To improve thrombolytic effect, a fusion protein SFH composed of staphylokinase (SAK) and hirudin (HV) with blood coagulation factor Xa (FXa) recognition peptide as a linker, was designed. SFH showed improved thrombolytic effect and low bleeding in vivo. Two thrombus-targeting mechanisms might account for the above features of SFH. This study was designed to study the two thrombus-targeting mechanisms of SFH. ELISA and immunohistochemistry assay were used to study the improved thrombus selectivity of SFH and the results showed that SFH, compared with SAK, displayed higher affinity for thrombin and thrombin-rich thrombus. To verify the thrombus-targeting release of anticoagulant activity of SFH, FH-a derivative of HV with only FXa recognition sequence at N terminus of HV was designed and used in animal tests. In inferior vena cava thrombosis model, FH showed equal antithrombotic effect as HV, indicating that HV could be successfully released from FH by FXa cleavage in vivo. More importantly, no prolongation of plasma TT, APTT and PT were found in FH group, but significant prolongations were discovered in HV group. This revealed that the anticoagulant activity of FH was released in thrombus-targeting way and limited in the vicinity of the thrombus, and this could be extrapolated to SFH. In conclusion, the high thrombus affinity and thrombus-targeting release of anticoagulant activity of SFH assigned low bleeding risk to SFH.
Animals ; Anticoagulants ; pharmacology ; Factor X ; pharmacology ; Hirudins ; biosynthesis ; genetics ; Metalloendopeptidases ; biosynthesis ; genetics ; Mice ; Rats ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Thrombolytic Therapy ; methods ; Thrombosis ; drug therapy ; Vena Cava, Inferior

Autoimmune pancreatitis (AIP) is an autoimmune-mediated abnormal chronic inflammatory disorder and is often misdiagnosed as pancreatic neoplastic lesions. With in-depth studies of this disease in recent years, it has been taken seriously by hepatobiliary physicians and surgeons. This article summarizes the clinical features, diagnostic criteria, and treatment methods for autoimmune pancreatitis at the present stage, so as to provide clinicians with diagnosis and treatment experience to reduce clinical misdiagnosis.