Preparation of Bone Marrow Dendritic Cells with TNF-? and The Immune Response Against Malignant Pancreatic Cell by Dendritic Cell Vaccine / 中国普外基础与临床杂志

Objective To study the method of obtaining a large number of dendritic cells (DC). To study the specific cytotoxicity T lymphocyte (CTL) effect against tumor cells initiated by DC pulsed with peptide of cancer cell. Methods Development of cells with cytologic features of DC in bone marrow cultures supplemented with granulocyte macrophage-colony stimulus factor (GM-CSF) and IL-4. Determining the DC phenotype and the specific structure by electronic microscopy. The CTL effect against pancreatic carcinoma leading by the DC pulsed with tumor cells lysate in vitro was observed. Results A large number of typical DC was proliferated by supplementing with GM-CSF and IL-4 cytokines. DC had specific cell appearance and structure, and highly expressed various cell surface molecules. TNF-? had the ability of stimulating DC mature, the mature DC had the enhancing abilities of antigen presenting and IL-12 self-secreting, as well as, expressed higher levels of CD54, MHC-Ⅱ and CD86 molecules than control group (P

Construction and appraise of SKP2 gene shRNA expression plasmid / 中国普通外科杂志

Objective To construct and appraise a series of shRNA eukaryotic expression plasmid which contains vary spots of human SKP2 gene.Methods Three pairs of hairpin-like oligonucleotide sequences specific for human SKP2 gene were designed and synthesized. They were cloned into the eukaryotic expression pSIREN-RetroQ plasmid. The recombinated plasmids were confirmed by PCR and sequencing.Results The shRNA sequences were constructed successfully, and could be inserted into the eukaryotic expression vector pSIREN-RetroQ.Conclusions Three shRNA transcripting plasmids pSIREN-RetroQ are successfully constructed, and form a basis for research in cancer gene target therapy against SKP2 gene.