HLA Class I Expressions on Peripheral Blood Mononuclear Cells in Colorectal Cancer Patients

Objective： To investigate the expression change of human leukocyte antigen （HLA） class I on human peripheral blood mononuclear ceils （PBMCs） at both mRNA and protein levels, and to evaluate its roles in the development of colorectal cancer （CRC）. Methods： In the present study, 50 patients with CRC, 35 patients with benign colorectal lesion and 42 healthy volunteers were enrolled. Expression levels of HLA class I mRNA and protein were determined using real-time quantitative reverse transcription PCR （RT-PCR） and flow cytometry analysis, respectively. Results： The expression levels of HLA class I mRNA and proteins were not influenced by age and gender. The relative ratios of HLA class I mRNA were 0.99±0.27 in healthy controls, 0.76±0.19 in benign patients, and 0.48±0.21 in CRC patients. Mean fluorescence intensities of HLA class I were 145.58±38.14 in healthy controls, 102.05±35.98 in benign patients and 87.44±34.01 in CRC patients. HLA class I on PBMCs was significantly down-regulated at both mRNA and protein levels in patients with stage III and IV CRC. CRC patients with lymph node metastasis also showed a decreased HLA class I expression at protein level. Conclusion： HLA class I expressions on PBMCs are associated with staging of CRC and lymph node metastasis. Monitoring the expression of HLA class I on PBMCs may provide useful information for diagnosis and metastasis judgement of CRC.

Detection of Replicative Form of HCV RNA in Peripheral Blood Leukocytes and Its Clinical Significance

Nested RT-PCR, done by using degenerated primer pair, was used to detect hepatitis C virus RNA (HCV RNA) in serum, plasma, liver and peripheral blood mononuclear cells (PBMC) of 30 patients with acute and chronic posttransfusion hepatitis C and 7 asymptomatic anti-HCV positive subjects. The results showed that the percentages of both the plus and minus strands of HCV RNA in PBMC of the patients with chronic hepatitis C was significantly higher than that with acute hepatitis C and asymptomatic anti-HCV positive subjects ( P<0.05-0.001). In 17 patients who were subjected to biopsy, the positive rate of the both strands of HCV RNA in PBMC of the patients with AH was lower than that of CAH ( P<0.05). In serum and plasma of all 37 cases, the minus strand of HCV RNA was not detected. Both plus and minus strands in liver of one patient with AH were positive , but the minus strand in PBMC negative. In 6 patients with CAH whose both strands in liver were positive,Both strands in PBMC in 5 patients were also found. The present data confirmed that PBMC of the patients with hepatitis C were infected by HCV and the longer the infection time, the bigger the possibility of PBMC infection by HCV. The patients with active liver disease (CAH) had higher positive rate of minus strands of HCV RNA in PBMC. The results suggested that HCV may not only infect PBMC but also replicate in PBMC , and that the occurrence of minus strand of HCV RNA is associated with activity of liver disease.

应用RT-PCR检测乳腺癌患者外周血CEA mRNA和CK-19 mRNA的研究及意义

目的:检测乳腺癌患者外周血中CEAmRNA和CK-19mRNA的表达,建立取材于外周血早期诊断乳腺癌微转移的方法。方法:65例乳腺癌和37例乳腺良性疾病患者的外周血,分离其有核细胞后进行细胞总RNA的抽提,运用巢式RT-PCR技术进行CEAmRNA和CK-19mRNA的检测。结果:以巢式RT-PCR终产物出现131bp为CEAmRNA阳性,460bp为CK-19mRNA阳性。65例乳腺癌患者CEAmRNA和CK-19mRNA分别有26例和24例表达,总阳性率分别为40.0%和36.92%。37例乳腺良性疾病者无1例CEAmRNA和CK-19mRNA表达,两组有统计学意义(P<0.001)。结论:RT-PCR可以用于临床检测乳腺癌患者外周血中微转移。

大肠癌患者外周血CEAmRNA、HNP1-3的检测及临床意义

目的:研究大肠癌患者外周血中CEAmRNA、HNP1-3的表达及临床意义。方法:采用NestedRT-PCR法检测外周血CEAmRNA、ELISA法检测血清HNP1-3在大肠癌患者术前组、术后组及正常对照组中的表达。结果:外周血CEAmRNA在大肠癌患者术前组、术后组及正常对照组的阳性率分别为54%、34%、5%,术前组和术后组相比存在显著差异(P=0.044),二者与正常对照组比较,有统计学差异(P<0.001、P=0.012);CEAmRNA与患者性别、年龄、肿瘤部位及分化程度无关,但与Dukes'分期、淋巴结转移有关;外周血CEAmRNA对大肠癌的诊断敏感性和特异性分别为54%、95%。大肠癌患者术前组血清HNP1-3浓度中位数为20.33ng/ml,正常对照组7.69ng/ml,术后组14.35ng/ml;术前组、术后组血清HNP1-3水平均明显高于正常对照组,差异有统计学意义(P<0.001、P=0.005);并且血清HNP1-3水平也与Dukes'分期、淋巴结转移有关;血清HNP1-3对大肠癌的诊断敏感性、特异性分别为62%、95%。联合检测外周血CEAmRNA与血清HNP1-3的敏感性(72%)及特异性(100%)均高于单一指标。结论:大肠癌患者外周血中CEAmRNA表达较正常对照组明显增高,且与Dukes'分期、淋巴结转移密切相关。HNP1-3在大肠癌患者血清中呈高表达,且具有较高的敏感性及特异性,有望成为一种新的大肠癌肿瘤标志物。联合检测外周血CEAmRNA表达及血清HNP1-3水平可提高检测敏感性和特异性,对发现大肠癌微转移可能有重要的临床意义。

外周血CEAmRNA、CEA及VEGF检测在诊断肿瘤微转移中的意义

目的探讨检测外周血癌胚抗原信使核糖核酸(CEAmRNA)表达、血清血管内皮细胞生长因子(VEGF)及血清癌胚抗原(CEA)水平在诊断恶性肿瘤微转移中的临床意义。方法36例经病理确诊的恶性肿瘤患者(其中13例临床确诊远处转移,23例无确切临床转移征象),17例健康体检对照者,采用巢式逆转录聚合酶链反应(nestRT-PCR)检测外周血CEAmRNA表达,并联合检测血清VEGF水平(酶联免疫吸附法,ELISA)及血清CEA的水平(化学发光法)。结果13例远处转移患者中CEAmRNA表达阳性11例(84.6%),23例未转移患者中CEAmRNA表达阳性6例(26.1%),17例健康对照组CEAmRNA表达均阴性,肿瘤转移病人外周血CEAmRNA表达明显高于无转移病人;血清CEA在远处转移患者与未转移患者中的差异无统计学意义;远处转移患者的VEGF水平高于未转移患者及健康对照者。联合检测外周血CEAmRNA表达及血清VEGF水平在肿瘤转移诊断中有更高的敏感性。结论联合检测外周血CEAmRNA表达及血清VEGF水平可能有助于恶性肿瘤微转移的早期诊断。

CEA分泌性肿瘤微循环检测体会

目的 检测肿瘤外周血肿瘤细胞，建立肿瘤微循环转移早期诊断的方法。方法 取ＣＥＡ分泌性肿瘤患者外周血，分离其有核细胞后进行细胞总ＲＮＡ 的抽提，运用ＲＴ－ ＰＣＲ技术进行基因检测。结果 以ＰＣＲ终产物出现１３１ ｂｐ 带定为ＣＥＡｍＲＮＡ阳性，结果２０ 例患者总阳性率为５５％ 。结论 ＲＴ－ ＰＣＲ是分子生物学领域的新技术，已成为肿瘤微循环检测的方法，具有灵敏度高的优点。

胃肠道癌患者外周血癌胚抗原信使核糖核酸的检测及其临床意义

目的:应用RT-PCR法检测胃肠道癌患者外周静脉血CEAmRNA,探讨与不同临床病理指标之间的关系,评估对临床治疗和预后的指导意义。方法:应用RT-PCR技术,选择特异性CEAmRNA引物,检测41例胃癌、大肠癌患者外周血CEAmRNA,同时定量检测外周血CEA和CA19-9糖蛋白。另抽取12例健康志愿者外周血标本作为对照。结果:41例胃肠癌患者外周血中CEAmRNA、CEA、CA19-9阳性率分别为26.8%、19.5%、14.6%。12例对照外周血中CEAmRNA、CEA、CA19-9均阴性。有淋巴结转移者外周血CEAmRNA阳性率(40%,10/25)高于无淋巴结转移者(6.25%,1/16),P=0.000。TNM分期IV期中外周血CEAmRNA阳性率63.6%(7/11),III期21.4%(3/14),I、II期6.3%(1/16),P=0.004。外周血CEAmRNA表达在不同年龄、性别、病理类型中无统计学差异(P>0.05)。结论:CEAmRNA、CEA和CA19-9在胃肠道癌患者中的阳性率,以CEAmRNA阳性率最高,随淋巴结转移数目、病期进展,阳性率更高,可能预后不良。