Objective： The aim of the present study is to explore the expression of a specific marker of breast cancer, small breast epithefial mucin （SBEM） mRNA, in peripheral blood and to investigate its significance. Method...Objective： The aim of the present study is to explore the expression of a specific marker of breast cancer, small breast epithefial mucin （SBEM） mRNA, in peripheral blood and to investigate its significance. Methods： The expressions of SBEM-mRNA in peripheral blood of 67 patients with breast cancer, 16 patients with benign breast disease, and 20 normal healthy volunteers were detected with nested reverse transcription-polymerase chain reaction （Nested-RT-PCR）. Results： SBEMomRNA was negative in healthy individuals and patients with benign breast tumor, but 50.7%（34/67） of breast cancer patients showed positive expression of SBEM-mRNA in peripheral blood, of whom 25%（2/8） were in stage Ⅰ, 45.8%（11/24） in stage Ⅱ, 43.75%（11/24） in stage Ⅲ and 73.7（14/19） in stage Ⅳ. The positive rate in stage Ⅳ was higher than that in stage Ⅰ, Ⅱ, Ⅲ （P〈0.05）. Expressions of SBEM-mRNA had no correlation with age, carcinoma size, pathological type, ER and PR state （P〉0.05）. Concision： SBEM-mRNA is specifically expressed in breast cancer and it may act as a marker for the detection of of breast cancer.展开更多
Objective: To study the clinical significance of human mammaglobin (hMAM) mRNA in peripheral blood of patients with breast cancer. Methods: The expression of hMAM mRNA was detected in peripheral blood from patients wi...Objective: To study the clinical significance of human mammaglobin (hMAM) mRNA in peripheral blood of patients with breast cancer. Methods: The expression of hMAM mRNA was detected in peripheral blood from patients with breast cancer, breast benign lesions, healthy volunteers by nested reverse transcription polymerase chain reaction (nested-RT-PCR) method. The possible correlations of hMAM mRNA expression with clinico-pathological parameters and related molecular markers such as p53, PCNA and HER-2 were analyzed. The hMAM mRNA changed in peripheral blood after chemotherapy was observed. Results: The positive rate of hMAM mRNA was 34.0% in peripheral blood of breast cancer, hMAM mRNA was not expressed in peripheral blood from other cancers, breast benign lesions or healthy volunteers. The expression of hMAM mRNA in peripheral blood was correlated with lymph nodes status and TNM stage. After 2-3 cycles adjuvant chemotherapy, In 13 of 17 cases, the positive hMAM mRNA turned to be negative, and there was statistical difference between pre-chemo- therapy and post-chemotherapy. Conclusion: hMAM mRNA has specific expression in breast cancer, the positive expression of hMAM mRNA in peripheral blood might predict hematogenous metastatic spreading of tumor cells, and hMAM mRNA may be potential biological marker for detecting breast cancer micrometastasis.展开更多
BACKGROUND:Hepatocellular carcinoma (HCC) is characterized by a multi-cause,multi-stage and multi-focus process of tumor progression.Its prognosis is poor and early diagnosis is of utmost importance.This study was und...BACKGROUND:Hepatocellular carcinoma (HCC) is characterized by a multi-cause,multi-stage and multi-focus process of tumor progression.Its prognosis is poor and early diagnosis is of utmost importance.This study was undertaken to investigate the dynamic expression of oncofetal antigen glypican-3 (GPC-3) and GPC-3 mRNA in hepatocarcinogenesis and to explore their early diagnostic value for HCC.METHODS:A hepatoma model was induced in male Sprague-Dawley rats with 0.05% 2-fluorenylacetamide and confirmed by hematoxylin and eosin staining and gamma-glutamyltransferase (GGT) expression.Total RNA was purified and transcribed into cDNA by reverse transcription.Fragments of the GPC-3 gene were amplified by nested RT-PCR,and confirmed by sequencing.GPC-3 was analyzed by immunohistochemistry,Western blotting or ELISA.RESULTS:Positive GPC-3 expression showed as brown granule-like staining localized in the cytoplasm.Histological examination of hepatocytes revealed three morphological stages of granule-like degeneration,atypical hyperplasia (precancerous),and cancer formation,with a progressive increase of liver total RNA and GGT expression.The incidence of liver GPC-3 mRNA and GPC-3,and serum GPC-3 was 100%,100% and 77.8% in the HCC group,100%,100%,and 66.7% in the precancerous group,83.3%,83.3%,and 38.9% in the degeneration group,and no expression in the liver or blood of the control group,respectively.There was a positive correlation between liver GPC-3 mRNA and total RNA level (r=0.475,P<0.05) or liver GPC-3 (r=1.0,P<0.001) or serum GPC-3 (r= 0.994,P<0.001).CONCLUSION:Abnormal oncofetal antigen GPC-3 and GPC-3 mRNA expression in hepatocarcinogenesis may be promising molecular markers for early diagnosis of HCC.展开更多
菜豆荚斑驳病毒（Bean pod mottle virus，BPMV）在美国东部和南部的大豆产区广泛分布。该病毒能够造成3％～52％的产量损失并使大豆品质降低，目前我国未见其分布。针对进口大豆种子的植物病毒检测，本文建立了半巢式RTPCR技术检测BPM...菜豆荚斑驳病毒（Bean pod mottle virus，BPMV）在美国东部和南部的大豆产区广泛分布。该病毒能够造成3％～52％的产量损失并使大豆品质降低，目前我国未见其分布。针对进口大豆种子的植物病毒检测，本文建立了半巢式RTPCR技术检测BPMV的方法。该方法采用Trizol快速提取大豆种子病毒总RNA，并根据BPMV的外壳蛋白编码基因设计特异性引物，经第一轮RT—PCR和第二轮半巢式PCR扩增．分别得到275bp和196bp大小的特异性基因片段。半巢式RT—PCR扩增产物的测序表明，该产物序列与BPMV的外壳蛋白编码基因存在91％～95％的高度同源性。检测灵敏度比较研究显示，DAS—ELISA与RT—PCR的检测灵敏度相近，半巢式RT—PCR的检测灵敏度比这2种方法高出10^3倍以上。展开更多
禽黄病毒病是新发的一种疫病,可引起鸭、鹅和鸡等家禽产蛋和采食量下降及死亡。本研究旨在建立一种快速诊断方法用于临床诊断及流行病学调查。根据GenBank发表的鹅黄病毒JS804株全基因序列,应用Primer Premier 5.0软件设计了2对特异性引...禽黄病毒病是新发的一种疫病,可引起鸭、鹅和鸡等家禽产蛋和采食量下降及死亡。本研究旨在建立一种快速诊断方法用于临床诊断及流行病学调查。根据GenBank发表的鹅黄病毒JS804株全基因序列,应用Primer Premier 5.0软件设计了2对特异性引物,建立了禽黄病毒套式RT-PCR检测方法。结果:该套式RT-PCR方法具有特异性强、敏感性高的特点,最低病毒检测量为101.89TCID50/0.1 mL,比普通RT-PCR方法敏感性高1 000倍。应用该方法对江苏地区疑似禽黄病毒病的70份鹅病料、4份鸭病料、12份鸡病料进行检测,总阳性率为58.14%,而用普通RT-PCR方法检测的阳性率仅为17.44%。结果表明,禽黄病毒套式RT-PCR检测方法具有快速、特异、敏感的特点,可用于禽黄病毒感染的临床诊断和流行病学调查。展开更多
基金This work was supported by the Natural Science Foundation of Guangxi Province (No. 0640161).
文摘Objective： The aim of the present study is to explore the expression of a specific marker of breast cancer, small breast epithefial mucin （SBEM） mRNA, in peripheral blood and to investigate its significance. Methods： The expressions of SBEM-mRNA in peripheral blood of 67 patients with breast cancer, 16 patients with benign breast disease, and 20 normal healthy volunteers were detected with nested reverse transcription-polymerase chain reaction （Nested-RT-PCR）. Results： SBEMomRNA was negative in healthy individuals and patients with benign breast tumor, but 50.7%（34/67） of breast cancer patients showed positive expression of SBEM-mRNA in peripheral blood, of whom 25%（2/8） were in stage Ⅰ, 45.8%（11/24） in stage Ⅱ, 43.75%（11/24） in stage Ⅲ and 73.7（14/19） in stage Ⅳ. The positive rate in stage Ⅳ was higher than that in stage Ⅰ, Ⅱ, Ⅲ （P〈0.05）. Expressions of SBEM-mRNA had no correlation with age, carcinoma size, pathological type, ER and PR state （P〉0.05）. Concision： SBEM-mRNA is specifically expressed in breast cancer and it may act as a marker for the detection of of breast cancer.
文摘Objective: To study the clinical significance of human mammaglobin (hMAM) mRNA in peripheral blood of patients with breast cancer. Methods: The expression of hMAM mRNA was detected in peripheral blood from patients with breast cancer, breast benign lesions, healthy volunteers by nested reverse transcription polymerase chain reaction (nested-RT-PCR) method. The possible correlations of hMAM mRNA expression with clinico-pathological parameters and related molecular markers such as p53, PCNA and HER-2 were analyzed. The hMAM mRNA changed in peripheral blood after chemotherapy was observed. Results: The positive rate of hMAM mRNA was 34.0% in peripheral blood of breast cancer, hMAM mRNA was not expressed in peripheral blood from other cancers, breast benign lesions or healthy volunteers. The expression of hMAM mRNA in peripheral blood was correlated with lymph nodes status and TNM stage. After 2-3 cycles adjuvant chemotherapy, In 13 of 17 cases, the positive hMAM mRNA turned to be negative, and there was statistical difference between pre-chemo- therapy and post-chemotherapy. Conclusion: hMAM mRNA has specific expression in breast cancer, the positive expression of hMAM mRNA in peripheral blood might predict hematogenous metastatic spreading of tumor cells, and hMAM mRNA may be potential biological marker for detecting breast cancer micrometastasis.
基金supported by grants-in-aid from the Projects of Medical Science (H200925)the Natural Science Foundation of Jiangsu Province,China (BK2008187)
文摘BACKGROUND:Hepatocellular carcinoma (HCC) is characterized by a multi-cause,multi-stage and multi-focus process of tumor progression.Its prognosis is poor and early diagnosis is of utmost importance.This study was undertaken to investigate the dynamic expression of oncofetal antigen glypican-3 (GPC-3) and GPC-3 mRNA in hepatocarcinogenesis and to explore their early diagnostic value for HCC.METHODS:A hepatoma model was induced in male Sprague-Dawley rats with 0.05% 2-fluorenylacetamide and confirmed by hematoxylin and eosin staining and gamma-glutamyltransferase (GGT) expression.Total RNA was purified and transcribed into cDNA by reverse transcription.Fragments of the GPC-3 gene were amplified by nested RT-PCR,and confirmed by sequencing.GPC-3 was analyzed by immunohistochemistry,Western blotting or ELISA.RESULTS:Positive GPC-3 expression showed as brown granule-like staining localized in the cytoplasm.Histological examination of hepatocytes revealed three morphological stages of granule-like degeneration,atypical hyperplasia (precancerous),and cancer formation,with a progressive increase of liver total RNA and GGT expression.The incidence of liver GPC-3 mRNA and GPC-3,and serum GPC-3 was 100%,100% and 77.8% in the HCC group,100%,100%,and 66.7% in the precancerous group,83.3%,83.3%,and 38.9% in the degeneration group,and no expression in the liver or blood of the control group,respectively.There was a positive correlation between liver GPC-3 mRNA and total RNA level (r=0.475,P<0.05) or liver GPC-3 (r=1.0,P<0.001) or serum GPC-3 (r= 0.994,P<0.001).CONCLUSION:Abnormal oncofetal antigen GPC-3 and GPC-3 mRNA expression in hepatocarcinogenesis may be promising molecular markers for early diagnosis of HCC.
文摘菜豆荚斑驳病毒（Bean pod mottle virus，BPMV）在美国东部和南部的大豆产区广泛分布。该病毒能够造成3％～52％的产量损失并使大豆品质降低，目前我国未见其分布。针对进口大豆种子的植物病毒检测，本文建立了半巢式RTPCR技术检测BPMV的方法。该方法采用Trizol快速提取大豆种子病毒总RNA，并根据BPMV的外壳蛋白编码基因设计特异性引物，经第一轮RT—PCR和第二轮半巢式PCR扩增．分别得到275bp和196bp大小的特异性基因片段。半巢式RT—PCR扩增产物的测序表明，该产物序列与BPMV的外壳蛋白编码基因存在91％～95％的高度同源性。检测灵敏度比较研究显示，DAS—ELISA与RT—PCR的检测灵敏度相近，半巢式RT—PCR的检测灵敏度比这2种方法高出10^3倍以上。
文摘禽黄病毒病是新发的一种疫病,可引起鸭、鹅和鸡等家禽产蛋和采食量下降及死亡。本研究旨在建立一种快速诊断方法用于临床诊断及流行病学调查。根据GenBank发表的鹅黄病毒JS804株全基因序列,应用Primer Premier 5.0软件设计了2对特异性引物,建立了禽黄病毒套式RT-PCR检测方法。结果:该套式RT-PCR方法具有特异性强、敏感性高的特点,最低病毒检测量为101.89TCID50/0.1 mL,比普通RT-PCR方法敏感性高1 000倍。应用该方法对江苏地区疑似禽黄病毒病的70份鹅病料、4份鸭病料、12份鸡病料进行检测,总阳性率为58.14%,而用普通RT-PCR方法检测的阳性率仅为17.44%。结果表明,禽黄病毒套式RT-PCR检测方法具有快速、特异、敏感的特点,可用于禽黄病毒感染的临床诊断和流行病学调查。
