Objective To generate cytokine induced killer (CIK) by inducing lymphocytes cells from peripheral blood in vitro , and to study the phenotype and biological activities of CIK. Methods Lymphocytes cells were isolated fresh from peripheral blood of healthy donors by Ficoll Hypaque density centrifugation, and the cells obtained were resuspended in RPMI medium consisting of 10% fetal calf serum at 37℃, 5%CO 2 for 2h. On day 0, the nonadherent cells were activated with IFN ? (1 000U/ml) and the following days were stimulated with CD3mAb (3 75?g/ml) and rhIL 2 (600U/ml). Phenotype of CIK were analysed by FACS. The proliferation of CIK was tested using 3 H Thymidine, and the killing experiment using MTT tests. Results The proliferation peak of CIK was at day 20, and declined at day 30. The percentages of the cytotoxicity of CIK cells for stomach cancer cell line MGC803 and liver cancer cell line Bel7402 was 65 45%?5 68% and 68 37%?3 93% respectively. CIK cells expressed the phenotypes of CD3CD56, CD3, CD54, CD28CD54, CD11a, HLA DR and CD28. Conclusion IFN ?, CD3mAb and rhIL 2 can induce strong proliferation of peripheral lymphocytes to produce CIK cells expressing the phenotype of CD3CD56, CD3, CD54, CD28CD54, CD11a, HLA DR, CD28, which have strong cytotoxicity on tumor cells.

Malignant tumor is one of the principle diseases which threatens the health of human being, and its incidence rate has been dramatically increased in recent years.Modern medical model has transformed from biomedical model to the model of "biology-psychology-society",from experience-based medicine to evidence-based medicine (EBM).At the same time,individualization is also one of important principles of tumor combined therapy.EBM and individualization treatment are not to opposed to each other but to complement each other. This article expounded the relationship of EBM and individualization treatment in tumor therapy,and showed that both of these two principles must be grasped arid carried out in clinical work.

Biochemotherapy refers to the combination of biotherapy and chemotherapy,and it has been rapidly developed since its emergence.Biochemotherapy has brought both new therapies and new therapeutic ideas for the treatment of malignant tumors.In this article,we reviewed the recent advances in tumor biochemotherapies,including expansion of the indications,retaining of therapy even when tumor progressed(unique to biochemotherapy),and reversal of chemotherapy resistance by targeted therapy,the predicting response to tumor biochemotherapy,recent clinical trials of immunotherapy-based chemotherapy,and evaluation criteria for assessment of biochemotherapy response,and so on.

Objective To investigate the possibility of using 5-aminolevulinic acid(5-ALA)induced protoporphyrin IX(PpIX)for photodynamic therapy(PDT)of nasopharyngeal carcinoma(NPC)cell line CNE2 cultured in vitro.Methods CNE2 cells cultured were incubated in a medium containing various concentrations of 5-ALA(0.01,0.05,0.1,0.25,0.5,1.0,2.0,4.0mmol/L)for 6 hours,then illuminated with different light doses(20,10,5,2J/cm2)using a semiconductor laser at 630nm.After 6,12,24 hours incubation with 5-ALA-PDT,the survival rates of CNE2 cells were analyzed by MTT assay.Results 5-ALA-PDT may effectively kill the CNE2 cells.The killing degree was positively correlated with the incubation time after irradiation,concentration of 5-ALA and the dosage of radiation(P

Objective To investigate the therapeutic mechanisms for differentiated thyroid cancer by using CIK and IL-2, to find out the better adjunctive clinical therapies for thyroid cancer patients after operation, and to evaluate the effects of cytokine-induced kill cell (CIK) and IL-2 on the secretion of thyroxine by thyroid papillary carcinoma. Methods The samples of thyroid papillary carcinoma were taken from the excised tissues of patient with thyroid cancer, and then dispersed with collagenase and trypsin for culturing. The carcinoma cells were then seeded in 24-well cell culture plates at 37℃, 5% CO2 and 95% humidity for 3 days. At the fourth day, the cells in the wells were separately stimulated four times with different dosage of CIK and IL-2, and the stimulation lasted for 72 hours each time. 12 days later, the solution of T3, T4 was detected with radio-immunity kits, TSH was detected with immuno-radiation kits, and the cell proliferation was detected with mono-nuclear cell direct cytotoxicity assay. Results The thyroid cancer cells did not respond to IL-2 in median and low concentrations, but responded to IL-2 in a higher concentration which may depress the secretary function of thyroid cancer cells. IL-2 of high concentrations can obviously decrease the hormone secretion, such as Thyroxine and Thyrotropin, of papillary carcinoma, and improve the CIK's ability of killing cancer. CIK can kill the cancer cells only when companied with IL-2. Conclusion IL-2 of high concentrations can't inhibit the proliferation of thyroid cancer cells, but can depress the secretary function of thyroid cancer cells, which is different from the killing mechanism of CIK.

Objective The purpose of present study was to compare the efficacy of topical dimethyl sulfoxide(DMSO),intralesional hyaluronidase(HAase)and intralesional HAase plus topical DMSO on skin damage caused by docetaxel extravasation in a rat model.Methods After the establishment of animal model of docetaxel extravasation on each hind limb in 30 SD rats,the animals were assigned randomly into 6 groups with 5 rats for each group:topical DMSO group(TD),intralesional HAase group(IH),IH plus TD group(IH+TD),topical normal saline group(tNS),intralesional NS group(iNS),and control group(C).The extent of skin damage and the healing time were observed and compared.Results The extent of skin damage was different among the 6 groups:severer skin damage was seen in tNS,iNS and C groups compared with TD,IH and IH+TD groups.The healing time was significantly shorter in TD group than in both tNS and C groups [(19.10?2.13)d vs(23.70?2.41)d and(25.70?2.26)d,P

Objective To establish a gemcitabine-resistant human nasopharyngeal carcinoma(NPC)cell line and study its characteristics.Methods The drug-resistant cell line CNE2/Gem was established by a procedure of treating the human NPC cells CNE2 with gemcitabine by pulse drug selection and continuous stepwise selection.The IC50 and resistance index(RI)to several commonly used anti-tumor drugs were tested by MTT assay.Fluorescence activated cell analysis(FACS)was employed for determining the cell cycle and concentration of fluorescence dye rhodamine 123 within the cells.Cell growth curve,doubling time and cell morphology were measured and observed.Results Duplication time of CNE2 and CNE2/Gem was 17.13h and 23.13h,respectively,as evaluated by the growth curve.The IC50 to gemcitabine increased from 2.84?1.63?g/ml in CNE2 to 42.95?7.53?g/ml in CNE2/Gem as tested by MTT assay at 48～72h exposure,the RI was 23.61(P

Objective To investigate the anticancer activity of dendritic cells(DC)transfected by recombinant adeno-associated virus(rAAV)-mediated carcinoembryonic antigen(CEA)gene in colorectal cancer treatment.Methods The recombinant plasmid rAAV/CEA was constructed and the rAAV/CEA virus stock was prepared.DC precursors were isolated from peripheral blood mononuclear cells(PBMC)by density gradient centrifugation and transfected with rAAV/CEA virus stock.The CEA expression,chromosome integration and efficiency of rAAV/CEA virus were determined by PCR,Southern blot and flow cytometery.DC precursor cells were induced by granulocyte macrophage colony-stimulating factor(GM-CSF),interleukin-4(IL-4)and tumor necrosis factor-?(TNF-?)to mature DC.The mature DC and T cells were mixed,then cultured with interleukin-2(IL-2)and interleukin-7(IL-7)resulting in generation of cytotoxic T lymphocyte(CTL),whose lethal effects on cancer cells were determined by 51chromium(51Cr)release assay.3H incorporation method was employed to detect the influence of rAAV/CEA transfected DC in the proliferation of activated T cells.Results rAAV medicated CEA gene was transferred into DC precursor successfully and the expression was high.Over 90% DC precursors expressed CEA protein.The viral gene was integrated into the DC chromosome.rAAV/CEA transfected DC could induce specific CTL which could specifically recognize and kill CEA-positive colorectal cancer LoVo cell line,and showed restrictive activity of anti-MHC class I antigen,while it was not able to kill the CEA-negative cancer cells.Conclusion The present study confirms that rAAV/CEA transfected DC vaccine can induce anti-tumor activity in vitro.It paves the way for further study on CEA as a target antigen for gene therapy on colorectal cancer.

Objective To explore the inhibitory effects of sorafenib combined with cisplatin (DDP) on human hepatocellular carcinoma cells HepG2 in vitro, and the possible underlying mechanisms. Methods The HepG2 cells were divided to 4 groups as control group, sorafenib group, cisplatin group and sorafenib-cisplatin group. Sorafenib and cisplatin were used in single agent or in combination against HepG2 in vitro. The inhibitory effects of sorafenib and/or cisplatin on proliferation of HepG2 were determined by MTT assay at 24h, 48h, 72h and 96h after the addition of the drugs to HepG2 culture. Cell cycle at 24h time point and apoptosis at 48h time point were examined by flow cytometry (FCM). At 24h time point, cells were labeled by Rhodamine123 and mitochondrial transmembrane potential (??m) was determined by FCM, while caspase-3 activity was assessed by caspase-3 colorimetric assay. Results MTT assay showed that sorafenib and cisplatin, when used as a single agent or in combination, could inhibit the growth of HepG2 cells and induce apoptosis in vitro, while synergistic effect was noted when lower doses of sorafenib and DDP were used in combination. It was also found that combined use of sorafenib and cisplatin arrested HepG2 cells at G0/G1 and G2 phase as shown by cell cycle analysis, and the highest apoptosis rate appeared in sorafenib-cisplatin combination group (P

Objective To observe and compare the anti-angiogenic effect of sorafenib, unfractionated heparin(UFH)and low molecular weight heparin (LMWH), singly used or in combination, with chick chorioallantoic membrane assay. Methods Sixty 8d-developed chick embryos were randomly divided into 6 groups (10 each): control group, sorafenib group, UFH group, LMWH group, sorafenib+UFH group, and sorafenib+LMWH group. Drugs were given respectively and incubation continued. The dosage of drugs for each embryo was 20 nmol of sorafenib, 5U of UFH and 5U of LMWH, respectively, and H-DMEM was applied to the embryo in control group in the same volume (2?l). After continuing incubation for another 72h, the anti-angiogenic effect in each group was observed and the vascular number in the chick chorioallantoic membrane was counted. Results Vascular number in single drug groups (sorafenib group: 8.9?1.9; UFH group: 7.6?2.3; LMWH group: 6.9?1.3) was significantly smaller than that in the control group (26.8?2.3, P