30 patients (48%) were stage I (early

stage), and the remaining 34 patients were (52%) stages II, III or IV (late stage) of the disease. Table 1 characteristics for the patients included in this study characteristic   No. a(N= 63) % Age/years(Median,range)   58 (37-76)   Sex         Male 40 63.5   Female 23 36.5 Tobacco use         Current 22 35   Former 12 19   Never 29 46 Histology       VE-821   Adenocarcinoma 34 54   Squamous cell carcinoma 20 32   Othersb 9 14 Stage         StageI 30 48   StageII 11 17   StageIII 17 27   StageIV c 5 8 Lymph node metastasis         N0 42 67   N1/N2 21 33 Pleural invasion         Negative 43 68   Positive 20 32 Lymphovascular invasion         Negative 51 81   Positive 12 9 Histologic differentiation         Well/Moderate 30 48   Poor 26 41   not availabled 7 11 a Number for all except age. b Include 2 Large cell

carcinoma, 2 Carcinoid, 1 malignant clear cell sugar tumor, 1 Sarcomatoid carcinoma, and 3 malignancy , but type undetermined. cStage IV was found incidentally during the operation or only for biopsy d Include Carcinoid, malignant clear cell Ulixertinib mw sugar tumor, Sarcomatoid carcinoma, multiple primary lung cancer. Isolation and identification of tumor-associated macrophages In our study, 71 NSCLC samples were collected and TAMs were successful isolated from all samples. However, cell number of TAMs isolated from 8 NSCLC was inadequate for gene expression analysis, and excluded from this study. So TAMs from 63 NSCLC were finally analyzed. The successful rate was 89%(63/71). Each sample weight ranged from 10 mg to 200 mg and the cell number of TAMs collected ranged from 5 × 105 to 1 × 107 per 100 mg tumor tissue. TAMs from lung CH5183284 mw cancer tissue had an irregular shape and projections (Figure 1A). To confirm that the cell isolated from the lung cancer tissue were TAMs without contamination by fibroblasts or tumor cells, staining for the macrophage specific marker CD68 was performed. Over ninety-five percent of the cells stained positively Morin Hydrate for each randomly selected patient

(Figure 1B). Figure 1 Characterization of tumor-associated macrophage. A. Representative cell morphology of tumor-associated macrophages, TAM, fibroblast and lung tumor cell. B. Immunofluorescent was used to distinguish macrophage, fibroblast and lung tumor cell with antibodies targeting CD68 (red), nuclei stained with DAPI (blue). Original magnification, × 400. The mRNA expression levels of IL-10, cathepsin B and cathepsin S in normal macrophages We performed a time course study to show the expression level of IL-10, cathepsin B and cathepsin S in monocytes changes after culture in medium with rhM-CSF. Our study showed the expression level of IL-10, cathepsin B and cathepsin S showed no significant changes in the time dependent study. (All p > 0.05) (Figure 2A). We also performed dose depedent study of rhM-CSF to see whether the expression level of IL-10, cathepsin B and cathepsin S were affected or not.