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PCR检测HBV DNA与HBV血清标志物常见模式和Pre-S2相互关系研究 被引量:9
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作者 刘树林 邹菊贤 常天恩 《重庆医学》 CAS CSCD 2001年第2期100-101,共2页
目的 探讨乙型肝炎病毒 (HBV)DNA与六种常见HBV血清免疫学标志物模式、前S2 蛋白抗原 (Pre -S2 )之间的相互关系及其临床检测意义。方法 采用PCR法对HBVDNA、ELISA法对HBV血清免疫学标志物、Pre -S2 同时进行检测。结果 1484例六种... 目的 探讨乙型肝炎病毒 (HBV)DNA与六种常见HBV血清免疫学标志物模式、前S2 蛋白抗原 (Pre -S2 )之间的相互关系及其临床检测意义。方法 采用PCR法对HBVDNA、ELISA法对HBV血清免疫学标志物、Pre -S2 同时进行检测。结果 1484例六种常见模式HBVDNA阳性率依次为 89 8% >5 5 6 % >2 1 8% >7 5 % >7 1% >6 8% ,即模式 (1) >(3) >(2 ) >(5 ) >(6 )>(4) ;而HBV血清免疫学标志阳性例数依次为 (2 ) >(1) >(3) >(5 ) >(4) >(6 )。 (2 )、(3)种模式HBVDNA与Pre S2阳性率比较P <0 0 1,其余 (1)、(4)~ (6 )种模式HBVDNA与Pre-S2 阳性检出率无显著性差异 (P >0 0 5 )。结论 HBV血清免疫学标志物、Pre -S2、HBVDNA的检测 ,三者缺一不可 ,ELISA法检测HBV血清免疫学标志物、Pre -S2 ,只是HBV的表型指标 ,提供HBV感染的间接证据。而PCR -HBVDNA的检测是HBV感染与否的直接证据。因此它们各自有其独特的临床检测意义。 展开更多
关键词 聚合酶链反应 乙型肝炎病毒 s2蛋白抗原 pre-s2 DNA
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乙肝病毒S基因Pre-S区突变的人肝癌细胞HepG2稳定株构建及其生物学行为变化 被引量:4
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作者 郑羽飘 钱宝鑫 +4 位作者 覃琴 骆莹 朱争艳 高英堂 王凤梅 《山东医药》 CAS 2019年第2期36-39,共4页
目的构建稳定过表达乙肝病毒(HBV) S基因Pre-S区突变(Pre-S1缺失突变、Pre-S2缺失突变)的HepG2细胞株,并观察该突变对HepG2细胞生物学行为的影响。方法以NCBI中HBV JX661479. 1的Pre-S/S片段序列信息为模板,设计并确定Pre-S1突变、Pre-S... 目的构建稳定过表达乙肝病毒(HBV) S基因Pre-S区突变(Pre-S1缺失突变、Pre-S2缺失突变)的HepG2细胞株,并观察该突变对HepG2细胞生物学行为的影响。方法以NCBI中HBV JX661479. 1的Pre-S/S片段序列信息为模板,设计并确定Pre-S1突变、Pre-S2突变的核苷酸序列,采用全基因合成法获得目的片段并定向导入带有FLAG标签的p LVX慢病毒质粒载体(p Lenti-CMV-3FLAG-PGK-Puro),PCR、双酶切、Sanger测序技术检测重组质粒中目的片段的准确性。将HepG2细胞随机分为5组,空白对照组不处理,p LVX-vector组、野生型组、Pre-S1突变组、Pre-S2突变组分别感染p LVX-vector、p LVX-Pre-S/S、p LVX-Pre-S1 mut/S、p LVX-Pre-S2 mut/S慢病毒液;利用嘌呤霉素筛选HepG2稳定株,采用Western blotting法鉴定目的蛋白,分别采用克隆形成、细胞划痕试验观察HepG2细胞稳定株增殖、迁移能力。结果构建的p LVX-Pre-S/S、p LVX-Pre-S1 mut/S、p LVX-Pre-S2 mut/S重组表达质粒PCR电泳产物与理论值相符,经EcoRⅠ和Bam HⅠ双酶切后电泳产物与理论值相符; Sanger测序结果显示,p LVXPre-S1 mut/S、p LVX-Pre-S2 mut/S突变型除突变序列外,其他序列与HBV S基因野生型序列完全相同。空白对照组HepG2细胞全部死亡,其余组HepG2细胞部分存活。在野生型组、Pre-S1突变组、Pre-S2突变组43 k D分子量位置检测到目的条带表达,而p LVX-vector组无法检测到相应条带。与其他组比较,Pre-S2突变组HepG2的第14天克隆形成数增多、细胞相对迁移距离增大(P均<0. 05)。结论成功构建了HBV Pre-S/S基因野生型及Pre-S突变型的HepG2细胞稳定株,HBV的S基因Pre-S2缺失突变导致肝癌HepG2细胞增殖及迁移能力增强。 展开更多
关键词 乙肝病毒 s基因pre-s突变 肝癌 HEPG2细胞 细胞增殖 细胞迁移
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Construction of exogenous multiple epitopes of helper T lymphocytes and DNA immunization of its chimeric plasmid with HBV pre-S2/S gene 被引量:2
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作者 Wen-JunGao Xiao-MouPeng +4 位作者 Dong-YingXie Qi-FengXie Zhi-LiangGao Ji-LuYao Wen-JunGao 《World Journal of Gastroenterology》 SCIE CAS CSCD 2004年第20期2979-2983,共5页
AIM: To design and construct an exogenous multiple epitope of helper T lymphocytes (HTL), and to evaluate its effect on anti-HBs response through DNA immunization. METHODS: Artificial HTL epitope, PADRE and four other... AIM: To design and construct an exogenous multiple epitope of helper T lymphocytes (HTL), and to evaluate its effect on anti-HBs response through DNA immunization. METHODS: Artificial HTL epitope, PADRE and four other HTL epitopes from different proteins were linked together using splicing by overlap extension to generate exogenous multiple epitopes of HTL, MTE5. pcMTE5 and pcHB were generated by cloning MTE5 and fragments of HBV pre-S2/S gene into mammalian expression plasmid pcDNA3. Four chimeric plasmids were constructed by cloning MTE5 into the region of pre-S2 gene (Barn HI), 5' terminal of S gene (HincII, Xba I) and 3' terminal of S gene (Acc I) of pcHB respectively. BALB/c mice were used in DNA immunization of the recombinant plasmids. Anti-HBs was detected using Abbott IMx AUSAB test kits.RESULTS: The sequences of MTE5 and the 6 constructs ofrecombinant plasmids were confirmed to be correct by DNA sequencing. The anti-HBs response of the coinoculation of pcHB and pcMTE5 was much higher than that of the inoculation of pcHB only (136.7+69.1 mIU/mL vs 27.6+17.3 mIU/mL, P<0.01, t =-6.56). Among the 4 chimeric plasmids, only the plasmid in which MTE5 was inserted into the pre-S2 region had good anti-HBs response (57.54±7.68 mIU/mL), and had no significant difference compared with those of pcHB and the co-inoculation of pcHB and pcMTE5. CONCLUSION: Exogenous multiple epitopes of HTL had immune enhancement when they were co-inoculated with pre-S2/S gene or inoculated in the chimeric form at a proper site of pre-S2/S gene of HBV. It might suggest that it was possible to improve hepatitis B vaccine using exogenous multiple epitopes of HTL. The antibody responses were very low using DNA immunization in the study. Thus, the immune enhancement effect of exogenous multiple epitopes of HTL has to be confirmed and the effect on overcoming the drawback of the polymorphism of HLA II antigens should also be evaluated after these chimeric plasmids are expressed in mammalian cell lines. 展开更多
关键词 外生性 多发性 抗原 辅助机车T 淋巴细胞 DNA 免疫学 质粒 HBV pre-s2/s基因 HTL 基因表达
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Hepatitis B virus pre-S2 start codon mutations in Indonesian liver disease patients 被引量:3
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作者 Andi Utama Marlinang Diarta Siburian +11 位作者 Ismail Fanany Mariana Destila Bayu Intan Rama Dhenni Tri Shinta Kurniasih Syafruddin AR Lelosutan Wenny Astuti Achwan Nasrul Zubir Arnelis Benyamin Lukito Irawan Yusuf Laurentius Adrianus Lesmana Ali Sulaiman 《World Journal of Gastroenterology》 SCIE CAS CSCD 2012年第38期5418-5426,共9页
AIM: To identify the prevalence of pre-S2 start codon mutations and to assess their association with liver disease progression. METHODS: The mutations were identified by direct sequencing from 73 asymptomatic carriers... AIM: To identify the prevalence of pre-S2 start codon mutations and to assess their association with liver disease progression. METHODS: The mutations were identified by direct sequencing from 73 asymptomatic carriers, 66 chronic hepatitis (CH), 66 liver cirrhosis (LC) and 63 hepatocellular carcinoma (HCC) patients. Statistical significances were determined using Fisher's exact test, χ 2 test, and t -test analyses whenever appropriate. Pre-S mutation as a risk factor for advanced liver disease was estimated by unconditional logistic regression model adjusted with age, sex, and hepatitis B e antigen (HBeAg). P < 0.05 was considered significant. RESULTS: Mutation of the hepatitis B virus (HBV) pre-S2 start codon was found in 59 samples from 268 subjects (22.0%), with higher prevalence in patients with cirrhosis 27/66 (40.9%) followed by HCC 18/63 (28.6%), chronic hepatitis 12/66 (18.2%) and asymptomatic carriers 2/73 (2.7%) (P < 0.001). Logistic regression analysis showed that pre-S2 start codon mutation was an independent factor for progressive liver disease. Other mutations, at T130, Q132, and A138, were also associated with LC and HCC, although this was not statistically significant when adjusted for age, sex, and HBeAg. The prevalence of pre-S2 start codon mutation was higher in HBV/B than in HBV/C (23.0% vs 19.1%), whilst the prevalence of T130, Q132, and A138 mutation was higher in HBV/C than in HBV/B. The prevalence of pre-S2 start codon mutation was higher in LC (38.9%) and HCC (40.0%) than CH (5.6%) in HBeAg(+) group, but it was similar between CH, LC and HCC in HBeAg(-) group. CONCLUSION: Pre-S2 start codon mutation was higher in Indonesian patients compared to other Asian countries, and its prevalence was associated with advanced liver disease, particularly in HBeAg(+) patients. 展开更多
关键词 Hepatitis B virus pre-s2 start codon Liver disease Hepatitis B e antigen seroconversion Indonesia
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Pre-S1、S2抗原检测在乙型肝炎临床诊断中的作用及价值 被引量:3
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作者 李青峰 毕雷 +2 位作者 尚鹏程 汪波 简玲 《中国医药科学》 2013年第14期119-120,共2页
目的探讨Pre-S1、S2抗原检测在乙型肝炎临床诊断中的作用及价值。方法回顾性分析2010年6月~2012年6月笔者所在医院就诊的Pre-S1、Pre-S2抗原检测为阳性和两项检测结果均为阳性的患者各50例临床资料,根据检查方法分为Pre-S1组、Pre-S2... 目的探讨Pre-S1、S2抗原检测在乙型肝炎临床诊断中的作用及价值。方法回顾性分析2010年6月~2012年6月笔者所在医院就诊的Pre-S1、Pre-S2抗原检测为阳性和两项检测结果均为阳性的患者各50例临床资料,根据检查方法分为Pre-S1组、Pre-S2组与观察组。所有患者均进行HBV-DNA含量检测,>1.0×103copiese/mL即为阳性,比较三组患者HBV-DNA检出率。结果观察组HBV-DNA检出率为94.0%,明显高于Pre-S1组(86.0%)与Pre-S2组(80.0%),差异具有统计学意义(P<0.05)。结论临床上检测Pre-S1、S2抗原的含量可以为乙肝病毒的复制提供可靠地依据,如果在同一个样本中检测出Pre-S1、S2两种抗原,则可以说明乙肝病毒已经复制。 展开更多
关键词 乙型肝炎病毒 pre-s1抗原 Pre—s2抗原 复制
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Establishment and preliminery use of hepatitis Bvirus preS1/2 antigen assay 被引量:14
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作者 CHEN Kun, HAN Bao Guang, MA Xian Kai, ZHANG He Qiu, MENG Li, WANG Guo Hua, XIA Fang, SONG Xiao Guo and LING Shi Gan 《World Journal of Gastroenterology》 SCIE CAS CSCD 1999年第6期550-552,共3页
关键词 HEPATITIs B VIRUs Pres1/*!s2 antigen ELIsA HEPATITIs B E antigen/analysis
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HBsAg与HBsAb双阳性慢性乙肝患者Pre-S/S区基因突变研究
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作者 王海坚 林建萍 +4 位作者 郑霜 邵伟 李超 童郁 戴显宁 《浙江实用医学》 2021年第2期111-114,共4页
目的探讨HBsAg与HBsAb双阳性慢性乙肝患者S区、Pre-S1区、Pre-S2区基因的突变情况。方法选择浙江省温州市人民医院2018年3月-2020年3月64例HBsAg/HBsAb双阳性慢性乙肝患者(双阳组),同期选择72例HBsAg单阳性慢性乙肝患者(对照组),分析两... 目的探讨HBsAg与HBsAb双阳性慢性乙肝患者S区、Pre-S1区、Pre-S2区基因的突变情况。方法选择浙江省温州市人民医院2018年3月-2020年3月64例HBsAg/HBsAb双阳性慢性乙肝患者(双阳组),同期选择72例HBsAg单阳性慢性乙肝患者(对照组),分析两组S区、Pre-S1区、Pre-S2区氨基酸突变情况。结果两组S区N端(aa1-99)、C端(aa170-226)、"a"决定簇第一环(aa124-137)差异有统计学意义(P<0.01),且C基因型突变率较B基因型高。双阳组中7例发生S区无义突变(χ^(2)=6.213,P<0.05),6例Pre-S区碱基大片段缺失(χ^(2)=5.104,P<0.05),双阳组杂合突变率(54.4%)较高(χ^(2)=10.186,P<0.01)。结论发生Pre-S区碱基大片段缺失,S区无义突变、氨基酸的N端、C端、"a"决定簇第一环氨基酸的突变可能与HBsAg与HBsAb双阳性现象的产生有关。 展开更多
关键词 慢性乙型肝炎病毒 HBsAg/HBsAb双阳性 s pre-s1区 pre-s2 基因型 氨基酸突变
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血清LDH、TPS、CEA和β2-MG在非霍奇金淋巴瘤诊断中的价值 被引量:7
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作者 陈维 魏涛 《分子影像学杂志》 2015年第3期257-259,共3页
目的研究血清乳酸脱氢酶(LDH)、组织多肽特异性抗原(TPS)、癌胚抗原(CEA)和β2-微球蛋白(β2-MG)在非霍奇金淋巴瘤(NHL)诊断中的价值。方法分别采用速率法和酶联免疫吸附法测定62例NHL患者、8例淋巴结良性病患者及40例健康体检者血清LDH... 目的研究血清乳酸脱氢酶(LDH)、组织多肽特异性抗原(TPS)、癌胚抗原(CEA)和β2-微球蛋白(β2-MG)在非霍奇金淋巴瘤(NHL)诊断中的价值。方法分别采用速率法和酶联免疫吸附法测定62例NHL患者、8例淋巴结良性病患者及40例健康体检者血清LDH、TPS、CEA和β2-MG水平。结果(1)NHL组血清LDH、TPS、CEA和β2-MG水平明显升高,与淋巴结良性病组和健康体检组比较差异有统计学意义(P<0.05);淋巴结良性病组和健康体检组比较,四种肿瘤标志物水平差异无统计学意义(P>0.05);(2)血清LDH、TPS、CEA和β2-MG水平在NHL中的高度恶性组、中度恶性组、低度恶性组之间差异无统计学意义(P>0.05)。结论血清肿瘤标志物LDH、TPS、CEA和β2-MG的检测,对NHL的诊断具有重要参考价值。 展开更多
关键词 非霍奇金淋巴瘤 乳酸脱氢酶 组织多肽特异性抗原 癌胚抗原 β2-微球蛋白
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HBV前S_2抗原、前S_2抗体与其病毒标志物的关系 被引量:8
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作者 游景韩 《现代诊断与治疗》 CAS 2001年第4期202-204,共3页
目的 探讨乙型肝炎患者前S2 抗原、抗体与HBVM、HBV DNA之间关系。方法 对 14 6例乙型肝炎患者用ELISA法检测前S2 抗原、前S2 抗体及乙型肝炎病毒标志物 (HBVM ) ;用聚合酶链反应 (PCR)法检测乙型肝炎病毒HBV DNA。结果 急性肝炎、... 目的 探讨乙型肝炎患者前S2 抗原、抗体与HBVM、HBV DNA之间关系。方法 对 14 6例乙型肝炎患者用ELISA法检测前S2 抗原、前S2 抗体及乙型肝炎病毒标志物 (HBVM ) ;用聚合酶链反应 (PCR)法检测乙型肝炎病毒HBV DNA。结果 急性肝炎、慢性肝炎、肝硬变患者前S2 抗原检出率分别为 8.33 %、71.96 %、77.0 7% ;前S2 抗体的检出率分别为 75 %、9.34%、3 .7%。前S2 抗原在HBV DNA、HBeAg阳性病例中检出率明显高于阴性组 (P <0 .0 0 1)。前S2 抗体阳性病例HBV DNA、HBeAg均为阴性。 结论 前S2 抗原的检出意味着病毒有复制或有传染性。前S2 抗体的出现标志着病毒被清除 ,在慢性肝病中检出并不意味病情稳定 ,同时发现该抗体可以在急性乙型肝炎急性期及血清HBVM全部阴性的患者中检出。 展开更多
关键词 乙型肝炎 HBV s2抗原 s2抗体 病毒标志物
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Procedure for preparing peptide-major histocompatibility complex tetramers for direct quantification of antigen-specific cytotoxic T lymphocytes 被引量:16
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作者 Xian-Hui He Li-Hui Xu Yi Liu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第27期4180-4187,共8页
AIM: To establish a simplified method for generating peptide-major histocompatibility complex (MHC) class I tetramers.METHODS: cDNAs encoding the extracellular domain of human lymphocyte antigen (HLA)-A*0201 he... AIM: To establish a simplified method for generating peptide-major histocompatibility complex (MHC) class I tetramers.METHODS: cDNAs encoding the extracellular domain of human lymphocyte antigen (HLA)-A*0201 heavy chain (A2) and β2-microglobulin (132m) from total RNA extracted from leukocytes of HLA-A2+ donors were doned into separate expression vectors by reverse transcription-polymerase chain reaction. The recombinant A2 and 132m proteins were expressed in ~/a oo/i^uain BL21(DE3) and recovered from the inclusion body fraction. Soluble A2 proteins loaded with specific antigen peptides were refolded by dilution from the heavy chain in the presence of light chain 132m and HLA-A2-restricted peptide antigens. The refolded A2 monomers were biotinylated with a commercial biotinylation enzyme (BirA) and purified by low pressure anion exchange chromatography on a Q-Sepharose (fast flow) column.The tetramers were then formed by mixing A2 monomers with streptavidin-PE in a molar ratio of 4:1. Flow cytometry was used to confirm the expected tetramer staining of CD8^+ T cells.RESULTS: Recombinant genes for HLA-A*0201 heavy chain (A2) fused to a BirA substrate peptide (A2-BSP) and mature β2m from HLA-A2+ donor leukocytes were successfully doned and highly expressed in E. coli, Two soluble monomeric A2-peptide complexes were reconstituted from A2-BSP in the presence of 132m and peptides loaded with either human cytomegalovirus pp65495-503 peptide (NLVPMVATV,NLV; designated as A2-NLV) or influenza virus matrix protein Mp58-66 peptide (GILGFVFTL, GIL; designated as A2-GIL). Refolded A2-NLV or A2-GIL monomers were biotinylated and highly purified by single step anion exchange column chromatography. The tetramers were then formed by mixing the biotinylated A2-NLV or A2-GIL monomers with streptavidin-PE, leading to more than 80% multiplicationas revealed by SDS-PAGE under non-reducing, unboiled conditions. Flow cytometry revealed that these tetramers could specifically bind to CD8^+ T cells from a HLA-A2^+ donor,but failed to bind to those from a HLA-A2- donor.CONCLUSION: The procedure is simple and efficient for generating peptide-MHC tetramers. 展开更多
关键词 Cloning Molecular HLA-A antigens HLA-A2 antigen Humans Recombinant Fusion Proteins Research support Non-U.s. Gov't T-Lymphocytes Cytotoxic beta 2-Microglobulin
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BST-2在pSS患者唇腺、外周血淋巴细胞中的表达及增殖作用 被引量:1
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作者 张硕 高尔涵 +3 位作者 达林泰 巴格那 杨文博 孙明启 《国际检验医学杂志》 CAS 2020年第5期544-547,共4页
目的探讨原发性舍格伦综合征(pSS)患者唇腺及外周血B淋巴细胞中骨髓基质细胞抗原2(BST-2)的表达情况及其在pSS中的作用。方法将2017年9月至2018年3月在内蒙古自治区人民医院接受治疗的40例pSS患者作为pSS组;将同期体检的30例健康者作为... 目的探讨原发性舍格伦综合征(pSS)患者唇腺及外周血B淋巴细胞中骨髓基质细胞抗原2(BST-2)的表达情况及其在pSS中的作用。方法将2017年9月至2018年3月在内蒙古自治区人民医院接受治疗的40例pSS患者作为pSS组;将同期体检的30例健康者作为对照组。采用实时荧光定量PCR检测纳入研究者的唇腺组织及外周血B淋巴细胞中BST-2的表达水平。免疫组织化学法检测唇腺组织内BST-2的表达情况,并分析BST-2对淋巴细胞增殖的影响。结果pSS组的外周血B淋巴细胞和唇腺组织中BST-2的表达水平明显高于对照组(P<0.05);pSS患者唇腺组织中浸润的局灶性淋巴细胞和少数周围导管上皮内BST-2表达量较高,而在对照组中仅在导管上皮细胞内存在散在性少量表达。连续切片的pSS患者唇腺组织染色显示,pSS患者的唇腺组织内CD19染色阳性细胞和BST-2阳性细胞基本一致。结论pSS患者唇腺组织和外周血B淋巴细胞BST-2水平明显增高;BST-2通过对腺体内B淋巴细胞的浸润及增殖,参与pSS的发病及进展。 展开更多
关键词 唇腺组织 CD19 B淋巴细胞 骨髓基质细胞抗原2 舍格伦综合征
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Identification of the epitopes on HCV core protein recognized by HLA-A2 restricted cytotoxic T lymphocytes 被引量:11
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作者 Hong-Chao Zhou De-Zhong Xu Xue-Ping Wang Jing-Xia Zhang Ying-Huang Yong-Ping Yan Yong Zhu Bo-Quan Jin Department of Epidemiology,the Fourth Military Medical University,Xi’an 710033,Shaanxi Province,ChinaDepartment of Immunology,the Fourth Military Medical University,Xi’an 710033,Shaanxi Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第4期583-586,共4页
AIM: To identify hepatitis C virus(HCV) core protein epitopes recognized by HLA-A2 restricted cytotoxic T lymphocyte (CTL). METHODS: Utilizing the method of computer prediction followed by a 4h(51)Cr release assay con... AIM: To identify hepatitis C virus(HCV) core protein epitopes recognized by HLA-A2 restricted cytotoxic T lymphocyte (CTL). METHODS: Utilizing the method of computer prediction followed by a 4h(51)Cr release assay confirmation. RESULTS: The results showed that peripheral blood mononuclear cells (PBMC) obtained from two HLA-A2 positive donors who were infected with HCV could lyse autologous target cells labeled with peptide &quot;ALAHGVRAL (core 150-158)&quot;. The rates of specific lysis of the cells from the two donors were 37.5% and 15.8%, respectively. Blocking of the CTL response with anti-CD4 mAb caused no significant decrease of the specific lysis. But blocking of CTL response with anti-CD8 mAb could abolish the lysis. CONCLUSION: The peptide (core 150-158) is the candidate epitope recognized by HLAA2 restricted CTL. 展开更多
关键词 Amino Acid sequence Antibodies Viral B-LYMPHOCYTEs Cell Line Epitope Mapping HLA-A2 antigen HEPACIVIRUs Hepatitis C Humans Peptide Fragments Predictive Value of Tests Research support Non-U.s. Gov't T-Lymphocytes Cytotoxic Viral Core Proteins
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SOX方案联合榄香烯治疗进展期伴转移胃癌患者的疗效及对血清MMP2、CA242的影响 被引量:4
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作者 沈预程 吉浩明 +2 位作者 陈国栋 王彧 谢锦华 《癌症进展》 2019年第17期2034-2037,共4页
目的探讨替吉奥/奥沙利铂(SOX)方案联合榄香烯治疗进展期伴转移胃癌患者的疗效及对血清基质金属蛋白酶2(MMP2)、糖类抗原242(CA242)的影响。方法根据随机数字表法将120例进展期伴转移胃癌患者分为观察组和对照组,每组60例。对照组患者接... 目的探讨替吉奥/奥沙利铂(SOX)方案联合榄香烯治疗进展期伴转移胃癌患者的疗效及对血清基质金属蛋白酶2(MMP2)、糖类抗原242(CA242)的影响。方法根据随机数字表法将120例进展期伴转移胃癌患者分为观察组和对照组,每组60例。对照组患者接受SOX方案治疗,观察组患者接受榄香烯联合SOX方案治疗。比较两组患者的近期疗效及血清MMP2、CA242水平变化情况,并对两组患者的生存情况和治疗期间不良反应发生情况进行比较。结果观察组患者的治疗有效率、疾病控制率均高于对照组(P﹤0.05)。治疗后,观察组与对照组患者的血清MMP2、CA242水平均明显低于本组治疗前,且观察组治疗前后血清MMP2、CA242水平的差值均明显高于对照组,差异均有统计学意义(P﹤0.01)。两组患者治疗期间不良反应发生情况比较,差异无统计学意义(P﹥0.05)。观察组患者的无进展生存情况和总生存情况均优于对照组(P﹤0.05)。结论榄香烯联合SOX化疗方案治疗进展期伴转移胃癌患者能够下调血清MMP2、CA242水平,延缓疾病进展,延长患者的生存时间。 展开更多
关键词 进展期胃癌 奥沙利铂 替吉奥 榄香烯 基质金属蛋白酶2 糖类抗原242
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神经胶质抗原2胶质细胞与中枢神经系统疾病的研究进展
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作者 贺嫣琪 王东钰 +3 位作者 张凯 姜媛 程智刚 王云姣 《医学综述》 CAS 2023年第23期5341-5346,共6页
神经胶质抗原2(NG2)胶质细胞是兼具神经元和胶质细胞特性的特殊细胞,其细胞表面特异性表达NG2硫酸软骨素蛋白多糖。NG2胶质细胞是脑内区别于神经元、成熟少突胶质细胞、星形胶质细胞和小胶质细胞的细胞亚群,广泛存在于发育和成年的哺乳... 神经胶质抗原2(NG2)胶质细胞是兼具神经元和胶质细胞特性的特殊细胞,其细胞表面特异性表达NG2硫酸软骨素蛋白多糖。NG2胶质细胞是脑内区别于神经元、成熟少突胶质细胞、星形胶质细胞和小胶质细胞的细胞亚群,广泛存在于发育和成年的哺乳动物中枢神经系统(CNS)中,参与调节CNS的重建、神经网络、轴突生长和多种CNS疾病。CNS失衡在帕金森病、阿尔茨海默病等神经系统疾病的发病机制中发挥重要作用。因此,深入研究NG2与CNS疾病的相关性,可以为疾病的治疗提供新思路。 展开更多
关键词 阿尔茨海默病 多发性硬化症 帕金森病 神经胶质抗原2胶质细胞 髓鞘 硫酸软骨素蛋白多糖
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p63、Bcl-2和PCNA蛋白在B细胞非霍奇金淋巴瘤中的表达及意义 被引量:3
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作者 张玉娜 高玉彤 杨晶 《山东医药》 CAS 2012年第11期25-27,共3页
目的探讨p63、Bcl-2和增殖细胞核抗原(PCNA)蛋白在B细胞非霍奇金淋巴瘤(B-NHL)发生、发展中的作用。方法应用免疫组化染色法检测p63、Bcl-2及PCNA蛋白在50份B-NHL组织和20份反应性增生淋巴组织中的表达,分析各指标间及与B-NHL临床病理... 目的探讨p63、Bcl-2和增殖细胞核抗原(PCNA)蛋白在B细胞非霍奇金淋巴瘤(B-NHL)发生、发展中的作用。方法应用免疫组化染色法检测p63、Bcl-2及PCNA蛋白在50份B-NHL组织和20份反应性增生淋巴组织中的表达,分析各指标间及与B-NHL临床病理参数的关系。结果 p63、Bcl-2和PCNA蛋白在B-NHL组织中阳性表达率分别为60%、78%和80%,在反应性增生淋巴组织中的阳性表达率分别为5%、35%和35%,两两比较P均<0.05;p63与Bcl-2、PCNA蛋白表达均呈正相关,r=0.453、0.800(P<0.001或0.01);三指标与患者年龄、B-NHL病理类型等临床病理参数均无明显关系(P均>0.05)。结论 p63可能与Bcl-2、PCNA协同参与B-NHL的发生,具体机制尚需进一步研究。 展开更多
关键词 淋巴瘤 非霍奇金 B细胞 P63蛋白 B细胞淋巴瘤/白血病-2 增殖细胞核抗原
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Effect of a cancer vaccine prepared by fusions of hepatocarcinoma cells with dendritic cells 被引量:26
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作者 Juan Zhang~1 Jin-Kun Zhang~2 Shao-Hong Zhuo~3 Hai-Bin Chen~2 1 Clinical Laboratory,The First Affiliated Hospital of Shantou University Medical College,Shantou 515041,Guangdong Province,China2 Cancer Pathology Laboratory,Shantou University Medical College,Shantou 515031,Guangdong Province,China3 Department of Gastroenterology,Third Municipal Hospital of Shantou,Shantou 515073,Guangdong Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第5期690-694,共5页
AIM: To prepare a cancer vaccine (H(22)-DC) expressing high levels of costimulatory molecules based on fusions of hepatocarcinoma cells (H(22)) with dendritic cells (DC) of mice and to analyze the biological character... AIM: To prepare a cancer vaccine (H(22)-DC) expressing high levels of costimulatory molecules based on fusions of hepatocarcinoma cells (H(22)) with dendritic cells (DC) of mice and to analyze the biological characteristics and induction of specific CTL activity of H(22)-DC. METHODS: DCs were isolated from murine spleen by metrizamide density gradient centrifugation, purified based on its characteristics of semi-adhesion to culture plates and FcR-,and were cultured in the medium containing GM-CSF and IL-4. A large number of DC were harvested. DCs were then fused with H(22) cells by PEG and the fusion cells were marked with CD11c MicroBeads. The H(22)-DC was sorted with Mimi MACS sorter. The techniques of cell culture, immunocytochemistry and light microscopy were also used to test the characteristics of growth and morphology of H(22)-DC in vitro. As the immunogen, H(22)-DC was inoculated subcutaneously into the right armpit of BALB/C mice, and their tumorigenicity in vivo was observed. MTT was used to test the CTL activity of murine spleen in vivo. RESULTS: DC cells isolated and generated were CD11c+ cells with irregular shape, and highly expressed CD80, CD86 and CD54 molecules. H22 cells were CD11c- cells with spherical shape and bigger volume, and did not express CD80, CD86 and CD54 molecules.H(22)-DC was CD11c+ cells with bigger volume, being spherical, flat or irregular in shape, and highly expressed CD80, CD86 and CD54 molecules, too. H(22)-DC was able to divide and proliferate in vitro, but its activity of proliferation was significantly decreased as compared with H(22) cells and its growth curve was flatter than H(22) cells. After subcutaneous inoculation over 60 days, H(22)-DC showed no tumorigenecity in mice, which was significantly different from control groups (P【0.01). The spleen CTL activity against H(22) cells in mice implanted with fresh H(22)-DC was significantly higher than control groups (P 【 0.01). CONCLUSION: H(22)-DC could significantly stimulate the specific CTL activity of murine spleen, which suggests that the fusion cells have already obtained the function of antigen presenting of parental DC and could present H(22)specific antigen which has not been identified yet, and H(22)-DC could induce antitumor immune response; although simply mixed H(22) cells with DC could stimulate the specific CTL activity which could inhibit the growth of tumor in some degree, it could not prevent the generation of tumor. It shows that the DC vaccine is likely to become a helpful approach in immunotherapy of hepatocarcinoma. 展开更多
关键词 Cancer Vaccines Animals antigens CD antigens CD80 antigens CD86 Cell Fusion Dendritic Cells Integrin alphaXbeta2 Intercellular Adhesion Molecule-1 Liver Neoplasms Experimental control Male Membrane Glycoproteins MICE Mice Inbred BALB C Research support Non-U.s. Gov't spleen
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Effects of endostatin on expression of vascular endothelial growth factor and its receptors and neovascularization in colonic carcinoma implanted in nude mice 被引量:17
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作者 Yun-HeJia Xin-ShuDong Xi-ShanWang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2004年第22期3361-3364,共4页
AIM:To investigate the antiangiogenic effects of endostatin on colonic carcinoma cell line implanted in nude mice and its mechanism. METHODS:Nude mice underwent subcutaneous injection with LS-174t colonic carcinoma ce... AIM:To investigate the antiangiogenic effects of endostatin on colonic carcinoma cell line implanted in nude mice and its mechanism. METHODS:Nude mice underwent subcutaneous injection with LS-174t colonic carcinoma cell line to generate carcinoma and were randomly separated into two groups.Mice received injection of vehicle or endostatin every day for two weeks. After the tumor was harvested,the tumor volumes were determined,and the expressions of CD34,VEGF and FIk-1 were examined by immunohistochemical method. RESULTS:Tumor volume was significantly inhibited in the endostatin group(84.17%)and tumor weight was significantly inhibited in the endostatin group(0.197±0.049) compared to the control group(1.198±0.105)(F=22.56, P=0.001),microvessel density(MVD)was significantly decreased in the treated group(31.857±3.515)compared to the control group(100.143±4.290)(F=151.62,P<0.001). Furthermore,the expression of FIk-1 was significantly inhibited in the treated group(34.29%) ompared to the control group(8.57%)(X^2=13.745,P=0.001).However no significant decrease was observed in the expression of vascular endothelial growth factor(VEGF)between these two groups(X^2=0.119,P=0.730). CONCLUSION:Endostatin can inhibit tumor growth and angiogenesis by blocking Vegf/FIk-1 pathway.This experiment provides the theory basis for developing a new anti-carcinoma drug through studying the properties of anti-angiogenesis inhibitors. 展开更多
关键词 Angiogenesis Inhibitors Animals antigens CD34 Cell Line Tumor Colonic Neoplasms ENDOsTATINs MICE Mice Nude Neovascularization Pathologic Research support Non-U.s. Gov't Vascular Endothelial Growth Factor A Vascular Endothelial Growth Factor Receptor-2 Xenograft Model Antitumor Assays
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EBNA2基因在霍奇金淋巴瘤中的表达及临床意义
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作者 王佳选 李冰玉 李迅 《新疆医科大学学报》 CAS 2020年第3期287-290,共4页
目的探讨EB病毒核心抗原2(EBNA2)基因在EB病毒(EBV)阳性霍奇金淋巴瘤(HL)患者的表达及临床意义。方法采用巢式聚合酶链反应(PCR)技术检测36例EBV阳性HL患者的全血及蜡块标本中EBNA2基因的表达,分析其在EBV阳性HL中的临床意义。结果全血... 目的探讨EB病毒核心抗原2(EBNA2)基因在EB病毒(EBV)阳性霍奇金淋巴瘤(HL)患者的表达及临床意义。方法采用巢式聚合酶链反应(PCR)技术检测36例EBV阳性HL患者的全血及蜡块标本中EBNA2基因的表达,分析其在EBV阳性HL中的临床意义。结果全血和蜡块样本中各检出EBNA2基因12例(33.3%)和3例(8.3%)。在全部HL患者全血标本中,EBNA2基因在男性和女性患者中各检出5例和7例;在Ann Arbor分期为Ⅰ~Ⅱ和Ⅲ~Ⅳ期患者中各检出4例和8例;在早期患者无和有不良预后因素组中各检出1例和3例;在晚期患者低危、中危和高危组中各检出3例、5例和0例;在结节硬化型、富于淋巴细胞型、混合细胞型、结节性淋巴细胞为主型和淋巴细胞减少型中各检出5例、3例、3例、1例和0例;在有和无巨大肿块患者中各检出0例和12例。结论EBV阳性HL患者全血标本中EBNA2基因的检测可能优于蜡块标本;EBNA2基因的表达与EBV阳性HL患者的性别、Ann Arbor分期、病理组织学类型、有无巨大肿块和风险评分差异均无统计学意义。 展开更多
关键词 霍奇金淋巴瘤 EB病毒 巢式PCR EB病毒核心抗原2
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SARS-CoV-2 δ变异株S抗原含量检测方法的建立及验证
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作者 陈可芝 钟子辉 +7 位作者 王笑天 慕容健昌 张梅 赖文龙 李东 甘建辉 刘萌萌 刘建凯 《中国生物制品学杂志》 CAS CSCD 北大核心 2023年第4期458-463,共6页
目的 建立严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)δ变异株S抗原含量的检测方法,并进行验证。方法 采用δ变异株重组表达的受体结合域(receptor binding domain,RBD)蛋白分别免疫山... 目的 建立严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)δ变异株S抗原含量的检测方法,并进行验证。方法 采用δ变异株重组表达的受体结合域(receptor binding domain,RBD)蛋白分别免疫山羊和家兔,制备抗RBD多克隆抗体。以制备的两种抗RBD多克隆抗体为包被抗体和一抗,HRP标记的羊抗兔IgG抗体为酶标抗体,建立用于检测S抗原含量的双抗体夹心ELISA法。棋盘滴定法确定包被抗体和一抗,同时优化3种抗体稀释度。验证方法的准确性、精密性、线性范围及专属性。采用建立的方法检测SARS-CoV-2 δ株灭活疫苗(Vero细胞)原液及其中间品的S抗原含量。结果 兔抗RBD多克隆抗体及羊抗RBD多克隆抗体的效价分别为1∶32 000和1∶64 000,蛋白浓度分别为2.26和5.41 mg/mL。最佳包被抗体为羊抗RBD多克隆抗体,一抗为兔抗RBD多克隆抗体,包被抗体、一抗及酶标抗体最佳稀释度分别为1∶4 000、1∶8 000和1∶16 000。40、20、10 U/mL3种浓度S抗原内部参考品重复6次检测结果的回收率为83.5%~103.0%;重复6次测定3批SARS-CoV-2 δ株灭活疫苗(Vero细胞)原液抗原含量的CV均<7.5%,2名实验员对3批上述疫苗原液重复检测3次结果的CV均<7.5%;S抗原含量理论值在10~40 U/mL范围内,与测定值呈良好的线性关系,线性回归方程为:y=0.942 3 x+0.049 2,R~2=0.995 4,定量限为10 U/mL;建立的方法与甲型肝炎灭活疫苗(人二倍体细胞)、四价流感病毒裂解疫苗、Vero细胞宿主蛋白、Vero细胞培养上清液等无交叉反应。SARS-CoV-2 δ株灭活疫苗(Vero细胞)原液及中间品3次检测结果的CV均<15.0%。结论 建立的双抗体夹心ELISA法具有良好的准确性、精密性、专属性,可用于SARS-CoV-2 δ株灭活疫苗(Vero细胞)原液及中间品中S抗原的定量检测。 展开更多
关键词 严重急性呼吸综合征冠状病毒2 灭活疫苗 s抗原 酶联免疫吸附试验
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HBV新型免疫原的设计、合成及免疫原性研究 被引量:29
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作者 吴玉章 刘茂昌 +1 位作者 贾正才 邹丽云 《第三军医大学学报》 CAS CSCD 北大核心 2000年第10期919-923,共5页
目的 探讨如何将表位构建成有效免疫原。方法 以Pre S( 2 ) 蛋白两优势性B细胞表位和HBcAg两Th细胞表位为基础 ,设计、合成了 2种MAP多肽 ,以此两MAP肽为免疫原免疫兔和小鼠 ,观察体液免疫反应。结果 此两种MAP多肽均比Pre S( 2 ) ... 目的 探讨如何将表位构建成有效免疫原。方法 以Pre S( 2 ) 蛋白两优势性B细胞表位和HBcAg两Th细胞表位为基础 ,设计、合成了 2种MAP多肽 ,以此两MAP肽为免疫原免疫兔和小鼠 ,观察体液免疫反应。结果 此两种MAP多肽均比Pre S( 2 ) 蛋白诱发更高抗体滴度 ;Th细胞表位引入可显著增强反应强度。结论 增加肽抗原结构复杂性可提高其免疫原性 ,其表位排列以B细胞表位在赖氨酸核心外侧为佳 。 展开更多
关键词 乙型肝炎 HBV MAP B细胞表位 Th细胞表位
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