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Serum concentrations of MMP-2 and TIMP-2 were measured using enzyme-linked immunosorbent assay ELISA kits (R&D Systems, Abingdon, England) according to the manufacturer's instructions. The serum SCH772984 solubility dmso samples were diluted 10-fold before measurement of MMP-2 and 50-fold before determination of TIMP-2 levels. The intra-assay coefficient of variation (CV%) of MMP-2 is reported by the manufacturer as 5.8% at a mean concentration of 18.9 ng/mL, SD=1.1. The CV% of TIMP-2 is referred to be 4.4% at a mean concentration of 1.23 ng/mL, SD=0.054. The cut-off values of serum MMP-2 (188 ng/mL) and TIMP-2 (85 ng/mL) levels correspond to the highest accuracy (minimal false-negative and false-positive results) and were determined using Microsoft Office Excel software. Fisher's exact probability test and Fisher-Freeman-Halton test were used to assess the Selleckchem Dabrafenib correlations between the expression of the analyzed parameters and clinicopathological parameters of tumor [29]. The levels of proteins tested did not follow a normal distribution based on Kolmogorov-Smirnov and Shapiro-Wilk tests, therefore nonparametric statistical analyses were used. The Mann-Whitney U-test was employed to compare two analyzed groups, whereas differences between more than two groups were compared using Kruskal-Wallis test. The post hoc Dwass-Steele-Critchlow-Fligner test was conducted to assess which groups were different, if statistical differences were found. The correlations were determined by the Spearman method, Ritonavir whereas the Kaplan and Meier method was used for the calculation of the survival curves. The log-rank test for univariate analyses of survival and the Cox proportional hazards model for multivariate analyses were employed. Serum levels of proteins tested were presented as the median and range. The differences were considered statically significant when p