To compare the effects of ticagrelor combined with prourokinase and reteplase thrombolysis on cardiac function and blood-related indexes in patients with ST-segment elevation myocardial infarction(STEMI),280 patients ...To compare the effects of ticagrelor combined with prourokinase and reteplase thrombolysis on cardiac function and blood-related indexes in patients with ST-segment elevation myocardial infarction(STEMI),280 patients hospitalized between January 2017 and December 2021 were randomly assigned into two groups:the experimental group and the control group,each comprising 140 cases.The control group received ticagrelor combined with prourokinase thrombolysis,while the experimental group received ticagrelor combined with reteplase thrombolysis.The impact of these treatments on cardiac function and blood-related indexes in STEMI patients was assessed.Results revealed that CK-MB,AST,and LDH levels significantly decreased after 7 d of treatment compared to pre-treatment levels,with the experimental group exhibiting lower levels compared to the control group.Additionally,NT-pro BNP levels decreased in both groups after 24 h of treatment,with the experimental group showing a greater reduction compared to the control group.Furthermore,LVEF and E/A values significantly increased after 7 d of treatment in both groups,while LVEDd and LVESd values notably decreased,indicating improved cardiac function in the experimental group compared to the control group.Following treatment,levels of PT,TT,and APTT increased in both groups,while FIB levels decreased significantly,with noticeable differences between the two groups.The incidence of major adverse cardiovascular events(MACE)in the experimental group was significantly lower compared to the control group,although the rate of mild bleeding did not significantly differ.In conclusion,the combination of ticagrelor and reteplase demonstrated superior efficacy in reducing myocardial enzyme and NT-pro BNP levels,improving cardiac function,and enhancing safety compared to ticagrelor and prourokinase.These findings suggested that ticagrelor combined with reteplase thrombolysis held promise for clinical application.展开更多
An rCHO cell line expressing recombinant human prourokinase （pro-UK） at the level of 5μg/ 10^6cells/d was cultivated on Cytopore cellulose porous microcarriers in a 7.5L Biostat CT stirred tank reactor. A periodic ...An rCHO cell line expressing recombinant human prourokinase （pro-UK） at the level of 5μg/ 10^6cells/d was cultivated on Cytopore cellulose porous microcarriers in a 7.5L Biostat CT stirred tank reactor. A periodic pressure oscillation of 0.04 MPa and 0.04 Hz was adopted to introduce a physical stimulus on the rCHO cells and to improve mass transfer characteristic between cells and medium in the process of porous microcarrier CHO cell culture. Compared to constant pressure culture, the oscillation culture didn＇t influence specific cell growth rate significantly, but could enhance the pro-UK specific production by 10% - 40%, and reduce production of lactate by 10% - 30%. In the perfusion culture of recombinant CHO cell with serum-free medium for 67 days, cell density could reach 2.64×10^7/ml, the maximal prourokinase concentration in harvested supernatant was about 118mg/L, a total of 21.1 grams of prourokinase was produced in 313 liters of supernatant. In conclusion, the perfusion cell culture with periodic pressure oscillation can enhance the production of recombinant protein and increase the reactor specific productivity.展开更多
CD-1, a genetically-engineered CHO cell line, was cultivated with a Biosilon microcarrier culture system.We successfully cultivated CD-1 cells to a very high density (over1×107cells/ml). Prourokinase was stably s...CD-1, a genetically-engineered CHO cell line, was cultivated with a Biosilon microcarrier culture system.We successfully cultivated CD-1 cells to a very high density (over1×107cells/ml). Prourokinase was stably secreted at about 180 IU/ 1e6 cells/24 h. Experiments showed that CD-1 cells growing on Biosilon microcarriers were able to spontaneously release from the microcarriers, then reatthch and proliferate on fresh microcarriers. This makes it very easy to scale up preduction. The microcarriers could be reused several times without affecting adhesion. proliferation and prourokinase secretion. With CMPECC membrane radial flow chromatography and MPG chromatography, the prourokinase in conditioned medium could be purified to a specific activity of 1×105 IU/mg of protein. The purification factor was about 600 fold, and approxiamately 90 % of the biological activity was recovered.展开更多
A recombinant prourokinase chimera was constructed by introduction of Lys-Gly-Asp-Trp-motif between Gly118 and Ile119 among the kringle domain. The structure of designed protein was predicted and simulated. The recomb...A recombinant prourokinase chimera was constructed by introduction of Lys-Gly-Asp-Trp-motif between Gly118 and Ile119 among the kringle domain. The structure of designed protein was predicted and simulated. The recombinant prourokinase chimera was produced in insect cell sf9 with baculovirus-expression vector and existed as active form. Chimera protein was purified by affinity chromatography coupled with antibody. The special activity of the chimera was 90,000 IU/mg detected by fibrin plate determination. It was also shown that chimera inhibited ADP-induced platelet aggregation in a concentration depenent manner. These results showed the prourokinase chimera exhibited not only high fibrinolytic activity but also had anti-thrombosis function.展开更多
Objective To study the duration of prourokinase gene expression in vein grafts and the role of the prourokinase gene in protecting vein grafts from neointimal hyperplasia.Methods Fifty-four Wistar rats were used in th...Objective To study the duration of prourokinase gene expression in vein grafts and the role of the prourokinase gene in protecting vein grafts from neointimal hyperplasia.Methods Fifty-four Wistar rats were used in this study. In each rat, the jugular vein was excised and distended for 30 minutes using a solution containing either Adv5-CMV (control group) or Adv5-CMV/ Pro-UK (treatment group). Next, the jugular vein was reversed and interposed into the divided carotid artery of the same rat. On the 14th day after transfection, vein grafts of the control group were collected in order to perform a fibrinolysis test for prourokinase (Pro-UK) activity. On the 2nd, 7th, 14th, 28th, and 60th day, the vein grafts of the treatment group were likewise collected in order to detect prourokinase activity. On the 28th day, the vein grafts of both groups were explanted to evaluate the 3H-TDR incorporation so that pathologic analysis could be performed.Results Pro-UK activity could not be detected in the control group, while in the treatment group, the Pro-UK activity could be detected from the 2nd day onwards, peaking on the 7th day and declining from the 14th day, but yet persisting at a low level for a further month. The amount of 3H-TDR incorporated in the control group was higher than that in the treatment group. Pathologic analysis demonstrated that vein grafts of both groups exhibited wall thickening, but that the degree of graft neointimal hyperplasia and reduction of the graft lumen was greater in the control group than that in the treatment group. The occlusion rate of grafts in the control group was 20%. All grafts in the treatment group were patent.Conclusions Pro-UK gene transfer before vein grafting in vitro results in a high level of gene expression in the vein graft from the 7th day to 14th day. And its gene expression in the vein graft could reduce neointimal hyperlasia in the vein graft.展开更多
BACKGROUND The combination of acute ST-segment elevation myocardial infarction(STEMI)and gastric ulcers poses a challenge to primary percutaneous coronary intervention(PPCI),particularly for young patients.The role of...BACKGROUND The combination of acute ST-segment elevation myocardial infarction(STEMI)and gastric ulcers poses a challenge to primary percutaneous coronary intervention(PPCI),particularly for young patients.The role of drug-coated balloons(DCBs)in the treatment of de novo coronary artery lesions in large vessels remains unclear,especially for patients with STEMI.Our strategy is to implement drug balloon angioplasty following the intracoronary administration of low-dose prourokinase and adequate pre-expansion.CASE SUMMARY A 54-year-old male patient presented to the emergency department due to chest pain on June 24,2019.Within the first 3 minutes of the initial assessment in the emergency room,the electrocardiogram(ECG)showed significant changes.There was atrial fibrillation with ST-segment elevation.Subsequently,atrial fibrillation terminated spontaneously and reverted to sinus rhythm.Soon after,the patient experienced syncope.The ECG revealed torsades de pointes ventricular tachycardia.A few seconds later,it returned to sinus rhythm.High-sensitivity tropon in I was normal.The diagnosis was acute STEMI.Emergency coronary angiography revealed subtotal occlusion with thrombus formation in the proximal segment of the left anterior descending artery.Considering the patient's age and history of peptic ulcer disease,after the intracoronary injection of prourokinase,percutaneous transluminal coronary angioplasty and cutting balloon angioplasty were conducted for thorough preconditioning,and paclitaxel drug-eluting balloon angioplasty was performed without any stents,achieving favorable outcomes.CONCLUSION A PPCI without stents may be a viable treatment strategy for select patients with STEMI,and further research is warranted.展开更多
Prourokinase(scu-PA),a thrombolytic agent,was inserted between Gly118 and Ile119 with foreign anti-thrombosis functional motif(Lys-Gly-Asp-Trp-motif)to construct a multi-functional chimeric molecule.The molecular mode...Prourokinase(scu-PA),a thrombolytic agent,was inserted between Gly118 and Ile119 with foreign anti-thrombosis functional motif(Lys-Gly-Asp-Trp-motif)to construct a multi-functional chimeric molecule.The molecular model of a chimera was simulated and pre-dicted.The recombinant chimeric protein was expressed by the baculovirus-insect cell expression system and puri-fied by affinity chromatography.The physico-chemical characteristics of the chimeric molecule were assayed.The thrombolytic activity was determined to be 90000 IU/mg of fibrinolytic special activity by the fibrin-plate method.The anti-thrombosis activities were also assayed with IC50 of 9.6 μM by an inhibition test of ADP-induced platelet aggregation.展开更多
文摘To compare the effects of ticagrelor combined with prourokinase and reteplase thrombolysis on cardiac function and blood-related indexes in patients with ST-segment elevation myocardial infarction(STEMI),280 patients hospitalized between January 2017 and December 2021 were randomly assigned into two groups:the experimental group and the control group,each comprising 140 cases.The control group received ticagrelor combined with prourokinase thrombolysis,while the experimental group received ticagrelor combined with reteplase thrombolysis.The impact of these treatments on cardiac function and blood-related indexes in STEMI patients was assessed.Results revealed that CK-MB,AST,and LDH levels significantly decreased after 7 d of treatment compared to pre-treatment levels,with the experimental group exhibiting lower levels compared to the control group.Additionally,NT-pro BNP levels decreased in both groups after 24 h of treatment,with the experimental group showing a greater reduction compared to the control group.Furthermore,LVEF and E/A values significantly increased after 7 d of treatment in both groups,while LVEDd and LVESd values notably decreased,indicating improved cardiac function in the experimental group compared to the control group.Following treatment,levels of PT,TT,and APTT increased in both groups,while FIB levels decreased significantly,with noticeable differences between the two groups.The incidence of major adverse cardiovascular events(MACE)in the experimental group was significantly lower compared to the control group,although the rate of mild bleeding did not significantly differ.In conclusion,the combination of ticagrelor and reteplase demonstrated superior efficacy in reducing myocardial enzyme and NT-pro BNP levels,improving cardiac function,and enhancing safety compared to ticagrelor and prourokinase.These findings suggested that ticagrelor combined with reteplase thrombolysis held promise for clinical application.
文摘An rCHO cell line expressing recombinant human prourokinase （pro-UK） at the level of 5μg/ 10^6cells/d was cultivated on Cytopore cellulose porous microcarriers in a 7.5L Biostat CT stirred tank reactor. A periodic pressure oscillation of 0.04 MPa and 0.04 Hz was adopted to introduce a physical stimulus on the rCHO cells and to improve mass transfer characteristic between cells and medium in the process of porous microcarrier CHO cell culture. Compared to constant pressure culture, the oscillation culture didn＇t influence specific cell growth rate significantly, but could enhance the pro-UK specific production by 10% - 40%, and reduce production of lactate by 10% - 30%. In the perfusion culture of recombinant CHO cell with serum-free medium for 67 days, cell density could reach 2.64×10^7/ml, the maximal prourokinase concentration in harvested supernatant was about 118mg/L, a total of 21.1 grams of prourokinase was produced in 313 liters of supernatant. In conclusion, the perfusion cell culture with periodic pressure oscillation can enhance the production of recombinant protein and increase the reactor specific productivity.
文摘CD-1, a genetically-engineered CHO cell line, was cultivated with a Biosilon microcarrier culture system.We successfully cultivated CD-1 cells to a very high density (over1×107cells/ml). Prourokinase was stably secreted at about 180 IU/ 1e6 cells/24 h. Experiments showed that CD-1 cells growing on Biosilon microcarriers were able to spontaneously release from the microcarriers, then reatthch and proliferate on fresh microcarriers. This makes it very easy to scale up preduction. The microcarriers could be reused several times without affecting adhesion. proliferation and prourokinase secretion. With CMPECC membrane radial flow chromatography and MPG chromatography, the prourokinase in conditioned medium could be purified to a specific activity of 1×105 IU/mg of protein. The purification factor was about 600 fold, and approxiamately 90 % of the biological activity was recovered.
文摘A recombinant prourokinase chimera was constructed by introduction of Lys-Gly-Asp-Trp-motif between Gly118 and Ile119 among the kringle domain. The structure of designed protein was predicted and simulated. The recombinant prourokinase chimera was produced in insect cell sf9 with baculovirus-expression vector and existed as active form. Chimera protein was purified by affinity chromatography coupled with antibody. The special activity of the chimera was 90,000 IU/mg detected by fibrin plate determination. It was also shown that chimera inhibited ADP-induced platelet aggregation in a concentration depenent manner. These results showed the prourokinase chimera exhibited not only high fibrinolytic activity but also had anti-thrombosis function.
文摘Objective To study the duration of prourokinase gene expression in vein grafts and the role of the prourokinase gene in protecting vein grafts from neointimal hyperplasia.Methods Fifty-four Wistar rats were used in this study. In each rat, the jugular vein was excised and distended for 30 minutes using a solution containing either Adv5-CMV (control group) or Adv5-CMV/ Pro-UK (treatment group). Next, the jugular vein was reversed and interposed into the divided carotid artery of the same rat. On the 14th day after transfection, vein grafts of the control group were collected in order to perform a fibrinolysis test for prourokinase (Pro-UK) activity. On the 2nd, 7th, 14th, 28th, and 60th day, the vein grafts of the treatment group were likewise collected in order to detect prourokinase activity. On the 28th day, the vein grafts of both groups were explanted to evaluate the 3H-TDR incorporation so that pathologic analysis could be performed.Results Pro-UK activity could not be detected in the control group, while in the treatment group, the Pro-UK activity could be detected from the 2nd day onwards, peaking on the 7th day and declining from the 14th day, but yet persisting at a low level for a further month. The amount of 3H-TDR incorporated in the control group was higher than that in the treatment group. Pathologic analysis demonstrated that vein grafts of both groups exhibited wall thickening, but that the degree of graft neointimal hyperplasia and reduction of the graft lumen was greater in the control group than that in the treatment group. The occlusion rate of grafts in the control group was 20%. All grafts in the treatment group were patent.Conclusions Pro-UK gene transfer before vein grafting in vitro results in a high level of gene expression in the vein graft from the 7th day to 14th day. And its gene expression in the vein graft could reduce neointimal hyperlasia in the vein graft.
基金Supported by Mianyang Health Commission 2019 Scientific Research Encouragement Project,No.201948.
文摘BACKGROUND The combination of acute ST-segment elevation myocardial infarction(STEMI)and gastric ulcers poses a challenge to primary percutaneous coronary intervention(PPCI),particularly for young patients.The role of drug-coated balloons(DCBs)in the treatment of de novo coronary artery lesions in large vessels remains unclear,especially for patients with STEMI.Our strategy is to implement drug balloon angioplasty following the intracoronary administration of low-dose prourokinase and adequate pre-expansion.CASE SUMMARY A 54-year-old male patient presented to the emergency department due to chest pain on June 24,2019.Within the first 3 minutes of the initial assessment in the emergency room,the electrocardiogram(ECG)showed significant changes.There was atrial fibrillation with ST-segment elevation.Subsequently,atrial fibrillation terminated spontaneously and reverted to sinus rhythm.Soon after,the patient experienced syncope.The ECG revealed torsades de pointes ventricular tachycardia.A few seconds later,it returned to sinus rhythm.High-sensitivity tropon in I was normal.The diagnosis was acute STEMI.Emergency coronary angiography revealed subtotal occlusion with thrombus formation in the proximal segment of the left anterior descending artery.Considering the patient's age and history of peptic ulcer disease,after the intracoronary injection of prourokinase,percutaneous transluminal coronary angioplasty and cutting balloon angioplasty were conducted for thorough preconditioning,and paclitaxel drug-eluting balloon angioplasty was performed without any stents,achieving favorable outcomes.CONCLUSION A PPCI without stents may be a viable treatment strategy for select patients with STEMI,and further research is warranted.
基金This work was supported by the National Natural Science Foundation of China(Grant No.30540071).
文摘Prourokinase(scu-PA),a thrombolytic agent,was inserted between Gly118 and Ile119 with foreign anti-thrombosis functional motif(Lys-Gly-Asp-Trp-motif)to construct a multi-functional chimeric molecule.The molecular model of a chimera was simulated and pre-dicted.The recombinant chimeric protein was expressed by the baculovirus-insect cell expression system and puri-fied by affinity chromatography.The physico-chemical characteristics of the chimeric molecule were assayed.The thrombolytic activity was determined to be 90000 IU/mg of fibrinolytic special activity by the fibrin-plate method.The anti-thrombosis activities were also assayed with IC50 of 9.6 μM by an inhibition test of ADP-induced platelet aggregation.
