Objective To investigate the immunophenotype and the cytotoxicity of cytokine-induced killer(CIK) against tumor cells in vitro.Methods Lymphocytes cells were isolated freshly from peripheral blood of healthy donors by Ficoll-Hypaque density centrifugation,and the cells obstained were induced by IFN?,IL-2 and CD3McAb.Phenotypes and cytotoxicity of CIK were analysed by FACS.Target cells were differentiated from effect cells by CFSE dying.The mortality of Target cells were determined by FACS.Results The maximum proliferation of CIK reached at the 22nd day.The phenotypes of CD3,CD11a,CD54,HLA-DR were expressed highly;CD25,CD28,CD69,FasL were expressed moderately on CIK.The expression of CD16 was not increased.CIK possessed the cytotoxicity against tumor cells of K562,HL-60,Hela,SMMC7721 and A375.Conclusion IFN?,CD3McAb,IL-2 can induce peripheral lymphocytes to produce CIK which own strongly proliferation and exert highly efficient cytotoxic effects on tumor cells.

Abstract Objective: To prepare high-purification and high-specific activity of recombinant human interleukin-6 (rhIL-6). Methods: The rhIL-6 was obstained from inclusion body expressed by IPTG-induced pT7.7hIL-6 expressed vector using extracting,denature and refolding techniques. The rhTL-6 was further purified by anionic-exchange chromatography. Activity of rhIL-6 was measured by 3H-TdR method. Results: After a single-step purification,the product purity reach 95% and it's specific activity was 3.0 x 10~8 U/mg. Conclusion:This scheme of puri-fication was an easy way requiring rhTL-6.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Biopsy ; CD56 Antigen ; metabolism ; DNA-Binding Proteins ; metabolism ; Gene Deletion ; Humans ; Keratin-7 ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Receptor, Epidermal Growth Factor ; genetics ; metabolism ; Small Cell Lung Carcinoma ; genetics ; metabolism ; pathology ; Synaptophysin ; metabolism ; Transcription Factors

Objective:To investigate the expression of cyclooxygenase-2 in primary hepatocellular carcinoma, in cancer surrounding tissues and normal liver tissue and its relationship with clinical pathological features.Methods:The expression of COX-2 was detected in 30 cases of hepatocellular carcinoma, 20 cases of cancer surrounding tissue and 10 normal liver tissue by flow cytometry (FCM) and immunohistochemistry (SP). The clinical data were analyzed retrospectively.Results:(1)The expression of COX-2 in the HCC tissue was significantly higher than in cancer surrounding tissues and normal liver tissue (P0.05).Conclusion:The hyperexpression of COX-2 in tissue can reflex the biological behavior of HCC,and have very important role in the development of HCC.The specificness of COX-2 protein expression make it to be new target of tumor diagnosis and treatment.These results provide a theoretical basis for the chemoprevention of hepatoma.

Objective To construct a recombinant expression vector of human interleukin-24(hIL-24) gene and express it in E.coli M15,and to evaluate the bioactivity of IL-24 fusion protein.Methods The human IL-24 cDNA fragment was amplified from plasmid by polymerase chain reaction(PCR),and sequenced.PQE/hIL-24 was constructed by gene rearrangement,then it was transformed into E.coli M15.The expression of the target protein was induced with IPTG and purified by Ni2+-NTA agarose column.The expressed recombinant hIL-24(rhIL-24) was identified by SDS-PAGE and Western blotting.Normal peripheral blood mononuclear cells(PBMCs) were cultivated with the expression protein for 48 and 72 h,the levels of IL-6,IFN-? and TNF-? of PBMCs stimulated with rhIL-24 were detected by ELISA.Results The recombinant prokaryotic expression vector PQE/IL-24 was constructed successfully and expressed stably in E.coli M15.At about 18 400 of molecular weight,there was an induced protein band.The levels of IFN-?,IL-6 and TNF-? in the group of cultivated with the expression protein were obviously higher than those in the groups without the expresson protein(P

Objective To get the protein expression of sFlt-1 by transfection and to investigate the effects of sFlt-1 gene transfection on the growth of K562 cells.Methods The recombinant plasmid pcDNA3-sFlt-1D4 was constructed.The recombinant plasmid pcDNA3-sFlt-1D4 was transfected into the bone marrow stromal cells by Lipofectamine 2000,which was identified by RT-PCR,ELISA and MTT.Results The transfection efficiency identified by flow cytometry was 9.27%.The protein expression of sFlt-1D4 was found in the culture supernatant 24 h,48 h and 4 weeks after transfetion by ELISA and the expression concentrations were(0.104?0.078),(0.158?0.022) and(0.171?0.069) ?g?L-1,respectively.The content of VEGF secreted by K562 culturing with transfectant cells culture supernatant was reduced compared with control.The inhibitoy rates on the proliferation of K562 cells via MTT assay were 9.41%?4.71%,23.63%?7.50%,and 33.13%?6.93%,respectively.Conclusion The bone marrow stromal cells transfected with recombinant plasmid pcDNA3-sFlt-1D4 could secrete sFlt-1D4 and inhibit the proliferation of K562 cells.

Gastrointestinal Stromal Tumors ; genetics ; pathology ; Genotype ; Humans

Objective To investigate the factors of effecting with yindex analysis of delivery for helical tomotherapy (HT).Methods Measuring γindex with the ArcCheck device for introduced errors in HT.The errors include setup errors in three-dimensional,the gantry angle error,calculating the dose in the phantom,low dose rate.All the results were compared with the 3％/3 mm and 2％/2 mm criteria.The effect of the accuracy in the application of kilovoltage computed tomography (KVCT) and mega-voltage computed tomography (MVCT) images in HT was also analyzed.Paired-t test method was used for difference compared.Results When the errors were introduced to the HT,theγpassing rate of left-right,superior-inferior,anterior-posterior direction dropped 2.7％,7.2％,3.6％ under the 3％/3 mm criteria (P =0.002,0.022,0.007),with 4.6％,15.7 ％,7.6％ under the 2％/2 mm criteria (P =0.001,0.003,0.002) respectively.There was no statistical significance for theγpassing between scanning the ArcCheek phantom with the KVCT and MVCT under the 3％/3 mm and 2％/2 mm criteria (98.6％ vs 98.7％,P =0.859 and 92.7％ vs 92.8％,P =0.984).Conclusions The errors of the setup position and machine paraments can lead to the dose delivery errors in HT,the quality accurance of machine and plan should be enhanced to minimize the dose errors.The results also showed that there is no difference of KVCT and MVCT image on the delivery of HT.

Objective:To investigate the sensitivity and the specificity of scorpions amplification refractory mutation system ( ARMS) in comparing with that of direct DNA sequencing in the detection of BRAF gene mutations in patients with papillary thyroid microcarcinoma.Methods:Direct sequencing and ARMS were used simultaneously to detect BRAF mutation status in 56 patients with PTMC.Results:BRAF mutations were identified in 46 cases with a mutation rate of 82.9%by ARMS,while in 18 cases with a mutation rate of 32.1%by direct sequencing.Besides,the sensitivity of ARMS was 100%and that of direct sequencing was 39.1%.There were significant differences of both mutation rate and sensitivity between two methods ( P<0.01 ).Conclusion: Compared to direct sequencing,ARMS gains a higher sensitivity in the detection of BRAF mutations in samples with tiny lesions.