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Elevating mitochondrial reactive oxygen species by mitochondria-targeted inhibition of superoxide dismutase with a mesoporous silica nanocarrier for cancer therapy

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Nano Research 2014年第8期7卷 1103-1115页

作者： Yi Zhang Zhengyan Hu Guiju Xu Chuanzhou Gao Ren'an Wu hanfa zou Key Lab of Separation Sciences for Analytical Chemistry National Chromatographic R&A Center Dalian Institute of Chemical Physics Chinese Academy of Sciences Dalian 116023 China University of Chinese Academy of Sciences Beijing 100049 China Institute of CancerStem Cell Dalian Medical University Dalian 116044 China

In the intrinsic pathway of apoptosis, stresses of mitochondrial reactive oxygen species （mitoROS） might be sensed as more effective signals than those in cytosol, as mitochondria are the major sources of reactive oxygen species （ROS） and pivotal components during cell apoptosis. Mitochondrial superoxide dismutase （SOD2） takes the leading role in eliminating mitoROS, and inhibition of SOD2 might induce severe disturbances overwhelming the mitochondrial oxidative equilibrium, which would elevate the intracellular oxidative stresses and drive cells to death. Herein, we report a general strategy to kill cancer cells by targeted inhibition of SOD2 using 2-methoxyestradiol （2-ME, an inhibitor for the SOD family） via a robust mitochondria-targeted mesoporous silica nanocarrier （mtMSN）, with the expected elevation of mitoROS and activation of apoptosis in HeLa cells. Fe304@MSN was employed in the mitochondria-targeted drug delivery and selective inhibition of mitochondrial enzymes, and was shown to be stable with good biocompatibility and high loading capacity. Due to the selective inhibition of SOD2 by 2-ME/mtMSN, enhanced elevation of mitoROS （132% of that with free 2-ME） was obtained, coupled with higher efficiency in initiating cell apoptosis （395% of that with free 2-ME in 4 h）. Finally, the 2-ME/mtMSN exhibited powerful efficacy in targeted killing of HeLa cells by taking advantage of both biological recognition and magnetic guiding, causing 97.0% cell death with only 2 Dg/mL 2-ME/mtMSN, hinting at its great potential in cancer therapy through manipulation of the delicate mitochondrial oxidative balance.

关键词： mitochondria reactive oxygen species apoptosis mesoporous silicananoparticles drug delivery

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Effects of splice sites on the intron retention in histamine H_3 receptors from rats and mice

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Journal of Genetics and Genomics 2009年第8期36卷 475-482页

作者： Wenyong Ding Lin Lin Feng Ren hanfa zou Ziyuan Duan Jianwu Dai Department of Biochemistry and Molecular Biology Dalian Medical University Dalian 116044 China Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100080 China National Chromatographic Research and Analysis Center Dalian Institute of Chemical Physics Chinese Academy of Sciences Dalian 116023 China

In the alternative splicing, intron retention, of histamine H3 receptors in rats and mice, the short transcript isoforms that are excised alternatively spliced introns are easily detected in a very low level in rats and are undetectable in mice using the regular PCR protocol. The retained introns have common 5＇ splice site and different 3＇ splice sites. The detailed mechanism for the special alternative splicing remains largely unclear. In this study, we developed a minigene splicing system to recapitulate natural alternative splicing of the receptors and investigated the effects of 5＇ and 3＇ splice sites on intron retention in HeLa cells. Mutating weak 5＇ and 3＇ splice sites of the alternatively spliced introns toward the canonical consensus sequences promoted the splicing of the corresponding introns in rat and mouse minigenes. The effect of splice site strength was context-dependent and much more sigiaificant for the 3＇ splice site of the longer alternative intron than for the 3＇ splice site of the shorter alternative intron and the common 5＇ splice sites; it was also more significant in the rat minigene than in the mouse minigene. Mutating the 3＇ splice site of the longer alternative intron resulted in almost complete splicing of the intron and made the corresponding isoform to become the nearly exclusive transcript in the rat minigene.

关键词： histamine H3 receptor (H3R) altemative splicing intron retention splice site minigene

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Comprehensive analysis of the N and C terminus of endogenous serum peptides reveals a highly conserved cleavage site pattern derived from proteolytic enzymes

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Protein & Cell 2012年第9期3卷 669-674页

作者： Fangjun Wang Jun Zhu Lianghai Hu Hongqiang Qin Mingliang Ye hanfa zou CAS Key Lab of Separation Sciences for Analytical Chemistry National Chromatographic Research and Analysis CenterDalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023China

The human serum proteome is closely associated with the state of the *** peptides derived from proteolytic enzymes cleaving on serum proteins are widely studied due to their potential application in disease-specific marker ***,the reproducibility of peptidome analysis of endogenous peptides is significantly influenced by the proteolytic enzymes within body fluids,thereby limiting the clinical use of the endogenous *** comprehensively investigated the N and C terminus of endogenous peptides using *** cleavage site patterns of the N and C terminus and adjacent sites from all the identified endogenous peptides were highly conserved under different sample preparation conditions,including long-term incubation at 37℃ and pretreatment with repeated freeze-thaw ***,a distinguishable cleavage site pattern was obtained when a different disease serum was *** conserved cleavage site pattern derived from proteolytic enzymes holds potential in highly specific disease diagnosis.

关键词： human serum endogenous peptide N and C termini disease diagnosis

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Large-scale phosphoprotein identification by Ti4+-IMAC coupled online RP-SCX-RPLC in plants

Large-scale phosphoprotein identification by Ti4+-IMAC coupl...

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第十四届全国植物基因组学大会

作者： Xu Wang hanfa zou Junxian He School of Life Sciences and State Key Laboratory of Agrobiotechnology The Chinese University of Hong Kong Key Laboratory of Separation Sciences for Analytical Chemistry National Chromatographic R&A CenterDalian Institute of Chemical PhysicsChinese Academy of Sciences

Large-scale protein phosphorylation analysis by MS is emerging as a powerful tool in plant signal transduction ***,our current understanding of the phosphorylation regulatory network in plants is still very *** establ... 详细信息

关键词： protein phosphorylation phosphoproteomics phosphorylation motif Ti4+-IMAC Arabidopsis thaliana

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A strategy with label-free quantification of the targeted peptides for quantitative peptidome analysis of human serum

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Science China Chemistry 2010年第4期53卷 759-767页

作者： ZHU Jun, WANG FangJun, DONG XiaoLi, YE MingLiang & zou hanfa Key Laboratory of Separation Sciences for Analytical Chemistry National Chromatographic Research and Analysis Center Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China Key Laboratory of Separation Sciences for Analytical Chemistry Chinese Academy of Sciences Dalian China National Chromatographic Research and Analysis Center Chinese Academy of Sciences Dalian China Dalian Institute of Chemical Physics Chinese Academy of Sciences Dalian China

Peptidomics draws more and more attention in discovering useful biomarkers for early diagnosis of disease. However, there is lack of efficient quantification strategy in peptidome analysis. In this study, a strategy with label-free quantification of the targeted endogenous peptides based on peak intensity using μUPLC-Q-TOF-MS/MS was developed for quantitative peptidome analysis of human serum. Different amounts of standard BSA tryptic digesting peptides were added into the same serum extracts for evaluation of the developed strategy, and it was observed that the average relative error of the targeted peptides was 6.42%, which was superior to the result obtained directly by commercially available software PLGS. It was also demonstrated that this quantification strategy could obviously increase the detection sensitivity of the peptide by DDA analysis. Then, this strategy was applied to comparatively analyze the peptides extracted from the serum of HCC or breast cancer patients and healthy individuals, respectively. Peptides with charge states up to 5 and molecular weight over 4000 can be reliably identified and quantified. This quantitative analysis method based on μUPLC-Q-TOF-MS/MS exhibited superior sensitivity than that by MALDI-TOF-MS commonly used in peptidome analysis. Finally, some interesting endogenous peptides related to corresponding diseases were successfully obtained.

关键词： peptidomics label-free quantification hepatocellular carcinoma breast cancer human serum

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MONOLITHIC CAPILLARY COLUMNS FOR EFFICIENT SEPARATION OF BIOLOGICAL SAMPLES

MONOLITHIC CAPILLARY COLUMNS FOR EFFICIENT SEPARATION OF BIO...

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第十三届北京分析测试学术报告会暨展览会(BCEIA 2009)

作者： hanfa zou Fangjun WANG Renan WU Mingliang YE CAS Key Laboratory of Separation Science for Analytical Chemistry National Chromatographic R&A Center Dalian Institute of Chemical PhysicsChinese Academy of Sciences

Monolithic columns have been widely applied in capillary liquid chromatography(μLC)and capillary electrochromatography(CEC)due to their easy preparations,low back pressure,convective mass transfer process and high se...

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The experimental study of configurating an immunoabsorption column for hepatitis B virus

The experimental study of configurating an immunoabsorption ...

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中华医学会第十六次全国病毒性肝炎及肝病学术会议

作者： Qingwei Gao hanfa zou Liang Kong Desheng Yuan Dallian Dalian No.6 People’s Hospital 116001

Objective To configure an immunoabsorption column for hepatitis B *** Being activated by epichlorohydrin,the human antibody HBs-IgG was bonded to the carrier of agarose *** configuration process was as follows:the synthesis of epoxide matrix,the synthesis and activation of amino matrix,the synthesis of aldehydic matrix,the synthesis of immunoabsorption matrix,the end capping and reduction of unbonded aldehydic,the blocking of unbonded mass and the filling of the *** The bonded rate of activa-

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色谱研究的最新重要进展

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色谱 2010年第1期28卷 100-101页

作者：邹汉法中国科学院大连化学物理研究所辽宁大连116023

细内径毛细管柱自由溶液用于分离宽范围DNA片段凝胶电泳是分离DNA最高效的方法,但将凝胶注入微纳分离通道或细内径毛细管柱内存在很大的技术难度。为了克服这一技术难题,美国University of Oklahoma的Shaorong Liu和University of Texas... 详细信息

细内径毛细管柱自由溶液用于分离宽范围DNA片段凝胶电泳是分离DNA最高效的方法,但将凝胶注入微纳分离通道或细内径毛细管柱内存在很大的技术难度。为了克服这一技术难题,美国University of Oklahoma的Shaorong Liu和University of Texas at Arlington的***等在无胶、无管壁改性和无外加电场的情况下，

关键词： DNA片段色谱分离通道毛细管柱凝胶电泳外加电场宽范围凝胶注

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涂布溶剂对涂敷型手性固定相手性识别能力的影响

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分析化学 2000年第7期28卷 879-882页

作者：陈小明杨利邹汉法中国科学院大连化学物理研究所!国家色谱研究分析中心!大连116011

以微晶纤维素为原料，在非均相乙酰化条件下反应合成了微晶纤维素三醋酸酯。分别以二氯甲烷、三氯甲烷作涂布溶剂，采用减压蒸发溶剂法将其涂布在硅胶上制备成适宜于高效液相色谱（ＧＰＬＣ）上用的涂敷型手性固定相（ＣＳＰｓ），得到... 详细信息

以微晶纤维素为原料，在非均相乙酰化条件下反应合成了微晶纤维素三醋酸酯。分别以二氯甲烷、三氯甲烷作涂布溶剂，采用减压蒸发溶剂法将其涂布在硅胶上制备成适宜于高效液相色谱（ＧＰＬＣ）上用的涂敷型手性固定相（ＣＳＰｓ），得到的ＣＳＰｓ对反－２，３－二苯基环氧乙烷和吡喹酮均具有较好的手性识别能力。在二氯甲烷中添加一定比例的芳香族化合物、有机酸等作溶剂也得到了具有不同手性识别能力的ＣＳＰｓ，其中苯酚、硝基苯的影响尤为突出。

关键词：溶剂涂敷微晶纤维素二醋酸酯手性识别

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微透析高效液相色谱法研究金属酶与小分子的相互作用

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化学学报 2003年第1期61卷 95-99页

作者：郭明孔亮厉欣邹汉法中国科学院大连化学物理研究所国家色谱研究中心

将微透析(microdialysis)高效液相色谱法(highperformanceliquidchromatography,HPLC)联用技术应用于定量研究超氧化物歧化酶(superoxidedismutase)与组氨酸和组氨酸 -金属离子配合物的相互作用.实验结果表明:Cu2 Zn2 SOD酶不仅与组氨酸,而且还可以与组氨酸 Co(Ⅱ)和组氨酸 Ni(Ⅱ)相互作用,Cu2 Zn2 SOD酶中的部分Cu(Ⅱ)和Zn(Ⅱ)被诱导和置换出来,并相应地影响了酶的活性;组氨酸 -金属离子配合物的作用比组氨酸强,酶活性降低也更明显;酶中结合的外来金属离子的量比从酶上被诱导和顶替下来的金属离子的量要少。

关键词：高效液相色谱法金属酶小分子微透析配合物超氧化物歧化酶相互作用金属离子

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