Objective: The aim of this study was to detect micrometastases in bone marrow of primary breast cancer patients, and compare with other clinical parameters. Methods: Cytokeratin 19 (CK-19) gene mRNA expression was det...Objective: The aim of this study was to detect micrometastases in bone marrow of primary breast cancer patients, and compare with other clinical parameters. Methods: Cytokeratin 19 (CK-19) gene mRNA expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot hybridization. Human breast cancer cell line T47D was mixed with bone marrow cells in different proportions. The positive detection rate was compared among RT-PCR, Southern blotting and immunohistochemistry (IHC) methods. Results: Cytokeratin 19 gene was expressed in all 6 positive control samples while the expression was not seen in 8 negative control samples. In all 54 patients 14 cases were CK-19 positive (25.9%) by RT-PCR, another positive signal was obtained in 5/54 (9.3%) of bone marrow samples by Southern blotting. The total positive cases are 19/54 (35.2%). CK-19 IHC+ cells were detected at a dilution of one T47D cell in 5×104 bone marrow cells, while the sensitivity detected by PCR and Southern blot hybridization was at 1∶5×105 and 1∶1×106, respectively. This demonstrates that RT-PCR and Southern blotting was at least 20 times more sensitive than the IHC method. The micrometastases positive rate of the larger tumor size group (>5.0 cm) was significantly (P<0.05) greater than that of the smaller tumor size group (0–2.0 cm). Conclusion: detection of micrometastases in bone marrow by RT-PCR and Southern blotting, using CK-19 as a biological marker, is highly sensitive and it is a method to be used for anticipating the prognosis of breast cancer patients.展开更多
Hepatitis B virus(HBV)infection is a leading cause of chronic hepatitis,liver cirrhosis,and hepatocellular carcinoma worldwide.Due to the shared modes of transmission,coinfection with HBV and human immunodeficiency vi...Hepatitis B virus(HBV)infection is a leading cause of chronic hepatitis,liver cirrhosis,and hepatocellular carcinoma worldwide.Due to the shared modes of transmission,coinfection with HBV and human immunodeficiency virus(HIV)is not uncommon.It is estimatedthat 10%of HIV-infected patients worldwide are coinfected with HBV.In areas where an HBV vaccination program is implemented,the HBV seroprevalence has declined significantly.In HIV/HBV-coinfected patients,HBV coinfection accelerates immunologic and clinical progression of HIV infection and increases the risk of hepatotoxicity when combination antiretroviral therapy(cART)is initiated,while HIV infection increases the risk of hepatitis events,cirrhosis,and end-stage liver disease related to chronic HBV infection.With the advances in antiviral therapy,concurrent,successful longterm suppression of HIV and HBV replication can be achieved in the cART era.To reduce the disease burden of HBV infection among HIV-infected patients,adoption of safe sex practices,avoidance of sharing needles and diluent,HBV vaccination and use of cART containing tenofovir disoproxil fumarate plus emtricitabine or lamivudine are the most effective approaches.However,due to HIV-related immunosuppression,using increased doses of HBV vaccine and novel approaches to HBV vaccination are needed to improve the immunogenicity of HBV vaccine among HIV-infected patients.展开更多
Objective： To study the effects of Shenggu injection （生骨注射液,SGI） on mRNA expression of vascular endothelia growth factor （VEGF） in rat osteoblasts in vitro and to explore its possible molecular mechanisms in...Objective： To study the effects of Shenggu injection （生骨注射液,SGI） on mRNA expression of vascular endothelia growth factor （VEGF） in rat osteoblasts in vitro and to explore its possible molecular mechanisms in promoting fracture healing. Methods： Rat osteoblasts cultured in vitro were stimulated with SGI according to the protocol. The expression levels of VEGF mRNA in the cells in every group were examined by reverse-transcriptase ploymerase chain reaction （RT-POR）. Results： When osteoblasts were stimulated with different concentrations of SGI for 5 days, the expression of VEGF mRNA peaked with 1 mg/ml SGI on the 5th day. When treated with 1 mg/ml SGI from the 1st to the 5th day, the expression of VEGF mRNA increased gradually with the increase of culturing time. Conclusion： SGI could promote significantly the expression of VEGF mRNA in rat osteoblasts in vitro. The levels of expression of VEGF mRNA changed along with different concentrations and stimulating time of SGI.展开更多
文摘Objective: The aim of this study was to detect micrometastases in bone marrow of primary breast cancer patients, and compare with other clinical parameters. Methods: Cytokeratin 19 (CK-19) gene mRNA expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot hybridization. Human breast cancer cell line T47D was mixed with bone marrow cells in different proportions. The positive detection rate was compared among RT-PCR, Southern blotting and immunohistochemistry (IHC) methods. Results: Cytokeratin 19 gene was expressed in all 6 positive control samples while the expression was not seen in 8 negative control samples. In all 54 patients 14 cases were CK-19 positive (25.9%) by RT-PCR, another positive signal was obtained in 5/54 (9.3%) of bone marrow samples by Southern blotting. The total positive cases are 19/54 (35.2%). CK-19 IHC+ cells were detected at a dilution of one T47D cell in 5×104 bone marrow cells, while the sensitivity detected by PCR and Southern blot hybridization was at 1∶5×105 and 1∶1×106, respectively. This demonstrates that RT-PCR and Southern blotting was at least 20 times more sensitive than the IHC method. The micrometastases positive rate of the larger tumor size group (>5.0 cm) was significantly (P<0.05) greater than that of the smaller tumor size group (0–2.0 cm). Conclusion: detection of micrometastases in bone marrow by RT-PCR and Southern blotting, using CK-19 as a biological marker, is highly sensitive and it is a method to be used for anticipating the prognosis of breast cancer patients.
基金Supported by Centers for Disease Control,Taiwan,Grant No.DOH 102-DC-1401
文摘Hepatitis B virus(HBV)infection is a leading cause of chronic hepatitis,liver cirrhosis,and hepatocellular carcinoma worldwide.Due to the shared modes of transmission,coinfection with HBV and human immunodeficiency virus(HIV)is not uncommon.It is estimatedthat 10%of HIV-infected patients worldwide are coinfected with HBV.In areas where an HBV vaccination program is implemented,the HBV seroprevalence has declined significantly.In HIV/HBV-coinfected patients,HBV coinfection accelerates immunologic and clinical progression of HIV infection and increases the risk of hepatotoxicity when combination antiretroviral therapy(cART)is initiated,while HIV infection increases the risk of hepatitis events,cirrhosis,and end-stage liver disease related to chronic HBV infection.With the advances in antiviral therapy,concurrent,successful longterm suppression of HIV and HBV replication can be achieved in the cART era.To reduce the disease burden of HBV infection among HIV-infected patients,adoption of safe sex practices,avoidance of sharing needles and diluent,HBV vaccination and use of cART containing tenofovir disoproxil fumarate plus emtricitabine or lamivudine are the most effective approaches.However,due to HIV-related immunosuppression,using increased doses of HBV vaccine and novel approaches to HBV vaccination are needed to improve the immunogenicity of HBV vaccine among HIV-infected patients.
文摘Objective： To study the effects of Shenggu injection （生骨注射液,SGI） on mRNA expression of vascular endothelia growth factor （VEGF） in rat osteoblasts in vitro and to explore its possible molecular mechanisms in promoting fracture healing. Methods： Rat osteoblasts cultured in vitro were stimulated with SGI according to the protocol. The expression levels of VEGF mRNA in the cells in every group were examined by reverse-transcriptase ploymerase chain reaction （RT-POR）. Results： When osteoblasts were stimulated with different concentrations of SGI for 5 days, the expression of VEGF mRNA peaked with 1 mg/ml SGI on the 5th day. When treated with 1 mg/ml SGI from the 1st to the 5th day, the expression of VEGF mRNA increased gradually with the increase of culturing time. Conclusion： SGI could promote significantly the expression of VEGF mRNA in rat osteoblasts in vitro. The levels of expression of VEGF mRNA changed along with different concentrations and stimulating time of SGI.
