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Atomic Force Microscopy-based Cell Nanostructure for Ligand-conjugated Quantum Dot Endocytosis

Acta Biochimica et Biophysica Sinica 2006年第9期38卷 646-652页

作者： Yun-Long PAN~1* Ji-Ye CAI~2 Li QIN~3 Hao WANG~4 1 The First Affiliated Hospital of jinan University,guangzhou 510632,china 2 college of Life Science and Technology,jinan University,guangzhou 510632,china 3 department of histology and embryology,medical college of jinan University,guangzhou 510632,china 4 Information and Optoelectronic Science and Engineering.South china Normal University,{14. 510631,china The First Affiliated Hospital of jinan Universityguangzhou 510632 China college of Life Science and Technology Jinan UniversityGuangzhou 510632 China department of histology and embryology Medical College of Jinan UniversityGuangzhou 510632 China School for Information and Optoelectronic Science and Engineering South China Normal UniversityGuangzhou 510631 China

While it has been well demonstrated that quantum dots (QDs) play an important role inbiological labeling both in vitro and in vivo,there is no report describing the cellular nanostructure basis ofreceptor-mediated ***,nanostructure evolution responses to the endocytosis of transferrin(Tf)-conjugated QDs were characterized by atomic force microscopy (AFM).AFM-based nanostructureanalysis demonstrated that the Tf-conjugated QDs were specifically and tightly bound to the cell receptorsand the nanostructure evolution is highly correlated with the cell membrane receptor-mediated ***,confocal microscopic and flow cytometry results have demonstrated the specificity anddynamic property of Tf-QD binding and *** found that the internalization of Tf-QD is linearlyrelated to ***,while the nanoparticles on the cell membrane increased,the endocytosis was stillvery active,suggesting that QD nanoparticles did not interfere sterically with the binding and function ***,ligand-conjugated QDs are potentially useful in biological labeling of cells at a nanometerscale.

关键词： quantum dot endocytosis nanostructure atomic force microscopy

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JTE-522-induced apoptosis in human gastric adenocarinoma cell line AGS cells by caspase activation accompanying cytochrome C release,membrane translocation of Bax and loss of mitochondrial membrane potential

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World Journal of Gastroenterology 2002年第2期8卷 217-223页

作者： Hong-Liang Li Xiao-Hong Li Jun-Hua Lü Xian-Da Ren,department of Pharmacology,jinan University Pharmacy college,guangzhou 510632,Guangdong Province,china Dan-Dan Chen,department of Cardiology,First Affiliated Hospital,Zhongshan University,guangzhou 510089,Guangdong Province,china Hai-Wei Zhang,department of Pathology,jinan University medical college,guangzhou 510632,Guangdong Province,china Cun-Chuan Wang,department of laparoscopic surgery,First Affiliated Hospital,jinan University medical college,guangzhou 510632,Guangdong Province,china department of Pharmacology Jinan University Pharmacy College Guangzhou 510632 Guangdong Province China. tsam@***

AIM: To investigate the role of the mitochondrial pathway in JTE-522-induced apoptosis and to investigate the relationship between cytochrome C release, caspase activity and loss of mitochondrial membrane potential (Deltapsim). METHODS: Cell culture, cell counting, ELISA assay, TUNEL, flow cytometry, Western blot and fluorometric assay were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanism. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Caspases 8 and 9 were activated during apoptosis as judged by the appearance of cleavage products from procaspase and the caspase activities to cleave specific fluorogenic substrates. To elucidate whether the activation of caspases 8 and 9 was required for the apoptosis induction, we examined the effect of caspase-specific inhibitors on apoptosis. The results showed that caspase inhibitors significantly inhibited the apoptosis induced by JTE-522. In addition, the membrane translocation of Bax and cytosolic release of cytochrome C accompanying with the decrease of the uptake of Rhodamin 123, were detected at an early stage of apoptosis. Furthermore, Bax translocation, cytochrome C release, and caspase 9 activation were blocked by *** and Z-IETD-CHO. CONCLUSION: The present data indicate a crucial association between activation of caspases 8, 9, cytochrome C release, membrane translocation of Bax, loss of Deltapsim and JTE-522-induced apoptosis in AGS cells.

关键词： Adenocarcinoma Stomach Neoplasms Amino Acid Chloromethyl Ketones Anti-Inflammatory Agents, Non-Steroidal Apoptosis Benzenesulfonates Caspases inhibitors Cyclooxygenase Inhibitors Cysteine Proteinase Inhibitors Cytochrome c Group Enzyme Activation Humans In Situ Nick-End Labeling Membrane Potentials Mitochondria Oxazoles Proto-Oncogene Proteins Proto-Oncogene Proteins c-bcl-2 Research Support, Non-U.S. Gov't Tumor Cells, Cultured bcl-2-Associated X Protein

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Proteomics-based Identification of Proteins with Altered Expression Induced by 12-O-Tetradecanoylphorbol 13-acetate In Nasopharyngeal Carcinoma CNE2 Cells

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Acta Biochimica et Biophysica Sinica 2005年第2期37卷 97-106页

作者： Pei-Zhou JIANG Ming GAN Hua HUANG Xin-Ming SHEN Shuang WANG Kai-Tai YAO Cancer Research Institute.Southern medical University,guangzhou 510515,china department of histology and embryology.Southern medical University,guangzhou 510515,china department of Parasitology,Zhongshan medical college.SUN Yat-sen University,guangzhou 510080.china Cancer Research Institute Southern Medical UniversityGuangzhou 510515 China department of histology and embryology Southern Medical UniversityGuangzhou 510515 China department of Parasitology Zhongshan Medical College SUN Yat-sen UniversityGuangzhou 510080 China

Nasopharyngeal carcinoma(NPC)is a malignancy with high incidence in Southern china andSouth-East *** studies indicate that chemical carcinogen promoters,such as 12-O-tetradecanoylphorbol-13-acetate(TPA),are important ... 详细信息

关键词： 12-O-tetradecanoyl-phorbol-13-acetate nasopharyngeal carcinoma proteomics apoptosis

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Harmine induces apoptosis in HepG2 cells via mitochondrial signaling pathway

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Hepatobiliary & Pancreatic Diseases International 2011年第6期10卷 599-604页

作者： Ming-Rong Cao,Qiang Li,Zhi-Long Liu,Hui-Hui Liu,Wei Wang,Xiao-Li Liao,Yun-Long Pan and Jian-Wei Jiang department of General Surgery,First Affiliated Hospital,jinan University,guangzhou 510632,china department of Anesthesiology,First Affiliated Hospital,guangzhou University of TCM,guangzhou 510632,china department of Biochemistry,medical college of jinan University,guangzhou 510632,china department of General Surgery First Affliated Hospital Jinan University Guangzhou 510632 China department of Anesthesiology First Affliated Hospital Guangzhou University of TCM Guangzhou 510632 China department of Biochemistry Medical College of Jinan University Guangzhou 510632 China

BACKGROUND:Harmine has antitumor and antinociceptive effects,and inhibits human DNA *** no detailed data are available on the mechanisms of action of harmine in hepatocellular *** study aimed to investigate the effects of harmine on proliferation and apoptosis and the underlying mechanisms in the human hepatocellular carcinoma cell line ***:The proliferation of HepG2 cells was determined by the cell counting kit-8 (CCK-8) assay and the clone formation *** morphology of HepG2 cells was examined using fluorescence microscopy after Hoechst 33258 staining Annexin V/propidium iodide (PI) was used to analyze apoptosis and PI to analyze the cell *** blotting was used to assess expression of the apoptosis-regulated genes Bcl-2,Bax,Bcl-xl,Mcl-1,caspase-3,and caspase-9 Mitochondrial transmembrane potential (Ψ m) was determined using ***:Harmine inhibited the proliferation of HepG2 cells in a dose-dependent *** 33258 staining revealed nuclear fragmentation and chromosomal condensation,cell shrinkage,and attachment loss in HepG2 cells treated with *** percentage of the sub/G1 fraction was increased in a concentration-dependent manner,indicating apoptotic cell *** staining showed that harmine changed the cell cycle distribution,by decreasing the proportion of cells inG0/G1 and increasing the proportion in S and G2/*** induced apoptosis in a concentration-dependent manner,with rates of 20.0%,32.7% and 64.9%,***-1 revealed a decrease in Ψ *** of HepG2 cells was associated with caspase-3 and caspase-9 activation,down-regulation of Bcl-2,Mcl-1,and Bcl-xl,and no change in ***:Harmine had an anti-proliferative effect in HepG2 cells by inducing *** signal pathways were involved in the *** cancer-specific selectivity shown in this study suggested that harmine is a promising novel drug for human hepatocellular carcinoma.

关键词： hepatocellular carcinoma harmine Bcl-2 protein caspase-3 apoptosis

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Changes of NF-kB,p53,Bcl-2 and caspase in apoptosis induced by JTE-522 in human gastric adenocarcinoma cell line AGS cells:role of reactive oxygen species

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World Journal of Gastroenterology 2002年第3期8卷 431-435页

作者： Hong-Liang Li Xiao-Hong Li Yan-Qing L Chun-Ling Ye Xian-Da Ren,department of Pharmacology,jinan University Pharmacy college,guangzhou 510632,Guangdong,china Dan-Dan Chen,department of Cardiology,First Affiliated Hospital,Zhongshan University,guangzhou 510089,Guangdong,china Hai-Wei Zhang,department of Pathology,jinan University medical college,guangzhou 510632,Guangdong,china department of Pharmacology Jinan University Pharmacy College Guangzhou 510632 Guangdong China.

AIM: To identify whether JTE-522 can induce apoptosis in AGS cells and ROS also involved in the process, and to investigate the changes in NF-kB, p53, bcl-2 and caspase in the apoptosis process. METHODS: Cell culture, MTT, Electromicroscopy, agarose gel electrophoresis, lucigenin, Western blot and electrophoretic mobility shift assay (EMSA) analysis were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanisms. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Lucigenin assay showed the generation of ROS in cells under incubation with JTE-522. The increased ROS generation might contribute to the induction of AGS cells to apoptosis. EMSA and Western blot revealed that NF-kB activity was almost completely inhibited by preventing the degradation of IkBalpha. Additionally, by using Western blot we confirmed that the level of bcl-2 was decreased, whereas p53 showed a great increase following JTE-522 treatment. Their changes were in a dose-dependent manner. CONCLUSION: These findings suggest that reactive oxygen species, NF-kB, p53, bcl-2 and caspase-3 may play an important role in the induction of apoptosis in AGS cells after treatment with JTE-522.

关键词： I-kappa B Proteins Adenocarcinoma Apoptosis Benzenesulfonates Caspases Cell Division DNA-Binding Proteins Humans NF-kappa B Oxazoles Proto-Oncogene Proteins c-bcl-2 Reactive Oxygen Species Research Support, Non-U.S. Gov't Stomach Neoplasms Tumor Cells, Cultured Tumor Suppressor Protein p53

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Effect of heparin on apoptosis in human nasopharyngeal carcinoma CNE2 cells

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Cell Research 2001年第4期11卷 311-315页

作者： LI HONG LIANG , KAI HE YE , HAI WEI ZHANG , YING RU LUO , XIAN DA REN, AI HUA XIONG, RUI SITU department of Pharmacology ,Pharmacy college, department of Pathology medical college, jinan University, guangzhou 510632, china department of Pharmacology Pharmacy College Jinan University Guangzhou China department of Pathology medical college Jinan University Guangzhou China

In order to study the mechanism of the effect of heparin on apoptosis in carcinoma cells, the nasopha- ryngeal carcinoma cell line CNE2 was used to identify the effect of heparin on apoptosis associated with the expression of c-myc,bax, bcl-2 proteins by use of Hoechst 33258 staining, terminal deoxynucleotidyl transferasemediated dUTP nick-end labeling (TUNEL), agarose gel electrophoresis, and flow cytometry, as well as Western blot analysis. The results showed that heparin induced apoptosis of cNE2 cens including the morphologic changes such as reduction in the volume, and the nuclear chromatin condensation, as well as the 'ladder pattern' revealed by agarose gel electrophoresis of DNA in a concentration-dependent manner. The number of TUNEL-positive cells was dramatically increased to 33.6+1.2% from 2.8+0.3% by treat- ment with heparin in different concentrations (10~40 kU/L). The apoptotic index was increased to 32.5% from 3.5% by detecting SubG1 peaks on flow cytometry. Western blot analysis showed that levels of bcl-2, bax and c-myc were significantly overexpressed by treatment with the increase of heparin concentrations. These results suggest that heparin induces apoptosis of CNE2 cells, which may be regulated by differential expression of apoptosis-related genes.

关键词： Heparin agarose gel electrophoresis flow cytometry apoptosis protein kinase C(PKC)

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Preparation of CaS:Eu^(2+) Phosphor by Microwave Heating Method and its Luminescence

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Chinese Chemical Letters 2000年第9期11卷 843-846页

作者： Peng Jun LIU Ying Liang LIU department of Chemistry, jinan University, guangzhou 510632 department of Chemistry, Hainan Normal college, Haikou 571158 department of Chemistry Hainan Normal College Haikou 571158 China department of Chemistry Jinan University Guangzhou 510632 China

This is the first report of using the microwave heating technique to synthesize calcium sulphide activated by europium whose structure is determined as the face-centered cubic by conventional X-ray powder diffraction method. The phosphor has maximum excitation peaks located at 280 urn and 560 urn and the maximum emission of the phosphor is 630 nm. When the concentration of Eu^(2+) in CaS increases from I .0 × 10^(-5) to l.0 × 10^(-2) mole per mole host, the body colour of the calcium sulphide activated with europium changes from white, through light-red to pink to deep-red. The phosphor obtains the longest afterglow at the concentration of 0.1% Eu^(2+)doped and is a kind of good material excited by sunlight.

关键词： Microwave radiation heating calcium sulphide europium luminescence.

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Neutral sulfate berberine modulates cytokine secretion and increases survival in endotoxemic mice

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Acta Pharmacologica Sinica 2006年第9期27卷 1199-1205页

作者： Fei LI Hua-dong WANG~2 Da-xiang LU Yan-ping WANG Ren-bin QI Yong-mei FU Chu-jie LI department of Pathophysiology,medical college of jinan University,guangzhou 510632,china department of Pathophysiology Medical College of Jinan University Guangzhou China

Aim:Berberine is thought to be an immunomodulator,so the present study aimedto investigate the effect of berberine on mortality,lung and intestine injury inendotoxemic mice,and the mechanism of its ***:Mice were chal-lenged with lipopolysaccharide(LPS,28 mg/kg,ip),and neutral sulfate berbefinewas administrated *** was monitored every 12 h,and his-tology of the lungs and intestine as well as the plasma tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),interleukin-12(IL-12),IL-10,and nitric oxide(NO)levels were ***:Pretreatment with 50 mg/kg neutral sulfate ber-berine once a day for 5 days significantly decreased the mortality rate and attenu-ated tissue injury of the lungs and small intestine in mice challenged with *** stimulated a marked increase in plasma levels of TNF-α,IFN-γ,IL-12,IL-10,and *** administration of berberine significantly reduced plasma TNF-α,IFN-γ,and NO levels,but did not suppress plasma IL-12 levels in mice exposed ***,pretreatment with neutral sulfate berbefine augmented IL-10secretion stimulated by LPS in ***:Pretreatment with neutral sul-fate berberine attenuates tissue injury and improves survival in endotoxemic mice,which may be mediated,at least in part,by the inhibition of pro-inflammatorymediator production and upregulation of IL-10 *** findings mightprovide a new strategy for the treatment of endotoxemia.

关键词： lipopolysaccharide berberine tumor necrosis factor-α interleukin-10 interleukin-12 interferon-γ nitric oxide mice

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中药臭灵丹中3,5-二羟基-6,7,3′,4′-四甲氧基黄酮对人鼻咽癌CNE细胞凋亡的影响及机制

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生物化学与生物物理进展 2011年第3期38卷 254-261页

作者：曹长姝沈伟哉李药兰王辉高明雅暨南大学医学院解剖学教研室广州510632 暨南大学药学院中药及天然产物研究所广州510632 暨南大学中药药效物质基础及创新药物研究所广东省重点实验室广州510632 暨南大学医学院微生物及免疫学教研室广州510632

中药臭灵丹中黄酮类化合物3,5-二羟基-6,7,3′,4′-四甲氧基黄酮(3,5-hydroxy-6,7,3′,4′-tetramethoxyflavone,HTMF)体外对多种肿瘤细胞有抑增殖作用,但机制尚未完全清楚.为探讨HTMF对人鼻咽癌CNE细胞凋亡的影响,用MTT法检测HTMF对CN... 详细信息

中药臭灵丹中黄酮类化合物3,5-二羟基-6,7,3′,4′-四甲氧基黄酮(3,5-hydroxy-6,7,3′,4′-tetramethoxyflavone,HTMF)体外对多种肿瘤细胞有抑增殖作用,但机制尚未完全清楚.为探讨HTMF对人鼻咽癌CNE细胞凋亡的影响,用MTT法检测HTMF对CNE细胞株的增殖抑制率,倒置显微镜观察HTMF作用于CNE细胞后细胞形态变化,Hoechst 33258荧光染色观察细胞核形态的变化,流式细胞仪检测细胞的凋亡率,Western blot法检测凋亡蛋白Caspase3和Caspase9的变化,流式细胞仪检测线粒体跨膜电位(△准m)值的改变,并用JC-1荧光染料染色,激光共聚焦显微镜观察线粒体膜电位变化.结果显示,分离自臭灵丹的HTMF呈浓度、时间双重依赖性显著抑制CNE细胞的增殖,诱导细胞凋亡,引起线粒体膜电位下降及凋亡蛋白Caspase3和Caspase9的活化.提示:3,5-二羟基-6,7,3′,4′-四甲氧基黄酮对人鼻咽癌CNE细胞的生长有显著的抑制作用,并可通过降低线粒体膜电位激活Caspase9,进而活化Caspase3诱导CNE细胞凋亡.

关键词： 3,5-二羟基-6 7 3’ 4’-四甲氧基黄酮(HTMT) CNE细胞细胞凋亡线粒体

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Binding activity of H-Ras is necessary for in vivo inhibition of ASK1 activity

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Cell Research 2004年第2期14卷 148-154页

作者： Jun DU~(1,2*), Shao Hui CAI~(3,4), Zhe SHI~3, Fumihiko NAGASE~1 ~1department of medical Technology, Nagoya University School of Health Sciences, Aichi 461-8673, Japan. ~2School of Pharmaceutical Science, Sun Yat-Sen University, guangzhou, 510089, china. ~3department of Immunology, Nagoya University Graduate and Faculty School of Medicine, Aichi, 466-8550, Japan. ~4college of Pharmacy, jinan University, guangzhou, 510632, china. department of medical Technology Nagoya University School of Health Sciences Aichi Japan School of Pharmaceutical Science Sun Yat-Sen University Guangzhou China department of Immunology Nagoya University Graduate and Faculty School of Medicine Aichi Japan college of Pharmacy Jinan University Guangzhou China

H-Ras is well known as one of the essential components of Ras/Raf/MEK/ERK cascade, which is a critical prosurvival signaling mechanism in most eukaryotic cells. Ras targets Raf/MEK/ERK cascade by integrating and transmitting extracellular signals from growth factor receptors to Raf, leading to the propagation of signals to modulate a serious of cellular survival events. Apoptosis signal-regulating kinasel (ASK1) serves as a general mediator of cell death because it is responsive to a variety of death signals. In this study, we found that H-Ras interacted with ASK1 to cause the inhibition of both ASK1 activity and ASK1-induced apoptosis in vivo, which was reversed only partially by addition of RafS621A, an antagonist of Raf, whereas MEK inhibitor, PD98059, and PI3K inhibitor, LY294002, did not disturb the inhibitory effect of H-Ras on ASK-1-induced apoptosis. Furthermore, by means of immunoprecipitate and kinase assays, we demonstrated that the interaction between H-Ras and ASK1 as well as the inhibition

关键词： H-Ras ASK1 apoptosis binding activity.

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