We produced low-molecular-weight heparin/protamine micro (nano) particles (LMW-H/P MPs·NPs) as a carrier for heparin-binding growth factors (GFs), such as fibroblast growth factor (FGF)-2 and various GFs in plate...We produced low-molecular-weight heparin/protamine micro (nano) particles (LMW-H/P MPs·NPs) as a carrier for heparin-binding growth factors (GFs), such as fibroblast growth factor (FGF)-2 and various GFs in platelet-rich plasma (PRP). A mixture of LMW-H (MW: approximately 5000 Da, 6.4 mg/ml) and protamine (MW: approximately 3000 Da, 10 mg/ml) at a ratio of 7:3 (vol:vol) yields a dispersion of micro (nano) particles (200 nm - 3 μm in diameter). The diluted LMW-H solution in saline (0.32 mg/ml) mixed with diluted protamine (0.5 mg/ml) at a ratio at 7:3 (vol:vol) resulted in soluble nanoparticles (approximately 100 nm in diameter). The generated NPs could be then stabilized by adding 2 mg/ml dextran (MW: 178-217 kDa) and remained soluble after lyophilization of dialyzed LMW-H /P NPs solution. The LMW-H/P MPs·NPs adsorb GFs, control their release, protect GFs and activate their biological activities. Furthermore, administration of GFs-containing F/P MPs·NPs exhibited significantly higher inductions of vascularization and fibrous tissue formation in vivo than GFs alone. LMW-H/P MPs·NPs can also efficiently bind to tissue culture plates and retain the binding of GFs. The LMW-H/P MPs·NP-coated matrix with various GFs or cytokines provided novel biomaterials that could control cellular activity such as proliferation and differentiation. Thus, LMW-H/P MPs·NPs are an excellent carrier for GFs and are a functional coating matrix for various kinds of cell cultures.展开更多
Semi-large scale liquid cultivation of transformed Escherichia coli (E. coli) in medium (100-200 ml) has been widely used for the acquisition of relatively large amounts of plasmid DNA (50-300 μg). However, this meth...Semi-large scale liquid cultivation of transformed Escherichia coli (E. coli) in medium (100-200 ml) has been widely used for the acquisition of relatively large amounts of plasmid DNA (50-300 μg). However, this method requires an expensive high-speed centrifugation apparatus to precipitate E. coli before lysis, which is both laborious and time-consuming. Here, we demonstrate a method for agar plate-based cultivation of bacteria that does not employ a high-speed centrifugation apparatus. This procedure proves to be simple and reproducible, yielding an average of 82 μg of plasmid DNA per experiment. It may therefore be valuable for cloning/transfection experiments under limited financial backgrounds.展开更多
We experienced two cases of lung injury resulting from fluted silastic drain extraction under reservoir-generated negative pressure suction. In the first case, a 67-year-old man underwent coronary artery bypass grafti...We experienced two cases of lung injury resulting from fluted silastic drain extraction under reservoir-generated negative pressure suction. In the first case, a 67-year-old man underwent coronary artery bypass grafting. A 19 Fr BLAKE drain was placed at the pericardial cavity, the mediastinum, and the left thoracic cavity. All three drains were connected to J-VAC reservoirs. After removing the drains (which maintained the negative pressure), subcutaneous emphysema and hemopneumothorax occur. A trocar catheter was inserted. Bleeding and the air leak terminated within a day. In the second case, a 73-year-old man underwent aortic valve replacement. Right pneumothorax occurred after the removal of the pleural cavity drain, which maintained the negative pressure generated by the reservoir. We inserted a trocar catheter, and the air leak terminated within a day. We hypothesized that the reservoir-generated negative pressure causes the pleura to enter the groove of the drain and become damaged during extraction. We stopped using a reservoir to connect to the fluted silastic drain placed at the pleural cavity and removed the negative pressure when extracting the pleural cavity drain. Since implementing this change, we have not experienced a similar lung injury in more than 500 cardiac surgery patients.展开更多
Cardiac papillary fibroelastoma is the third most frequent primary cardiac tumor, after myxoma and fibroma. A papillary fibroelastoma that arises from the papillary muscle is rare. We removed a papillary fibroelastoma...Cardiac papillary fibroelastoma is the third most frequent primary cardiac tumor, after myxoma and fibroma. A papillary fibroelastoma that arises from the papillary muscle is rare. We removed a papillary fibroelastoma located at the anterior papillary muscle of the tricuspid valve. The papillary muscle needed reconstruction. We used the loop technique to adjust the length of the papillary muscle and performed annuloplasty by using an artificial ring.展开更多
文摘We produced low-molecular-weight heparin/protamine micro (nano) particles (LMW-H/P MPs·NPs) as a carrier for heparin-binding growth factors (GFs), such as fibroblast growth factor (FGF)-2 and various GFs in platelet-rich plasma (PRP). A mixture of LMW-H (MW: approximately 5000 Da, 6.4 mg/ml) and protamine (MW: approximately 3000 Da, 10 mg/ml) at a ratio of 7:3 (vol:vol) yields a dispersion of micro (nano) particles (200 nm - 3 μm in diameter). The diluted LMW-H solution in saline (0.32 mg/ml) mixed with diluted protamine (0.5 mg/ml) at a ratio at 7:3 (vol:vol) resulted in soluble nanoparticles (approximately 100 nm in diameter). The generated NPs could be then stabilized by adding 2 mg/ml dextran (MW: 178-217 kDa) and remained soluble after lyophilization of dialyzed LMW-H /P NPs solution. The LMW-H/P MPs·NPs adsorb GFs, control their release, protect GFs and activate their biological activities. Furthermore, administration of GFs-containing F/P MPs·NPs exhibited significantly higher inductions of vascularization and fibrous tissue formation in vivo than GFs alone. LMW-H/P MPs·NPs can also efficiently bind to tissue culture plates and retain the binding of GFs. The LMW-H/P MPs·NP-coated matrix with various GFs or cytokines provided novel biomaterials that could control cellular activity such as proliferation and differentiation. Thus, LMW-H/P MPs·NPs are an excellent carrier for GFs and are a functional coating matrix for various kinds of cell cultures.
文摘Semi-large scale liquid cultivation of transformed Escherichia coli (E. coli) in medium (100-200 ml) has been widely used for the acquisition of relatively large amounts of plasmid DNA (50-300 μg). However, this method requires an expensive high-speed centrifugation apparatus to precipitate E. coli before lysis, which is both laborious and time-consuming. Here, we demonstrate a method for agar plate-based cultivation of bacteria that does not employ a high-speed centrifugation apparatus. This procedure proves to be simple and reproducible, yielding an average of 82 μg of plasmid DNA per experiment. It may therefore be valuable for cloning/transfection experiments under limited financial backgrounds.
文摘We experienced two cases of lung injury resulting from fluted silastic drain extraction under reservoir-generated negative pressure suction. In the first case, a 67-year-old man underwent coronary artery bypass grafting. A 19 Fr BLAKE drain was placed at the pericardial cavity, the mediastinum, and the left thoracic cavity. All three drains were connected to J-VAC reservoirs. After removing the drains (which maintained the negative pressure), subcutaneous emphysema and hemopneumothorax occur. A trocar catheter was inserted. Bleeding and the air leak terminated within a day. In the second case, a 73-year-old man underwent aortic valve replacement. Right pneumothorax occurred after the removal of the pleural cavity drain, which maintained the negative pressure generated by the reservoir. We inserted a trocar catheter, and the air leak terminated within a day. We hypothesized that the reservoir-generated negative pressure causes the pleura to enter the groove of the drain and become damaged during extraction. We stopped using a reservoir to connect to the fluted silastic drain placed at the pleural cavity and removed the negative pressure when extracting the pleural cavity drain. Since implementing this change, we have not experienced a similar lung injury in more than 500 cardiac surgery patients.
文摘Cardiac papillary fibroelastoma is the third most frequent primary cardiac tumor, after myxoma and fibroma. A papillary fibroelastoma that arises from the papillary muscle is rare. We removed a papillary fibroelastoma located at the anterior papillary muscle of the tricuspid valve. The papillary muscle needed reconstruction. We used the loop technique to adjust the length of the papillary muscle and performed annuloplasty by using an artificial ring.
