Pro-apoptotic effect on osteosarcoma SOSP-9607 cells by human recombinant caspase-6 fusion protein / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the pro-apoptotic effect of Her-2 targeted recombinant caspase-6 fusion protein on osteosarcoma SOSP-9607 cells.</p><p><b>METHODS</b>Recombinant immunocasp-6 was generated by sequential fusion of the genes of a signal peptide, a single-chain Her-2 antibody (e23sFv), a PEA translocation domain (PEA aa253-364) and an active caspase-6. The immunocasp-6 gene was cloned into pCMV plasmid to construct a kind of eukaryotic expression vector, i.e. pCMV-e23sfv-PE II-caspase-6 (abbr. pCMV-6) and transfected into SOSP-9607 cells. Murine xenograft models were randomly divided into two groups that received i.m. injections of liposome encapsulated pCMV-6 or pCMV alone. The tumor volume and weight of the nude mice and the tumor weight of the cured mice were observed and statistically analyzed. The morphological changes of the tumors were examined with HE staining, apoptotic morphology of the tumor was observed by TUNEL staining and the gene expression was analyzed by immunohistochemical staining.</p><p><b>RESULTS</b>The tumor growth of the mice in the treatment group was significantly slower than that of the control group (P = 0.001). The weight of the nude mice in the treatment group was significantly higher than that of the control group (P = 0.0002). The tumor weight of the mice in the treatment group was significantly lower than that of the control group (P = 0.0006). HE and TUNEL staining of the tumor of nude mice in the treatment groups showed typical characteristics of apoptosis, while normal structure was found in the control group. Furthermore, caspase-6 was not found in the tumor and muscle tissues in the control group, but only in the treatment group by immunohistochemistry.</p><p><b>CONCLUSION</b>Immunocasp-6 can selectively recognize and bind to and kill HER-2 positive osteosarcoma cells, therefore, to offer some foundation for the clinical treatment of osteosarcoma.</p>

Investigation of Apoptosis of the SGC7901 Cells Induced by the Expression of the Recombinant Gene of anti-HER2 ScFv/tBid / 中国生物工程杂志

Objetive: To investigate whether apoptosis of SGC7901 cells can be induced by the expression of the recombinant gene of anti-HER2 ScFv/tBid. Methods: The recombinant anti-HER2 ScFv/tBid gene was cloned into vector pCMV and the recombinant plasmid was transfected into SGC7901 cells. The gene expression was detected by RT-PCR and immunofluorescent staining. Cell counting was carried out to show the effect of the gene transfection on cell growth. At the same time, significant apoptotic peak was detected by flow cytometry in recombinant anti-HER2 ScFv/tBid gene transfected cells. Results: The fusion protein of anti-HER2 ScFv/tBid was observed in the cytoplasm of transfected SGC7901 cells. The transfected cells displayed typical cell growth inhibition and apoptosis. Conclusion: Fusion protein of anti-HER2 ScFv/tBid can induce apoptosis of SGC7901.

Anti-tumor immune response in vitro induced by fusion of Tca8113 cells with macrophages / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the therapeutic effectiveness of fusion tumor vaccine in tongue cancer treatment.</p><p><b>METHODS</b>Human macrophages fused with human tongue carcinoma cell line Tca8113 cell. The fusion cells were selected by magnetic cell sorting (MACS) and cultured. The biological properties of fusion cells and anti-tumor immune response in vitro induced by fusions were observed.</p><p><b>RESULTS</b>In contrast to Tca8113, the fused cells grew significantly slow in vitro. The expression of MHC I, II antigen of the fusion cells which was detected by flow cytometry (FCM) was higher than that of Tca8113. The fused cells significantly increased the proliferation of mixed lymphocyte and induced their cytotoxicity on parental Tca8113.</p><p><b>CONCLUSIONS</b>The fusion tumor vaccine of macrophages and OSCC cells increase in vitro immunogenicity significantly. This indicates that fusion tumor vaccine could be a new method of anti-tumor immunotherapy, which has important potentials for effective individualized human OSCC vaccine.</p>

Establishment and characteristics of a human chordoma cell line / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To present an established human chordoma cell line for chordoma research.</p><p><b>METHODS</b>The specimens pathologically identified as chordoma were cultured, using primary tissue culture in vitro. The surviving cells were analyzed by morphology, histochemical stain, cell cycling analysis, karyotype analysis, electron microscopic observation, heterotransplantation and study of invasive capacity in vitro.</p><p><b>RESULTS</b>The newly established cell line CM-319 has been maintained in continual cultures for over 100 generations in two years. Its morphological observation, histochemical staining properties, electron microscopic observation and heterotransplantation showed the common characteristics of chordoma. The doubling time of cells was about 33 hours. Cell cycle analysis showed: G(1) 55.6%, G(2) 21.9% and S 22.5%, G(2)/G(1) = 1.90. Chromosome analysis showed a hypotriploid feature and the success rate of heterotransplantation was 100%. It is capable of invasion in vitro.</p><p><b>CONCLUSION</b>CM-319, as a cell line derived from human chordoma cells, may serve for further studies of chordoma.</p>

Long-term result of guided nerve regeneration with an inert microporous polytetrafluoroethylene conduit / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the long-term outcome of Polytetrafluoroethylene (PTFE) conduit in nerve repair and to provide more evidence in view of its potential application to achieve a satisfactory functional recovery in clinical settings.</p><p><b>METHODS</b>Thirty-six Wistar rats had their right sciatic nerve transected and were repaired with either conventional microsuture technique (Control group, n=18) or a PTFE conduit with a gap of 5 mm left between the nerve stumps (PTFE group, n=18). At 6 and 9 months after the operation, electrophysiological assessment and measurement of gastrocnemius muscle weight were conducted and morphology of the regenerated nerves were studied with image analysis.</p><p><b>RESULTS</b>At 6 months postoperatively, the nerve conduction velocity recovered to 60.86% and 54.36% (P<0.05), and the gastrocnemius muscle weight recovered to 50.89% and 46.11% (P>0.05) in the Control group and the PTFE group respectively. At 9 months postoperatively, the recovery rate was 65.99% and 58.79% for NCV (P>0.05), and 52.56% and 47.89% for gastrocnemius muscle weight (P>0.05) in the Control group and the PTFE group respectively. Regenerated nerve fibers in the PTFE group had a regular round shape with no fragmentation, wrinkling or splitting of the myelin sheath. Image analysis revealed that the ratio of the myelin area to the total fiber area was larger at 9 months than at 6 months in both groups (P<0.01).</p><p><b>CONCLUSIONS</b>Microporous PTFE conduit may be an alternative for nerve repair allowing of guided nerve regeneration and functional recovery with no obvious adverse effect at long-term.</p>