Objective:This work aims determine the expression of the neurotensin (NTS) gene in hepatocellular carcinoma (HCC) subgrouping using immunohistochemical staining (IHC) as well as to evaluate the correlation between the activation of NTS/IL-8 pathway in HCC and inflammatory response in microenvironment and epithelial mesenchymal transition (EMT) in cancer and in the prognosis of patients. Methods:Tumor tissues and corresponding adjacent normal tissue were collected from 64 cases of HCC patients. The expression levels of NTS protein and multiple inflammation and EMT-related proteins, including IL-8, VEGF, MMP9, CD68, E-Cadherin,β-Catenin, and Vimentin, were examined in 64 cases of paraffin-embedded HCC tissues using the immunohistochemistry (IHC) staining method. The clinical outcome and overall survival (OS) among 64 cases of HCC patients were compared. Results:We found that the frequency of NTS-expressing tissues among all HCC samples was 17.19%(11/64). Significantly increased IL-8 protein was confirmed in 90.91%of NTS+HCC samples and was positively correlated with the levels of NTS protein in cancer tissues (P=0.036), which implied the dysfunctional activation of NTS/IL-8 pathway in HCC. The levels of VEGF and MMP9 were significantly correlated with the co-expression of NTS and IL-8 in HCC. Evident features of EMT, including decreased membrane expression of E-Cadherin and increased accumulation of cytoplasmicβ-Catemin and Vimentin, were found in NTS+IL-8+samples. The co-expression of NTS and IL-8 in cancer was significantly correlated with the clinical outcomes of patients, as the mortality rate of NTS+IL-8+HCC patients is 2.5-fold higher than that of others after surgery (P=0.022).Accordingly, the OS of NTS+IL-8+HCC patients significantly decreased (24.65±4.45 m vs. 75.79±16.32 m, P=0.013), and these patients are at a higher risk of death at an expected hazard ratio (HR) of 3.457. Conclusion:The NTS/IL-8 pathway is dysfunctionally activated in a subgroup of HCC samples. Highly expressed NTS is associated with increased inflammatory response in microenvironment, enhanced EMT in cancer, and worse prognosis in HCC patients.

Objective To explore the clinical effects of S-1 single agent chemotherapy for the patients undergoing radical resection of cholangiocarcinoma.Methods The clinical data of 51 patients receiving radical resection of cholangiocarcinoma who were admitted to the Second Affiliated Hospital of Zhejiang University from November 2011 to December 2013 were retrospectively analyzed.After radical resection of cholangiocarcinoma,25 patients receiving non-special treatment and 26 patients receiving S-1 single agent chemotherapy were divided into the operation group and chemotherapy group,respectively.S-1 was taken orally twice per day.Forty mg/once of S-1 was applied to patients with the body surface area ＜ 1.25 m2,50 mg/once of S-1 was applied to patients with the body surface area ≥ 1.25 m2 and ＜ 1.50 m2,and 60 mg/once of S-1 was applied to patients with the body surface area ≥ 1.50 m2.The 14 days usage and 7 days withdrawal of S-1 were used as one course of treatment.The standard usage of S-1 was 6-8 courses of treatment.All the patients were followed up by outpatient examination and telephone interview till December 1,2014.Count data were analyzed using the chi-square test.Measurement data with normal distribution were presented as (x) ± s and analyzed using the t test.Survival curve was drawn by the Kaplan-Meier method,and survival analysis was done using the Log-rank test.Results Twenty-six patients in the chemotherapy group finished the courses of chemotherapy without chemotherapy-related death,and 14 patients had chemotherapy adverse reactions with remission after discontinuation of S-1.All the 51 patients were followed up for 5-37 months with a median time of 19 months.The median overall survival time,1-,3-year overall survival rates,tumor-free median survival time and 1-,3-year tumor-free survival rates were 22 months (range,18-27 months),72.3％,42.9％,21 months (range,16-26 months),60.0％,55.0％ in the operation group and 32 months (range,29-35 months),84.6％,44.4％,26 months (range,21-31 months),76.9％,61.9％ in the chemotherapy group,respectively.There was a significant difference in the overall survival between the 2 groups (x2=6.032,P ＜ 0.05).There was no significant difference in the tumor-free survival between the 2 groups (x2=0.498,P ＞ 0.05).Conclusion S-1 single agent chemotherapy after radical resection of cholangiocarcinoma could improve the survival of patients,while no obvious advantages of inhibiting tumor recurrence is observed.

Objective:To highlight the developmental process of 3D cell culture technology system, which is more suitable for isolating and identifying lung cancer stem-like cells than 2D cell culture technology system, and to explore the application of 3D cell cultures in the evaluation of proliferation, apoptosis, invasion, and drug resistance of lung cancer. Methods:Cells (104/well) from the human lung adenocarcinoma cell lines A549 and RPMI 1640 were cultured in complete medium containing 10%fetal bovine serum. Cell suspension was cultured in a BME basal medium. A growth curve was drawn after 7 d of culture. The stem-like cell was identified through a mammosphere culture, drug resistance and invasion assay, and flow cytometry. Data of A549 cells cultured in 3D and 2D tra-ditional cell culture technologies were compared. Results: Cells from the 3D cell culture had higher tumor formation rates [(20.75 ± 0.85) d vs. (60.25 ± 1.49) d, P<0.01)] and tumor sphere formation (28.50%± 1.17%vs. 8.67%± 0.80%, P<0.01) than those from the 2D cell culture. Moreover, cells from 3D cell culture were more invasive and resistant to therapy (58.17%± 2.19%vs. 41.70%±5.81%in 48 h, P<0.01;33.27%±5.76%vs. 27.30%±4.25%in 72 h, P<0.01). Phenotype experimental results demonstrated that the CD44 and CD326 cells were double-positive, whereas the CD24 cell was negative. Conclusion:The proportion of stem-like cells in A549 cell line after 3D cell culture significantly increased compared with 2D cell culture. The 3D cell culture can promote the proliferation of lung cancer stem cells.