The study aimed to investigate the impact of intraclot recombinant tissue-type plasminogen activator （rt-PA） on perihematomal edema （PHE） development in patients with intracerebral hemorrhage （ICH） treated with ...The study aimed to investigate the impact of intraclot recombinant tissue-type plasminogen activator （rt-PA） on perihematomal edema （PHE） development in patients with intracerebral hemorrhage （ICH） treated with minimally invasive surgery （MIS） and the effects of intraclot rt-PA on the 30-day survival. We reviewed the medical records of ICH patients undergoing MIS between October 2011 and July 2013. A volumetric analysis was done to assess the change in PHE and ICH volumes at pre-MIS （T1）, post-MIS （T2） and day 10-16 （T3） following diagnostic computed tomographic scans （To）. Forty-three patients aged 52.8±11.1 years with （n=30） or without rt-PA （n=13） were enrolled from our institutional ICH database. The median rt-PA dose was 1.5 （1） mg, with a maximum dose of 4.0 mg. The ratio of clot evacuation was significantly increased by intraclot rt-PA as compared with controls （77.9%±20.4% vs. 64%±15%; P=0.046）. From TI to T2, reduction in PHE volume was strongly associ- ated with the percentage of clot evacuation （p=0.34; P=-0.027）. In addition, PHE volume was positively correlated with residual ICH volume at the same day （p ranging from 0.39-0.56, P〈0.01）. There was no correlation between the cumulative dose of rt-PA and early （T2） PHE volume （p=0.24; P=0.12） or de- layed （T3） PHE volume （p=0.19; P=0.16）. The 30-day mortality was zero in this cohort. In the selected cohort of ICH patients treated with MIS, intraclot rt-PA accelerated clot removal and had no effects on PHE formation. MIS aspiration and low dose of rt-PA seemed to be feasible to reduce the 30-day mor- tality in patients with severe ICH. A large, randomized study addressing dose titration and long-term outcome is needed.展开更多
Background Cigarette smoking has an influence on both arterial-type and venous-type thrombosis. However, little is known about the direct effect of cigarette smoke extract （CSE） on fibrinolytic activity of human umb...Background Cigarette smoking has an influence on both arterial-type and venous-type thrombosis. However, little is known about the direct effect of cigarette smoke extract （CSE） on fibrinolytic activity of human umbilical vein endothelial cells （HUVECs）. Most recently, simvastatin has been marked in its effect on endothelial cells protection and anticoagulation. In this study, the effect of CSE on the expression of tissue-type plasminogen activator （t-PA） and plasminogen activator inhibitor-l（PAl-1） in HUVECs was addressed. The role of simvastatin in CSE-induced fibrinolytic activity changes was investigated as well. Methods The fourth to fifth generation of HUVECs were incubated respectively with 0, 5%, 10% and 20% CSE for 6 hours or exposed to 5% CSE for 0, 4, 6, 8, 12, 24 hours to determine the expression changes of t-PA and PAl-1 protein. Meanwhile, cells were also accordingly exposed either to 5% CSE alone or simvastatin pre-treated and 5% CSE for 24 hours to assess the role of simvastatin in CSE-induced t-PA and PAl-1 protein and mRNA expression in HUVECs. RT-PCR and ELISA techniques were used for detecting the t-PA or PAl-1 mRNA and protein. Results After 6-hour exposure to CSE, the expression levels of t-PA protein in 10% and 20% CSE-treated groups reduced significantly （（0.0365±0.0083） ng/ml, （0.0255±0.0087） ng/ml） when compared with that of control group （（0.0660±0.0120） ng/ml） （P 〈0.05）. In contrast, the levels of PAl-1 protein in 5%, 10% and 20% CSE-treated groups increased remarkably （（13.3225±0.5680） ng/ml, （14.2675±1.5380） ng/ml, （14.4292±1.6230） ng/ml） when compared with that of control group （（8.5193_±0.7537）ng/ml） （P〈0.05）. After stimulation with 5% CSE for 0, 4, 6, 8, 12, 24 hours, the levels of PAl-1 protein increased over time and reached the peak at 24 hours （（14.6400±1.0651） ng/ml）, which was significantly higher than that of control group （（12.0656±0.6148） ng/ml） （P 〈0.05）. Additionally, CSE could up-regulate PAl-1 expression at both the mRNA and the protein levels. The levels of PAl-1 mRNA and protein increased significantly in 5% CSE-treated group （（8.8030±0.4745） ng/ml, （1.8155±0.0412） ng/ml） compared with those of control groups （（5.0588±0.2315） ng/ml, （1.3030±0.0647） ng/ml） （P 〈0.01）, and decreased after 2-hour simvastatin pre-treatment （（5.4875±0.3166） ng/ml, （1.3975-±0.0297） ng/ml） （P 〈0.01）. No significant difference was found at the levels of t-PA protein and mRNA （P 〉0.05）. Conclusions CSE inhibits the fibrinolytic activity of HUVECs in vitro. Simvastatin plays a protective role in CSE-induced fibrinolytic malfunction.展开更多
Translation reading frame of human plasminogen activator inhibitor 1 (PAI 1) cDNA was amplified from total RNA extracted from glomerular mesangial cells by using reverse transcription polymerase chain reaction (RT PCR...Translation reading frame of human plasminogen activator inhibitor 1 (PAI 1) cDNA was amplified from total RNA extracted from glomerular mesangial cells by using reverse transcription polymerase chain reaction (RT PCR) and inserted into plasmid pUC19 in a sense orientation. Sequencing results revealed that PAI 1 cDNA had new initial codon ATG, but no signal peptide sequences, and translation reading frame was in agreement with PAI 1 cDNA derived from human endothelial cells. PAI 1 cDNA determined by sequencing was inserted into prokaryotic expression plasmid pBV220, and recombinant plasmid pBV220 PAI 1 which had high level expression in Escherichia coli was obtained. Recombinant PAI 1 protein attained to approximately 45% of total bacterial proteins. Western blotting showed that there was a specific band in the region of 43.0 ku. Latent recombinant PAI 1 with 97% purity was obtained from inclusion bodies after denaturation, renaturation and purification by FPLC. Recombinant PAI 1 activated by treatment with 4.0 mol/L guanidinium hydrochloride had significant inhibition activity to u PA and shared bioactivity in common with natural PAI 1.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81171089 and No.30770751)Key Clinical Program of the Ministry of Health of China(2010)the Future Program of New Technology and New Business in Tongji Hospital,China(2012)
文摘The study aimed to investigate the impact of intraclot recombinant tissue-type plasminogen activator （rt-PA） on perihematomal edema （PHE） development in patients with intracerebral hemorrhage （ICH） treated with minimally invasive surgery （MIS） and the effects of intraclot rt-PA on the 30-day survival. We reviewed the medical records of ICH patients undergoing MIS between October 2011 and July 2013. A volumetric analysis was done to assess the change in PHE and ICH volumes at pre-MIS （T1）, post-MIS （T2） and day 10-16 （T3） following diagnostic computed tomographic scans （To）. Forty-three patients aged 52.8±11.1 years with （n=30） or without rt-PA （n=13） were enrolled from our institutional ICH database. The median rt-PA dose was 1.5 （1） mg, with a maximum dose of 4.0 mg. The ratio of clot evacuation was significantly increased by intraclot rt-PA as compared with controls （77.9%±20.4% vs. 64%±15%; P=0.046）. From TI to T2, reduction in PHE volume was strongly associ- ated with the percentage of clot evacuation （p=0.34; P=-0.027）. In addition, PHE volume was positively correlated with residual ICH volume at the same day （p ranging from 0.39-0.56, P〈0.01）. There was no correlation between the cumulative dose of rt-PA and early （T2） PHE volume （p=0.24; P=0.12） or de- layed （T3） PHE volume （p=0.19; P=0.16）. The 30-day mortality was zero in this cohort. In the selected cohort of ICH patients treated with MIS, intraclot rt-PA accelerated clot removal and had no effects on PHE formation. MIS aspiration and low dose of rt-PA seemed to be feasible to reduce the 30-day mor- tality in patients with severe ICH. A large, randomized study addressing dose titration and long-term outcome is needed.
文摘Background Cigarette smoking has an influence on both arterial-type and venous-type thrombosis. However, little is known about the direct effect of cigarette smoke extract （CSE） on fibrinolytic activity of human umbilical vein endothelial cells （HUVECs）. Most recently, simvastatin has been marked in its effect on endothelial cells protection and anticoagulation. In this study, the effect of CSE on the expression of tissue-type plasminogen activator （t-PA） and plasminogen activator inhibitor-l（PAl-1） in HUVECs was addressed. The role of simvastatin in CSE-induced fibrinolytic activity changes was investigated as well. Methods The fourth to fifth generation of HUVECs were incubated respectively with 0, 5%, 10% and 20% CSE for 6 hours or exposed to 5% CSE for 0, 4, 6, 8, 12, 24 hours to determine the expression changes of t-PA and PAl-1 protein. Meanwhile, cells were also accordingly exposed either to 5% CSE alone or simvastatin pre-treated and 5% CSE for 24 hours to assess the role of simvastatin in CSE-induced t-PA and PAl-1 protein and mRNA expression in HUVECs. RT-PCR and ELISA techniques were used for detecting the t-PA or PAl-1 mRNA and protein. Results After 6-hour exposure to CSE, the expression levels of t-PA protein in 10% and 20% CSE-treated groups reduced significantly （（0.0365±0.0083） ng/ml, （0.0255±0.0087） ng/ml） when compared with that of control group （（0.0660±0.0120） ng/ml） （P 〈0.05）. In contrast, the levels of PAl-1 protein in 5%, 10% and 20% CSE-treated groups increased remarkably （（13.3225±0.5680） ng/ml, （14.2675±1.5380） ng/ml, （14.4292±1.6230） ng/ml） when compared with that of control group （（8.5193_±0.7537）ng/ml） （P〈0.05）. After stimulation with 5% CSE for 0, 4, 6, 8, 12, 24 hours, the levels of PAl-1 protein increased over time and reached the peak at 24 hours （（14.6400±1.0651） ng/ml）, which was significantly higher than that of control group （（12.0656±0.6148） ng/ml） （P 〈0.05）. Additionally, CSE could up-regulate PAl-1 expression at both the mRNA and the protein levels. The levels of PAl-1 mRNA and protein increased significantly in 5% CSE-treated group （（8.8030±0.4745） ng/ml, （1.8155±0.0412） ng/ml） compared with those of control groups （（5.0588±0.2315） ng/ml, （1.3030±0.0647） ng/ml） （P 〈0.01）, and decreased after 2-hour simvastatin pre-treatment （（5.4875±0.3166） ng/ml, （1.3975-±0.0297） ng/ml） （P 〈0.01）. No significant difference was found at the levels of t-PA protein and mRNA （P 〉0.05）. Conclusions CSE inhibits the fibrinolytic activity of HUVECs in vitro. Simvastatin plays a protective role in CSE-induced fibrinolytic malfunction.
文摘Translation reading frame of human plasminogen activator inhibitor 1 (PAI 1) cDNA was amplified from total RNA extracted from glomerular mesangial cells by using reverse transcription polymerase chain reaction (RT PCR) and inserted into plasmid pUC19 in a sense orientation. Sequencing results revealed that PAI 1 cDNA had new initial codon ATG, but no signal peptide sequences, and translation reading frame was in agreement with PAI 1 cDNA derived from human endothelial cells. PAI 1 cDNA determined by sequencing was inserted into prokaryotic expression plasmid pBV220, and recombinant plasmid pBV220 PAI 1 which had high level expression in Escherichia coli was obtained. Recombinant PAI 1 protein attained to approximately 45% of total bacterial proteins. Western blotting showed that there was a specific band in the region of 43.0 ku. Latent recombinant PAI 1 with 97% purity was obtained from inclusion bodies after denaturation, renaturation and purification by FPLC. Recombinant PAI 1 activated by treatment with 4.0 mol/L guanidinium hydrochloride had significant inhibition activity to u PA and shared bioactivity in common with natural PAI 1.
