The fusion protein of Humanized mouse anti-human fibrin ScFv and the low molecuar weight urokinase (IIn-UK) contained seven disulfide bonds and formed inclusion body while expressing in normal E.coli strain.By coexpre...The fusion protein of Humanized mouse anti-human fibrin ScFv and the low molecuar weight urokinase (IIn-UK) contained seven disulfide bonds and formed inclusion body while expressing in normal E.coli strain.By coexpressing DsbC and using the special E.coli strain Origami(DE3) which was trxB/gor double mutant,the fusion protein IIn-UK was functionally expressed in the cytoplasm of E.coli. The expressed fusion protein in the soluble fraction was purified by using affinity chromatography specific against urokinase. The purified fusion protein could combine the thrombus in vitro ,and the specific activity of urokinase reached 80 000IU/mg fusion protein.The result showed that the fusion protein retained the activity of two moieties,and this study laid a foundation for further research of targeting thrombolytic agent.展开更多
AIM： To investigate the role of cryofibrinogen （CF） in active inflammatory bowel disease （IBD）. METHODS： CF was assayed in 284 subjects： 61 with active and 63 with inactive ulcerative colitis （UC）, 45 who had...AIM： To investigate the role of cryofibrinogen （CF） in active inflammatory bowel disease （IBD）. METHODS： CF was assayed in 284 subjects： 61 with active and 63 with inactive ulcerative colitis （UC）, 45 who had proctocolectomy, 35 with active and 20 with inactive Crohn＇s disease （CD）, 40 with other diseases and 20 healthy controls. Trypsin inhibitor （TI） and TI antibody （TI-Ab） were measured in plasma and CF complex by ELISA. RESULTS： CF in active UC was strikingly high compared with all other groups （x^2〈0.001）. Similarly, CF was significantly higher in active CD than in inactive CD or in controls （x^2〈0.01）. In UC, high CF and TI-Ab were associated with the need for operations. Further, high CF, CF/fibrinogen ratio, low TI and high TI-Ab in plasma were associated with disease activity or refractoriness to medication. Elevated CF was not associated with acute reactants like C-reactive protein and white blood cell counts except for erythrocyte sedimentation rate, suggesting that elevated CF was not a consequence of acute inflammation. CONCLUSION： Elevated CF in active IBD appears to be morbigenous. CF promotes IBD via two main mechanisms, quenching of TI （an anti-inflammatory substance） and impairing microvascular perfusion by forming protein aggregates. CF may also serve as a biomarker of chronic IBD. Additional studies are warranted to fully evaluate the role of CF in IBD and the outcome should contribute to a better understanding of the pathogenesis of IBD.展开更多
This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specifi...This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin αbβ3 by the two mAbs. Results.HI117 and SJ9A4(10μg/ml and 20μg/ml) induced evident specific Fg binding to human platelets,suggesting that the two mAbs evoked activation of platelet integrin αbβ3.Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αbβ3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. Conclusion.The anti platelet tetraspanin(CD9)mAbs,HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors.Three signaling pathways,i.e.thromboxane,secreted ADP, and cAMP pathways may be involved in the process,with protein kinase C activation presumably being the common step of the three pathways.展开更多
文摘The fusion protein of Humanized mouse anti-human fibrin ScFv and the low molecuar weight urokinase (IIn-UK) contained seven disulfide bonds and formed inclusion body while expressing in normal E.coli strain.By coexpressing DsbC and using the special E.coli strain Origami(DE3) which was trxB/gor double mutant,the fusion protein IIn-UK was functionally expressed in the cytoplasm of E.coli. The expressed fusion protein in the soluble fraction was purified by using affinity chromatography specific against urokinase. The purified fusion protein could combine the thrombus in vitro ,and the specific activity of urokinase reached 80 000IU/mg fusion protein.The result showed that the fusion protein retained the activity of two moieties,and this study laid a foundation for further research of targeting thrombolytic agent.
文摘AIM： To investigate the role of cryofibrinogen （CF） in active inflammatory bowel disease （IBD）. METHODS： CF was assayed in 284 subjects： 61 with active and 63 with inactive ulcerative colitis （UC）, 45 who had proctocolectomy, 35 with active and 20 with inactive Crohn＇s disease （CD）, 40 with other diseases and 20 healthy controls. Trypsin inhibitor （TI） and TI antibody （TI-Ab） were measured in plasma and CF complex by ELISA. RESULTS： CF in active UC was strikingly high compared with all other groups （x^2〈0.001）. Similarly, CF was significantly higher in active CD than in inactive CD or in controls （x^2〈0.01）. In UC, high CF and TI-Ab were associated with the need for operations. Further, high CF, CF/fibrinogen ratio, low TI and high TI-Ab in plasma were associated with disease activity or refractoriness to medication. Elevated CF was not associated with acute reactants like C-reactive protein and white blood cell counts except for erythrocyte sedimentation rate, suggesting that elevated CF was not a consequence of acute inflammation. CONCLUSION： Elevated CF in active IBD appears to be morbigenous. CF promotes IBD via two main mechanisms, quenching of TI （an anti-inflammatory substance） and impairing microvascular perfusion by forming protein aggregates. CF may also serve as a biomarker of chronic IBD. Additional studies are warranted to fully evaluate the role of CF in IBD and the outcome should contribute to a better understanding of the pathogenesis of IBD.
文摘This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin αbβ3 by the two mAbs. Results.HI117 and SJ9A4(10μg/ml and 20μg/ml) induced evident specific Fg binding to human platelets,suggesting that the two mAbs evoked activation of platelet integrin αbβ3.Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αbβ3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. Conclusion.The anti platelet tetraspanin(CD9)mAbs,HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors.Three signaling pathways,i.e.thromboxane,secreted ADP, and cAMP pathways may be involved in the process,with protein kinase C activation presumably being the common step of the three pathways.
