Objective To investigate the changes of immunity in patients with hepatocellular carcinoma (HCC) treated by high intensity focused ultrasound (HIFU). Methods HIFU system was used to treat 21 patients with mid-advanced HCC. The blood tests of cellular and humoral immunity related indexes were made before and on the 3rd, 7th, 14th, 21st day following the sonication. The histological changes of the target lesions were observed by light and electron microscope for the cases undergoing second stage operation. Results Light and electron microscope all revealed that HIFU could lead to irreversible changes in the tumor cells of the target lesions. There were not significant differences between the values of CD 3, CD 4, CD 8, CD 16, IgG, IgA, IgM,etc. before and after the ablation. Conclusions HIFU could destroy the tissues of HCC effectively. HIFU could not change the immunity of the patients with mid-advanced HCC remarkably. It may be ideal to unite immunotherapy and the other methods to get a sound clinical prognosis for HCC patients.

Objective To construct a fusion protein to transfect some cell lines that were difficult to be transfected such as neoplastic cells, nerve cells, stem cells. Methods PCR was performed to amplified protein transductions domain(PTD),G4S and streptavidin (Strep).Enzymatic digestion and ligation were used to construct pAYZ-PTD-Strep plasmid. Fusion protein was induced to express by AP5 medium and was isolated by E-tag affinity chromatography. Fusion protein was identified by Western blot. eGFP was trasfected into U937 cells by pAYZ-PTD-Strep. FACS was performed to detect transfection percentage. Results Fusion protein PTD-G4S-Strep was expressed as soluble protein.The concentration of fusion protein was about 0.7 mg/L,and purity was over 90 ％. The protein could carry plasmid into a suspended cell line, U937 cells. The transfection-efficiency of protein was higher than monometer PTD.Conclusion The protein PAYZ-PTD-Strep could carry macromolecules into blood tumor cells,and its biological activity may be expected to develop into a highly efficient and reliable transfection method.

Objective To identify multidrug resistance (MDR) associated cell surface antigen in hematologic neoplasia and to investigate the universality of membrane-relocated expression of this antigen in hematologic neoplasia.Methods The membrane antigen was isolated and precipitated by SDS-PAGE and co-immunoprecipitation (co-IP),then was identified by mass spectrum (MS).Specific siRNA was used to interfere with gene expression,laser confocal microsopy was used to validate the results involved in antigen information.FACS was performed to analyse relocated expression of the antigen in hematologic neoplasia.Results Co-IP and MS show that a nuclear factor PSF was the antigen of 5D12,a leukemia-MDR associated McAb,and this antigen could relocate on HL-60 cell membrane.A series of experiences further confirmed that PSF overexpressed on HL-60 cell membrane compared with HL-60/ADR.The binding percentages of 5D12 to many hematologic tumor cells were observed,HL-60 (78.56±0.76) ％,K562 (26.54±4.42) ％,Nomalwa (38.10±5.11) ％,U937 (64.03±7.96) ％,Jurkat (29.12±5.58) ％,Raji (74.92±3.41) ％,CEM (12.18±3.21) ％.Conclusion Nuclear protein,PSF relocalizes on cell surfaces in hematologic tumor cells and contributes to cell sensitivity.PSF is a potential target of MDR prediction in hematologic neoplasia.

Objective To explore the potential role and function of miR-30 a in myocardial fibrosis after myocardi-al infarction( MI) .Methods We constructed the AAV plasmid vector which carried the miR-30 a gene of rat.The recombinant plasmid was detected by gene sequencing , enzyme digestion and PCR .Virus was packaged into HEK293 cells and virus titer was determined after extraction and purification by PCR .PBS fluid, rAAV9-miR-30 a-NC and rAAV9-miR-30 a were transmited to rat hearts from PBS group , miR-30 a-NC group and miR-30 a group respectively through transcoronary infusion before anterior descending coronary artery ligation .Sham group was set up at the same time.After 4 weeks, heart function was monitored by serial echocardiography , including fractional shortening ( FS) , and left ventricular ejection fraction ( LVEF) .Masson staining was used to calculate collagen volume fraction ( CVF) .The expression of collagen ⅠandⅢwere detected by immunohistochemistry . The mRNA level of miR-30a, TGF-β1 and CTGF were detected by real-time PCR.The protein level of TGF-β1 and CTGF were detected by western blot analysis .Results The cardiac function of miR-30 a group was improved significantly compared with PBS group and miR-30a-NC group (P<0.05).The levels of CVF,collagenⅠ,Ⅲexpression and Collagen Ⅰ/Ⅲ ratio in miR-30 a group was significantly lower than PBS group and miR-30 a-NC group ( P<0.01 ) .The mRNA and protein level of TGF-β1 and CTGF in miR-30 a group were reduced signifi-cantly than PBS group and miR-30 a-NC group ( P<0.001 ) .Conclusions The overexpression of miR-30 a after MI may reduce the mRNA and protein level of TGF-β1 and CTGF, so as to suppress myocardial fibrosis and im-prove cardiac function.