The staining intensity was scored as: 0 (negative), 1 (weak), 2 (moderate) and 3 (strong). Raw data were converted to IHS by multiplying the quantity score (0-4) by the staining intensity score (0-3). Theoretically, the scores can EPZ015938 range from 0 to 12. An IHS of 9-12 was considered a strong immunoreactivity; 5-8, moderate; 1-4, weak; and 0, negative. In statistical analysis, COX-2 and VEGF-C scores were placed in a high expression group (strong and moderate immunoreactivity) and a low expression group

(weak and negative immunoreactivity). Immunoreactivity was scored by two independent researchers. LVD was detected by immunostaining for D2-40, according to the criteria of Masakau et al. [25]. First, areas with highly D2-40-positive see more vessels (hot spots) in peritumoral, intratumoral and normal tissue were identified, by scanning the sections at low magnification (×100);

then the number of D2-40 positive vessels was counted in five high-magnification fields (×400) for each case. The mean value for the five fields was calculated as the LVD for each tumor. To evaluate the impact of LVD on prognosis, we divided the 56 cases into two groups according to the mean LVD level. Statistical analysis Statistical analyses were performed with SPSS 11.5 software (SPSS Inc, Chicago, USA). The correlations among the expression of COX-2, VEGF-C, levels of LVD, and learn more clinicopathologic characteristics were calculated by Student’s t-test, chi-square correlation test and Spearman’s coefficient of correlation Amobarbital as appropriate. The Kaplan-Meier method was used to estimate

survival as a function of time, and survival differences were analyzed with the log-rank test. A multivariable test was performed to determine the factor correlated with survival length by Cox regression analysis. The statistical significance level was defined as P < 0.05. Results Patient information The 56 patients (35 males and 21 females) had a mean age of 56.2 (range 27-74) years. Twenty-six of the cases displayed weight loss, and 17 presented anemia with hemoglobin (HGB) < 90 g/l. Histological examination showed that 4 displayed well differentiated adenocarcinoma, 18 moderate and 34 poor. According to the sixth AJCC TNM classification, 16 patients were in stage I, 18 in stage II, 19 in stage III, and 3 in stage IV. Of the 56 patients, 39 (69.6%) had lymph node metastasis. Up to 2008, there were 32 patients in total that had died. COX-2, VEGF-C and D2-40 expression in gastric carcinoma Positive expression of COX-2 protein and VEGF-C showed as a yellow or brownish yellow stain in the cytoplasm of carcinoma cells (Figures 1 and 2). The expression rates of COX-2 and VEGF-C were 69.64% (39/56) and 55.36% (31/56), respectively, in gastric carcinoma. However, normal tissue showed no immunoreactivity for COX-2 and VEGF-C.