AIM： To explore the effects of ultrasound exposure combined with microbubble contrast agent （SonoVue） on the permeability of the cellular membrane and on the expression of plasrnid DNA encoding enhanced green fluor...AIM： To explore the effects of ultrasound exposure combined with microbubble contrast agent （SonoVue） on the permeability of the cellular membrane and on the expression of plasrnid DNA encoding enhanced green fluorescent protein （pEGFP） transfer into human umbilical vein endothelial cells （HUVECs）. METHODS： HUVECs with fluorescein isothiocyanatedextran （FD500） and HUVECs with pEGFP were exposed to continuous wave （1.9 MHz, 80.0 mW/cm^2） for 5 min, with or without a SonoVue. The percentage of FD500 taken by the HUVECs and the transient expression rate of pEGFP in the HUVECs were examined by fluorescence microscopy and flow cytornetry, respectively. RESULTS： The percentage of FDS00-positive HUVECs in the group of ultrasound exposure combined with SonoVue was significantly higher than that of the group of ultrasound exposure alone （24.0%± 5.5% vs 66.6% ± 4.1%, P 〈 0.001）. Compared with the group of ultrasound exposure alone, the transfection expression rate of pEGFP in HUVECs was markedly increased with the addition of SonoVue （16.1% ± 1.9% vs 1.5% ± 0.2%, P 〈 0.001）. No statistical significant difference was observed in the HUVECs survival rates between the ultrasound group with and without the addition of SonoVue （94.1% ± 2.3% vs 91.1% ± 4.1% ）. CONCLUSION： The cell membrane permeability of HUVECs and the transfection efficiency of pEGFP into HUVECs exposed to ultrasound are significantly increased after addition of an ultrasound contrast agent without obvious damage to the survival of HUVECs. This non- invasive gene transfer method may be a useful tool for clinical gene therapy of hepatic tumors.展开更多
Background and Objective:Percutaneous ultrasound-guided thermal ablation is one of the major treatment methods for liver cancer.Tumor location close to the diaphragm or gastrointestinal tract was regarded as the treat...Background and Objective:Percutaneous ultrasound-guided thermal ablation is one of the major treatment methods for liver cancer.Tumor location close to the diaphragm or gastrointestinal tract was regarded as the treatment contraindication before due to poor visibility of the tumor or increased risk of thermal injury to the adjacent organs.This study used artificial pleural effusion or ascites to extend the indications of thermal ablation for liver cancer.Methods:Artificial pleural effusion (20 cases) or ascites (36 cases) was performed in 56 difficult cases of percutaneous thermal ablation for liver tumors.The technical success rates, the rate of approaching the procedure goal, complications, and local treatment response were assessed.Results:The technical success rates were 95%(19/20) for artificial pleural effusion and 100% (36/36) for artificial ascites, the achieve purpose rates were 100%(19/19) and 91.7% (33/36), the complete ablation rates were 84.2% (16/19) and 93.9% (31/33), respectively.Coughing, transient hematuria, and subcutaneous effusion were observed in 3 patients after the procedure of artificial pleural effusion, and hydrothorax in the right chest occurred in 1 patient during the artificial ascites process.Conclusions:Thermal ablation with the use of artificial pleural effusion or ascites is a safe and effective treatment for liver tumors, and the technique can widen the indications of thermal ablation for liver tumors.展开更多
AIM:To assess the usefulness of contrast-enhanced ultrasound (CEUS) during follow-up after percutaneous ablation therapy for hepatocellular carcinoma (HCC).METHODS:A total of 141 patients with HCCs who received percut...AIM:To assess the usefulness of contrast-enhanced ultrasound (CEUS) during follow-up after percutaneous ablation therapy for hepatocellular carcinoma (HCC).METHODS:A total of 141 patients with HCCs who received percutaneous ablation therapy were assessed by paired follow-up CEUS and contrast-enhanced computed tomography (CECT).The follow-up scheme was designed prospectively and the intervals between CEUS and CECT examinations were less than 14 d.Both im-ages of follow-up CEUS and CECT were reviewed by radiologists.The ablated lesions were evaluated and classified as local tumor progression (LTP) and LTPfree.LTP was defined as regrowth of tumor inside or adjacent to the successfully treated nodule.The detected new intrahepatic recurrences were also evaluated and defined as presence of intrahepatic new foci.On CEUS and CECT,LTP and new intrahepatic recurrence both were displayed as typical enhancement pattern of HCC (i.e.,hyper-enhancing during the arterial phase and washout in the late phase).With CECT as the reference standard,the ability of CEUS in detecting LTP or new intrahepatic recurrence during follow-up was evaluated.RESULTS:During a follow-up period of 1-31 mo (median,4 mo),169 paired CEUS and CECT examinations were carried out for the 141 patients.For a total of 221 ablated lesions,266 comparisons between CEUS and CECT findings were performed.Thirty-three LTPs were detected on CEUS whereas 40 LTPs were detected on CECT,there was significant difference (P < 0.001).In comparison with CECT,the numbers of false positive and false negative LTPs detected on CEUS were 6 and 13,respectively;the sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and overall accuracy of CEUS in detecting LTPs were 67.5%,97.4%,81.8%,94.4% and 92.3%,respectively.Meanwhile,131 new intrahepatic recurrent foci were detected on CEUS whereas 183 were detected on CECT,there was also significant difference (P < 0.05).In comparison with CECT,the numbers of false positive and false negative intrahepatic r展开更多
AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endo...AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endothelial cells.METHODS: The KDR-TK fragment was extracted from pBluescript Ⅱ KDR-TK plasmid by enzymatic digestion with Xho I and Sal I. The enhanced green fluorescence protein (EGFP) carrier was extracted from pEGFP by the same procedure. The KDR-TK was inserted into the pEGFP carrier to construct pEGFP-KDR-TK. Using ultrasound irradiation and microbubble, pEGFP-KDR-TK was transferred into human umbilical vein endothelial cells (HUVECs). The transient infection rate was estimated by green fluorescent protein (GFP) expression. Transfected HUVECs, non-transfected HUVECs, and HepG2 cells were cultured in the presence of different concentrations of ganciclovir (GCV), and the killing efficacy of HSV-TK/GCV was analyzed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The recombinant pEGFP-KDR-TK was successfully constructed by inserting the KDR-TK fragment into the pEGFP carrier. Transfected HUVECs showed cytoplasmic green fluorescence, and the transient transfection rate was about 20.3%. Pools of G418-resistant cells exhibited a higher sensitivity to theprodrug/GCV compared to non-transfected HUVECs or non-transfected HepG2 cells, respectively. CONCLUSION: KDR promoter and the suicide gene/prodrug system mediated by diagnostic ultrasound combined with microbubble can significantly kill HUVECs. Such therapy may present a novel and attractive approach to target gene therapy on tumor vessels.展开更多
With an arid climate and shortage of water resources,the groundwater dependent ecosystems in the oasis-desert ecotone of the Shiyang River Watershed has been extremely damaged,and the water crisis in the oasis has bec...With an arid climate and shortage of water resources,the groundwater dependent ecosystems in the oasis-desert ecotone of the Shiyang River Watershed has been extremely damaged,and the water crisis in the oasis has become a major concern in the social and the scientific community.In this study,the degene-ration characteristics of the groundwater ecological function was identified and comprehensive evaluated,based on groundwater depth data,vegetation quadrat and normalized difference vegetation index(NDVI)from Landsat program.The results showed that(1)the suitable groundwater depth for sustainable ecology in the Shiyang River Watershed is about 2-4 m;(2)the terms of degenerative,qualitative and disastrous stages of the groundwater ecological function are defined with the groundwater depths of about 5 m,7 m and 10 m;(3)generally,the groundwater ecological function in the oasis-desert ecotone of the lower reaches of Shiyang River Watershed is weak with an area of 1397.9 km2 identified as the severe deterioration region,which accounted 74.7%of the total area.In the meantime,the percentages of the good,mild and moderate deterioration areas of groundwater ecological function are 3.5%,5.5%and 16.3%,respectively,which were mainly distributed in the Qingtu lake area and the southeastern area of the Shoucheng town;(4)the degradation and shrinkage of natural oasis could be attributed to the dramatic groundwater decline,which is generally caused by irrational use of water and soil resources.This study could provide theoretical basis and scientific support for the decision-making in environmental management and ecological restoration of the Shiyang River Watershed.展开更多
基金Supported by grants from the Nationl Natural Scientific Foundation of China, No.30300082, 30470467, and Scientific Foundation Committee of Guangdong Province, China, No.04009360
文摘AIM： To explore the effects of ultrasound exposure combined with microbubble contrast agent （SonoVue） on the permeability of the cellular membrane and on the expression of plasrnid DNA encoding enhanced green fluorescent protein （pEGFP） transfer into human umbilical vein endothelial cells （HUVECs）. METHODS： HUVECs with fluorescein isothiocyanatedextran （FD500） and HUVECs with pEGFP were exposed to continuous wave （1.9 MHz, 80.0 mW/cm^2） for 5 min, with or without a SonoVue. The percentage of FD500 taken by the HUVECs and the transient expression rate of pEGFP in the HUVECs were examined by fluorescence microscopy and flow cytornetry, respectively. RESULTS： The percentage of FDS00-positive HUVECs in the group of ultrasound exposure combined with SonoVue was significantly higher than that of the group of ultrasound exposure alone （24.0%± 5.5% vs 66.6% ± 4.1%, P 〈 0.001）. Compared with the group of ultrasound exposure alone, the transfection expression rate of pEGFP in HUVECs was markedly increased with the addition of SonoVue （16.1% ± 1.9% vs 1.5% ± 0.2%, P 〈 0.001）. No statistical significant difference was observed in the HUVECs survival rates between the ultrasound group with and without the addition of SonoVue （94.1% ± 2.3% vs 91.1% ± 4.1% ）. CONCLUSION： The cell membrane permeability of HUVECs and the transfection efficiency of pEGFP into HUVECs exposed to ultrasound are significantly increased after addition of an ultrasound contrast agent without obvious damage to the survival of HUVECs. This non- invasive gene transfer method may be a useful tool for clinical gene therapy of hepatic tumors.
基金National Natural Science Foundation of China (No.30970837)New Century Excellent Talent Supporting Program of the Chinese Ministry of Education (No.NCET-06-0723)
文摘Background and Objective:Percutaneous ultrasound-guided thermal ablation is one of the major treatment methods for liver cancer.Tumor location close to the diaphragm or gastrointestinal tract was regarded as the treatment contraindication before due to poor visibility of the tumor or increased risk of thermal injury to the adjacent organs.This study used artificial pleural effusion or ascites to extend the indications of thermal ablation for liver cancer.Methods:Artificial pleural effusion (20 cases) or ascites (36 cases) was performed in 56 difficult cases of percutaneous thermal ablation for liver tumors.The technical success rates, the rate of approaching the procedure goal, complications, and local treatment response were assessed.Results:The technical success rates were 95%(19/20) for artificial pleural effusion and 100% (36/36) for artificial ascites, the achieve purpose rates were 100%(19/19) and 91.7% (33/36), the complete ablation rates were 84.2% (16/19) and 93.9% (31/33), respectively.Coughing, transient hematuria, and subcutaneous effusion were observed in 3 patients after the procedure of artificial pleural effusion, and hydrothorax in the right chest occurred in 1 patient during the artificial ascites process.Conclusions:Thermal ablation with the use of artificial pleural effusion or ascites is a safe and effective treatment for liver tumors, and the technique can widen the indications of thermal ablation for liver tumors.
基金Supported by Public Welfare Research Special Project from Chinese Ministry of Health,No.2008-2-10Key Project from Shanghai Health Bureau,No.20114003+2 种基金Shanghai Talent Development Project from Shanghai Human Resource and Social Security Bureau,No.2012045National Natural Science Foundation of China,No.30970837Chinese Ministry of Education,NCET-06-0723
文摘AIM:To assess the usefulness of contrast-enhanced ultrasound (CEUS) during follow-up after percutaneous ablation therapy for hepatocellular carcinoma (HCC).METHODS:A total of 141 patients with HCCs who received percutaneous ablation therapy were assessed by paired follow-up CEUS and contrast-enhanced computed tomography (CECT).The follow-up scheme was designed prospectively and the intervals between CEUS and CECT examinations were less than 14 d.Both im-ages of follow-up CEUS and CECT were reviewed by radiologists.The ablated lesions were evaluated and classified as local tumor progression (LTP) and LTPfree.LTP was defined as regrowth of tumor inside or adjacent to the successfully treated nodule.The detected new intrahepatic recurrences were also evaluated and defined as presence of intrahepatic new foci.On CEUS and CECT,LTP and new intrahepatic recurrence both were displayed as typical enhancement pattern of HCC (i.e.,hyper-enhancing during the arterial phase and washout in the late phase).With CECT as the reference standard,the ability of CEUS in detecting LTP or new intrahepatic recurrence during follow-up was evaluated.RESULTS:During a follow-up period of 1-31 mo (median,4 mo),169 paired CEUS and CECT examinations were carried out for the 141 patients.For a total of 221 ablated lesions,266 comparisons between CEUS and CECT findings were performed.Thirty-three LTPs were detected on CEUS whereas 40 LTPs were detected on CECT,there was significant difference (P < 0.001).In comparison with CECT,the numbers of false positive and false negative LTPs detected on CEUS were 6 and 13,respectively;the sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and overall accuracy of CEUS in detecting LTPs were 67.5%,97.4%,81.8%,94.4% and 92.3%,respectively.Meanwhile,131 new intrahepatic recurrent foci were detected on CEUS whereas 183 were detected on CECT,there was also significant difference (P < 0.05).In comparison with CECT,the numbers of false positive and false negative intrahepatic r
基金New Century Distinguished Scholar Supporting Program of Ministry of Education (80000-3171404) The National Natural Science Foundation of China, No. 30300082, No. 30470467
文摘AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endothelial cells.METHODS: The KDR-TK fragment was extracted from pBluescript Ⅱ KDR-TK plasmid by enzymatic digestion with Xho I and Sal I. The enhanced green fluorescence protein (EGFP) carrier was extracted from pEGFP by the same procedure. The KDR-TK was inserted into the pEGFP carrier to construct pEGFP-KDR-TK. Using ultrasound irradiation and microbubble, pEGFP-KDR-TK was transferred into human umbilical vein endothelial cells (HUVECs). The transient infection rate was estimated by green fluorescent protein (GFP) expression. Transfected HUVECs, non-transfected HUVECs, and HepG2 cells were cultured in the presence of different concentrations of ganciclovir (GCV), and the killing efficacy of HSV-TK/GCV was analyzed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The recombinant pEGFP-KDR-TK was successfully constructed by inserting the KDR-TK fragment into the pEGFP carrier. Transfected HUVECs showed cytoplasmic green fluorescence, and the transient transfection rate was about 20.3%. Pools of G418-resistant cells exhibited a higher sensitivity to theprodrug/GCV compared to non-transfected HUVECs or non-transfected HepG2 cells, respectively. CONCLUSION: KDR promoter and the suicide gene/prodrug system mediated by diagnostic ultrasound combined with microbubble can significantly kill HUVECs. Such therapy may present a novel and attractive approach to target gene therapy on tumor vessels.
基金This research was supported by the National Key Research and Development Plan of China(No.2017YFC0406103)the National Natural Science Foundation of China(No.41902262)the Geological Survey Project of China(No.DD20190349).
文摘With an arid climate and shortage of water resources,the groundwater dependent ecosystems in the oasis-desert ecotone of the Shiyang River Watershed has been extremely damaged,and the water crisis in the oasis has become a major concern in the social and the scientific community.In this study,the degene-ration characteristics of the groundwater ecological function was identified and comprehensive evaluated,based on groundwater depth data,vegetation quadrat and normalized difference vegetation index(NDVI)from Landsat program.The results showed that(1)the suitable groundwater depth for sustainable ecology in the Shiyang River Watershed is about 2-4 m;(2)the terms of degenerative,qualitative and disastrous stages of the groundwater ecological function are defined with the groundwater depths of about 5 m,7 m and 10 m;(3)generally,the groundwater ecological function in the oasis-desert ecotone of the lower reaches of Shiyang River Watershed is weak with an area of 1397.9 km2 identified as the severe deterioration region,which accounted 74.7%of the total area.In the meantime,the percentages of the good,mild and moderate deterioration areas of groundwater ecological function are 3.5%,5.5%and 16.3%,respectively,which were mainly distributed in the Qingtu lake area and the southeastern area of the Shoucheng town;(4)the degradation and shrinkage of natural oasis could be attributed to the dramatic groundwater decline,which is generally caused by irrational use of water and soil resources.This study could provide theoretical basis and scientific support for the decision-making in environmental management and ecological restoration of the Shiyang River Watershed.
