Single immunoreactive endothelial cells or endothelial cell clusters separated from other micro-lymphatic vessels were counted as individual micro-lymphatic vessels. Endothelial staining in large vessels with tunica media and nonspecific staining of non-endothelial structures were excluded in micro-lymphatic vessels counts. The mean visual micro-lymphatic vessel density of VEFGR-3 staining was calculated as the average of six AZD8931 research buy counts (two hot spots and three microscopic fields). Micro-lymphatic vessel counts higher than the median micro-lymphatic vessel count

were taken as high LVD, and those that were lower than the median were taken as low LVD. Statistical analysis All calculations were done using the statistical software SPSS V.14.0 (Chicago, Illinois, USA). Data were shown as mean ± standard deviation. Spearman’s coefficient of correlation, Chi-squared tests, and

Mann-Whitney tests were used as appropriate. A multivariate model using logistic regression analysis was used selleck kinase inhibitor to evaluate statistical associations among variables. For all tests, a two-sided P-value less than 0.05 was considered to be significant. Selleck AICAR Hazard ratios (HR) and their corresponding 95% confidence intervals (95% CI) were computed to provide quantitative information about the relevance of the results of the statistical analysis. Results Basic clinical information and tumor characteristics Forty-six male and 14 female patients (mean age, 57.6 ± 10.4 years; range, 36-79 years) with ESCC treated by curative surgical resection were enrolled in the study. Of the 60 tumors, 15 were well differentiated, 27 were moderately differentiated, and 18 were poorly differentiated. Using the TNM staging system of the International Union Against Cancer (2009) [18], cases were classified as stage I (n = 9), stage II (n

= 11), and stage III (n = 40). Twenty-four of 60 patients had lymph node metastasis, according to surgery and pathology reports. Patient data were analyzed after a 5-year follow-up; information was obtained in 91.7% (55 of 60) of cases. The median overall survival was 26.9 ± 2.7 months (95% CI: 21.4-31.9 months), and the mean overall survival was 38.1 ± 6.5 months (95% CI: 27.6-52.0 months). The clinical characteristics of study samples are summarized in Table 1. Table 1 Association of NF-κB and isothipendyl Notch1 expression with clinical characteristics Clinicopathological feature NF-κB expression P-value Notch1 expression P-value   High Low   High Low   Gender               Male 21 25 0.451 22 24 0.887   Female 8 6   7 7   Age (years)               ≤ 60 17 23 0.201 23 17 0.058   > 60 12 8   6 14   Differentiation               Well 7 8 0.231 3 12 0.001   Moderate 16 11   10 17     Poor 6 12   16 2   TNM stages               I + II 8 12 0.361 10 10 0.855   III 21 19   19 21   Lymphatic metastasis               With 23 2 0.001 6 19 0.001   Without 6 29   23 12   LVD (VEGF-R3)               High 19 12 0.038 10 21 0.010   Low 10 19   19 10   Podoplanin               High 20 10 0.004 8 19 0.