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胚胎移植不同结局人群的游离DNA核小体印迹差异表达分析
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作者 梅凤琦 蔡晶 +1 位作者 何珮清 刘彦慧 《分子诊断与治疗杂志》 2023年第12期2206-2210,共5页
目的分析体外受精-胚胎移植(IVF-ET)治疗后不同结局患者的血浆游离DNA(cfDNA)核小体印迹和差异基因表达谱,探讨影响妊娠结局及胚胎植入的差异基因的生物进程及细胞组分。方法收集2021年5月至2021年10月在广东医科大学顺德妇女儿童医院... 目的分析体外受精-胚胎移植(IVF-ET)治疗后不同结局患者的血浆游离DNA(cfDNA)核小体印迹和差异基因表达谱,探讨影响妊娠结局及胚胎植入的差异基因的生物进程及细胞组分。方法收集2021年5月至2021年10月在广东医科大学顺德妇女儿童医院生殖中心的IVF-ET病例,根据胚胎植入后的结局分为失败组(n=5)和成功组(n=9),按胚胎植入前1天、植入后14天和植入后28天三个时间点分别检测患者血浆游离DNA,通过高通量测序对胚胎植入后不同结局和不同时间点的cfDNA核小体印迹和差异基因表达谱进行分析。结果血浆cfDNA核小体印迹中高表达基因的覆盖深度低于低表达基因,同一结局不同时间和同一时间不同结局的cfDNA核小体印迹的基因表达差异有统计学意义(P<0.05)。基于基因本体(GO)进行功能富集和信号通路分析发现,失败组在不同时间点的差异基因主要与免疫系统变化、发育调节和有机化合物的代谢相关,而成功组的与细胞发育和有机化合物的代谢相关。失败组差异表达基因在植入后14天的有机化合物代谢过程中表达下调,在植入后28天的免疫相关的生物过程表达上调。结论基于血浆游离DNA核小体印迹的差异基因表达分析可有效筛选和富集到胚胎植入后不同结局在不同时间点之间的差异表达基因和相关生物信号通路,为改善IVF-ET结局提供理论依据,利于研究和临床参考。 展开更多
关键词 胚胎移植 血浆游离DNA 小体印迹 差异基因表达
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哺乳动物配子发生过程中核的印迹作用
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作者 杨增明 谭景和 秦鹏春 《动物学杂志》 CAS CSCD 北大核心 1992年第1期53-55,共3页
很多无脊椎动物、部分鱼类和鸟类都可进行单性生殖。关于哺乳动物的单性生殖,长期以来人们进行了大量的研究。Pincus和shapiro报道用冷刺激法处理兔卵母细胞后,有的卵进一步发育成胚胎,超过植入期,并且获得了几例活的单性生殖个体。以后... 很多无脊椎动物、部分鱼类和鸟类都可进行单性生殖。关于哺乳动物的单性生殖,长期以来人们进行了大量的研究。Pincus和shapiro报道用冷刺激法处理兔卵母细胞后,有的卵进一步发育成胚胎,超过植入期,并且获得了几例活的单性生殖个体。以后,Hoppe及Illmensee报道,他们也得到了纯合子的小鼠。但是,这些实验至今一直未能被别人所重复,故其真实性有待进一步研究。除此之外,哺乳类中至今仍未见有单性生殖的个体,其原因何在? 展开更多
关键词 哺乳动物 配子发生 核印迹
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胶体金核壳型沙丁胺醇分子印迹聚合物的光谱特征分析 被引量:1
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作者 任慧鹏 管毓渝 +2 位作者 戴荣华 刘国艳 柴春彦 《光谱学与光谱分析》 SCIE EI CAS CSCD 北大核心 2016年第2期372-378,共7页
为了解决饲料和动物产品中沙丁胺醇残留现场快速检测的难题,开发以分子印迹技术为基础的快速检测沙丁胺醇的新方法,使用沙丁胺醇做为模板分子,甲基丙烯酸(methacrylic acid,MA)作为功能单体,以本体聚合法为基础合成常规SAL分子印迹聚合... 为了解决饲料和动物产品中沙丁胺醇残留现场快速检测的难题,开发以分子印迹技术为基础的快速检测沙丁胺醇的新方法,使用沙丁胺醇做为模板分子,甲基丙烯酸(methacrylic acid,MA)作为功能单体,以本体聚合法为基础合成常规SAL分子印迹聚合物(molecularly imprinted polymer,MIPs)和非分子印迹聚合物(non imprinted polymer NIPs)。在此基础上,以胶体金粒子为引发核,制备出新型的核壳型沙丁胺醇MIPs。应用紫外吸收光谱(UV spectra)、傅里叶红外光谱(IR spectra)和拉曼光谱(Raman spectru)、扫描电镜(scanning electron microscopy,SEM)等技术手段获得两种印迹物及各种相关化合物的光谱图、电镜图等表征图像。由实验结果可知,SAL和MA上的羧基形成稳定又容易洗脱的1∶1型氢键配合物,化学结合常数K=-0.245×10~6 L^2·mol^(-2)。与MA的—COOH中氢原子形成氢键的可能结合位点是SAL C—O中的氧原子。MIPs与MA中—OH的吸收峰比较可知,前者明显红移;证明SAL作为模板分子与MA之间发生特定结合。未洗脱MIPs的C—O的伸缩振动产生的吸收峰红移;即能量损失明显,可知MA中—COOH的氢原子如果要生成氢键,可能的结合位点就是SAL分子内C—O中的氧原子。MIPs和NIPs中C—C,C—O,—OH等吸收明显的官能团峰型大致相同。将MIPs洗脱掉作为模板分子的SAL后,留下了含有特殊且确定结构官能团化学及空间构成均与SAL高度匹配的空穴,可与待测液中的目标检测分子SAL发生特异性识别和专一结合作用。而胶体金核壳型MIPs与常规MIPs相比,除具有以上相同特点外,其表面更加松散,表面孔穴明显增多。由此增加了吸附目标分子的有效面积,具有更优良的吸附性能。这两种印迹物的合成及光谱特征分析为建立基于分子印迹技术的快速检测SAL新方法奠定了理论和实践基础。 展开更多
关键词 沙丁胺醇 壳型分子印迹 红外光谱 扫描电镜 快速检测
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基于SI-ATRP技术制备磁性甲基对硫磷分子印迹聚合物及其吸附性能 被引量:1
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作者 焦琳娟 徐先燕 +1 位作者 吴晓莹 赵存元 《环境化学》 CAS CSCD 北大核心 2020年第1期89-100,共12页
基于DFT计算,优选出甲基对硫磷(MP)和功能单体甲基丙烯酸(MAA),4-乙烯基吡啶(4-VP)的配比为1∶2∶1,采用表面引发原子转移自由基聚合(SI-ATRP)技术,制备了核壳式磁性甲基对硫磷分子印迹聚合物(Fe3O4@MPIPs).通过扫描电子显微镜(SEM)、... 基于DFT计算,优选出甲基对硫磷(MP)和功能单体甲基丙烯酸(MAA),4-乙烯基吡啶(4-VP)的配比为1∶2∶1,采用表面引发原子转移自由基聚合(SI-ATRP)技术,制备了核壳式磁性甲基对硫磷分子印迹聚合物(Fe3O4@MPIPs).通过扫描电子显微镜(SEM)、透射电子显微镜(TEM)、傅立叶变换红外光谱仪(FTIR)、X-射线衍射仪(XRD)和振动样品磁强计(VSM)对该磁性印迹聚合物进行了表征,并结合磁固相萃取(M-SPE)技术和气相色谱(GC)研究了其对MP的吸附行为,结果表明,Fe3O4@MPIPs对模板分子MP具有良好的特异性识别作用,在30 min内快速达到吸附平衡,最大吸附量为11.5 mg·g^-1;与乐果和马拉硫磷相比,Fe3O4@MPIPs对MP的选择性系数分别为4.57和5.10,相对选择性系数分别为4.11和4.18.气相色谱检测结果表明,该磁性印迹聚合物可用于土豆样品中MP的快速分离富集,其加标回收率为87.4%—99.4%,RSD为3.6%—4.5%;重复使用5次后,Fe3O4@MPIPs回收率仍在90.3%以上,吸附量仍保持在第1次吸附量的82%以上. 展开更多
关键词 壳式磁性分子印迹聚合物 表面引发原子转移自由基聚合 甲基对硫磷 功能单体 DFT计算
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亲水性尿苷分子印迹聚合物的制备及分子选择性识别
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作者 罗爱芹 刘赟 吴俐斐 《北京理工大学学报》 EI CAS CSCD 北大核心 2011年第7期878-882,共5页
研究亲水性核-壳结构分子印迹聚合物的制备并考察其吸附、分离性能.以尿苷为模板分子,通过回流沉淀聚合方法(DPP)制备亲水性的核-壳结构分子印迹聚合物微球.研究了模板分子UR与功能单体MMA之间的相互作用,考察了分子印迹聚合物对模板分... 研究亲水性核-壳结构分子印迹聚合物的制备并考察其吸附、分离性能.以尿苷为模板分子,通过回流沉淀聚合方法(DPP)制备亲水性的核-壳结构分子印迹聚合物微球.研究了模板分子UR与功能单体MMA之间的相互作用,考察了分子印迹聚合物对模板分子的吸附动力学、吸附平衡以及吸附选择性,评价了该分子印迹聚合物作为高效液相色谱柱填料时对尿苷及其类似物的分离能力.UR分子印迹聚合物对UR的吸附符合准一级动力学方程;Freund lich等温方程能更好地拟合分子印迹聚合物对印迹分子尿苷的等温吸附数据;UR分子印迹色谱柱能够实现UR及其类似物的分离,对尿苷具有选择性识别性能. 展开更多
关键词 尿苷 亲水性分子印迹-壳结构聚合物 吸附动力学 等温线 分子识别
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磁性分子印迹聚合物核壳微球的制备及应用 被引量:23
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作者 杨卫海 吴瑶 +3 位作者 张轶 卫晨 严守雷 王清章 《化学进展》 SCIE CAS CSCD 北大核心 2010年第9期1819-1825,共7页
分子印迹技术是综合高分子化学、生物化学等学科发展起来的一门边缘学科。通过分子印迹技术制备的聚合物具有吸附选择性好、色谱效率高、便于功能设计等优点,在色谱分离、固相萃取、传感器、药物控释等领域得到了广泛的应用。磁性聚合... 分子印迹技术是综合高分子化学、生物化学等学科发展起来的一门边缘学科。通过分子印迹技术制备的聚合物具有吸附选择性好、色谱效率高、便于功能设计等优点,在色谱分离、固相萃取、传感器、药物控释等领域得到了广泛的应用。磁性聚合物微球是近年发展起来的一种新型多功能材料,已广泛应用于生物分离、药物控释、疾病诊断等领域。在磁性粒子表面进行分子印迹制备的磁性分子印迹聚合物核壳微球,兼有良好的超顺磁性和高选择吸附性两大优点,具有广阔的应用前景。本文重点综述了磁性分子印迹聚合物核壳微球的制备方法以及在化学分析、生物分离和药物控释方面的应用进展,并指出了该领域研究存在的问题及今后的发展方向。 展开更多
关键词 分子印迹 磁性分子印迹聚合物壳微球 制备 应用
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AS患者PBMCMCP-1蛋白表达及其临床意义 被引量:1
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作者 刘健 陈立强 +3 位作者 陈平 李海珠 梁结玲 梁洁玲 《医学检验与临床》 2013年第3期14-15,11,共3页
目的:研究单核细胞趋化因子蛋白-1(MonocyteChemoattractantProtein-1,MCP-1)在动脉粥样硬化(Atherosclerosis,AS)患者外周血单个核细胞(peripheralbloodmononuclearcells,PBMC)蛋白表达及其与疾病发生、发展的关系。方法:... 目的:研究单核细胞趋化因子蛋白-1(MonocyteChemoattractantProtein-1,MCP-1)在动脉粥样硬化(Atherosclerosis,AS)患者外周血单个核细胞(peripheralbloodmononuclearcells,PBMC)蛋白表达及其与疾病发生、发展的关系。方法:用蛋白质印迹法(Westernblotting)检测20例As患者和20例健康者PBMCMCP-1的蛋白表达。结果:AS患者PBMCMCP-1蛋白表达明显高于健康者,差异有统计学意义(P〈O.05o结论:MCP-1表达与As存在一定相关性,在As发病过程中起重要的作用。 展开更多
关键词 动脉粥样硬化MCP-1外周血单个细胞蛋白质印迹
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核-壳式银-分子印迹聚合物的制备及其SERS性能研究 被引量:1
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作者 常立民 赵媛媛 李欣 《吉林师范大学学报(自然科学版)》 2016年第2期7-11,共5页
采用表面分子印迹技术在银微球表面包覆一层印迹聚合物(Ag@MIPs).利用扫描电子显微镜(SEM)、透射电子显微镜(TEM)及X射线衍射仪(XRD)等手段对所制备Ag@MIPs进行表征.对Ag@MIPs的SERS性能研究表明,Ag@MIPs的SERS强度明显高于银微球,且对... 采用表面分子印迹技术在银微球表面包覆一层印迹聚合物(Ag@MIPs).利用扫描电子显微镜(SEM)、透射电子显微镜(TEM)及X射线衍射仪(XRD)等手段对所制备Ag@MIPs进行表征.对Ag@MIPs的SERS性能研究表明,Ag@MIPs的SERS强度明显高于银微球,且对罗丹明B(RhB)的检测限可达10-13mol/L,Ag@MIPs的竞争吸附性结果表明Ag@MIPs对RhB具有优越的选择特异性. 展开更多
关键词 表面增强拉曼散射技术 -壳式银-分子印迹聚合物 表面分子印迹技术
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Detection of eukaryotic translation initiation factor 4E and its clinical significance in hepatocellular carcinoma 被引量:2
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作者 Xiao-Lin Wang Hong-Pei Cai +1 位作者 Jun-Hui Ge Xiao-Feng Su 《World Journal of Gastroenterology》 SCIE CAS CSCD 2012年第20期2540-2544,共5页
AIM:To study the expression of eukaryotic translation initiation factor 4E(eIF4E),which is closely correlated with malignant tumors,and its relationship to prognosis in hepatocellular carcinoma. METHODS:Western blotti... AIM:To study the expression of eukaryotic translation initiation factor 4E(eIF4E),which is closely correlated with malignant tumors,and its relationship to prognosis in hepatocellular carcinoma. METHODS:Western blotting was performed to quantify the elF4E protein expression in the normal human liver cell line L02 and the hepatoma cell lines Hep3B, HepG2,and Huh7.Forty-six hepatocellular carcinoma samples with complete clinical data were obtained from Changzheng Hospital during the period of December 2008 to July 2009.The expression of eIF4E in the tumor samples and their adjacent tissues were detected by immunohistochemistry.The relationship between the test results and hepatocellular carcinoma(HCC) prognosis was statistically analysed by using a COX proportional hazard model. RESULTS:Western blotting analysis showed that there were distinct eIF4E protein bands in all three of the hepatoma cell lines.In particular,the HepG2 cell line had the highest level of eIF4E protein expression.The L02 cell group had a low eIF4E expression.Immunohistochemical assay showed that there were 32 cases in which the tumour tissue expression was higher than their adjacent tissues,accounting for 69.57%.There were also 14 cases in which the tumour tissue expression was lower or no significant difference was found, accounting for 30.43%.COX proportional hazards model analysis showed that HCC prognosis was related to the depth of invasion,the overexpression of eIF4E and p53, possibly as independent HCC prognostic predictors. CONCLUSION:In summary,eIF4E expression is associated with liver cancer,and patients with high eIF4E expression levels have a higher risk of recurrence. 展开更多
关键词 Hepatocellular carcinoma Eukaryotic translation initiation factor 4E Western blotting IMMUNOHISTOCHEMISTRY PROGNOSIS
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Heme oxygenase-1 prevents liver fibrosis in rats by regulating the expression of PPAR_γ and NF-_κB 被引量:17
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作者 Hui Yang Long-Feng Zhao +3 位作者 Zhong-Fu Zhao Yan Wang Jing-Jing Zhao Li Zhang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2012年第14期1680-1688,共9页
AIM:To investigate the effects of heme oxygenase(HO)-1 on liver fibrosis and the expression of peroxisome proliferator-activated receptor gamma(PPARγ) and nuclear factor-kappa B(NF-κB) in rats.METHODS:Sixty Wistar r... AIM:To investigate the effects of heme oxygenase(HO)-1 on liver fibrosis and the expression of peroxisome proliferator-activated receptor gamma(PPARγ) and nuclear factor-kappa B(NF-κB) in rats.METHODS:Sixty Wistar rats were used to construct liver fibrosis models and were randomly divided into 5 groups:group A(normal,untreated),group B(model for 4 wk,untreated),group C(model for 6 wk,untreated),group D [model for 6 wk,treated with zinc protoporphyrin Ⅸ(ZnPP-Ⅸ) from week 4 to week 6],group E(model for 6 wk,treated with hemin from week 4 to week 6).Next,liver injury was assessed by measuring serum alanine aminotransferase(ALT),aspartate aminotransferase(AST) and albumin levels.The degree of hepatic fibrosis was evaluated by measuring serum hyaluronate acid(HA),type Ⅳ collagen(Ⅳ-C) and by histological examination.Hydroxyproline(Hyp) content in the liver homogenate was determined.The expres-sion levels of alpha-smooth muscle actin(α-SMA) in liver tissue were measured by real-time quantitative polymerase chain reaction(RT-PCR).The expression levels of PPARγ and NF-κB were determined by RT-PCR and Western blotting.RESULTS:The expression of HO-1 increased with the development of fibrosis.Induction of HO-1 by hemin significantly attenuated the severity of liver injury and the levels of liver fibrosis as compared with inhibition of HO-1 by ZnPP-Ⅸ.The concentrations of serum ALT,AST,HA and Ⅳ-C in group E decreased compared with group C and group D(P < 0.01).Amount of Hyp and α-SMA in the liver tissues in group E decreased compared with group C(0.62 ± 0.14 vs 0.84 ± 0.07,1.42 ± 0.17 vs 1.84 ± 0.17,respectively,P < 0.01) and group D(0.62 ± 0.14 vs 1.11 ± 0.16,1.42 ± 0.17 vs 2.56 ± 0.37,respectively,P < 0.01).The expression of PPARγ at levels of transcription and translation decreased with the development of fibrosis especially in group D;and it increased in group E compared with groups C and D(0.88 ± 0.15 vs 0.56 ± 0.19,0.88 ± 0.15 vs 0.41 ± 0.11,respectively,P < 0.01).The expression of NF-κB increased with the development of fibrosis especially in group D;and it decreased in group E compared with groups C and D(1.43 ± 0.31 vs 1.89 ± 0.29,1.43 ± 0.31 vs 2.53 ± 0.54,respectively,P < 0.01).CONCLUSION:Our data demonstrate a potential mechanism that HO-1 can prevent liver fibrosis by enhancing the expression of PPARγ and decreasing the expression of NF-κB in liver tissues. 展开更多
关键词 Heme oxygenase-1 Peroxisome proliferator-activated receptor gamma Nuclear factor-kappa B Liver fibrosis HEMIN
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The quantitative expressions of lymphangiogenic factors and metastasis in human colorectal cancers 被引量:2
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作者 Jianyu He Yuanfang Lv 《The Chinese-German Journal of Clinical Oncology》 CAS 2011年第3期170-173,共4页
Objective: The purpose of this paper was to study the expression levels of newly described lymphatic endothelial markers – LYVE-1, Prox-1, podoplanin and 5’-nucleotidase, and their correlation with metastasis of hu... Objective: The purpose of this paper was to study the expression levels of newly described lymphatic endothelial markers – LYVE-1, Prox-1, podoplanin and 5’-nucleotidase, and their correlation with metastasis of human colorectal cancers. Methods: Tumor and corresponding tumor-side normal tissue samples were obtained from resected specimens immediately after operation. Expression level of each factor was determined by quantitative real-time PCR (RT-PCR) and Western blot technique. Results: Expression levels of lymphatic endothelial markers LYVE-1, Prox-1, podoplanin and 5’-nucleotidase were significantly different in tumor and tumor-side normal groups. Expression levels of Prox-1 and podoplanin were higher in patients with positive lymph node metastasis than those without metastasis. LYVE-1, but not 5’-nucleotidase expression level was higher in both cancer and normal groups. Conclusion: These results indicate that combined quantitative analysis of lymphangiogenic markers LYVE-1, Prox-1 and podoplanin in colorectal cancer specimens may be useful in predicting metastasis of colorectal cancer to regional lymph nodes. However, the role of 5’-nucleotidase in predicting metastasis of colorectal cancer still remains to be further analyzed. 展开更多
关键词 colorectal cancer lymphatic endothelial markers quantitative real-time PCR (RT-PCR)
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Construction of recombinant plasmid pEGFP-C2-L539fs/47 and its expression in HEK293 cells 被引量:2
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作者 Lue Ying Zhang Aifeng +6 位作者 Han Wenqi Li Guoliang Zhang Junbo Gao Jie Pan Junqiang Zhang Yong Sun Chaofeng 《Journal of Medical Colleges of PLA(China)》 CAS 2012年第3期125-133,共9页
Objective:To reconstruct pEGFP-C2-L539fs/47,a HERG nonsense mutant in eukaryotic expression plasmid,and observe the fusion protein expressed in HEK293 cells(human embryo kidney cells).Methods:After double digestion of... Objective:To reconstruct pEGFP-C2-L539fs/47,a HERG nonsense mutant in eukaryotic expression plasmid,and observe the fusion protein expressed in HEK293 cells(human embryo kidney cells).Methods:After double digestion of pcDNA3-L539fs/47 and pEGFP-C2-HERG with sbf I and Eco91 I,the small product fragment,from pcDNA3-L539fs/47,was subcloned into the big fragment of pEGFP-C2-HERG under T4 ligase.pEGFP-C2-L539fs/47 was identified by agarose gel electrophoresis and sequencing.pcDNA3-L539fs/47 and pEGFP-C2-L539fs/47 were transiently transfected into HEK293 cells by Lipofect,respectively.The expression of fusion protein in HEK293 cells was detected through immunofluorescence,laser confocal imaging scanning in vivo,Western blot and PCR.Results:Mutation region cDNA fragment(about 1 kb) and target vector fragment(about 7.2 kb) were ligated after purification and gel recovery.Agarose gel electrophoresis and sequencing successfully demonstrated eukaryotic expression plasmid pEGFP-C2-L539fs/47,constructed approximately 8.2 kb,sequencing consistent with template gene.The transfection efficiency of recombinant plasmid by fluorescence microscopy was more than60%.Western blot analysis detected pcDNA3-L539fs/47 expression of the protein size 60 KD,the expression of pEGFP-C2 fusion protein size of approximately 90 KD.The L539fs/47 gene expression in HEK293 cells was significant by PCR analysis.Confocal laser imaging showed that pEGFP-C2-L539fs/47 protein was successfully expressed in cytoplasm and cytomembrane of HEK293 cells.Conclusion:pEGFP-C2-L539fs/47 containing the HERG gene mutant was successfully constructed by double digestion method and expressed fusion protein in HEK293 cells,which laid a foundation for the further study on L539fs/47. 展开更多
关键词 HERG gene Nonsense mutations Eukaryotic expression vector PEGFP HEK293 cells
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Effects of CpG-ODNs on phenotype and function of monocyte-derived dendritic cells in chronic hepatitis B 被引量:1
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作者 Xiao-Xing Xiang Xia-Qiu Zhou Jun-Xue Wang Qing Xie Xiong Cai Hong Yu Hui-Juan Zhou 《World Journal of Gastroenterology》 SCIE CAS CSCD 2011年第43期4825-4830,共6页
AIM:To study the effects of synthetic nonmethylated CpG-containing oligodeoxynucleotides(CpG-ODNs) ,either alone or combined with recombinant Hepatitis B surface antigen(HBsAg) polypeptide,on the phenotype,function,an... AIM:To study the effects of synthetic nonmethylated CpG-containing oligodeoxynucleotides(CpG-ODNs) ,either alone or combined with recombinant Hepatitis B surface antigen(HBsAg) polypeptide,on the phenotype,function,and intracellular signaling pathways of monocyte-derived dendritic cells(DCs) in patients with chronic hepatitis B(CHB) .METHODS:Peripheral blood monocytes isolated from CHB patients and healthy volunteers were induced to be dendritic cells by recombinant human granulocyte-monocyte colony stimulating factor and interleukin-4.The DCs were then treated with CpG-ODNs,CpGODNs/HBsAg,or tumor necrosis factor(TNF)-αfor 18 h.The expression of surface molecules including HLA-DR,CD86,and CD1a in DCs were detected by flow cytometry,and the expression of signal transducers and activators of transcription(STAT1,3,4,5,6) and suppressors of cell signaling(SOCS1,3) were determined by Western blotting assay.In addition,the capacity of DCs to stimulate allogeneic T lymphocytes and the amount of IL-12p70 released from DCs were measured.RESULTS:In the DCs derived from patients with CHB,treatment with TNF-α,CpG-ODNs,or CpG-ODNs/HBsAg,as compared to the vector control,significantly increased the expression of HLA-DR,stimulated the release of IL-12p70,and enhanced the capacity of DCs to stimulate allogenic T lymphocytes.The expressions of STAT1/4/6 and SOCS1/3,but not STAT3/5,were upregulated by TNF-α,CpG-ODNs,and CpG-ODNs/HBsAg.In addition,the expression of CD1a was upregulated only in the presence of both CpG-ODNs and HBsAg.CONCLUSION:The treatment with CpG-ODNs,either alone or combined with HBsAg,has a remarkable stimulatory effect on the impaired phenotype and function of DCs in CHB,possibly by regulating the expression of STAT1,4,6 and SOCS1,3. 展开更多
关键词 Chronic hepatitis B Dendritic cell CpG oligodeoxynucleotides Hepatitis B surface antigen Signal transduction
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Identification of the Epitopes of Monoclonal Antibodies against P74 of Helicoverpa armigera Nucleopolyhedrovirus
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作者 Limin Liao Dianhai Hou +5 位作者 Huachao Huang Manli Wang Fei Deng Hualin Wang Zhihong Hu Tao Zhang 《Virologica Sinica》 SCIE CAS CSCD 2013年第6期360-367,共8页
P74 is a per os infectivity factor of baculovirus.Here,we report the production of three monoclonal antibodies (mAbs),denoted as 20D9,20F9 and 21E1,raised against P74 of Helicoverpa armigera nucleopolyhedrovirus (Hear... P74 is a per os infectivity factor of baculovirus.Here,we report the production of three monoclonal antibodies (mAbs),denoted as 20D9,20F9 and 21E1,raised against P74 of Helicoverpa armigera nucleopolyhedrovirus (HearNPV),and the identification of their recognition epitopes.The full-length P74,without the transmembrane domains at the C-terminus,was first divided into three segments (N,M and C,respectively),based on the proposed cleavage model for the protein,which were then expressed individually.Western blot analyses revealed specific cross-reactions with the N fragment,for both 20D9 and 21E1.Extensive truncation,followed by prokaryotic expression,of the P74 N fragment was then performed in order to screen for linear epitopes of P74.The recognition regions of 20D9 and 21E1 were revealed to be localized at R144-T153 and T199-C219,respectively.In addition,immunofluorescence microscopy indicated that 20D9 and 20F9 could recognize native P74 in HearNPV-infected cells.These findings will facilitate further investigations of the proteolytic processing of HearNPV P74,and of its involvement in virus-host interactions. 展开更多
关键词 HEARNPV P74 Linear epitope Monoclonal antibody
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