Objective To study the in vitro anti-tumor activity of dendritic cells (DCs) loading with antigen produced by radiofrequency ablation of tumor lysate in situ combined with cytokine-induced killer cells (CIK).Methods CIK ceils derived from BALB/C mouse spleen and DCs derived from bone marrow were prepared,and experimental model of murine colon carcinoma were established for radiofrequency ablation.The supernatant of tumor tissue in situ lysis after repeated freezing and thawing were tested by lowry protein quantitative statutory,amounting to a final concentration of 5 μg/ml,then load to the first 5 days of culture DCs (Ag-DC),2 days later,co-cultured with CIK cells after the first seven days of culture 48 h (Ag-DC-CIK).Flow cytometry was used to analyze costimulatory molecules on the surface of the cells,and CCK-8 assay to detect in vitro cytotoxic activity.Results The DCs loading with antigen resulted in an increase in the proportion of CD86 + CD11 c +,MHC Ⅱ + CD11 c + and MHC Ⅱ + CD80 + cells.The main effector cells of CIK cells were CD3 + NK1.1 + cells.The percentage of CD3 + NK1.1 + cells was 1.45％ on the first day of the culture ; while when they had been cultured for 7 days,the percentage CD3 + NK1.1 + significantly increased to 36.9％.The cytotoxicity of Ag-DC-CIK cells toward C26 cells was much more efficient than that of DC-CIK,CIK cells.The cytotoxic activity of the former was significantly lower than the latter and the same target ratio.When the ratios of effector cells to target cells were 5 ∶ 1,the cytotoxic activity of Ag-DC-CIK cells against C26 cells was (74.9 ± 3.5) ％,; while the DC-CIK was (71.2 ± 2.1) ％ and the CIK cells was (68.7 ± 2.9) ％.The difference was statistically significant(F =7.007,P =0.007).When the ratios of effector cells to target cells were 10 ∶ 1,the cytotoxic activity of Ag-DC-CIK cells against C26 cells was (82.3 ± 4.5) ％,while the DC-CIK cells was (77.1 ± 5.1) ％,and the CIK cells was (72.7 ± 2.8) ％.The difference was statistically significant (F =7.727,P =0.005).When the ratios of effector cells to target cells were 20 ∶ 1,the cytotoxic activity of Ag-DC-CIK cells against C26 cells was (83.2 ± 1.9) ％,while the DC-CIK cells was (77.2 ± 4.2) ％,and the CIK cells was (73.0 ± 2.6) ％.The difference was statistically significant (F =16.594,P =0.000).Conclusion DCs loading with antigen produced by radiofrequency ablation of tumor in situ pyrolysis products can improve in vitro cytotoxic activity combined with CIK cells,which can provide a new comprehensive cancer treatment strategy.

ObjectiveTo investigate the relationships between T and NK cells in peripheral blood and the relapse or metastasis of malignant tumors.MethodsFlow cytometry was used to test the percentages ofT and NK cells in peripheral blood of 48 malignant tumor patients with relapse or metastasis (relapse or metastasis group) and 48 malignant tumor patients without relapse or metastasis(control group).ResultsCompared with control group,the ratio of Th/Ts decreased,CD4+/CD25+ raised and the activities of NK cells decreased in relapse or metastasis group (0.95 ±0.52 vs.1.35 ±0.43,7.15 ±3.81 vs.6.01 ±2.67,0.2053 ±0.1135 vs.0.2501 ±0.0745)(P＜0.01 or ＜0.05).ConclusionsTesting the T and NK cells in peripheral blood of malignant tumor patients regularly can know their immune status and find the relapse or metastasis early.It is easy and useful in patients' follow-up.

Objective To evaluate the predictive value of tumor metabolic indexes measured by 18F-FDG PET/CT in recurrence of stage Ⅰ NSCLC after surgery.Methods A total of 85 patients (44 males,41 females,age (62.46± 10.38) years) in Shanghai Renji Hospital with stage Ⅰ NSCLC,who underwent 18F-FDG PET/CT and subsequent surgical resection,were retrospectively enrolled from April 2006 to December 2011.Gender,age,tumor size,pathology,SUVmax,MTV and TLG of the primary tumor were selected as variables.ROC curve analysis was used to analyze the cut off value.The prognostic significance of parameters for recurrence-free survival (RFS) was evaluated by univariate and multivariate analyses.Survival analysis was analyzed by Kaplan-Meier method.Results During follow-up period,tumor recurrence occurred in 21 patients (24.7％,21/85) and 11 patients (12.9％,11/85) died.The median follow-up period was 44 months.The median values of SUVmax,MTV and TLG were 4.100,3.048 cm3 and 7.970,respectively.Cut off values of SUVmax,MTV and TLG were 7.115,4.701 cm3 and 12.015 according to ROC curve analysis.Univariate Cox analysis showed that SUVmax(x2 =22.091),MTV (x2 =4.941) and TLG(x2 =10.488) were associated with RFS(all P＜0.05).But gender,age,tumor size,and pathology were not independent risk factors of recurrence (x2=0.248-3.888,all P＞0.05).Multivariate Cox analysis revealed that SUVmax(=16.902,HR=15.426,P＜0.05) and TLG (x2=6.029,HR=4.054,P＜0.05) were independent prognostic factors for recurrence.Kaplan-Meier survival analysis showed that the period of RFS in high SUVmax (＞ 7.115) group (x2=32.545,P＜0.05) and in high TLG (＞12.015) group (x2=12.665,P＜0.05) were lower than those in low SUVmax group and low TLG group.Conclusion The SUVmax and TLG measured by 18F-FDG PET/CT have significant value for predicting the recurrence of stage Ⅰ NSCLC.

Objective To compare treatment response according to the PERCIST1.0,RECIST1.1,EORTC,and WHO criteria in patients with colorectal liver metastases (CLM) who received neoadjuvant chemotherapy.Methods A total of 41 CLM patients (27 males,average age 68.48 years;14 females,average age 62.43 years) from January 2010 to September 2013 were included in this retrospective study.PET/CT scan was performed before chemotherapy and after 4-6 cycles′ chemotherapy.The baseline and the sequential follow-up 18F-FDG PET/CT of each patient were evaluated according to the PERCIST1.0,RECIST1.1,EORTC,and WHO criteria.The response was categorized into 4 levels including CR,PR,SD,PD.PET/CT images were used for both metabolic and anatomic evaluation.The concurrent diagnostic CT or MRI images (performed within 1 week of PET/CT) were also utilized when needed.The agreements of criteria were analyzed using Kappa test.The response rate (RR) and disease control rate (DCR) were compared using χ2 test.Results The RR and DCR according to the PERCIST1.0,EORTC and RECIST1.1 criteria were 31.71％(13/41) and 63.41％(26/41),31.71％(13/41) and 60.98％(25/41),17.07％(7/41) and 68.29％(28/41),respectively.The general comparison of PERCIST1.0 and RECIST1.1,EORTC and RECIST1.1 criteria showed good agreements (κ values: 0.711,0.689).Significant difference was not found in the DCR(χ2=2.000,P>0.05) but found in the RR(χ2=6.000,P<0.05) between PERCIST1.0 and RECIST1.1.Difference of DCR between EORTC and RECIST1.1 was not significant(χ2=3.000,P>0.05),while the RR had significant difference(χ2=6.000,P<0.05).The RR and DCR according to WHO criterion were 12.20％(5/41) and 70.73％(29/41),which had a good consistency with those according to PERCIST1.0 criteria (κ=0.629).Significant statistical difference was not found in the DCR(χ2=3.000,P>0.05) but found in the RR(χ2=8.000,P<0.05) between PERCIST1.0 and WHO criteria.Conclusions In evaluating CLM treatment response,anatomical criteria and metabolic criteria have a good consistency.But metabolic criteria are more sensitive for RR evaluating.

Objective To evaluate the significance of AFP-IgM, this is one of new tumor markers, in the diagnosis of primary hepatocellular carcinoma (PHC). Methods The contents of AFP-IgM and AFP in serum of 103 healthy subjects, 74 patients suffered primary hepatic carcinoma, 27 patients affected by liver cirrhosis and 63 patients affected by chronic hepatitis were detected by means of enzyme linked immunosorbent assay and electrochemiluminescence. No-PHC is comprised of liver cirrhosis,chronic hepatitis and health subjects as control group. Results The area under ROC curve of AFP was larger than that of AFP-IgM (0.85 vs 0.72, Z=3.21) and the best cut-off value of AFP-IgM and AFP was 3×105-AU/L and 10 ug/L respectively, which was determined by ROC curve. Under the cut-off value, the sensitivity of AFP- lgM and AFP for PHC were 64.9% and 79.7%, and the specificity were 75.6% and 80.3%, yet their efficacies were similar. However, for early diagnosis of liver cancer (stage Ⅰ and Ⅱ), the area under ROC curve of AFP-IgM was larger than that of AFP (0.91 vs 0.82,Z=1.73). The sensitivity of AFP-IgM andAFP were 94.4% and 72. 2%, and the specificity were 81.9% and 79.9%. The differences of AFP-IgMand AFP for early diagnosis of liver cancer were statistically significant. When both of the test results combined AFP-IgM with AFP are positive, it can be diagnosed as liver cancer. The specificity of combineddetermination of the two forms was 89.1%, and the efficacy was 79. 0%. Conclusions Both of thesensitivity and specificity of the AFP-IgM test were higher than that of the AFP for early diagnosis of livercancer. We also found that combined determination of the two forms significantly increased the specificityand the positive predictive value for the diagnosis of PHC, thus AFP-IgM was of especially significance forearly diagnosis of liver cancer.

Objective: To investigate the effect of IFN-γ on the expression of IL-33 in colon cancer CT26 cells. Methods: CT26 cells were treated with IFN-γ and IFN-γ combined with PKA inhibitor H89, respectively, and a negative control group (NC, untreated) was set up at the same time. The mRNA expression levels of PKA, CREB and IL-33 in CT26 cells were detected by qRT-PCR. The expression levels of PKA, CREB, p-CREB and IL-33 proteins in CT26 cells were determined by western blot. The localization of CREB protein in CT26 cells was analyzed by the immunofluorescence confocal microscopy. Results: The relative expression levels of PKA, CREB and IL-33 mRNA in the IFN-γ-treated group were 2.50±0.11, 3.10±0.08 and 2.80±0.22, respectively, which were significantly higher than those in the NC group (P＜0.05). The relative expression levels of PKA, CREB and IL-33 mRNA in the IFN-γ combined with H89 treatment group were 0.21±0.02, 0.59±0.05 and 0.35±0.04, respectively, which were significantly lower than those in the NC group and IFN-γ-treated group (P＜0.05). The expression levels of PKA, CREB and IL-33 proteins detected by western blot were consistent with that of mRNA. Immunofluorescence confocal results showed that the expression level of CREB in the IFN-γ-treated group was significantly higher than that in the NC group, and that the expression level of CREB in the IFN-γ combined with H89 treatment group was significantly lower than that in the IFN-γ-treated group. Conclusion IFN-γ may induce the expression of IL-33 in colon cancer CT26 cells via the PKA-CREB pathway.