The effect of nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres in repairing the rabbits bone defects / 实用医学杂志

Objective To explore the effect of nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres in repairing large bone defects of rabbit femoral condyle. Methods Animal models of bone defects were induced in 21 New Zealand white rabbits by drilling holes in bilateral femoral lateral condyles , and the rabbits were equally divided into 3 groups:group A as the control group with the defects untreated , group B treated by filling with nano-hydroxyapatite/collagen scaffolds (NHAC), and group C treated by filling with the nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres (ADM-PLGA-NHAC). At week 12 after implanting , the rabbits were all sacrificed for the implanted scaffolds , which were then examined by X-ray , and Micro-CT 3D reconstruction and in histology for evaluation of the new bone formation. Results X-ray, Micro-CT and the measurement and analysis of BMD indicated thatthere was no significant differencein the new bone formation between group B and group C (P > 0.05). The histological examination revealed that. 12 weeks after operation an evident number of new born bones were seen on the implanted scaffolds in groups B and C , while very few were seen scattering in group A. Conclusion The nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres is effective in repairing bone defect without influencing the prosthetic process.

Overexpression of bcl-2 protects hepatoma cell line HCC-9204 from ethanol-induced apoptosis / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the effect of overexpression of bcl-2 on ethanol-induced apoptosis of primary hepatocellular carcinoma (HCC) cells.</p><p><b>METHODS</b>The retrovirus expression vector pDOR-SB containing human bcl-2 cDNA was introduced into a human HCC cell line HCC-9204 by liposome-mediated transfection. pDOR-transfected and non-transfected HCC-9204 cells were used as controls. The expression of Bcl-2 protein by transfected HCC-9204 cells was detected by the immunohistochemical method. Then the cells were cloned with the limited dilution method continually until a monoclonal cell strain whose positive rate of Bcl-2 protein was 100% detected by flow cytometry was obtained. The killing rates of cells were detected by Methabenzthiazuron assay after the treatment of 6% ethanol for 6 h. The extent of apoptosis was analyzed by transferase-mediated dUTP nick end labeling (TUNEL) staining and flow cytometry.</p><p><b>RESULTS</b>Most of the pDOR-SB-transfected cells demonstrated Bcl-2 positive signals, while no signal was found in the controls. The positive rate of Bcl-2 protein detected by flow cytometry in the obtained monoclonal cell strain, which was named HCC-bcl2, was 100% after the cells had been cloned 3 times continually. The killing rate, TUNEL index and the scale of sub-G1 apoptotic peak in HCC-bcl2 cells were all significantly lower than those in the control cells.</p><p><b>CONCLUSION</b>Overexpression of Bcl-2 protein suppresses ethanol-induced apoptosis of the HCC cell line HCC-9204.</p>

Ethanol-induced apoptosis of hepatoma cell line HCC-9204 and its relationship to Bax and Bcl-2 proteins / 细胞与分子免疫学杂志

Aim To explore the ethanol-induced apoptosis effect on hepatocellular carcinoma(HCC) cells and its relationship to the expression of apoptosis associated genes, bax and bcl-2. Methods The cytotoxic effect of 20～ 100 mL/ L ethanol on HCC cell line HCC-9204 was tested by thiazolyl-blue (MTT) assay. Then apoptosis of HCC-9204 cells was induced with 60 mL/ L of ethanol for 6 h. The morphological change, DNA breakage and the change of DNA content of different cell cycles of the apoptotic cells were detected by May-Grunwald Giemsa(MGG) staining, and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay and flow cytometer respectively. The changes of expression level of Bax and Bcl-2 proteins were detected by immunocytochemical staining and image analysis. Results The higher the concentration of ethanol was, the stronger the cytotoxic effect on HCC-9204 cells was. 60 mL/ L of ethanol could lead to obviously morphological apoptotic changes of HCC-9204 cells, and majority of the cells were TUNEL positive by TUNEL labeling assay. Typical apoptotic sub G1 peak was observed by flow cytometer. The level of Bax protein expression increased significantly after induced with 60 mL/ L of ethanol for 6 h, no expression of Bcl 2 were found before and after induced with ethanol. Conclusion Low dose of ethanol can induce apoptosis of HCC-9204 cells obviously, and occurance of the apoptosis is related to the increase of the level of Bax protein expression.

The Anti-Hepatoma Effect of Superantigen Staphylococcal Enterotoxin A Targeted by Monoclonal Antibody / 中国肿瘤生物治疗杂志

Objective: To prepare the conjugate of supcrantigen (SAg) staphylococcal enterotoxin A (SEA) and monoclonal antibody (McAb) against human hepatocellular carcinoma HAbl8 F(ab' )_(2) fragment and to investigate the anti-human hepatoma effect of peripheral blood mononuclear cells (PBMC) targeted by HAbl8 F(ab' )i-SEA. Methods: McAb HAbl8 was extracted and its F(ab' )_(2) fragment was prepared with papain; the conjugate HAblS F(ab' )_(2)-SEA was prepared with N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP); eveny collected peak after purification was identified with gel chromatography; the activity of antibody in the conjugate was identified with immunohistocheinical ABC method; the anti-hepatoma effect of PBMC targeted by HAbl8 F(ab' )_(2)-SEA was observed with MTT method. Results: The conjugate HAbl8 F(ab' )_(2)-SEA was prepared successfully and it had obvious effect of targeting PBMC to kill hepatoma cells, and this effect is correlated positively with the dose of HAbl8 F(ab')_(2)-SEA. Control groups had no such effect. Conclusion: Targeting therapy of hepatoma with McAb-SAg conjugate might be a kind of hopeful model of hepatoma im-munotherapy.