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
研究了pH和温度对重组毕赤酵母表达retep lase(rPA)发酵过程中的降解现象。在低pH(4.5)诱导条件下,由于蛋白酶活性高,rPA降解严重,表达量几乎为0,提高诱导pH至6.5,蛋白酶活性降低,rPA的表达量最高达到122 m g/L。通过将诱导温度从30...研究了pH和温度对重组毕赤酵母表达retep lase(rPA)发酵过程中的降解现象。在低pH(4.5)诱导条件下,由于蛋白酶活性高,rPA降解严重,表达量几乎为0,提高诱导pH至6.5,蛋白酶活性降低,rPA的表达量最高达到122 m g/L。通过将诱导温度从30°C降低到20°C,rPA的表达量从153.2 m g/L提高到207.9 m g/L。降低温度能减少细胞死亡率,从而减少了发酵液中蛋白酶的释放,降低了蛋白酶的活性,抑制了对目的蛋白rPA的降解。另外,低温使胞内AOX酶活性提高,从而增强了rPA的表达。展开更多
The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte. The expression of foreign genes in sporophte has been proved. In this research, the rec...The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte. The expression of foreign genes in sporophte has been proved. In this research, the recombinant expression in gametophyte was investigated by particle bombardment with the rt-PA gene encoding the recombinant human tissue-type plasminogen activator (Reteplase), which is a thrombolytic agent for acute myocardial infarction (AMI). Transgenic gametophytes were selected by their resistance to herbicide phosphiothricin (PPT), and proliferated in an established bubble column photo-bioreactor. According to the results from quantitative ELISA, Southern blotting, and fibrin agarose plate assay (FAPA) for bioactivity, it was showed that the rt-PA gene had been inte-grated into the genome of gametophytes of L. japonica, and the expression product showed the ex-pected bioactivity, implying the proper post-transcript modification in haploid gametophyte.展开更多
[Objective] To establish an efficient, convenient and economical method for site-directed mutagenesis. [Method] The target mutation was introduced into primers designed by DNAMAN5.0 software. Through overlap extension...[Objective] To establish an efficient, convenient and economical method for site-directed mutagenesis. [Method] The target mutation was introduced into primers designed by DNAMAN5.0 software. Through overlap extension PCR for twice obtained the mutation gene which of the full length of the recombinant Human Tissue type plasminogen activator （Reteplase）. The mutation gene cloned it into pEASY- blunt simple cloning vector for sequencing. [Result] The sequencing results showed that three site mutations were fully consistent with the expected results （10~ site had been added a base-pair of A, C had been changed into G at 137~ site, G had been changed into A at 686~ site）.Three site mutations were introduced by using overlap extension PCR on one-step. The overall rate of obtaining the mutant sites was 100%. Site-directed mutagenesis will clone the recombinant Human Tissue type plas- minogen activator and laid the basis for the functional study. [Conclusion] Site-directed mutagenesis was successfully implemented based on the overlap extension PCR which is an efficient, convenient and economical DNA-directed mutagenesis method.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.
文摘研究了pH和温度对重组毕赤酵母表达retep lase(rPA)发酵过程中的降解现象。在低pH(4.5)诱导条件下,由于蛋白酶活性高,rPA降解严重,表达量几乎为0,提高诱导pH至6.5,蛋白酶活性降低,rPA的表达量最高达到122 m g/L。通过将诱导温度从30°C降低到20°C,rPA的表达量从153.2 m g/L提高到207.9 m g/L。降低温度能减少细胞死亡率,从而减少了发酵液中蛋白酶的释放,降低了蛋白酶的活性,抑制了对目的蛋白rPA的降解。另外,低温使胞内AOX酶活性提高,从而增强了rPA的表达。
基金Supported by the National High Technology Research and Development Program of China (Grant Nos. 20060110A4013 and 2006AA100311)National Natural Science Foundation of China (Grant No. 40506030)Innovative Key Project of the Chinese Academy of Sciences (Grant No. KZCX2-YW-209)
文摘The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte. The expression of foreign genes in sporophte has been proved. In this research, the recombinant expression in gametophyte was investigated by particle bombardment with the rt-PA gene encoding the recombinant human tissue-type plasminogen activator (Reteplase), which is a thrombolytic agent for acute myocardial infarction (AMI). Transgenic gametophytes were selected by their resistance to herbicide phosphiothricin (PPT), and proliferated in an established bubble column photo-bioreactor. According to the results from quantitative ELISA, Southern blotting, and fibrin agarose plate assay (FAPA) for bioactivity, it was showed that the rt-PA gene had been inte-grated into the genome of gametophytes of L. japonica, and the expression product showed the ex-pected bioactivity, implying the proper post-transcript modification in haploid gametophyte.
基金Supported by National Natural Science Foundation of China(31160032)~~
文摘[Objective] To establish an efficient, convenient and economical method for site-directed mutagenesis. [Method] The target mutation was introduced into primers designed by DNAMAN5.0 software. Through overlap extension PCR for twice obtained the mutation gene which of the full length of the recombinant Human Tissue type plasminogen activator （Reteplase）. The mutation gene cloned it into pEASY- blunt simple cloning vector for sequencing. [Result] The sequencing results showed that three site mutations were fully consistent with the expected results （10~ site had been added a base-pair of A, C had been changed into G at 137~ site, G had been changed into A at 686~ site）.Three site mutations were introduced by using overlap extension PCR on one-step. The overall rate of obtaining the mutant sites was 100%. Site-directed mutagenesis will clone the recombinant Human Tissue type plas- minogen activator and laid the basis for the functional study. [Conclusion] Site-directed mutagenesis was successfully implemented based on the overlap extension PCR which is an efficient, convenient and economical DNA-directed mutagenesis method.
