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胶体金免疫层析法检测猪PRVgE抗体方法的建立及应用 总被引：1，自引：0，他引：1
利用PCR技术从猪伪狂犬病毒基因组中克隆gE抗原表位基因,并将其插入到载体pET-22b（＋）中,构建成原核表达质粒pET-22b（＋）-gE,使其在E．coli BL21（DE3）中以IVFG诱导表达,经斑点杂交证实表达产物具有抗原性．表达产物纯化后作为抗原,结合胶体金标记技术,运用双抗原夹心法于国内首先建立了猪PRVgE抗体检测的免疫层析试纸条．该方法操作简单灵敏度高、特异性好,适合猪伪狂犬的临床诊断． 相似文献
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采用S-P免疫组织化学的方法,配对检测了37例大肠癌组织及同一病人的正常粘膜上皮中p53蛋白的表达情况。结果表明,p53蛋白表达阳性19例,阴性18例,阳性率为51.4％,而配对的大肠正常粘膜检测结果均呈p53阴性;p53蛋白表达与病人性别、年龄、Dukes分期等因素无明显相关性,而与淋巴结转移、浸润深度和不同分化程度等存在明显相关性。提示p53蛋白的检测可以作为大肠癌病人病情诊断和预后分析的辅助手段。 相似文献
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[目的]探讨E-钙粘素(E-Cad)表达与大肠癌发生发展及预后的关系.[方法]采用S.P免疫组织化学染色技术,检测30例大肠腺瘤,60例大肠癌组织E-Cad的表达情况.[结果]E-Cad的表达率在大肠腺瘤中为87.10％,显著高于大肠癌中的55.00％(P相似文献
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[目的]探讨CD44v6及nm23-H1蛋白的表达与大肠癌的临床病理意义以及其对大肠癌预后判断的价值.[方法]应用免疫组化技术对52例大肠癌进行上述指标的表达及对比研究.[结果]大肠癌中CD44v6及nm23-H1的阳性表达率分别为65.4％及36.5％.CD44v6与分化程度有关,而nm23-Hl与分化程度无关.二者与肿瘤临床Duke's分期及淋巴结转移密切相关(P<0.05).[结论]CD44v6及nm23-H1的联合检测对判断大肠癌预后有参考价值. 相似文献
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[目的]了解胃癌患者血清中抗核抗体谱的特点.[方法]以鸡卵核为抗原,应用免疫印迹法对18份胃癌术前患者以及22份正常对照组(年龄和性别分布均无差异)的血清进行分析.[结果]胃癌术前患者血清中特异识别5条抗原条带的抗核抗体的阳性率显著高于正常对照组,靶抗原的蛋白分子质量及胃癌术前组血清中特异识别这5条抗原条带的抗核抗体的阳性率分别为:69KD(44.4％,8/18),62KD(50％,9/18),58KD(33.9％,7/18),44KD(22.2％,4/18),37KD(38.9％,7/18).[结论] 胃癌患者体内存在抗核抗体,其相对的靶抗原有多种. 相似文献
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目的从昆明鼠睾丸中克隆Bmi1基因,构建真核表达载体,并转染支持细胞,以便用作培养精原干细胞（SSCs）的滋养层.方法以5日龄昆明鼠为材料,提取小鼠睾丸组织中总RNA后,以RT-PCR技术克隆小鼠睾丸Bmi1基因,构建真核表达载体,并转染TM4细胞（睾丸支持细胞株）,在转染后40 h进行免疫荧光鉴定.结果成功克隆小鼠睾丸Bmi1基因的cDNA,测序正确;免疫荧光细胞染色显示,转染后的支持细胞中有Bmi1蛋白表达.结论本研究为以转染了Bmi1基因的支持细胞作饲养层培养SSCs奠定了基础. 相似文献
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[目的]探讨抗增殖核蛋白的单克隆抗体(Ki-67)在肝细胞肝癌(HCC)中的表达情况及临床意义.[方法]采用免疫组织化学方法检测51例HCC病人术后存档的石蜡标本中Ki-67的表达情况.[结果]HCC中Ki-67阳性表达率58.8％.Ki-67表达阳性率与肿瘤大小、门脉癌栓和肿瘤分级相关,与肝硬化、病灶数目、包膜、AFP及HBsAg不相关,Ki-67阳性表达组HCC病人术后无瘤生存期均显著低于阴性表达组.[结论]Ki-67对HCC的浸润转移均起促进作用,是HCC预后的重要标志物. 相似文献
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目的:通过检测慢性移植物抗宿主病(c GVHD)病人血清中自身抗体的表达来发现和鉴定新的用于诊断和疾病监测的标志物。创新点:我们创新性地应用了包含核仁形成区抗原成分的自身抗体谱对各实验组进行了超过30种自身抗体的筛查,首次发现anti-Ro52抗体可能与肝脏cGVHD有关。方法:联合应用间接免疫荧光、线性免疫印迹和酶联免疫吸附试验(ELISA)对各实验组进行自身抗体谱筛查,采用统计学软件对实验结果和临床数据进行比对分析。结论:我们的研究证实异基因造血干细胞移植后各组病人都可能出现以核仁型抗核抗体(ANA)为主的自身抗体表达增高,并且anti-Ro52抗体在肝脏cGVHD病人血清中显著增高,然而自身抗体与c GVHD的活动性、严重程度和预后均无明显相关性。我们的研究结果表明,自身抗体在预测cGVHD方面的价值有限。 相似文献
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Evaluation of ST13 gene expression in colorectal cancer patients 总被引：2，自引：0，他引：2
DONG Qing-hua 《Journal of Zhejiang University. Science. B》2005,6(12):1170-1175
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Ruan WJ Lin J Xu EP Xu FY Ma Y Deng H Huang Q Lv BJ Hu H Cui J Di MJ Dong JK Lai MD 《Journal of Zhejiang University. Science. B》2006,7(11):929-932
Insulin-like growth factor binding-protein-7 （IGFBP7） was obtained from our previous colonic adenocarcinoma （CRC） and normal mucosa suppression subtraction hybridization （SSH） cDNA libraries. By RT-PCR and immunohistochemistry, we found that IGFBP7 was overexpressed in CRC tissue compared to normal tissue. However, our in vitro experiments performed in 10 CRC cell lines showed that IGFBP7 expressed only in SW480 and Caco2 cell lines, which implied an underlying reversible regulatory mechanism. Using methylation-specific PCR （MSP） and bisulfite sodium PCR （BSP）, we found that its expression was associated with DNA hypomethylation of exonl. This was further supported by the in vitro study which showed restored IGFBP7 expression after demethylation agent 5-aza-2＇-deoxycytidine treatment. Correlation analysis between IGFBP7 expression and prognosis indicated that overexpression of IGFBP7 in CRC tissue correlated with favourable survival. Investigation of the functional role of IGFBP7 through transfection studies showed that IGFBP7 protein could inhibit growth rate, decrease colony formation activity, and induce apoptosis in RKO and SW620 cells, suggesting it a potential tumor suppressor protein in colorectal carcinogenesis. In conclusion, our study clearly demonstrated that IGFBP7 plays a potential tumor suppressor role against colorectal carcinogenesis and its expression is associated with DNA hypomethylation of exon 1. 相似文献
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Construction and characterization of a cDNA library from human liver tissue with chronic hepatitis B 总被引：3，自引：0，他引：3
Chen XH Chen Z Yao HP Chen F Zhu HH Zhou HJ 《Journal of Zhejiang University. Science. B》2005,6(4):288-294
INTRODUCTION Chronic hepatitis B virus (HBV) infection is aserious clinical problem because of its wide distribu-tion and possible adverse consequences, such as he-patic decompensation, cirrhosis and/or primary livercancer (PLC). The natural course of chronic HBVinfection is characterized by a series of hepatitic flaresor exacerbations and remissions (Ganem and Prince,2004). The severity, extent, duration and frequency ofhepatic histopathological changes in hepatitic flaresare d… 相似文献
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LIU Ai-jun FURUSATO Bungo RAVINDRANATH Lakshmi CHEN Yong-mei SRIKANTAN Vasanta MCLEOD David G. PETROVICS Gyorgy SRIVASTAVA Shiv 《Journal of Zhejiang University. Science. B》2007,8(12)
Objective: To investigate molecular alterations associating with prostate carcinoma progression and potentially provide information toward more accurate prognosis/diagnosis. Methods: A set of laser captured microdissected (LCM) specimens from 300 prostate cancer (PCa) patients undergoing radical prostatectomy (RP) were defined. Ten patients representing "aggressive" PCa, and 10 representing "non-aggressive" PCa were selected based on prostate-specific antigen (PSA) recurrence,Gleason score, pathological stage and tumor cell differentiation, with matched patient age and race between the two groups.Normal and neoplastic prostate epithelial cells were collected with LCM from frozen tissue slides obtained from the RP specimens.The expressions of a panel of genes, including NPY, PTEN, AR,AMACR, DD3, and GSTP1, were measured by quantitative real-time RT-PCR (TaqMan), and correlation was analyzed with clinicopathological features. Results: The expressions of AMACR and DD3 were consistently up-regulated in cancer cells compared to benign prostate epithelial cells in all PCa patients, whereas GSTP1 expression was down regulated in each patient. NPY, PTEN and AR exhibited a striking difference in their expression patterns between aggressive and non-aggressive PCas (P=0.0203, 0.0284, and 0.0378, respectively, Wilcoxon rank sum test). The lower expression of NPY showed association with "aggressive" PCas based on a larger PCa patient cohort analysis (P=0.0037,univariate generalized linear model (GLM) analysis). Conclusion: Despite widely noted heterogeneous nature of PCa, gene expression alterations of AMACR, DD3, and GSTP1 in LCM-derived PCa epithelial cells suggest for common underlying mechanisms in the initiation of PCa. Lower NPY expression level is significantly associated with more aggressive clinical behavior of PCa; PTEN and AR may have potential in defining PCa with aggressive clinical behavior. Studies along these lines have potential to define PCa-associated gene expression alterations and likely co-regulation of genes/pathways critical in the biology of PCa onset/progression. 相似文献
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目的白细胞介素15（interleukin-15,IL-15）与炎症反应的关系。方法在含兔IL-15全长编码cDNA的质粒上缺失了部份片段．构建成竞争性RT—PCR的竞争子（competitor）。研究炎症反应后IL-15基因的转录情况。结果竞争性RT-PCR结果显示,IL-15与炎症反应密切相关,且脂多糖（LPS）增加了兔子内脏器官（心脏,肺,肾脏,脾脏和肝脏）中IL-15mRNA的表达。结论LPS可以增强IL-15的免疫反应。 相似文献
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LIU Ai-jun FURUSATO Bungo RAVINDRANATH Lakshmi CHEN Yong-mei SRIKANTAN Vasanta MCLEOD David G. PETROVICS Gyorgy SRIVASTAVA Shiv 《Journal of Zhejiang University. Science. B》2007,8(12):853-859
Objective： To investigate molecular alterations associating with prostate carcinoma progression and potentially provide information toward more accurate prognosis/diagnosis. Methods： A set of laser captured microdissected （LCM） specimens from 300 prostate cancer （PCa） patients undergoing radical prostatectomy （RP） were defined. Ten patients representing ＂aggressive＂ PCa, and 10 representing ＂non-aggressive＂ PCa were selected based on prostate-specific antigen （PSA） recurrence, Gleason score, pathological stage and tumor cell differentiation, with matched patient age and race between the two groups. Normal and neoplastic prostate epithelial cells were collected with LCM from frozen tissue slides obtained from the RP specimens. The expressions of a panel of genes, including NPY, PTEN, AR, AMACR, DD3, and GSTP1, were measured by quantitative real-time RT-PCR （TaqMan）, and correlation was analyzed with clinicopathological features. Results： The expressions of AMACR and DD3 were consistently up-regulated in cancer cells compared to benign prostate epithelial cells in all PCa patients, whereas GSTP1 expression was down regulated in each patient. NPY, PTEN and AR exhibited a striking difference in their expression patterns between aggressive and non-aggressive PCas （P=0.0203, 0.0284, and 0.0378, respectively, Wilcoxon rank sum test）. The lower expression of NPY showed association with ＂aggressive＂ PCas based on a larger PCa patient cohort analysis （P=0.0037, univariate generalized linear model （GLM） analysis）. Conclusion： Despite widely noted heterogeneous nature of PCa, gene expression alterations ofAM,4CR, DD3, and GSTP1 in LCM-derived PCa epithelial cells suggest for common underlying mechanisms in the initiation of PCa. Lower NPY expression level is significantly associated with more aggressive clinical behavior of PCa; PTEN and AR may have potential in defining PCa with aggressive clinical behavior. Studies along these lines have potential to define PCa-associated gene expression alterations and likely co-regulation of genes/pathways critical in the biology of PCa onset/progression. 相似文献
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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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泛素结合酶UFC1是一个新鉴定的类似泛素结合酶E2的基因。目前,对UFC1的功能研究报道还比较少,有待更进一步的研究和探讨。采用半定量RT-PCR技术分别检测一系列的乳腺细胞系MCF7、HBL100、MDA-MB231和MDA-MB-453中UFC1基因的转录水平,结果显示,在正常的HBL100乳腺细胞系中的表达最高,而在乳腺癌细胞系MCF7,MDA-MB231,MDA-MB-453中的表达明显降低。UFC1在乳腺细胞系中的差异表达的结果为将UFC1作为新的靶分子引入乳腺癌的临床预防和治疗提供实验依据。 相似文献