Ki-67 index of the endothelium cells of the micro lymphatic vessels (Ki67%) was calculated according to Wulff et al [22]. Statistical Analysis Correlations between podoplanin, VEGFR-3, LYVE-1

and the vessel numbers as continuous variables were used to assess CD31-positive vessel counts with the Spearman rank correlation test. Categorical data were compared by the χ2 or Fishers’ exact probability test. Distribution was normal or with Mann Whitney U test buy I-BET-762 if the sample distribution was asymmetrical. The relationship between lymph vessel variables and lymph node status was analyzed by one-way ANOVA, followed by the Neuman-Keuls test. Overall survival intervals were determined as the time period from initial diagnosis to the time of death. Overall survival analyses were done using the Kaplan-Meier method. The comparison between survival

functions for different strata was assessed with the log-rank statistic. selleck compound Multivariate analysis of prognostic factors was this website done using Cox’s regression model. Differences were considered significant when P ≤ 0.05. All statistical analyses were done using the statistical package spss13.0. Results CD31, VEGFR-3, LYVE-1, VEGF-C Expression in NSCLC Numerous intratumoral and peritumoral vessels could be observed in each NSCLC tumor irrespective of histologic grade and pathologic stage. CD31 was positive in endothelial cell plasma in micro vessels, appeared yellow granular. Micro vessels of tumor tissues were

mainly located at intra-tumor and peritumoral area. However, large blood vessels with muscular coat were also positive stained for CD31 (Fig. 1a). VEGFR-3 showed an expression similar to CD31. VEGFR-3 positive vessels included not only dilated and irregular thin-walled lymphatic vessels, but also blood vessels containing erythrocytes and large blood vessels with smooth muscle (Fig. 1b). LYVE-1 was positively stained in endothelial cell plasma and plasma membrane in micro vessels, appeared yellow granular (Fig. 1c). However, few LYVE-1 positive vessels were large blood vessels with smooth muscle, and tumor embolus were observed in Prostatic acid phosphatase their muscular layer and lumen (Fig. 1d). VEGF-C positive substance in tumor tissue was yellow fine granular, mainly located in tumor cell plasma. Positive cells were dispersed, limited locally or in small patches (Fig. 1e). In the para-tumor normal bronchia, VEGF-C expression was dispersed in columnar epithelium cells (Fig. 1f). Figure 1 Immunohistochemical analysis of different markers. Podoplanin Expression in NSCLC Podoplanin expression was mainly present in thin-walled (lymphatic) structures. Podoplanin was positive in endothelial cell plasma in thin-walled lymph vessel, appeared yellow granular.