Of the 41 T-NHL patients, 23 were males and 18 were females. The mean age was 48.34 ± 16.19 years. According to the WHO classification, the histological types of the specimens in our study included peripheral T cell lymphoma, not otherwise characterized (32 cases), extranodal NK/T cell lymphoma, www.selleckchem.com/products/Adriamycin.html nasal type (5 cases), anaplastic large cell lymphoma (2 cases), and angioimmunoblastic T cell lymphoma (2 cases). Method Immunohistochemical Staining The avidin-biotin complex

method was used to detect the CCR7 (anti-CCR7, 1:300 dilution; Epitomics Inc.), MMP-2 (anti-MMP-2, 1:250 dilution; Zhong Shan Inc., Beijing), and MMP-9 (anti-MMP-9, 1:250 dilution; Zhong Shan Inc., Beijing). The formalin-fixed, paraffin-embedded tissues were deparaffinized and subsequently heated in a microwave oven with EDTA buffer. After preincubation with hydrogen peroxide, an avidin/biotin blocking kit, and rabbit serum, the primary antibodies were applied overnight in the wet box at 4°C, and then

incubated with the secondary antibodies (rabbit anti-goat biotinylated; 1:200 dilution, ZhongShan Inc., Beijing) for about 50min. At last avidin-biotin complex was added, and enzyme activity was visualized with diaminobenzidine. Counterstaining was done with hematoxylin. For the negative controls, only the secondary antibodies were used. A negative control was done for every lymphoma and reactive lymph node sample (n = 60). For the positive controls, formalin-fixed, paraffin-embedded tissue samples of the human spleen were applied. Evaluation of Immunohistochemical Staining Immunohistochemical staining was independently evaluated by four authors, blinded to patient outcome and all clinicopathologic this website findings. The immunohistochemical staining was analyzed according to staining index, which was calculated by multiplying the score for staining intensity (0, absent, no color in tumor cells; 1, weak, pale yellow in tumor cells;

2, intermediate, yellow in tumor cells; 3, strong staining, brown yellow in tumor cells) with the score for percentage of stained tumor cells (0, positive cells account for 0%-10%; 1, 11%-25%; 2, 26%-50%; 3, >50%). The staining index value ranges from 0 to 9. The specimens grouped by staining index value as – (<2), + (2-4), ++ (5-7), +++ (8-9). The slide of ++ or higher than ++ was classified as high expression. Otherwise, the slide was classified as low expression. Thalidomide The slides were usually evaluated by four observers. The final classification of a slide was determined by the value agreed to by a majority of observers. In vitro Experimentation Materials Cell Culture The human cutaneous T cell lymphoma cell line Hut78 and the adult T lymphocytic leukemia/lymphoma Jurkat cell line were inoculated into cellular culture boards with improved 1640 medium supplemented with 10% fetal bovine serum (Hyclone, Inc., USA), 100 units/mL penicillin, 100 μg/mL streptomycin (Cambrex, East Rutherford, NJ), and 1 mmol/L L-glutamine.