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find Keyword "聚合酶链反应" 74 results
  • Joint Detection of hTERT mRNA and GGT mRNA-H Expressions in Peripheral Blood of Small Hepatocellular Carcinoma Patients and Its Clinical Significance

    ObjectiveTo investigate the expressions and clinical significance of human telomerase reverse transcriptase (hTERT) mRNA and γglutamyl transpeptidase mRNA-H (GGT mRNA-H) in the peripheral blood of small hepatocellular carcinoma (HCC) patients. MethodsThe expressions of hTERT mRNA and GGT mRNA-H were detected in the peripheral blood of thirty patients with small HCC by RT-PCR, eighteen patients with benign liver diseases, and twelve normal volunteers. ResultsThe positive rate of hTERT mRNA and GGT mRNA-H expression in patients with small HCC were 80.0% (24/30) and 46.7%(14/30), respectively. In patients with hepatitic cirrhosis the positive rate of hTERT mRNA expression was 33.3% (6/18), while the expression of GGT mRNA was not detected. Both the expressions of hTERT mRNA and GGT mRNA-H were negative in all normal volunteers. The combination analysis of hTERT mRNA and GGT mRNA-H expression achieved positive rate of 86.7% in the diagnosis of small HCC, which was significantly higher than the positive rate of AFP (26.7%), Plt;0.05. ConclusionThe hTERT mRNA and GGT mRNA-H are significantly expressed in small HCC patients, the combination analysis of hTERT mRNA and GGT mRNA-H seems to be useful in the early diagnosis of small HCC.

    Release date:2016-09-08 10:46 Export PDF Favorites Scan
  • Mutation and Expression of β-Catenin Gene in Breast Hyperplasia and Breast Cancer

    Objective To investigate the role of β-catenin gene in breast tumorigenesis by detecting mutation and expression of β-catenin gene in breast hyperplasia and breast cancer. Methods Mutation and expression of β-catenin gene in 42 breast cancer, 15 simple hyperplasia and 15 atypical hyperplasia were detected by polymerase chain reaction-single strand conformation polymorphism and immunohistochemistry. Results Normal expression of β-catenin occurred in tissue of breast simple hyperplasia. The rate of abnormal expression of β-catenin in tissue of breast atypical hyperplasia and breast cancer were 26.7% (4/15) and 59.5% (25/42), respectively, which were higher than that of simple hyherplasia tissue (P<0.05). And there was a markedly difference between the atypical hyperplasia tissue and breast cancer tissue (P<0.05). Mutation of β-catenin gene wasn’t detected in this three kinds of tissues. Conclusion Abnormal expression of β-catenin plays an important role in human breast tumorigenesis, reason of abnormal expression of β-catenin isn’t mutation of β-catenin gene. Expression of β-catenin can be regulated by other mechanisms.

    Release date:2016-08-28 04:08 Export PDF Favorites Scan
  • Experimental Study on Changes of Expression of AFP mRNA in Human Hepatocellular Carcinoma Tissues after Oral Xeloda Therapy

    【Abstract】Objective To explore the changes of expression of AFP mRNA in human hepatocellular carcinoma (HCC) tissues after oral Xeloda therapy.Methods Total RNA was extracted from HCC tissue samples collect after operation and nested reverse transcription polymerase chain reaction (RT-nested PCR) assay was performed to determine the expression of AFP mRNA in this study.Results The final product of AFP mRNA amplified by RT-PCR was 174 bp and by RT-nested PCR was 101 bp. The AFP mRNA is positive in 12 of 21 patients (positive rate 57.14%) amplified by RT-nested PCR assay in Xeloda treatment group which is much lower than control group: 18 of 20 patients (positive rate 90.00%),P<0.05.The serum AFP value of Xeloda treatment group 〔(23.2±12.8) μg/L〕 is much lower than that of control group 〔(39.6±24.3) μg/L〕 four weeks after operation (P<0.05). However, There was no difference between two groups in serum AFP value before operation.Conclusion Xeloda can effectively suppress the expression of AFP mRNA in human HCC tissues and lower it’s product serum AFP value.The clinical application of Xeloda in HCC patients deserve further study.

    Release date:2016-08-28 04:44 Export PDF Favorites Scan
  • Expression and Significance of the Multidrug Resistance-Associated Protein Gene in Primary Hepatocellular Carcinomas

    Objective To study the expression and significance of multidrug resistance-associated protein (MRP) gene in hepatocellular carcinoma (HCC). Methods Reverse transcription polymerase chain reaction (RT-PCR) assay was used to detect the expression of MRP mRNA in 25 fresh specimens of the primary HCC and its surrounding liver tissues. Immunohistochemistry LSAB technique was adopted to test MRP in 60 HCC specimens. The drug sensitivity was also tested by flow cytometry.Results The positive expression rates of MRP mRNA and MRP protein in primary HCC were 44.00%(11/25) and 45.00%(27/60) respectively. All the intensity of expression was low, but significant higer than its surrouding liver tissues (P<0.05). The intensity and expression rate of MRP protein in 5 recurrent HCC had a tendency to increase. There was a correlation between the expression of MRP mRNA and MRP protein in 25 patients using RT-PCR and immunohistochemistry assay (Plt;0.05). Detected by flow cytometry, the average sensitivity of drugs in vitro of 60 HCC sp-cimens were 5-FU (15.80±7.63)%,DDP(18.45±9.59)%,ADM(17.95±7.99)%,MMC(16.60±8.69)% and CTX(17.40±10.14)%. Only 5FU and ADM were significantly affected by the expression of MRP protein (Plt;0.05).Conclusion The expression of MRP in primary HCC may be one of the important mechanisms of the intrinsic and acquired drug resistance in HCC. To study the expression of MRP could give a predictive value in HCC chemotherapy.

    Release date:2016-08-28 04:47 Export PDF Favorites Scan
  • CONTRAST STUDY ON DIAGNOSIS OF LYMPH NODES METASTASIS BY CONVENTIONAL PATHOLOGY AND GENETIC DETECTION

    Objective To evaluate the potential of specific mRNA marker keratin 19(K19) to detect micrometastasis by reverse transcriptase polymerase chain reaction (RT-PCR) .Methods One hundred and ninty four regional lymph nodes harvested from 6 cases of benign diseases, 4 cases of breast carcinoma, 5 cases of gastric carcinoma and 12 cases of colorectal carcinoma patients were examined by conventional pathology and amplifying tissue specific K19 mRNA by RT-PCR separately, then the two methods were compared with each other. Results None of the 34 lymph nodes which were pathological metastasis-negative from benign diseases expressed K19 mRNA by RT-PCR, all of the 28 regional lymph nodes which were pathological metastasis-positive from malignant cases showed trains of K19 mRNA by RT-PCR. Of the 132 lymph nodes which were pathological metastasis-negative from malignant cases, 11 lymph nodes were detected with micrometastasis by genetic diagnosis.Conclusion Genetic diagnosis of lymph node micrometastasis is more sensitive than conventional pathology and has diagnostic value and merits further study.

    Release date:2016-08-28 05:30 Export PDF Favorites Scan
  • mRNA EXPRESSION OF nm23-H1 GENE IN HUMAN LIVER TUMOR ASSAYED BY REVERSE TRANSCRIPTIONPOLYMERASE CHAIN REACTION

    To investigate the mRNA expression of nm23-H1 gene in human liver tumor. In tumor and corresponding nontumoral liver specimens from 20 patients, nm23-H1 mRNA were examined by reverse transcriptionpolymerase chain reaction (RT-PCR) method with specific primers. Results: The primers designed in this study could amplified nearly entire coding sequence of nm23-H1 gene. All the samples showed positive expression of nm23-H1 mRNA, indicating there was no expression loss or obvious alteration. Conclusions: The achievement of RT-PCR method lays foundation for quantitative gaugement of nm23-H1 mRNA in liver tumor.

    Release date:2016-08-29 09:20 Export PDF Favorites Scan
  • A STUDY ON BACTERIAL DNA IN CHOLESTERAL GALLSTONES AND ITS CLINICAL SIGNIFICANCE BY NP-PCR

    The aim of the this study was to search for bacterial DNA sequences in cholesterol gallstones with negative bacterial culture by NP-PCR technique. Bacterial gene fragments were amplified in vitro from DNA which were extracted from cholesterol gallstones in gallbladder for identifying the existence of bacteria. The gallbladder gallstones of 30 patients were analysed. Bacterial DNA was found in the stones of 26 patients, indicating that most cholesterol gallstones harbor bacterial DNA.

    Release date:2016-08-29 03:19 Export PDF Favorites Scan
  • OVEREXPRESSION OF THE MDRl GENE EXPRESSION IN HUMAN PRIMARY HEPATOCELLU-LAR CARCINOMAS

    We have devised a highly sensitive, specific, and quantitative assay for multidrug resistance (mdr1) mRNA expression based on the reverse transcription-polymerase chain reaction (RT-PCR). mdr1 mRNA levels were detected in 30 human primary hepatocellular carcinoma (PHC) tissue and adjacent liver tissue. Five of the patients had received chemotherapy before hepatectomy. The results show that the level of expression of mdr1 gene is higher in tumor tissue than in adjacent liver tissue. mdr1 gene is overexpressed in PHC after chemotherapy. Furthermore, mdr1 gene expression in the treated tumor adjacent liver tissue is higher than that in untreated tumor adjacent liver tissue. Our results indicated that overexpression of mdr1 gene may be responsible for the intrinsic and acquired drug resistance of PHC.

    Release date:2016-08-29 03:25 Export PDF Favorites Scan
  • 风湿性二尖瓣狭窄心房颤动患者HCN4基因cDNA序列测定及mRNA的表达

    摘要: 目的 通过分析风湿性心脏病二尖瓣狭窄患者心房肌组织超级化激活环核苷酸调控通道基因家族4(HCN4)基因表达与心房颤动发生的关系,为探讨心房颤动发生的机制奠定理论基础。 方法 52例风湿性二尖瓣狭窄患者,根据是否合并心房颤动将其分为两组,实验组:38例,男18例,女20例;年龄26~68岁,平均年龄46.47岁;均合并心房颤动。对照组:14例,男6例,女8例;年龄21~62岁,平均年龄42.93岁;不合并心房颤动。提取并逆转录两组患者心房肌组织中HCN4基因的总核糖核酸(RNA),应用SYBR GreenⅠ荧光染料, 建立检测 HCN4基因信使RNA(mRNA)的实时荧光定量聚合酶链反应 (PCR)法,并对PCR产物测序进行分析。根据标准曲线计算出两组心房肌组织中HCN4基因 mRNA含量,并以HCN4基因mRNA和内参β肌动蛋白(β-actin)含量的比值作为评价HCN4基因mRNA表达水平指标。 结果 测定HCN4基因cDNA 序列同源性为100%。建立的实时荧光定量 PCR方法在103~107拷贝数/μl的标准品梯度稀释范围内r为0.999。实验组HCN4基因mRNA与β-actin含量的相对表达值比值与对照组比较明显升高(1.323±1.226 vs. 0.116±0.192,P<0.05)。 结论 实时荧光定量PCR对HCN4基因mRNA能进行准确定量,HCN4基因的过度转录表达提示其可能参与了调控风湿性二尖瓣狭窄心房颤动的发生过程。

    Release date:2016-08-30 06:01 Export PDF Favorites Scan
  • Measurement of the Forkhead Box P3 Gene Expression Levels in Esophageal Squamous Cell Carcinoma by Realtime Quantitative Reverse Transcriptionpolymerase Chain Reaction

    Objective To detect the expression of forkhead box P3 (FOXP3 )gene in esophageal squamous cell carcinoma(ESCC) and provide a new basis for immunotherapy of esophageal cancer. Methods Based on fluorescent TaqMan methodology, a realtime quantitative reverse transcription polymerase chain reaction (RT-PCR) for detecting the expression of FOXP3 was set up. In this method, a cloning vector pMD 18-T-FOXP3 was constructed as a standard plasmid. The specific expression of FOXP3 in 42 patients with ESCC and 30 healthy controls were measured by using GeneAmp 7500 Sequence Detection Systems. Results FOXP3 mRNA copy number in ESCC was significantly higher than that in healthy control tissue [(72.20±23.10)×104copy/μg RNA vs.(0.68±0.34)×104 copy/μg RNA;Plt;0.05]. Conclusion A realtime quantitative RT-PCR method for detecting the expression of FOXP3 gene in ESCC has been successfully established. The expression level of FOXP3 is increased in ESCC compare with healthy controls.

    Release date:2016-08-30 06:05 Export PDF Favorites Scan
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