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1.
增殖细胞核抗原(proliferating cell nuclear antigen,PCNA),也称周期蛋白或DNA聚合酶的辅助蛋白,是真核细胞合成所必需的核蛋白,在DNA复制中起重要作用。前期实验发现日本七鳃鳗肝脏cDNA文库的表达序列标签(Expressed Sequence Tag,EST)中存在与高等脊椎动物pcna基因同源的序列。提取日本七鳃鳗(Lampetra japonica)肝脏组织RNA,通过RT-PCR方法扩增七鳃鳗pcna基因,对其进行生物信息学分析,并将Lj-pcna基因成功构建到pGFP-N2真核表达载体上,重组质粒PGFP-N2-Lj-pcna转染人Hela细胞,荧光显微镜下观察有荧光蛋白的表达。日本七鳃鳗pcna基因的真核表达载体成功构建和转染,为探讨七鳃鳗pcna基因功能研究及其它七鳃鳗相关研究提供条件。 相似文献
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目的 :探讨慢性肝病病毒病原学 ,为临床提供准确的治疗依据。方法 :应用ELISA及PCR等方法对 71例慢性肝病患者血清进行甲、乙、丙、丁、戊等病毒性肝炎检测。结果 :乙肝占6 6 .2 % ,丙肝占 2 .8% ,乙丙混合型肝炎占 9.9% ,乙丁混合型肝炎占 4 .2 % ,未定型肝炎占 16 .9% ,未发现甲、戊型肝炎。结论 :乙型肝炎病毒是慢性肝病 (尤其是肝硬化和肝癌 )的主要病原。 相似文献
3.
利用常规病毒分离方法，自死亡病人肺组织病料剖检样品中成功分离到了强致病性SARS-Coy肺组织毒。根据已发表的SARS-Coy Tor2株序列，设计并合成了38条覆盖全长基因组的PCR引物。应用RT-PCR技术从实验感染的Vero E6细胞上清中成功地扩增出了各相应的cDNA重叠片段，分别克隆至pGEM-T载体后，构建了SARS-Coy肺组织毒全基因组cDNA文库。SARS-Coy肺组织毒全基因组cDNA文库的构建为进一步研究SARS-Coy的分子生物学特性奠定了基础。 相似文献
4.
To construct a hepatic stellate cells (HSCs) subtracted cDNA library to find differentially expressed genes in normal mice and mice infected with Schistosomajaponicum (S. japonicum). Suppression subtractive hybridization (SSH) was used. The cDNA fragments of normal mouse were compared to those of schistosoma-infected mice to find differentially expressed genes. Then differentially expressed cDNA fragments were directly inserted into T/A cloning vector to set up the subtractive library. Amplification of the library was carried out with transformation of DH5α. The amplified library contained more than 400 positive bacterial clones, which were then hybridized with forward and backward subtracted probes for differential screening, One hundred positive bacterial clones were randomly selected for sequencing and BLAST analysis, Finally, virtual Northern Blot confirmed such differential expression. The subtracted cDNA library of differentially expressed genes of HSCs was constructed successfully, the library is efficient and lays foundation for screening and cloning new and specific genes of schistosomiasis. 相似文献
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6.
目的：探讨血清总胆汁酸测定在肝病各组中的阳性率，提示总胆汁酸测定应列为肝功能检查项目。方法：东瓯公司总胆汁酸试剂盒，用奥林巴斯AU640自动生化分析仪，双酶双波长测定。结果：总胆汁酸在下列各组肝病：急性肝炎、慢性活动性肝炎、慢性迁延性肝炎、肝硬化和肝癌的阳性率依次为100％、92％、15％、100％和100％，与同时测定ALT对比，其阳性率依次为100％、64％、27％、21％和36％。结论：总胆汁酸（TBA）是一项高敏感性肝功试验项目，应列为常规开展。 相似文献
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[目的]观察双环醇对慢性乙型病毒性肝炎患者肝纤维化程度以及肝组织内转化生长因子β1(TGF-β1)和Ⅰ、Ⅲ型胶原含量的影响.[方法]20例慢性乙型病毒性肝炎患者以双环醇75~150 mg/d治疗24周,治疗前后分别肝活检进行常规病理学检查,并应用免疫组化技术和多媒体彩色病理图文分析系统观察双环醇治疗前后肝组织内TGF-β1和Ⅰ、Ⅲ型胶原含量的变化.[结果]慢性乙型病毒性肝炎肝纤维化患者经双环醇治疗后,肝组织炎症和纤维化程度明显改善,肝组织内TGF-β1和Ⅰ、Ⅲ型胶原含量均较治疗前明显降低,差异有显著性(P<0.05).[结论]双环醇治疗可以减轻慢性乙型病毒性肝炎患者肝脏炎症反应,同时明显降低慢性乙型病毒性肝炎患者肝组织内TGF-β1和Ⅰ、Ⅲ型胶原的含量,从而发挥抗纤维化作用. 相似文献
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乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)的慢性感染,与慢性病毒性肝炎、肝硬化、肝细胞癌(HCC)的发病密切相关,其发病机理涉及到很多基因的共同参与.病毒基因的复制和表达受到肝细胞中蛋白质因子的调节,肝炎病毒基因编码的蛋白与肝细胞中的蛋白能够结合,肝炎病毒蛋白在肝细胞中的表达对于肝细胞的基因表达谱产生影响,也可能是病毒性肝炎、肝硬化、肝细胞癌发病机制的重要机制.酵母单杂交技术、酵母双杂交技术、基因芯片技术、抑制性消减杂交技术、噬菌体展示技术、蛋白质分离纯化与基因克隆化的反向遗传学技术等,在肝炎病毒基因调控、肝炎病毒蛋白结合蛋白的研究、肝炎病毒蛋白反式激活靶基因的研究中都有重要的应用,是促进慢性病毒性肝炎发病机理研究,探索病毒性肝炎新型治疗技术和治疗方法的有效途径. 相似文献
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Appressorium formation is an important event in establishing a successful interaction between the rice blast fungus, Magnaporthe oryzae, and its host plant, rice. An understanding of molecular events occurring in appressorium differentiation will give new strategies to control rice blast. A quick and reliable method to extract total RNA from appressorium is essential for studying gene expression during appressorium formation and its mechanism. We found that duplicate film is an efficient substratum for appressorium formation, even when inoculated with high density conidia. When inoculated with conidia at 1×106ml-1, the percentages of conidium germination and appressorium formation were (97.98±0.67)% and (97.88±0.45)%, respectively. We applied Trizol before appressorium collection for total RNA isolation, and as much as 113.6 pg total RNA was isolated from the mature appressoria at 24 h after inoculation. Functional analysis of two genes, MNH6 and MgATG1, isolated from the cDNA subtractive library, revealed that the quantity of RNA was good enough to construct a cDNA (complementary DNA) library or a cDNA subtractive library. This method may be also applicable for the appressorium RNA isolation of other pathogenic fungi in which conidia differentiate into appressoria in the early stages of host infection. 相似文献
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目的:探讨乙型肝炎患者机体抗氧化能力的变化及其与病情的关系.方法: 门诊乙型肝炎患者共124例和健康者30例,均经放射免疫方法检测后加以确诊.乙型肝炎患者分为三组:乙型肝炎病毒复制组、乙型肝炎病毒恢复组、乙型肝炎合并肝癌组.患者组和对照组均经7150型全自动生化分析仪测定血清ALT、AST含量,应用改良TBA微量法测定血清丙二醛(MDA),以黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)、Cu/Zn-SOD及Mn-SOD水平的变化.结果: 乙型肝炎病毒复制组、乙型肝炎病毒恢复组、乙型肝炎合并肝癌组三组患者血清MDA水平均显著高于对照组(P<0.01),而血清SOD、Cu/Zn-SOD及Mn-SOD水平均显著低于对照组(P<0.01);乙型肝炎病毒复制组、乙型肝炎合并肝癌组患者血清MDA水平均显著高于乙型肝炎病毒恢复组(P<0.01),而血清SOD、Cu/Zn-SOD及Mn-SOD水平均显著低于乙型肝炎病毒恢复组(P<0.01);乙型肝炎合并肝癌组患者血清MDA水平均显著高于乙型肝炎病毒复制组(P<0.01),而血清SOD、Cu/Zn-SOD及Mn-SOD水平均显著低于乙型肝炎病毒复制组(P<0.01).但血清MDA水平与ALT、AST水平(r=0.151),血清SOD、Cu/Zn-SOD及Mn-SOD水平与ALT、AST水平(r=-0.218)均无相关性(P>0.05).结论: 乙型肝炎患者出现明显抗氧化能力的异常.通过血清MDA、SOD、Cu/Zn-SOD及Mn-SOD水平的检测,可为乙型肝炎的临床防治及疗效观察提供一定的依据. 相似文献
11.
已度过围手术期的肝移植患者,肝功能再次异常的原因主要包括原发病的复发,特别是乙型和丙型肝炎的复发、新病毒感染,特别是巨细胞病毒的感染、慢性排斥反应、排斥药物中毒所致肝损害等.临床诊断除常规检测肝功能外,主要依靠病毒学检测及病理诊断.内科治疗方法包括手术前的抗病毒治疗、预防呼吸道病毒感染、定期检测抗排斥药物的浓度、及时更换抗排斥药物等. 相似文献
12.
Yue-qiu Luo Xiao-xing Wu Zong-xin Ling Yi-wen Cheng Li Yuan Charlie Xiang 《Journal of Zhejiang University. Science. B》2014,15(10):900-906
To conduct a systematic review of group studies assessing the association of serum vitamin D status with the severity of liver fibrosis in chronic hepatitis C patients using meta-analysis. The relevant research literatures were identified by searching PubMed and EMBASE databases prior to October 2013 with no restrictions. We included group studies that reported odds ratio (OR) estimates with 95% confidence intervals (CIs) or a mean with standard deviation (SD) for the association between serum vitamin D status and the severity of liver fibrosis in chronic hepatitis C patients. Approximately 8321 participants from several countries were included in this analysis. Six studies on serum vitamin D status and the severity of liver fibrosis were included in this meta-analysis. ORs with 95% CIs were extracted from four studies and the pooled ORs were 0.866 (95% CI, 0.649 to 1.157). The means with SDs were extracted from three studies and the pooled means were −0.487 (95% CI, −0.659 to −0.315). There was statistically significant heterogeneity among the mean data extracted studies (P=0.029; I
2=71.8%) but not among the OR data extracted studies (P=0.061; I
2=55.6%). Finally, results from the mean data extracted studies suggest that lower serum vitamin D is a risk factor for the severity of liver fibrosis in chronic hepatitis C patients. However, there is no conclusive evidence on this association because of inconsistencies between the OR data extracted studies and the mean data extracted studies. 相似文献
13.
目的：克隆蚓激酶基因并在GENEBANK中进行序列分析．方法：采用RT—PCR技术，以蚯蚓总．RNA为模板进行扩增，克隆蚓激酶基因、用Blast软件对基因进行同源性分析．结果：克隆了一个cDNA片段，与GENEBANK中蚓激酶基因序列最高同源性为99％．结论：本方法实用可行，同源性分析表明克隆的cDNA片段具备完整的蚓激酶编码区，为下一步进行蚓激酶的表达研究奠定了良好的基础． 相似文献
14.
Character of HBV （hepatitis B virus） polymerase gene rtM204V／I and rtL180M mutation in patients with lamivudine resistance 总被引：2，自引：0，他引：2
Objectives: To investigate the relationship between HBV (hepatitis B virus) polymerase gene 180 and 204 sites mutation and lamivudine resistance. Methods: One hundred forty-one patients with lamivudine resistance after lamivudine treatment and 60 chronic hepatitis B patients without lamivudine treatment were enrolled in this study. The serum HBV DNA mutation was analyzed by sequence detection via polymerase chain reaction (PCR). The sequences of the same patient were analyzed before and after lamivudine treatment. Results: One hundred and nine lamivudine resistance patients had HBV YMDD (tyrosine-methionine-aspartate-aspartate) mutation. Among them, 45 patients had rtL180M/M204V mutation (41.28%), 28 patients had rtL 180M/M204I mutation (25.70%) and 36 patients had rtM204I mutation (33.02%). There were 6 patients with rtL180M mutation in 32 lamivudine resistance patients. Sixty chronic hepatitis patients without lamivudine treatment had no mutations. Conclusions: HBV mutations, which play an important role in lamivudine resistance usually locate at polymerase gene 204 site; 180 site mutation was also observed in these patients. Evaluation of the anti-virus therapy by surveillance of the two sites mutations is of importance. 相似文献
15.
肝病患者血清总胆汁酸测定的临床意义 总被引：1，自引：0，他引：1
伏小阳 《泰州职业技术学院学报》2010,10(3):27-28
目的探讨肝脏疾病临床诊断中血清总胆汁酸（TBA）测定结果的临床意义。方法在日立7600型全自动生化分析仪上检测272例肝病患者（急性肝炎98例、慢性活动性肝炎54例、肝硬化71例和肝癌49例）的血清TBA、ALT,并与健康对照组比较。结果各种肝病患者的TBA、AIT均显著升高。急性肝炎、慢性活动性肝炎TBA与ALT升高显著,敏感性高,但肝硬化、肝癌两项指标阳性率结果差异明显（χ^2=60．553,P〈0．001）：TBA阳性率高达90％以上,而ALT阳性率仅为20％左右。结论TBA测定对于监测慢性肝病,尤其是肝硬化的早期诊断价值很大,是一项很具实用价值高敏威性的肝功能诊断指标。 相似文献
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郭钢 《岳阳职业技术学院学报》2010,25(1):87-89
目的通过观察乙型肝炎病毒感染者腮腺组织中HBsAg、HBcAg和HBVDNA的表达情况,研究乙肝血清学指标与它们的相关性。方法免疫组化检测乙肝血清指标与腮腺组织中HbsAg、HbcAg和HBVDNA的相关性。结果23例腮腺组织中HBsAg阳性11例,阳性率为47．83％;HBcAg阳性10例,阳性率为43．48％。其中HBsAg和HBcAg同时阳性8例,占34．78％。14例免疫组化阳性患者检出HBVDNA9例,阳性率为64．29％,乙肝血清学指标与腮腺组织中HBsAg、HBcAg检测结果高度正相关（x^2=3．305,P〉0．05）,其Kendall等级相关系数rs=0．661（P〈0．05）。结论腮腺组织对HBV有较强的亲和力．唾液中HBV的出现可能源于受染的唾液腺组织,含HBV的唾液是乙型肝炎生活接触性传播的媒介。 相似文献
17.
抑癌基因PTEN载体构建及在LOVO细胞中的表达 总被引：1，自引：0，他引：1
目的:构建抑癌基因PTEN的表达载体并在LOVO细胞中表达。方法:从人外周血中提取总RNA,通过RT-PCR扩增出PTEN基因片段,将PTEN基因连入pLenti6/V5载体。测序鉴定后,经脂质体转染入LOVO细胞,对细胞株进行RT-PCR和Western blot检测。结果:DNA测序证实pLenti6/V5-PTEN表达载体为阳性克隆;该表达载体转染LOVO细胞后上调了PTEN mRNA和蛋白的表达。结论:成功构建了pLen-ti6/V5-PTEN表达载体,为进一步研究PTEN在LOVO细胞中的功能奠定了基础。 相似文献
18.
目的：探讨NF-кB在肝癌介入治疗后肿瘤组织和癌旁肝细胞中的表达及其变化特点。方法：建立VX2肝癌模型兔59只,分为A组（肝癌组）29只,B组（肝癌经导管肝动脉栓塞介入组）30只、C组（对照组）20只,免疫组化检测各组肝癌及癌旁肝组织中的NF-кB表达。结果：A组肝癌组织与癌旁肝组织NF-κB阳性表达率分别为72.4%（21/29）、37.9%（11/29）,肝癌介入治疗后肝癌组织和癌旁组织NF-κB阳性表达率82.4%（24/30）、60%（18/30）,C组NF-κB阳性表达率为10%（2/20）,各组与对照比较,除A组癌旁肝组织外差异均有统计学意义（P〈0.05）,其余两两比较差异无统计学意义（P〉0.05）。结论：治疗前癌旁组织,其内NF-кB的表达水平尚处于较低水平,介入治疗后的癌旁肝组织的脂肪变性及炎性反应等因素可能会导致NF-κB的表达增加。 相似文献
19.
数字图书馆的服务朝着个性化的方向发展,现有的个性化服务模式主要是信息分类定制服务、信息推送服务、信息智能式代理服务等。在此基础上,探究数字图书馆环境下,个性服务模式中馆员行为能力要发挥作用,馆员的技术能力、用户研究沟通能力、知识能力是提升个性化服务质量的关键。 相似文献
20.
本文分析了高校图书馆信息服务现状和信息服务在图书馆工作中的地位，阐述了高校图书馆信息服务的内容和任务，提出了深化高校图书馆信息服务工作的举措。 相似文献