1.Study on The Expression of Coagulation Factor Mediated by Recombinant AAV Serotype 8
Zuowu CAO ; Guangui LI ; Yuqiang LIANG ; Dongmei HE
Progress in Biochemistry and Biophysics 2006;0(01):-
Recombinant AAV serotype 8 (rAAV8) vector is new and promising for gene delivery,which transduces muscular and hepatic cells highly efficiently. The transduction varies with the administration routes. It was observed that the rAAV8 virus injected intraperitoneally had been delivered to many different tissues through the blood circulation and resulted in extensive and prolonged gene expression. The interest gene (human coagulation factor Ⅸ) has been highly expressed via the rAAV8 vector injected intraperitoneally into mice at the dose of 5?1010 gc/ mouse. Considerable expression was detected in mouse plasma at two weeks after the administration. Expression peak up to 1 000% level compared to the UCRP occurred between 1~2 months after administration, and then the expression declined gradually but obvious expression persisted at 4 months post administration. APTT test proved the clotting activity of the recombinant hFⅨ in mouse plasma. Immunohistochemical assay showed that the human coagulation factor has been expressed significantly in several organs including liver, kidney, heart and muscles (abdominal and hindleg). These suggest that the rAAV8 virus injected intraperitoneally has been delivered via blood circulation and transduced different cells of multi-organs, and the product of the gene mediated by the rAAV8 vector is biologically active.
2.Role of hyperpolarization-activated cyclic nucleotide-gated cation nonselective channel in spontaneous contrac-tion of smooth muscles in human isolated ureters
Guangui FENG ; Qian LIU ; Bishao SUN ; Jingzhen ZHU ; Zhou LONG ; Longkun LI
Journal of Regional Anatomy and Operative Surgery 2015;(2):119-122
Objective To investigate effects of hyperpolarization-activated cyclic nucleotide-gated cation nonselective channel in the hu-man ureter on the spontaneous contraction of smooth muscles. Methods Four HCN subtypes were detected in human ureteral tissue using reverse transcription polymerase chain reaction,Western blotting and immunohistochemical. ZD7288,the HCN blocker, was used to observe the changes of ureteral muscle contraction amplitude and frequency by applying the ureteral smooth muscle strip test in vitro. Results HCN1-4 isoforms were all identified in human ureter using reverse transcription-polymerase chain reaction and Western blotting. Through the immunohistochemical,HCN channel was found mostly in the urothelium layer and muscular layer of human ureteral wall. ZD7288 significantly decreased the bladder excitation. Conclusion All 4 HCN channel hypotypes exist in the human ureter, and affect the ureteral excitation.