2可能是植物细胞中ABA信号转导途径的重要成分。     

p16与原发性肝细胞癌的研究进展

p16基因又称肿瘤抑制基因(MTS1),是近年来发现的一个重要的抑癌基因,在细胞周期中发挥重要的调节作用.现对p16基因的结构、p16蛋白的作用途径以及该基因突变与原发性肝细胞癌的关系进行综述.     

NADPH氧化酶在油酸诱导的H9c2心肌细胞凋亡中的作用

本研究旨在探讨NADPH氧化酶活化在油酸诱导的H9c2心肌细胞损伤中的作用.以体外培养H9c2心肌细胞为研究对象,用不同浓度(200,400,800μmol/L)油酸刺激H9c2心肌细胞(24,48,72 h),采用MTT法检测细胞活力;流式细胞术检测细胞内活性氧和凋亡指标;免疫印迹法检测细胞内Caspase 3,Bax,Bcl-2变化.结果表明,油酸刺激明显抑制细胞增殖、细胞内活性氧水平明显增加、Cleaved-caspase 3蛋白水平明显增加、Bcl-2/Bax蛋白水平明显降低;NADPH氧化酶抑制剂DPI预处理明显改善油酸所致H9c2心肌细胞增殖抑制、活性氧增加和凋亡.本研究结果提示NADPH氧化酶活化参与油酸所致H9c2心肌细胞凋亡.     

NADPH氧化酶活化介导七叶皂苷诱导人神经瘤母细胞凋亡

本研究旨在探究NADPH氧化酶活化所致氧化胁迫在七叶皂苷诱导SH-SY5Y神经母细胞瘤凋亡中的重要作用.以体外培养SH-SY5Y细胞作为研究对象,采用MTT法测定细胞增殖活力,流式细胞术测定细胞周期、细胞凋亡和活性氧指标,Western Blot测定凋亡蛋白Caspase 3变化.结果表明,七叶皂苷处理致显著细胞增殖抑制(P 0. 05)、细胞内活性氧水平明显增加(P 0. 05)、细胞周期阻滞于S期(P 0. 05)、细胞凋亡数目增加、Caspase 3蛋白切割水平显著增加(P 0. 05); NADPH氧化酶抑制剂DPI预处理明显逆转七叶皂苷所致的细胞增殖抑制、活性氧增加、周期阻滞和凋亡(P 0. 05).提示NADPH氧化酶活化所致氧化胁迫参与七叶皂苷所引起SH-SY5Y细胞的凋亡.     

miR-194-5p通过Wnt5a抑制肝星状细胞活化

为研究miR-194-5p对肝星状细胞(HSCs)活化的影响及作用机制,采用CCK-8、β-半乳糖苷酶染色、Transwell、划痕、集落形成实验和RT-qPCR、Western blot分别检测miR-194-5p对LX-2细胞生物学特性、细胞活化标志物和靶基因表达的影响,并用生物信息学和双荧光素酶报告实验确定miR-194-5p靶基因.结果显示：miR-194-5p抑制LX-2细胞增殖、细胞集落形成、迁移能力以及LX-2细胞中活化标志物的表达,促进LX-2细胞衰老;Wnt5a为miR-194-5p的直接靶基因.研究表明miR-194-5p通过靶定Wnt5a抑制LX-2细胞活化,miR-194-5p和Wnt5a可能是治疗肝纤维化新的靶点.     

p38MAPK信号传导通路及其与细胞凋亡

丝裂原活化蛋白激酶(MAPKs)是广泛表达的丝氨酸／酪氨酸激酶，在哺乳动物细胞多种信号转导通路中起重要作用，MAPKs有三个主要家族：ERKs。JNKs和p38MAPKs。p38信号通路是MAPK通路的一重要分支，它在炎症、细胞应激、凋亡、细胞周期和生长等多种生理和病理过程中起重要作用。四种已知的p38异构体包括p38a、p38p、p38y和p386。多年来已发现p38MAPK通路可以由应激包括高渗、热休克、放射线和其他应激反应活化。因此，p38MAPK通路参与了多种刺激引起的信号级联反应，表明它在引起多种细胞反应中起重要作用，并且，p38在细胞凋亡和多样性变化中也显示调节效应。     

慢性粒细胞白血病急变与bcr／abl，p53,p16基因相关性研究

应用RT-PCR、PCR-SSCP方法对33例慢性粒细胞白血病（Chronic myeloid leukemia,CML）进行了bcr/abl,p53,p16基因检测，发现33例CML中32例bcr/abl基因阳性（97.0%）。22例慢性期无p53基因突变，而11例急变组中一例外显子5和一例外显子7发生p53基因突变，且均发生于急粒变组，2例出现p16基因纯合缺失，且均发生在急淋变组，33例CML均未见p16基因突变，结果提示：bcr/abl基因与CML发生有关，而p53基因突变可能与CML急粒变有关，p16纯合缺失可能与CML急淋变有关。     

p70S6K结构与功能研究进展

p70核糖体蛋白S6激酶（p70S6K)是mTOR的主要下游作用靶物,在翻译起始、蛋白合成、细胞周期、细胞迁移等方面具有重要作用。本文综述了p70S6K的结构和功能,这对于了解p70S6K以及其调控的相关信号通路分子具有重要意义。     

ASPP家族对p53诱导细胞凋亡发挥调节作用的研究进展

ASPP(apoptosis stimulating protein of p53)即p53凋亡刺激蛋白家族,是科学家们近年来发现的新的蛋白家族.该家族包含有三个成员:ASPP1,ASPP2和iASPP.它们均可以与p53相结合,从而正向或负向地调节p53诱导细胞凋亡的作用,成为肿瘤治疗的新靶点.     

盐及低温胁迫对油菜ROS和抗氧化酶活性的影响

以超强抗寒冬油菜陇油6号和抗寒性较弱的天油2号为材料,探究在盐胁迫和低温胁迫下活性氧(ROS)和抗氧化酶活性的变化以及烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶、促分裂原活化蛋白(MAP)激酶级联途径对ROS和抗氧化酶变化的影响.结果表明,盐胁迫和低温胁迫均增加了油菜w(H2O2)和c (·OH),且伴随着超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性的升高;经二苯基碘、U0126和二甲基硫脲预处理后再胁迫,与单独胁迫相比,陇油6号和天油2号油菜中w(H2O2)、c (·OH)和SOD、POD、CAT、APX活性均有所下降,表明MAP激酶级联途径、NADPH氧化酶均参与了盐和低温胁迫对ROS和抗氧化酶活性的诱导过程.     

14Mev中子的~(64)Zn(n,2n)~(63)Zn反应截面测量

本文用活化法分别以~(56)Fe(n,P)~(56)Mn和~(63)Cu(n,2n)~(62)Cu反应截面为标准,测量了14.61Mev中子的~(64)Zn(n,2n)~(63)Zn反应截面,其值分别为190.1±5.3mb和196.3±7.7mb。又以~(27)Al(n,P)~(27)Mg反应截面为标准,测量了14.13Mev、14.61Mev和14.85Mev三个能点的~(64)Zn(n,2n)~(63)Zn反应戡面,其值分别为116.0±8.7mb、189.4±14.2mb和197.2±14.8mb。活化生成核发射的γ射线用高纯锗(HPGc)γ谱议进行测量。     

O(3 P)与C2H2反应机理的量子化学研究

用量子化学计算方法对O(^3P)与C2H2的反应进行了研究．在HF／6—311 G(d，p)，HF／6—311 G(3df，3pd)，MP2／6—311 G(d，p)和MP2／6—311 G(3df，3pd)计算水平上，优化了反应势能面上各驻点的几何结构．通过内禀反应坐标(IRC)计算和振动分析．对反应过渡态进行了确认，并确定了反应机理．     

40Ar/39Ar同位素体系及其在地学上的应用

40Ar/39Ar同位素体系大体经历了3个技术阶段常规 40Ar/39Ar定年技术、40Ar/39Ar分步加热定年技术和40Ar/39Ar激光显微探针定年技术.研究表明可以通过大气中40Ar/39Ar比值随时间的变化窥探地球去气作用的时间和强度;40Ar/39Ar法不仅有助于研究岩浆冷却过程和花岗岩侵位机制等岩浆动力学的历史和壳幔演化,而且对研究热液成矿年龄也是很有效方法之一;在一定的地质条件下,沉积地层时代越老,40Ar/39Ar值越高;当今40Ar/39Ar年代法已为全球环境演变研究提供了一种强有力的工具.     

Involvement of NADPH oxidase NtrbohD in the rapid production of H2O2 induced by ABA in cultured tobacco cell line BY-2

The mechanisms for the production of hydrogen peroxide （H2O2） induced by abscisic acid （ABA） were investigated in suspension culture cells of tobacco BY-2 cells. The results showed that the immediate generation of H2O2, which was mainly derived from super-oxide dismutase-catalyzed dismutation of superoxide radical, was significantly induced by ABA. Furthermore, treatment of the cultured tobacco cells with ABA resulted in a time-dependent quick increase in plasma membrane （PM） NADPH oxidase activity, which coin- cided on time and magnitude with the elevation in ABA-induced accumulation of H2O2. Moreover, these enhanced effects were pro- nouncedly inhibited by two NADPH oxidase inhibitors, diphenylene iodonium and imidazole, suggesting that PM NADPH oxidase is involved in the rapid accumulation of H2O2 in cultured tobacco cells. In addition, analysis of the expression level of NtrbohD, a PM NADPH oxidase gene in tobacco, by RT-PCR and protein gel blot revealed that the gene at both mRNA and protein levels was upregulated by ABA, indicating that NtrbohD participates in the ABA-stimulated rapid production of H2O2 in tobacco culture cells. Taken together, these findings suggest that ABA induces the rapid accumulation of reactive oxygen species via NADPH oxidase in sus-pension culture cells of tobacco, and that NADPH oxidase and H2O2 appear to be important components in ABA signal transduction pathway in plants.     

The voltage dependence of NADPH oxidase reveals why phagocytes need proton channels

The enzyme NADPH oxidase in phagocytes is important in the body's defence against microbes: it produces superoxide anions (O2-, precursors to bactericidal reactive oxygen species). Electrons move from intracellular NADPH, across a chain comprising FAD (flavin adenine dinucleotide) and two haems, to reduce extracellular O2 to O2-. NADPH oxidase is electrogenic, generating electron current (I(e)) that is measurable under voltage-clamp conditions. Here we report the complete current-voltage relationship of NADPH oxidase, the first such measurement of a plasma membrane electron transporter. We find that I(e) is voltage-independent from -100 mV to >0 mV, but is steeply inhibited by further depolarization, and is abolished at about +190 mV. It was proposed that H+ efflux mediated by voltage-gated proton channels compensates I(e), because Zn2+ and Cd2+ inhibit both H+ currents and O2- production. Here we show that COS-7 cells transfected with four NADPH oxidase components, but lacking H+ channels, produce O2- in the presence of Zn2+ concentrations that inhibit O2- production in neutrophils and eosinophils. Zn2+ does not inhibit NADPH oxidase directly, but through effects on H+ channels. H+ channels optimize NADPH oxidase function by preventing membrane depolarization to inhibitory voltages.     

紫龙金对BGC-823细胞增殖的抑制作用及对p16~(INK4a)启动子活性的影响

利用记数法和Brdu脉冲标记法研究了紫龙金对细胞增殖的影响,western blot检测结果表明:紫龙金(3,4mg.mL-1)处理可明显抑制人胃癌BGC-823细胞的生长、提高BGC-823细胞中p16的表达水平.进一步利用含有p16INK4a启动子片段(-967~-165区域)及荧光素酶报告系统的载体pGL3-Basic-p16N研究了紫龙金对p16INK4a启动子活性的影响及其作用的分子机制,结果表明,紫龙金可能通过提高p16INK4a启动子活性而促进p16的表达,从而抑制细胞的增殖.     

L-Fuzzy拓扑空间中的T21/3分离性

在LF拓扑空间中定义T_(2~(1/3)),ST_(2~(1/3))和层T_(2~(1/3))分离性,讨论与其他分离性的关系,论证了它们是L-好的推广,并研究了它们的一些性质.     

Optical Properties of Ru(bipy)_2(dppx)~(2 ) in Different Environments

0Introduction Withtheincreasinginterestinginthestudyoftheinter actionbetweenpolypyridylruthenium(Ⅱ)complexesandDNA,ligandsderivedfromvariousmodificationof2,2′bipyridine(bipy)and1,10phenanthroline(phen)havebeen developed.Oneofthemostinterestingobservationsisthedis coveryofthemolecular“lightswitch”complexes[1].Theyarenotphotoluminescentinwaterbutdoemitinnonaqueoussol ventsorinthepresenceofDNA.Atfirst,thecomplexesRu(bipy)2(dppz)2 andRu(phen)2(dppz)2 (dppz=dipyrido[3,2a:2′,3′c]phena zine…     

Involvement of NADPH oxidase NtrbohD in the rapid production of H2O2 induced by ABA in cultured tobacco cell line BY-2

The mechanisms for the production of hydrogen peroxide (H₂O₂) induced by abscisic acid (ABA) were investigated in suspension culture cells of tobacco BY-2 cells. The results showed that the immediate generation of H₂O₂, which was mainly derived from superoxide dismutase-catalyzed dismutation of superoxide radical, was significantly induced by ABA. Furthermore, treatment of the cultured tobacco cells with ABA resulted in a time-dependent quick increase in plasma membrane (PM) NADPH oxidase activity, which coincided on time and magnitude with the elevation in ABA-induced accumulation of H₂O₂. Moreover, these enhanced effects were pronouncedly inhibited by two NADPH oxidase inhibitor, diphenylene iodonium and imidazole, suggesting that PM NADPH oxidase is involved in the rapid accumulation of ₂O₂ in cultured tobacco cells. In addition, analysis of the expression level of NtrbohD, a PM NADPH oxidase gene in tobacco, by RT-PCR and protein gel blot revealed that the gene at both mRNA and protein levels was upregulated by ABA, indicating that NtrbohD participates in the ABA-stimulated rapid production of H₂O₂ in tobacco culture cells. Taken together, these findings suggest that ABA induces the rapid accumulation of reactive oxygen species via NADPH oxidase in suspension culture cells of tobacco, and that NADPH oxidase and H₂O₂ appear to be important components in ABA signal transduction pathway in plants.