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Hemophilia B is ideal model to asses the validity of gene therapy for its pathogenesis and is characteristic.Previously, we have used plasmid that containingl6 bp RBE element derived from the adeno-associated virus, CMV as promoter and hFIX for the target gene, at the presence of Rep donor plasmid, we delivered hFIX expression gene to transgenic mice by hydrodynamic injection.We did observed the long-term express-ion of hFIX protein and phenomenon of integration.In the present experiment, we used a liver specific element as a substitute of CMV promoter, the same way injected into mice carried human AAVS1 locus.We use plasmid of RBE-CMV-hFIX as control.Plasma hFIX levels were both peaked at 1 day after injection, RBE-HCR-hAAThFIX group was 2326.3±224.5ng/mL and RBE-CMV-hFIX group was 2201.8±447.4ng/mL, there was no significant difference, after partial hepatectomy (PH), both group dropped sharply, at day 120, RBE-HCRhAAT-hFIX group was 76.6±53.4 ng/mL and RBE-CMV-hFIX group was 22.4±11.3 ng/mL, RBE-HCRhAAT-hFIX group was almost 4fold than RBE-CMV-hFIX group (p=0.0063).The results of PCR and RT-PCR revealed that the exotic DNA could distribute and transcribe in all examined organs (heart, liver, spleen, lung, and kidney) 1 day after injection, but it only could be identified in liver and heart over time.Site specific integration was detected by nested-PCR.Transient acute liver damage induced by rapid intravenous plasmid injection was found by detection of trans-aminate levels and liver histological figures, which was rapidly repaired within 10 days.The results shows liver-specific element would lead to more efficient and long-term expression of hFIX protein.