003), age (P =0.034), AFP (P <0.001), tumor number (P =0.02), and TNM stage (P =0.009). IDH2 expression correlated with HBsAg (P =0.015), AFP (P <0.001), and tumor differentiation (P =0.015) (Additional file 2: Table S2). Other clinical characteristics were not directly related to the expression of 5-hmC or IDH2. selleckchem Table 1 Summary of the correlations of 5-hmC and IDH2 protein expression with clinicopathological features in the training cohort (N = 318) Clinicopathological indexes   No. of patients No. of patients   5-hmC Low 5-hmC High P† IDH2

Low IDH2 High P† Sex Female 18 36 0.007 28 26 0.765   Male 141 123   131 133   Age(year) ≤50 55 65 0.247 60 60 1.000   >50 104 94   99 99   HBsAg Negative 30 26 0.556 28 28 1.000   Positive 129 133   131 131   HCV Negative 158 158 1.000 157 159 0.156   Positive 1 1   2 0   AFP ≤20 83 37 <0.001 58 62 0.644   >20 76 122   101 97 XMU-MP-1 solubility dmso   γ-GT(U/L) ≤54 87 81 0.500 78 90 0.178   >54 72 78   81 69   Liver cirrhosis No 32 26 0.384 23 35 0.081   Yes 127 133   136 124   Tumor number Single 131 134 0.652 134 131 0.652   Multiple 28 25   25 28   Tumor size(cm) ≤5 97 108 0.197 99 106 0.412   >5 62 51   60 53   Tumor encapsulation Complete 94 88 0.496 93 89 0.650   None

65 71   66 70   Microvascular invasion Absent 113 107 0.466 106 114 0.331   Present 46 52   53 45   Tumor differentiation I + II 129 115 0.063 113 131 0.017   III + IV 30 44   46 28   TNM stage I 98 93 0.567 93 98 0.567   II + III 61 66   66 61   Abbreviations: HBsAg, hepatitis B surface antigen; AFP, α-fetoprotein; γ-GT, γ-glutamyl

C646 mouse transferase; TNM, tumor-node-metastasis. †A P-value < 0.05 was considered statistically significant. P-values were calculated using the Pearson chi-square test. Boldface type indicates significant values. Association between combined 5-hmC and IDH2 expression and outcome in the training cohort By the last Adenosine triphosphate follow-up in the training cohort (November 2011), 47.2% (150/318) of the patients had suffered a recurrence and 36.5% (116/318) had died. The 1-, 3-, and 5-year OS rates in the cohort were 83.6%, 67.6%, and 63.5% and the cumulative recurrence rates were 32.7%, 46.9%, and 52.8%, respectively. Additionally, we found that the 1-, 3-, and 5-year survival rates of the 5-hmC High patients were significantly higher than those of the 5-hmC Low group (87.4% vs. 79.9%, 77.4% vs. 57.9%, and 73.0% vs. 54.1%, respectively) (Figure 2a). Similarly, the 5-hmC Low patients had a poorer prognosis at 1, 3, and 5 years, with higher cumulative recurrence rates than the 5-hmC High patients (40.3% vs. 25.2%, 56.6% vs. 37.1%, and 61.6% vs. 44.0%, respectively) (Figure 2b). We also discovered that the 1-, 3-, and 5-year survival rates of the IDH2 High patients were significantly higher than those of the IDH2 Low group (93.7% vs.