Taking Shinshu University of Japan for example,we attempted to introduce the postgraduate clinical education system for new oral surgeons by analyzing the management of clinical training programs,studies of oral maxillofacial and associated medicine,and practices of scientific research in the department of oral surgery.Their experiences may be useful for us to improve our medical continuing education for young oral surgery residences.

Objective To study plasmid-mediated transfer,plasmid replicon typing,and genetic environment of blaNDM-1 gene in Enterobacteraerogenes(E.aerogenes).Methods E.aerogenes HN-NDM0711 was used as the subject of this research,the transferable properties of plasmid were analyzed by conjugation testing,conjugant was performed stability testing,plasmid type was determined by PCR-based replicon typing (PBRT),downstream and upstream of blaNDM-1 were sequenced using chromosome walking method,genetic context was analyzed by BLASTN and BALSTP,as well as annotated using Vector NTI 11.5.1 software,sequence pipeline graph was made,the sequence was submitted to Genbank through software Banklt.Results The conjugation testing of E.aerogenes pHN-NDM0711 was positive,after positive conjugant was conducted 4-day passage,minimal inhibitory concentrations (MICs) of imipenem and meropenem to all the cloned strains didn't change,blaNDM-1 were all positive.The replicon type was IncA/C;blaNDM-1 gene was localized between ISAba14 and IS91,at upstream of the blaNDM-1,class 1 integron and Tn3 transposon were identified,class 1 integron contained a new mosaic structure of a drug-resistant resistance gene cassette.Conclusion E.aerogenes pHN-NDM071 1,bearing blaNDM-1 gene in IncA/C plasmid,derived from gene recombination under different antimicrobial selection pressure.Antimicrobial use in clinical,industrial and agricultural area should be strictly controlled,so as to reduce the emergence of such bacteria.

Objective To recognize the relationship between high cutantous artery branches and descending branch of lateral femoral circumflex artery artery,and to investigate the blood supply of anterolateral thigh flap in clinical.Methods Retrospective analysis 152 cases of anterolateral thigh flap from November,2003 to December,2016.It contains cutting with descending branchs in 99 cases,cutting with lateral branchs in 43 cases,the union of them in 8 cases and high cutantous artery branches in 2 cases.Results The flaps survival in 147 cases,cutting necrosis in 3 cases and partly necrosis in 2 cases.Descending and lateral branchs were both dominance neurovascular bundle of vastus lateralis muscle,independently or commonly dominate the skin of anterolateral thigh,occurrence rate was 33.5％.Conclusion The descending branch of lateral femoral circumflex artery include lateral branch,all of which are the neurovascular bundles,supply the anterolateral thigh muscle,and divid into some perforator branches crossing the musle to the anterolateral thigh skin subsequently,which constitute "the anatomical functional unit".High cutantous artery branche is the one of all of the branches,its origin lies hight and arise from lateral branch.

Objective:To investigate the ability of the anti-PD-1(scFv)/hIL-15 fusion protein(PD-15) to specifically bind to PD-1 in vitro and the effect of the combination of PD-15 with GF-hIL-15Ra-K562(G15Ra-K562) feeder cells to expand NK/T cells. Methods:Overlap PCR was used to construct G15Ra expression vector. pMXs-G15Ra-IP was transfected into K562 by electroporation. G15Ra-K562 feeder cell lines were obtained by limiting dilution method. pUC57-PD-15 was constructed by digestion and ligation. Lipofectamine? 2000 was used to transiently transfect pUC57-PD-15 into HEK293T cells and the conditioned medium containing PD-15 fusion protein was obtained. Density gradient centrifugation was used to obtain human peripheral blood mononuclear lymphocytes(PBMC), and CFSE staining was used to mark active proliferating cells. Flow cytometry was used to detect the ability of PD-15 to specifically bind to PD-1 and its effect on the proliferation of human PBMC and the proportion of different subpopulations of lymphocytes.Results:The feeder cells G15Ra-K562 with high expression of fusion protein G15Ra was successfully constructed. The addition of hIL-15 can increase the ability of G15Ra-K562 to expand human PBMC by more than 5 times( P<0.05). PD-15 fusion protein has PD-1 specific binding ability( P<0.001), combined with G15Ra-K562 can efficiently expand human peripheral blood-derived NK/T cells in vitro( P<0.05). The cells expanded by PD-15 and G15Ra-K562 are mainly natural killing cell, CD8 + T and CD4 + T cells. Conclusions:The PD-15 fusion protein can specifically target the PD-1 molecule and has a strong human peripheral blood-derived NK/T cell expansion ability when combined with G15Ra-K562 feeder cells. These results shed light on selective expansion of PD-1 + lymphocytes in vitro.

Objective: To investigate the function of anti-PD-1 (scFv)/IL-15/IL-15Rα-sushi fusion protein (PD-S15) to specifically bind to PD-1 in vitro and to explore its effect on NK/T cell proliferation. Methods: The human anti-PD-1 (scFv) gene sequence and human IL-15/IL-15Rα-sushi fusion gene sequence were synthesized chemically. The recombinant expression plasmid pUC57-PD-S15 was constructed by enzyme digestion and ligation of the two target genes, and then transiently transfected into HEK293T cells by lipofectamineTM 2000. The supernatants of cell culture medium were acquired, and the expression of PD-S15 fusion protein in cell culture supernatants was detected by Wb assay. PBMCs and TILs were cultured in mediums with different proportion of PD-S15/X-VIVOTM15, respectively. Then, the capacity of PD-S15 fusion protein to bind to PD-1 in vitro and its effect on the proliferation of PBMCs and the proportion of CD3+CD8+, CD3+CD4+ and CD3-CD56+ subsets were detected by flow cytometry. The effect of PD-S15 fusion protein on the proliferation of TILs was detected by cytometry. Results: The successful construction of pUC57-PD-S15 eukaryotic expression plasmid was confirmed by double enzyme digestion and sequencing, and then successfully transfected into HEK293T cells. The relative molecular weight of the target protein was approximately 55 000, and was in line with expectations. PD-S15 fusion protein could specifically combine with PD-1 in vitro (P<0.05) and stimulate NK/T cell proliferation (P<0.05). Compared with classical TILs culture method, the efficiency of activation and amplification of T cells in vitro by PD-S15 culturemethodwasbetter (P<0.01). Conclusion: PD-S15 fusion protein can specifically target PD-1 and rapidly expand NK/T cells in vitro, which lays a foundation for the selective expansion of CD8+PD-1+ antigen-specific T lymphocytes from tumor tissues and even peripheral blood.