Objective To explore the level and main functions of P-type ATPase7B in hepatic cells of patients with hepatolenticular degeneration(HLD).Methods The hepatic cells from 5 normal controls and 9 patients with HLD were cultured in vitro. P-type ATPase7B levels in hepatic cells were examined and compared by SDS-PAGE and Western-blot techniques.Results Compared with the controls, 9 patients displayed various changes of electrophoresis strip. Almost normal strips at 155?103 were found in 3 cases, no strip was found in 1 case, and thinner and lighter strips were showed in the remain 5 cases. 6 cases presented abnormal specific reaction strips.Conclusion Mutations of gene ATPase7B in HLD patients cause change of P-type ATPase7B in quantity and quality, thus leads to dysmetabolism of copper.

BACKGROUND:Chronic venous insufficiency is a major health problem worldwide. Clinical treatments include venous valve repair and venous segment containing valve transplantation. However, these are invasive procedures, and the supply of vein containing valves is limited. Significant progress in the fields of tissue engineering and regenerative medicine has been made towards the creation of tissue engineered vascular grafts for the repair of damaged or malformed vessels. It has been reported that using tissue engineering, a tissue engineered vein containing valves constructed with self-derived endothelial cells and al ogeneic acellular matrices can provide the complex physiological valve structure and mechanical stability, but this elicited an immunogenic response.
OBJECTIVE:To create a viable and functional vein containing valves, which has the ability to grow, repair, and imitate natural tissues.
METHODS:Bone marrow mesenchymal stem cells were obtained from Beagle dogs by density gradient centrifugation and adherence methods. Bone marrow mesenchymal stem cells were cultured in vitro. Fol owing isolation and culture the cells were examined using flow cytometry and identified by direct induction towards the osteogenic and adipogenic lineages. We fabricated biodegradable venous scaffold containing valves using the method of injection molding combined with thermal y induced phase separation. Based on the self-made cast, a three-dimensional biodegradable vein scaffold containing valves was constructed from poly(lactic-co-glycolic acid). Morphological structure was tested. Bone marrow mesenchymal stem cells were used as seed cells to be seeded onto the lumen of the tissue engineered vein scaffold containing valves in vitro and then incubated for 2 weeks.
RESULTS AND CONCLUSION:Scanning electron microscopy images showed that the scaffold demonstrated sufficient porosity. Cultured cells expressed mesenchymal cellmarkers, CD44 and CD29, but did not express hematopoietic cellmarkers, CD34 and CD45 at the same time point. Scaffolds were nontoxic to cells and were favorable for the growth and migration of bone marrow mesenchymal stem cells. cells attached on the surface of poly(lactic-co-glycolic acid) scaffolds formed a confluent layer after incubation. The cellular constructs were tested in vitro, and the valve leaflets were functional y capable of opening and closing when stimulated. These results suggested that the tissue engineered vein containing valves have been successful y constructed by using a three-dimensional poly(lactic-co-glycolic acid) scaffold and bone marrow mesenchymal stem cells as seed cells. Tissue engineered vein containing valves is potential y useful for the substitution and regeneration of vein valves.

OBJECTIVETo investigate the effect of activator protein-1 (AP-1) decoy-oligodeoxynucleotides (Decoy-ODNs) on the expression of fibroblast alpha2 type I collagen, so as to explore the gene therapy of pathologic scar.

METHODSDecoy-ODNs targeting AP-1 were designed and synthesized. NIH3T3 cells were transfected by cationic liposomes. The distribution of Decoy-ODNs in the cells was investigated. The inhibiting effects of Decoy-ODNs on AP-1 were determined by electrophoretic mobility shift assay (EMSA). And the effects of Decoy-ODNs on the collagen synthesis in the cells were analyzed by RT-PCR.

RESULTSAP-1 Decoy-ODNs could competitively inhibit the AP-1 in vitro activity. Cationic liposomes could play roles by effectively transfecting Decoy-ODNs into the plasma and nucleus. The mRNA expression of fibroblast alpha2 type I collagen decreased evidently after 24 hours of Decoy-ODNs action.

CONCLUSIONDecoy-ODNs could inhibit the mRNA expression of fibroblast alpha2 type I collagen by antagonizing AP-1.

Animals ; Collagen Type I ; biosynthesis ; Fibroblasts ; drug effects ; metabolism ; Mice ; NIH 3T3 Cells ; Oligodeoxyribonucleotides ; genetics ; pharmacology ; RNA, Messenger ; metabolism ; Transcription Factor AP-1 ; genetics ; Transfection