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.

Objective A comparability analysis was performed between the various parameters of the self-developed dry-type blood cell analyzer with the reference instrument (Sysmex XT-1800i).Methods In line with the evaluation program of blood cells issued by ICSH,the detection results,six parameters as HCT,Hb,GRAN (granulocytes),LM (lymphatic and monocytes),WBC and PLT,were compared between the two instruments.Some indexes of the subject instrument such as precision,comparability,outlier points,bias,etc.were analyzed and compared so as to calculate the equation of linear regression and the expected bias intervals.Results The intra and inter CV of the subject instrument about six parameters were respectively less than 5％ and 6％,so it showed that the instrument had good precision.Correlation analysis showed that the classification results of HCT,Hb,GRAN and WBC had a good correlation (r =0.991,0.972,0.986,0.975,P ＜0.01),while that of LM and PLT were slightly lower (r =0.952,0.942,P ＜0.01) ; As to the medical decision level,HCT values were 14％ and 70％,WBC 0.5 × 109/L and 3 ×109/L,and PLT 10 × 109/L,50 × 109/L and 100 × 109/L,and the acceptable bias of these values all fell in the confidence intervals; the values of the rest items were slightly higher than the high value of the confidence interval of expected bias.Conclusions The subject instrument produces the same results in the test of its parameters as the reference instrument Sysmex XT-1800i and therefore the two instruments are comparable.The subject instrument is particularly suitable for field rapid detection.

This study was aimed to establish the thin layer chromatography (TLC) identification and content determination of Tibetan medicineJi-Ni De-Xie (JNDX). TLC was used to identifyBerberis kansuensisSchneid,Terminalia chebula Retz.,Phyllanthus emblicaandCurcuma LongaL. The high performance liquid chromatography (HPLC) was used to determine the content of berberine hydrochloride. The results showed that the TLC identification had a good resolution with clear spots and no interference from the negative reference. The content of berberine hydrochloride showed a good linear relationship with the peak area in the range of 0.05-0.45 mg·mL-1. The regression equation wasY = 40 679X-130.56. The average recovery was 99.07%. It was concluded that the established method was easy to operate with good repeatability. It can effectively control the quality of Tibetan medicine JNDX.

Objective To study the CT number and noise of HT MVCT on different dose rate and scanning thickness.Methods The CT number of different relative electron density were measured in the MVCT image of Cheesephantom with tissue substitute plugs scanned with different dose rate and slice thickness.The physical density corresponding to the CT number was plotted as the image value to density table (IVDT).The noise was measured in the MVCT image of Cheesephantom with solid water plugs scanned with different dose rate and slice thickness.Results There was a significant different of the CT number of the plugs with different dose rates (P =0.000),it shows a positive correlation between the varied CT number and density (R2 =0.846),there is larger impact on the high density number.There was still a significant effect on the noise with different dose rate (P =0.000 density),the noise increase as the dose rate decrease.There was no significant effect on IVDT (P =1.000) and noise (P =0.667) with different slice thickness.Conclusions The CT number and the noise vary with the dose rate,the QA of MVCT should be performed regularly to assure the quality of image and the accuracy of dose calculating on MVCT in adaptive.