Coexpression of Schwann cell marker GFAP and myoepithelial cell marker alpha-SMA in salivary adenoid cystic carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To observe the expression of Schwann cell marker GFAP and myoepithelial cell marker alpha-SMA in salivary adenoid cystic carcinoma (ACC), and to evaluate the relationship of GFAP, alpha-SMA and perineural invasion in ACC.</p><p><b>METHODS</b>Immunohistochemical SABC method, double-label immunofluorescence and CLSM were used to detect the expression of GFAP and alpha-SMA proteins in salivary ACC tissue samples.</p><p><b>RESULTS</b>In salivary ACC tissue samples, both GFAP and alpha-SMA proteins were positive, which were coexpressed in cytoplasm of the same onco-myoepithelial cells.</p><p><b>CONCLUSIONS</b>There may be Schwann cell differentiation in onco-myoepithelial cell of salivary ACC, and it may be the pathological base of perineural invasion in salivary ACC.</p>

Preliminary study on anti-periodontitis immunization with DNA vaccine / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To observe the protection against periodontal bone loss in the Sprague-Dawley (SD) rats periodontitis model, with the recombined plasmid pcDNA3.1+/kgpcd as DNA gene vaccine.</p><p><b>METHODS</b>PcDNA3.1+/kgpcd was delivered into rats by submandibular gland-targeted injection. The anti-KGPcd sIgA in saliva was measured by indirect ELISA method. Immunohistochemistry staining was used to assess the protection in the animal model.</p><p><b>RESULTS</b>The level of specific anti-KGPcd sIgA in saliva of the experimental group was significantly higher than that of control group. HE staining showed that immunization with recombined plasmid pcDNA3.1+/kgpcd could protect or minimize tissue destruction caused by subsequent P. gingivalis challenge in the rat model.</p><p><b>CONCLUSIONS</b>The results indicate that pcDNA3.1+/kgpcd was a good candidate for anti-periodontitis gene vaccine and could provide protection against Porphyromonas gingivalis-caused periodontitis in rat lesion model.</p>

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>

Construction of eukaryotic expression vector for KGPcd gene from Porphyromonas gingivalis and expression in mammalian cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aimed at constructing secretory eukaryotic expression vector of KGPcd gene encoding whole amino acid residues of mature KGPcd from Porphyromonas gingivalis and investigating the transcription and expression of recombined plasmid VR1020/KGPcd in mammalian cells.</p><p><b>METHODS</b>Eukaryotic expression plasmid VR1020/KCPcd was constructed by using molecular cloning methods. Then, the VR1020/KGPcd was transfected into mammalian cell COS7 with Lipofectamine 2000 according to the manufacturer's instruction. The transcription of VR1020/KGPcd was assayed by reverse transcription polymerase chain reaction (RT-PCR). The expression product of VR1020/KGPcd was analyzed by using indirect immunofluorescence. The protein secretion in cultural medium was detected by ELISA method.</p><p><b>RESULTS</b>It proved that the VR1020/KGPcd could be transcribed and translated into transfected COS7 cells. The expressed targeted protein could be secreted into cultural supernatant and could be detected by ELISA.</p><p><b>CONCLUSION</b>The eukaryotic expression plasmid of VR1020/KGPcd was constructed successfully and its product can be expressed in mammalian cells. The results indicated that the recombinant plasmid has antigenicity and may be acted as candidate gene vaccine. This laid a basis for its use as gene vaccine candidates in the development of anti-periodontitis and paved the way for further study.</p>

A study of injectable autogenous tissue-engineered bone / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the utilization of carrier for delivering osteoblasts and creating autogenous bone tissue in ectopic site of animal via injection.</p><p><b>METHODS</b>Bone marrow cells harvested from iliac bone of New Zealand rabbits were induced to differentiate into marrow stromal osteoblasts. The osteoblasts were mixed with 1.5% alginate sodium solution to generate osteoblasts/alginate composites with final cellular density of 4 x 10(9)/L. Calcium chloride was used as cross-linking agent to gel aqueous alginate solution. The marrow stromal osteoblasts/alginate composites were injected into the dorsal subcutaneous tissue of rabbits with autogenous cells transplantation. The samples were examined with X-ray and histological analysis.</p><p><b>RESULTS</b>Four, eight and twelve weeks after injection, the hard knobbles were easily palpated under the dorsal skin of animals. On X-ray photograph the samples showed calcified image with more density than surrounding soft tissue, new bone formation was observed in the osteoblasts/alginate composites in histological analysis. The osteogenesis was in association with regenerated hematopoietic bone marrow.</p><p><b>CONCLUSIONS</b>These results demonstrate that new bone tissue could be created through the injection of alginate sodium treated with autogenous marrow stromal osteoblasts.</p>

CLONING AND SEQUENCING ANALYSIS OF GINGIPAIN K OF PORPHYROMONAS GINGIVALIS / 微生物学通报

The desired DNA product of KGPcd and KGP-hag was obtained from the total DNA of Porphyromonas gingivalis by PCR with two pairs of gene specific primers. The segment of KGPcd and KGP-hag (about 1.5kb and 1.6kb) was inserted into pGEM-T easy Vector. The double-stranded DNA of the postitive clone was analyzed by restriction endonuclease mapping and DNA sequenceing. The sequences of KGPcd and KGP-hag were consistent with those of the references appeared. The proteins of KGPcd and KGP-hag will be obtained for further study.