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富硒麦芽粉对黄曲霉毒素诱导外周血白细胞DNA非程序合成的影响 被引量:10
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作者 诸亚君 陈望秋 +2 位作者 曹禄森 于树玉 肖平 《营养学报》 CAS CSCD 北大核心 1990年第3期248-252,共5页
本文报告了富硒麦芽粉阻断黄曲霉毒素(AFB_1)诱导血白细胞DNA非程序合成(Unsheduled DNA Synthesis,UDS)的作用。 大鼠以富硒麦芽粉Se lppm饲养,可使外周血白细胞经AFB_1 6×10^(-7)M体外诱导的UDS值明显降低。大剂量AFB_1(1mg/kg)... 本文报告了富硒麦芽粉阻断黄曲霉毒素(AFB_1)诱导血白细胞DNA非程序合成(Unsheduled DNA Synthesis,UDS)的作用。 大鼠以富硒麦芽粉Se lppm饲养,可使外周血白细胞经AFB_1 6×10^(-7)M体外诱导的UDS值明显降低。大剂量AFB_1(1mg/kg)一次灌胃,或以每次250μg/kg,二周内10次灌胃的大鼠,如预先饲以富硒麦芽饲料,亦能明显降低其白细胞UDS值。 正常人每日食用300μg硒的硒麦芽粉强化饼干,一年后,可明显降低AFB_1所致人外周血UDS值,麦芽饼干组UDS值为2.56。而富硒麦芽饼干组UDS值为0.35。 本实验提示:富硒麦芽有阻断致癌物AFB_1对外周白细胞UDS的诱发作用。实验中富硒麦芽无毒性但具有硒的生物效应,可望在低硒或肝癌高发区作为防病防癌食品添加剂推广。 展开更多
关键词 黄曲霉毒素 白细胞dna 合成
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外周血白细胞线粒体DNA水平对结直肠癌患者的诊断价值
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作者 王艺晓 张哲 +2 位作者 张静瑜 杨光 王莉 《中国肿瘤外科杂志》 CAS 2021年第4期406-409,415,共5页
目的探讨结直肠癌(CRC)患者外周血白细胞线粒体DNA(mtDNA)水平与临床病理特征的关系及诊断价值。方法选取2018年5月至2020年5月,空军军医大学第二附属医院收治的CRC患者164例,设为CRC组;另选同期体检健康者160人为对照组,检测两组外周... 目的探讨结直肠癌(CRC)患者外周血白细胞线粒体DNA(mtDNA)水平与临床病理特征的关系及诊断价值。方法选取2018年5月至2020年5月,空军军医大学第二附属医院收治的CRC患者164例,设为CRC组;另选同期体检健康者160人为对照组,检测两组外周血白细胞mtDNA水平,分析外周血白细胞mtDNA水平与CRC患者临床病理特征的关系。Logistic回归分析影响CRC发生的相关因素,并以受试者工作特征曲线(ROC)分析外周血白细胞mtDNA对CRC的诊断价值。结果肿瘤分化程度低、有淋巴结转移、发生远处转移的患者外周血白细胞mtDNA水平低于肿瘤分化程度高、无淋巴结转移、未发生远处转移的患者(P<0.05)。Logistic回归分析显示,外周血白细胞mtDNA低表达、结肠息肉史、溃疡性结肠炎史、肿瘤家族史及梭杆菌门、拟杆菌门肠道菌群占比高是影响CRC发生的危险因素(P<0.05)。ROC曲线分析显示,外周血白细胞mtDNA水平对CRC的灵敏度为82.94%、特异度为75.83%、准确度为80.56%、曲线下面积(AUC)为0.815。结论外周血白细胞mtDNA在CRC患者中呈低表达,且与肿瘤分化程度、淋巴结转移、远处转移密切相关,同时外周血白细胞mtDNA水平是影响CRC发生的因素,对临床诊断CRC具有一定价值。 展开更多
关键词 结直肠癌 外周血白细胞线粒体dna 临床病理特征 诊断价值
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Targeting hepatitis B virus antigens to dendritic cells by heat shock protein to improve DNA vaccine potency 被引量:7
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作者 Qin-Long Gu Xue Huang +3 位作者 Wen-Hong Ren Lei Shen Bing-Ya Liu Si-Yi Chen 《World Journal of Gastroenterology》 SCIE CAS CSCD 2007年第44期5911-5917,共7页
AIM: To investigate a novel DNA vaccination based upon expression of the HBV e antigen fused to a heat shock protein (HSP) as a strategy to enhance DNA vaccine potency.METHODS: A pCMV-HBeAg-HSP DNA vaccine and a c... AIM: To investigate a novel DNA vaccination based upon expression of the HBV e antigen fused to a heat shock protein (HSP) as a strategy to enhance DNA vaccine potency.METHODS: A pCMV-HBeAg-HSP DNA vaccine and a control DNA vaccine were generated. Mice were immunized with these different construct. Immune responses were measured 2 wk after a second immunization by a T cell response assay, CTL cytotoxicity assay, and an antibody assay in C57BL/6 and BALB/c mice. CT26-HBeAg tumor cell challenge test in vivo was Performed in BALB/c mice to monitor anti-tumor immune responses.RESULTS: In the mice immunized with pCMV-HBe-HSP DNA, superior CTL activity to target HBV-positive target cells was observed in comparison with mice immunized with pCMV-HBeAg (44% ± 5% vs 30% ± 6% in E: T 〉 50:1, P 〈 0,05), ELISPOT assays showed a stronger T-cell response from mice immunized with pCMV-HBe- HSP than that from pCMV-HBeAg immunized animals when stimulated either with MHC class I or class Ⅱ epitopes derived from HBeAg (74% ± 9% vs 31% ± 6%, P 〈 0.01). ELISA assays revealed an enhanced HBeAg antibody response from mice immunized with pCMV- HBe-HSP than from those immunized with pCMV-HBeAg. The lowest tumor incidence and the slowest tumor growth were observed in mice immunized with pCMV- HBe-HSP when challenged with CT26-HBeAg.CONCLUSION: The results of this study demonstrate a broad enhancement of antigen-specific CD4^+ helper,CD8^+ cytotoxic T-cell, and B-cell responses by a novel DNA vaccination strategy. They also proved a stronger antigen-specific immune memory, which may be superior to currently described HBV DNA vaccination strategies for the treatment of chronic HBV infection. 展开更多
关键词 Hepatitis B virus antigen Dendritic cell Heat shock protein dna vaccine
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Preparation and Detection of Extended DNA Fibers of Chinese Cabbage
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作者 李九欢 王彦华 +2 位作者 李晓峰 申书兴 轩淑欣 《Agricultural Science & Technology》 CAS 2012年第3期517-519,591,共4页
Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei w... Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei were lysed by SDS to release DNA, and DNA fibers were dragged and extended with a coverslip. [Result] The results of Fiber-FISH with genomic DNA and 25S rDNA as probes showed that DNA fiber size as long as about 1.93 Mb could be measured and the number of 25S rDNA copies region were estimated to be 258 and 687 in Chinese cabbage genome. DNA fibers prepared by this method showed equally spread parallel thread with clear background, and were suitable for FISH analysis. [Conclusion] The study would accelerate Chinese cabbage genome mapping and organization analysis. 展开更多
关键词 Chinese cabbage Nuclei isolation Extended dna fibers (EDFs) Fluorescence in situ hybridization (FISH) 25S rdna
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Enhancing cellular immune response to HBV M DNA vaccine in mice by codelivery of interleukin-18 recombinant 被引量:10
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作者 陈建忠 朱海红 +1 位作者 刘克洲 陈智 《Journal of Zhejiang University Science》 CSCD 2004年第4期467-471,共5页
Objective:To investigate the effect of interleukin-18 (IL-18) on immune response induced by plasmid encoding hepatitis B virus middle protein antigen and to explore new strategies for prophylactic and therapeutic HBV ... Objective:To investigate the effect of interleukin-18 (IL-18) on immune response induced by plasmid encoding hepatitis B virus middle protein antigen and to explore new strategies for prophylactic and therapeutic HBV DNA vaccines.Methods:BALB/c mice were immunized with pCMV-M alone or co-immunized with pcDNA3-18 and pCMV-M and then their sera were collected for analysing anti-HBsAg antibody by ELISA;splenocytes were isolated for detecting specific CTL response and cytokine assay in vitro.Results:The anti-HBs antibody level of mice co-immunized with pcDNA3-18 and pCMV-M was slightly higher than that of mice immunized with pCMV-M alone,but there was not significantly different (P>0.05).Compared with mice injected with pCMV-M, the specific CTL cytotoxity activity of mice immunized with pcDNA3-18 and pCMV-M was significantly enhanced (P<0.05) and the level of IFN-γ in supernatant of splenocytes cultured with HBsAg in vitro was significantly elevated (P<0.05) while the level of IL-4 had no significant difference (P>0.05).Conclusion:The plasmid encoding IL-18 together with HBV M gene DNA vaccines may enhance specific TH1 cells and CTL cellular immune response induced in mice, so that IL-18 is a promising immune adjuvant. 展开更多
关键词 INTERLEUKIN-18 Hepatitis B virus dna vaccines Immune response
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Cell polarity protein Par3 complexes with DNA-PK via Ku70 and regulates DNA double-strand break repair 被引量:2
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作者 Longhou Fang YiGuo Wang +6 位作者 Dan Du Guang Yang Tim Tak Kwok Siu Kai Kong Benjamin Chen David J Chen Zhengjun Chen 《Cell Research》 SCIE CAS CSCD 2007年第2期100-116,共17页
The partitioning-defective 3 (Par3), a key component in the conserved Par3/Par6/aPKC complex, plays fundamental roles in cell polarity. Herein we report the identification of Ku70 and Ku80 as novel Par3-interacting ... The partitioning-defective 3 (Par3), a key component in the conserved Par3/Par6/aPKC complex, plays fundamental roles in cell polarity. Herein we report the identification of Ku70 and Ku80 as novel Par3-interacting proteins through an in vitro binding assay followed by liquid chromatography-tandem mass spectrometry. Ku70/Ku80 proteins are two key regulatory subunits of the DNA-dependent protein kinase (DNA-PK), which plays an essential role in repairing double-strand DNA breaks (DSBs). We determined that the nuclear association of Par3 with Ku70/Ku80 was enhanced by γ-irradiation (IR), a potent DSB inducer. Furthermore, DNA-PKcs, the catalytic subunit of DNA-PK, interacted with the Par3/Ku70/Ku80 complex in response to IR. Par3 over-expression or knockdown was capable of up- or downregulating DNA-PK activity, respectively. Moreover, the Par3 knockdown cells were found to be defective in random plasmid integration, defective in DSB repair following IR, and radiosensitive, phenotypes similar to that of Ku70 knockdown cells. These findings identify Par3 as a novel component of the DNA-PK complex and implicate an unexpected link of cell polarity to DSB repair. 展开更多
关键词 cell polarity DSB repair dna-PK Ku70/Ku80/Par3
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DNA methylation in hepatocellular carcinoma 被引量:17
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作者 Iris Tischoff Andrea Tannapfel 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第11期1741-1748,共8页
As for many other tumors,development of hepatocellular carcinoma(HCC)must be understood as a multistep process with accumulation of genetic and epigenetic alterations in regulatory genes,leading to activation of oncog... As for many other tumors,development of hepatocellular carcinoma(HCC)must be understood as a multistep process with accumulation of genetic and epigenetic alterations in regulatory genes,leading to activation of oncogenes and inactivation or loss of tumor suppressor genes(TSG).In the last decades,in addition to genetic alterations,epigenetic inactivation of(tumor suppressor) genes by promoter hypermet hylation has been recognized as an important and alternative mechanism in tumorigenesis.In HCC,aberrant methylation of promoter sequences occurs not only in advanced tumors, it has been also observed in premalignant conditions just as chronic viral hepatitis B or C and cirrhotic liver. This review discusses the epigenetic alterations in hepatocellular carcinoma focusing DNA methylation. 展开更多
关键词 Hepatocellular carcinoma dna methylation Histone modification Tumor suppressor genes
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Interleukin-12 as a Genetic Adjuvant Enhances Hepatitis C Virus NS3 DNA Vaccine Immunogenicity 被引量:5
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作者 Malihe Naderi Atefeh Saeedi +4 位作者 Abdolvahab Moradi Mishar Kleshadi Mohammad Reza Zolfaghari Ali Gorji Amir Ghaemi 《Virologica Sinica》 SCIE CAS CSCD 2013年第3期167-173,共7页
Hepatitis C virus (HCV) chronic infection is a worldwide health problem, and numerous efforts have been invested to develop novel vaccines. An efficient vaccine requires broad immune response induction against viral p... Hepatitis C virus (HCV) chronic infection is a worldwide health problem, and numerous efforts have been invested to develop novel vaccines. An efficient vaccine requires broad immune response induction against viral proteins. To achieve this goal, we constructed a DNA vaccine expressing nonstructural 3 (NS3) gene (pcDNA3.1-HCV-NS3) and assessed the immune response in C57BL/6 mice. In this study, the NS3 gene was amplified with a nested-reverse transcriptase-polymerase chain reaction (RT-PCR) method using sera of HCV-infected patients with genotype 1a. The resulting NS3 gene was subcloned into a pcDNA3.1 eukaryotic expression vector, and gene expression was detected by western blot. The resultant DNA vaccine was co-administered with interleukin-12 (IL-12) as an adjuvant to female C57BL/6 mice. After the final immunizations, lymphocyte proliferation, cytotoxicity, and cytokine levels were assessed to measure immune responses. Our data suggest that co-administration of HCV NS3 DNA vaccine with IL-12 induces production of significant levels of both IL-4 and interferon (IFN)-γ (p<0.05). Cytotoxicity and lymphocyte proliferation responses of vaccinated mice were significantly increased compared to control (p<0.05). Collectively, our results demonstrated that co-administration of HCV NS3 and IL-12 displayed strong immunogenicity in a murine model. 展开更多
关键词 Hepatitis C virus (HCV) NS3 INTERLEUKIN-12 dna vaccine
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Ultrasensitive detection of DNA and protein markers in cancer cells
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作者 Irina V.Smolina Natalia E.Broude 《Cancer Biology & Medicine》 SCIE CAS CSCD 2015年第3期143-149,共7页
Cancer cells differ from normal cells in various parameters, and these differences are caused by genomic mutations and consequential altered gene expression. The genetic and functional heterogeneity of tumor cells is ... Cancer cells differ from normal cells in various parameters, and these differences are caused by genomic mutations and consequential altered gene expression. The genetic and functional heterogeneity of tumor cells is a major challenge in cancer research, detection, and effective treatment. As such, the use of diagnostic methods is important to reveal this heterogeneity at the single-cell level. Droplet microfluidic devices are effective tools that provide exceptional sensitivity for analyzing single cells and molecules. In this review, we highlight two novel methods that employ droplet microfluidics for ultrasensitive detection of nucleic acids and protein markers in cancer cells. We also discuss the future practical applications of these methods. 展开更多
关键词 Microfluidic droplets rolling circle amplification (RCA) peptide nucleic acids (PNA) cell-surface protein marker
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Modulation of cellular and humoral immune responses to anHIV-1 DNA vaccine by interleukin-12 and interleukin-18 DNA immunization
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作者 孙永涛 王福祥 +5 位作者 孙永年 徐哲 王临旭 刘娟 白雪帆 黄长形 《Journal of Medical Colleges of PLA(China)》 CAS 2004年第4期205-210,共6页
Objective: To investigate the effect of interleukin-12 (IL-12) and interleukin-18 (IL-18)DNA immunization on immune response induced by HIV-1 DNA vaccine and to explore new strategies for therapeutic HIV DNA vaccine. ... Objective: To investigate the effect of interleukin-12 (IL-12) and interleukin-18 (IL-18)DNA immunization on immune response induced by HIV-1 DNA vaccine and to explore new strategies for therapeutic HIV DNA vaccine. Methods: The recombinant expression vector pCI-neoGAG was constructed by inserting HIV Gag gene into the eukaryotic expression vector pCI-neo. Balb/c mice were immunized with pCI-neoGAG alone or co-immunized with the DNA encoding for IL-12 or IL-18.Anti-HIV antibody and IFN-γ were tested by ELISA,and splenocytes were isolated for detecting antigen-specific lymphoproliferative responses and specific CTL response by MTT assay and LDH assay respectively. Results: The anti-HIV antibody titers of mice co-immunized with pCI-neoGAG and the DNA encoding for IL-12 or IL-18 were lower than that of mice immunized with pCI-neoGAG alone(P<0.01). In contrast, the IFN-γ level of mice co-immunized with pCI-neoGAG and the DNA encoding for IL-12 or IL-18 was higher than that of mice immunized with pCI-neoGAG alone (P<0.01).Furthermore, compared with mice injected with pCI-neoGAG alone, the specific CTL cytotoxity activity and antigen-specific lymphoproliferative responses of mice immunized with pCI-neoGAG and the DNA encoding for IL-12 or IL-18 were significantly enhanced respectively (P<0.01). Conclusion: The DNA encoding for IL-12 or IL-18 together with HIV DNA vaccine may enhance specific Th-1 responses and cellular immune response elicited in mice. Hence, the DNA encoding for IL-12 or IL-18 are promising immune adjuvants for HIV-1 DNA vaccine. 展开更多
关键词 HIV dna vaccination INTERLEUKIN-12 INTERLEUKIN-18
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Immunization with Cytomegalovirus Envelope Glycoprotein M and Glycoprotein N DNA Vaccines can Provide Mice with Complete Protection against a Lethal Murine Cytomegalovirus Challenge 被引量:1
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作者 Huadong Wang Yanfeng Yao +3 位作者 Chaoyang Huang Quanjiao Chen Jianjun Chen Ze Chen 《Virologica Sinica》 SCIE CAS CSCD 2013年第3期174-182,共9页
Human cytomegalovirus virions contain three major glycoprotein complexes (gC I, II, III), all of which are required for CMV infectivity. These complexes also represent major antigenic targets for anti-viral immune res... Human cytomegalovirus virions contain three major glycoprotein complexes (gC I, II, III), all of which are required for CMV infectivity. These complexes also represent major antigenic targets for anti-viral immune responses. The gC II complex consists of two glycoproteins, gM and gN. In the current study, DNA vaccines expressing the murine cytomegalovirus (MCMV) homologs of the gM and gN proteins were evaluated for protection against lethal MCMV infection in a mouse model. Humoral and cellular immune responses, spleen viral titers, and mice survival and body-weight changes were examined. The results showed that immunization with gM or gN DNA vaccine alone was not able to offer good protection, whereas co-immunization with both gM and gN induced an effective neutralizing antibody response and cellular immune response, and provided mice with complete protection against a lethal MCMV challenge. This study provides the first in vivo evidence that the gC II (gM-gN) complex may be able to serve as a protective subunit antigen for future HCMV vaccine development. 展开更多
关键词 CYTOMEGALOVIRUS Envelope glycoprotein complex gM/gN dna vaccine
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人颈动脉粥样硬化斑块组织及外周血白细胞相对端粒长度的检测及相关性分析 被引量:6
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作者 陈宇 刘继斌 +3 位作者 刘鹏 叶志东 张伟丽 惠汝太 《中国分子心脏病学杂志》 CAS 2012年第1期27-31,共5页
目的研究人颈动脉粥样硬化斑块组织与外周血白细胞的DNA相对端粒长度是否有相关性,外周血白细胞能否反映血管组织的相对端粒长度,从而为外周血白细胞端粒长度与动脉粥样硬化的相关性研究提供依据。方法分别取12例颈动脉内膜剥脱术患者... 目的研究人颈动脉粥样硬化斑块组织与外周血白细胞的DNA相对端粒长度是否有相关性,外周血白细胞能否反映血管组织的相对端粒长度,从而为外周血白细胞端粒长度与动脉粥样硬化的相关性研究提供依据。方法分别取12例颈动脉内膜剥脱术患者的颈动脉内膜斑块组织及外周血,提取DNA,采用实时荧光定量PCR(qRT-PCR)的方法检测相对端粒长度,并进行相关性分析。结果人颈动脉斑块的平均相对端粒长度(Relative telomere length,RTL)为0.56±0.12(平均值±标准差),显著高于外周血白细胞的平均相对端粒长度(0.45±0.13)(P=0.038)。多元线性回归方法校正年龄、性别、体重指数、血脂水平、高血压、糖尿病等心血管病危险因素,表明来自同一患者颈动脉斑块组织和外周血白细胞的RTL具有显著的相关性(标化相关系数β=0.87;P=0.0001);外周血白细胞RTL与年龄呈负相关(标化相关系数β=-0.704,P<0.001);颈动脉斑块样本RTL与年龄呈负相关(标化相关系数β=-0.528,P=0.002)。结论人颈动脉斑块组织与外周血白细胞的相对端粒长度具有相关性,外周血白细胞是替代血管组织研究端粒长度与心血管疾病关系的一个重要指标。 展开更多
关键词 端粒长度 实时荧光定量PCR 颈动脉斑块 外周血白细胞dna
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Connective tissue growth factor reacts as an IL-6/STAT3-regulated hepatic negative acute phase protein 被引量:3
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作者 Olav A Gressner Ieva Peredniene Axel M Gressner 《World Journal of Gastroenterology》 SCIE CAS CSCD 2011年第2期151-163,共13页
AIM:To investigate the mechanisms involved in a possible modulator role of interleukin(IL) -6 signalling on CYR61-CTGF-NOV(CCN) 2/connective tissue growth factor(CTGF) expression in hepatocytes(PC) and to look for a r... AIM:To investigate the mechanisms involved in a possible modulator role of interleukin(IL) -6 signalling on CYR61-CTGF-NOV(CCN) 2/connective tissue growth factor(CTGF) expression in hepatocytes(PC) and to look for a relation between serum concentrations of these two parameters in patients with acute inflammation. METHODS:Expression of CCN2/CTGF,p-STAT3,p-Smad 3/1 and p-Smad2 was examined in primary freshly isolated rat or cryo-preserved human PC exposed to various stimuli by Western blotting,electrophoretic mobility shift assay(EMSA) ,reporter-gene-assays and reversetranscriptase polymerase chain reaction. RESULTS:IL-6 strongly down-regulated CCN2/CTGF protein and mRNA expression in PC,enhanceable by extracellular presence of the soluble IL-6 receptor gp80,and supported by an inverse relation between IL-6 and CCN2/CTGF concentrations in patients'sera.The inhi-bition of TGFβ1 driven CCN2/CTGF expression by IL-6 did not involve a modulation of Smad2(and Smad1/3) signalling.However,the STAT3 SH2 domain binding peptide,a selective inhibitor of STAT3 DNA binding activity,counteracted the inhibitory effect of IL-6 on CCN2/CTGF expression much more pronounced than pyrrolidine-dithiocarbamate,an inhibitor primarily of STAT3 phosphorylation.An EMSA confirmed STAT3 binding to the proposed proximal STAT binding site in the CCN2/CTGF promoter. CONCLUSION:CCN2/CTGF is identified as a hepatocellular negative acute phase protein which is downregulated by IL-6 via the STAT3 pathway through interaction on the DNA binding level. 展开更多
关键词 HEPATOCYTES INTERLEUKIN-6 Connective tissue growth factor STAT3 Liver fibrosis Acute phase reaction
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Epigenetic Repression of SATB1 by Polycomb Group Protein EZH2 in Epithelial Cells 被引量:1
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作者 Chih-chuan Liang 《Chinese Medical Sciences Journal》 CAS CSCD 2010年第4期199-205,共7页
Objective To study the regulatory mechanism of SATB1 repression in cells other than T cells or erythroid cells, which have high expression level of SATB1. Methods HeLa epithelial cells were treated with either histone... Objective To study the regulatory mechanism of SATB1 repression in cells other than T cells or erythroid cells, which have high expression level of SATB1. Methods HeLa epithelial cells were treated with either histone deacetylase inhibitor (HDACi) trichostatin A (TSA) or DNA methylation inhibitor 5-Aza-C before detecting SATB1 expression. Luciferase reporter system was applied to measure effects of EZH2 on SATB1 promoter activity. Over-expression or knockdown of EZH2 and subsequent quantitative reverse transcription-polymerase chain reaction were performed to determine the effect of this Polycomb group protein on SATB1 transcription. Chromatin immunoprecipitation (ChIP) assay was applied to measure enrichment of EZH2 and trimethylated H3K27 (H3K27me3) at SATB1 promoter in HeLa cells. K562 cells and Jurkat cells, both having high-level expression of SATB1, were used in the ChIP experiment as controls. Results Both TSA and 5-Aza-C increased SATB1 expression in HeLa cells. Over-expression of EZH2 reduced promoter activity as well as the mRNA level of SATB1, while knockdown of EZH2 apparently enhanced SATB1 expression in HeLa cells but not in K562 cells and Jurkat cells. ChIP assay results suggested that epigenetic silencing of SATB1 by EZH2 in HeLa cells was mediated by trimethylation modification of H3K27. In contrast, enrichment of EZH2 and H3K27me3 was not detected within proximal promoter region of SATB1 in either K562 or Jurkat cells. Conclusion SATB1 is a bona fide EZH2 target gene in HeLa cells and the repression of SATB1 by EZH2 may be mediated by trimethylation modification on H3K27. 展开更多
关键词 SATB 1 EZH2 Polycomb group protein gene silencing trimethylated H3K27
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Wortmannin induced apoptosis of leukemia cells by reducing PI3K/Akt
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作者 Xiaonan Wang Qing Wu +2 位作者 Liansheng Zhang Yiping Wu Yanwen Shu 《The Chinese-German Journal of Clinical Oncology》 CAS 2010年第12期734-738,共5页
Objective: We studied the effects of Wortmannin (WM) on the proliferation and apoptosis of leukemia cells, and explore the possible mechanisms. Methods: The human myeloid leukemia cell line K562 was treated with d... Objective: We studied the effects of Wortmannin (WM) on the proliferation and apoptosis of leukemia cells, and explore the possible mechanisms. Methods: The human myeloid leukemia cell line K562 was treated with different concentrations of WM, and then detected the activity of the cell proliferation by MTT assay, comet tail formation of cell DNA damage phenomenon by single cell gel electrophoresis, cell apoptosis byAnnexin V-FITC/PI double staining and the expression levels of total Akt, phoshorylated Akt, NF-KB and protein in K562 cell by Western blotting, RT-PCR test before and after WM. Results: WM inhibited cell proliferation of K562 in a concentration-dependent manner, with IC50 value for 24 h being 25 nmol/L. WM induced apoptosis of K562 cells in a concentration-dependent manner, and could induce the breakage of DNA strand of K562 cell. The rate of DNA tail and the tail length of experimental groups were significantly higher than that of control group. WM may inhibit the expression of phosphorylated Akt and NF-KB protein in a dose-dependent manner in both the protein and gene levels, but no significant effect on total Akt protein. Conclusion: WM inhibited cell proliferation and induced apoptosis in K562 and concentration-dependent manner. The possible mechanism may be involved in the regulation of survival signaling pathway, such as PI3K/Akt/NK-KB. 展开更多
关键词 WORTMANNIN K562 cell P-AKT NF-KB
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Effect of interleukin-18 polymorphisms-607 and -137 on clinical characteristics of prostate cancer patients
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作者 Shaojun Nong Yueping Zhang +4 位作者 Bin Cheng Chongsheng He Limin Ma Shujun Zhou Wenguang Li 《The Chinese-German Journal of Clinical Oncology》 CAS 2013年第4期188-193,共6页
Objective: The aim of this study was to determine whether the presence of IL-18 polymorphisms -137 G/C and -607 A/C was associated with grade, clinical stage, and survival in patients with prostate cancer. Methods: Th... Objective: The aim of this study was to determine whether the presence of IL-18 polymorphisms -137 G/C and -607 A/C was associated with grade, clinical stage, and survival in patients with prostate cancer. Methods: The study cohort included 126 patients with prostate cancer. Control group consisted of 125 samples from Chinese population. Genomic DNA was extracted from EDTA-anticoagulated peripheral blood leukocytes by the salting-out method. The genotyping of the two IL-18 polymorphisms was performed using predesigned TaqMan SNP Genotyping Assays. Results: The studied IL-18 gene polymorphisms did not influence susceptibility to prostate cancer in the analyzed group of patients (IL-18-607, P = 0.342; IL-18-137 P = 0.715) but may contribute to disease onset and aggressiveness. IL-18-607 CC genotype was significantly associated with higher tumor grade (P = 0.025) and stage (P = 0.001). IL-18-137 GG genotype was correlated with higher tumor grade (P = 0.018) and stage (P = 0.007). The Cox proportional hazard model showed that tuumor grade and stage grouping were independent prognostic factors but IL-18 polymorphism was not. Polymorphism variants in the IL-18 gene (IL-18-607 and IL-18-137) may be associated with a worse prognosis for prostate cancer. Conclusion: High levels of IL-18 production may play a major role in the growth, invasion and metastasis of prostate cancer. 展开更多
关键词 intedeukin (IL)-18 POLYMORPHISM prostate cancer clinical characteristics
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Identification of an kB-like motif at the 5' upstream region of human lymphotoxin gene
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作者 XURENER SHOUYUANZHAO 《Cell Research》 SCIE CAS CSCD 1994年第1期1-7,共7页
Lymphotoxin(LT) is a glycoprotein secreted by activated T cell. The expression of LT gene is mainly regulated at the level of transcription. By using human LT DNA as a probe, we carried out a RNA dot blotting test and... Lymphotoxin(LT) is a glycoprotein secreted by activated T cell. The expression of LT gene is mainly regulated at the level of transcription. By using human LT DNA as a probe, we carried out a RNA dot blotting test and found that the longer the time of Jurkat human Tlymphoma cells exposed to the PMA and PHA, the more endogenous LT mRNA could be produced. Results of gel retardation assay showed that the nuclear extract from Jurkat cells treated with PMA and PHA formed different DNA-protein complexes. Changes in complex patterns were observed at various time intervals of PMA and PHA induction. A specific protein-binding site was mapped out to be a 22-bp sequence at the 5’upstream regioll of human LT gene by DNase I footprinting analysis. This region was similar to the sequence recognized by the proteins of NFkB family The results of fragment competition and homology analysis indicated that the 22-bp sequence contains a kB-like motif only which is located at the base pairs -100 to -90 (5’-GGGGGCTTCCC-3’). Thus, the NF-kBlike factors were involved in the protein-DNA interaction.Furthermore, there were more than one retarded bands appearing in the gel retardation assay. It suggested that there may be several NF-kB-like factors involved in theregulation of LT gene transcription at the same site. 展开更多
关键词 human LT gene protein-dna interaction transcription regulation
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Relationships between blood leukocyte mitochondrial DNA copy number and inflammatory cytokines in knee osteoarthritis 被引量:4
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作者 Dong ZHAN Aree TANAVALEE +3 位作者 Saran TANTAVISUT Srihatach NGARMUKOS Steven WEDWARDS Sittisak HONSAWEK 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2020年第1期42-52,共11页
Osteoarthritis(OA)is a degenerative articular disorder manifested by cartilage destruction,subchondral sclerosis,osteophytes,and synovitis,resulting in chronic joint pain and physical disability in the elderly.The pur... Osteoarthritis(OA)is a degenerative articular disorder manifested by cartilage destruction,subchondral sclerosis,osteophytes,and synovitis,resulting in chronic joint pain and physical disability in the elderly.The purpose of this study was to investigate mitochondrial DNA copy number(mtDNACN)and inflammatory cytokines in primary knee OA patients and healthy volunteers.A total of 204 knee OA patients and 169 age-matched healthy volunteers were recruited.Their relative blood leukocyte mtDNACN was assessed by quantitative real-time polymerase chain reaction(qRT-PCR),and ten inflammatory cytokines in their plasma were detected by multiplex immunoassay.Blood leukocyte mtDNACN in the OA group was significantly lower than that in the control group.Leukocyte mtDNACN in the control group was negatively correlated with their age(r=−0.380,P<0.0001),whereas mtDNACN in the OA group was positively correlated with their age(r=0.198,P<0.001).Plasma interleukin-4(IL-4)and IL-6 were significantly higher in the knee OA group than in the control group.The plasma IL-6 level was positively correlated with blood leukocyte mtDNACN in the OA group(r=0.547,P=0.0014).IL-5 showed as a major factor(coefficient 0.69)in the second dimension of principle components analysis(PCA)-transformed data and was significantly higher in the OA group(P<0.001)as well as negatively correlated with mtDNACN(r=−0.577,P<0.001).These findings suggest that elevation of plasma IL-4 and IL-6 and a relative reduction in mtDNACN might be effective biomarkers for knee OA.IL-5 is a plausible factor responsible for decreasing blood leukocyte mtDNACN in knee OA patients. 展开更多
关键词 Inflammatory cytokine Blood leukocyte KNEE Mitochondrial dna copy number OSTEOARTHRITIS
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Immunogenicity analysis following human immunodeficiency virus recombinant DNA and recombinant vaccinia virus Tian Tan prime-boost immunization 被引量:7
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作者 LIU CunXia DU ShouWen +9 位作者 LI Chang WANG YuHang WANG MaoPeng LI Yi YIN RongLan LI Xiao REN DaYong QIN YanQing REN JingQiang JIN NingYi 《Science China(Life Sciences)》 SCIE CAS 2013年第6期531-540,共10页
This study assessed and compared the immunogenicity of various immunization strategies in mice using combinations of re- combinant DNA (pCCMp24) and recombinant attenuated vaccinia virus Tian Tan (rddVTT_ccMpe4). ... This study assessed and compared the immunogenicity of various immunization strategies in mice using combinations of re- combinant DNA (pCCMp24) and recombinant attenuated vaccinia virus Tian Tan (rddVTT_ccMpe4). Intramuscular immuniza- tion was performed on days 0 (prime) and 21 (boost). The immunogenicity of the vaccine schedules was determined by meas- uring human immunodeficiency virus (HIV)-specific binding antibody levels and cytokine (interleukin-2 and interleukin-4) concentrations in peripheral blood, analyzing lymphocyte proliferation capacity against HIV epitopes and CD4~/CD8+cell ratio, and monitoring interferon-gamma levels at different times post-immunization. The results showed that pCCMp24, rddVTT.ccMp24 and their prime-boost immunization induced humoral and cellular immune responses. The pCCMp24/ rddVTT.ccMp24 immunization strategy increased CD8+ T cells and induced more IFN-7-secreting cells compared with sin- gle-shot rDNA. The prime-boost immunization strategy also induced the generation of cellular immunological memory to HIV epitope peptides. These results demonstrated that prime-boost immunization with rDNA and rddVTT_ccMp24 had a tendency to induce greater cellular immune response than single-shot vaccinations, especially IFN-7 response, providing a basis for further studies. 展开更多
关键词 vaccinia virus Tian Tan human immunodeficiency virus recombinant dna PRIME-BOOST IMMUNOGENICITY
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Formation and repair of DNA-protein crosslink damage 被引量:6
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作者 Naeh L.Klages-Mundt Lei Li 《Science China(Life Sciences)》 SCIE CAS CSCD 2017年第10期1065-1076,共12页
DNA is constantly exposed to a wide array of genotoxic agents, generating a variety of forms of DNA damage. DNA-protein crosslinks(DPCs)—the covalent linkage of proteins with a DNA strand—are one of the most deleter... DNA is constantly exposed to a wide array of genotoxic agents, generating a variety of forms of DNA damage. DNA-protein crosslinks(DPCs)—the covalent linkage of proteins with a DNA strand—are one of the most deleterious and understudied forms of DNA damage, posing as steric blockades to transcription and replication. If not properly repaired, these lesions can lead to mutations, genomic instability, and cell death. DPCs can be induced endogenously or through environmental carcinogens and chemotherapeutic agents. Endogenously, DPCs are commonly derived through reactions with aldehydes, as well as through trapping of various enzymatic intermediates onto the DNA. Proteolytic cleavage of the protein moiety of a DPC is a general strategy for removing the lesion. This can be accomplished through a DPC-specific protease and and/or proteasome-mediated degradation.Nucleotide excision repair and homologous recombination are each involved in repairing DPCs, with their respective roles likely dependent on the nature and size of the adduct. The Fanconi anemia pathway may also have a role in processing DPC repair intermediates. In this review, we discuss how these lesions are formed, strategies and mechanisms for their removal, and diseases associated with defective DPC repair. 展开更多
关键词 dna-protein crosslinks nucleotide excision repair SPRTN Fanconi anemia
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