1 Characteristics of Study Population
Table 1. showed the characteristics of 328 hepatitis participants and the variability of each indicator between groups as superscripts. Among the 233 CHB participants, 64 participants developed LC (27.47%), and 107 participants developed comorbid LC and HCC (45.92%). The Hepatitis E group had the highest levels of AST, ALT, GGT, ALP, Fn, and most of the variations in expression compared to the other groups were statistically significant (P < 0.05).
Table 1
Characteristics of the Study Population
Characteristics | HCC (n = 107) | LC (n = 64) | CHB (n = 62) | CHC (n = 46) | HEV (n = 49) | H/F | P |
Male (%) | 57.01 | 57.81 | 66.13 | 47.83 | 77.55abd | 10.59 | 0.032 |
Age (years) | 57.00(50.00, 64.00) | 53.00(47.25,58.75)a | 48.00(35.75,60.25)a | 58.00(51.00,67.00)bc | 58.00(49.50,69.00)bc | 24.23 | < 0.001 |
Smoking (%) | 34.58 | 26.56 | 64.52ab | 34.78c | 32.65c | 23.19 | < 0.001 |
Alcohol drinking (%) | 25.23 | 12.50 | 62.90ab | 30.43c | 28.57c | 41.35 | < 0.001 |
Chronic hepatitis durations (years) | 19.00(13.00,23.00) | 12.00(9.00,17.75)a | 7.00(4.00,12.25)ab | 2.00(1.00,3.00)abc | 0.00(0.00,0.00)abcd | 241.70 | < 0.001 |
Anti-hepatitis (%) | 79.44 | 87.50 | 88.71 | 89.13 | 0.00abcd | 152.20 | < 0.001 |
AST (U/L) | 38.00(27.00,75.00) | 42.00(29.00,64.00) | 43.00(25.00,92.25) | 34.00(25.00,52.25) | 65.00(31.50,180.00)abcd | 13.47 | 0.009 |
ALT (U/L) | 30.00(25.00,43.00) | 37.00(22.00,63.00) | 33.00(19.75,78.25) | 29.50(16.50 ~ 56.25) | 78.00(30.50 ~ 235.50)abcd | 22.28 | < 0.001 |
ALP (U/L) | 118.90(84.00 ~ 197.20) | 94.90(67.00,134.70)a | 100.10(73.13,172.00) | 129.10(97.68,208.53)bc | 197.58(120.95,251.00)abcd | 40.32 | < 0.001 |
GGT (U/L) | 98.00(42.00,198.00) | 52.00(29.00,105.00)a | 80.00(36.75,174.25)b | 66.00(43.50,129.00)a | 132.00(91.00,195.00)abcd | 53.40 | < 0.001 |
Fn (mg/L) | 217.00(192.00,240.03) | 201.34(172.00,223.96)a | 219.09(182.93,245.91)b | 217.25(182.3,256.35)b | 251.00(225.30,283.68)abcd | 39.10 | < 0.001 |
CHE (U/L) | 6618.00(4855.00,8615.00) | 4777.00(3300.00,7495.00)a | 6996.50(4882.00,9171.50)b | 6071.00(3201.75,8392.75) | 6756.00(3933.50,8590.00) b | 14.78 | 0.005 |
FFA (mmol/L) | 0.51(0.34,0.83) | 0.60(0.27,0.97) | 0.51(0.33,0.74) | 0.45(0.23,0.64)ab | 0.44(0.22,0.68)b | 10.18 | 0.038 |
Urea (mmol/L) | 5.10(4.36,6.87) | 6.07(4.93,7.52)a | 4.89(3.31,6.17)ab | 5.10(3.95,7.25)b | 5.38(4.11,7.18)c | 18.80 | < 0.001 |
TCh (mmol/L) | 4.01(3.45,4.67) | 3.40(2.93,4.29)a | 4.07(3.49,4.66)b | 3.56(2.83,4.64)a | 4.11(3.24,5.11)b | 17.89 | 0.001 |
TG (mmol/L) | 0.98(0.76,1.34) | 0.99(0.68,1.34) | 1.31(1.00,1.86)ab | 1.03(0.75,1.62)c | 1.51(1.20 ~ 2.22)abd | 38.02 | < 0.001 |
HDL-C (mmol/L) | 1.05(0.80,1.39) | 0.95(0.63,1.26) | 1.00(0.68,1.32) | 0.95(0.74,1.27) | 0.90(0.45,1.19)a | 6.96 | 0.138 |
LDL-C (mmol/L) | 2.27(1.81,2.88) | 1.67(1.17,2.36)a | 2.23(1.75,2.90)b | 2.00(1.47,2.96)b | 2.18(1.70,3.09)b | 28.95 | < 0.001 |
sdLDL-C (mmol/L) | 0.50(0.40,0.71) | 0.33(0.20,0.52)a | 0.57(0.31,0.89)b | 0.45(0.27,0.64)a | 0.57(0.35,0.91)b | 27.58 | < 0.001 |
TCh/HDL-C ratio | 4.07(3.04,5.04) | 3.50(2.82,5.21) | 4.36(3.11,5.56) | 3.84(2.98,4.78) | 4.67(3.41,9.19)abd | 12.94 | 0.012 |
Lp(a) (mmol/L) | 36.00(14.00,70.00) | 45.6(11.63,88.25) | 72.10(28.03,211.00)ab | 72.55(42.20,164.68)ab | 80.10(34.90,203.60)ab | 37.84 | < 0.001 |
LDH (U/L) | 241.00(217.00,318.00) | 213.00(180.25,254.50)a | 228.50(190.00,260.50)a | 212.50(185.00,237.50)a | 249.00(193.50,307.00)bd | 23.63 | < 0.001 |
ApoA1 (g/L) | 1.15(0.93,1.36) | 1.06(0.80,1.25) | 0.98(0.83,1.17)a | 1.11(0.99,1.34)c | 1.10(0.68,1.28) | 14.26 | 0.007 |
ApoB (g/L) | 0.72(0.58,0.82) | 0.78(0.67,1.05)a | 0.95(0.84,1.09)ab | 0.69(0.56,0.90)bc | 0.82(0.66,1.02)acd | 44.32 | < 0.001 |
Note: a, compared with HCC; b, compared with LC; c, compared with CHB; d, compared with CHC. |
2 ApoB/ApoA1 is differentially expressed in serum of patients with various liver diseases and in patients with different stages of HCC and LC
ApoB/ApoA1 expression levels in the serum of individuals with various forms of liver diseases vary, as shown in Fig. 2A. The ApoB/ApoA1 expression levels in the HCC group [0.65 (0.47,0.85)], which was significantly lower than the LC group [0.80(0.63,1.07)] and CHB group [0.93(0.76,1.30)] (P < 0.05). ApoB/ApoA1 expression levels were 0.63(0.51,0.82), and 0.78(0.59,1.18) in the CHC group, and HEV group, respectively.
As shown in Fig. 2B, serum ApoB/ApoA1 levels in patients with advanced-stage hepatocellular carcinoma [0.89(0.71,1.26)] were significantly higher than those in patients with early-stage hepatocellular carcinoma [0.47(0.35,0.66)] and those in patients with middle-stage hepatocellular carcinoma [0.61(0.49,0.74)] (P < 0.05), and serum ApoB/ApoA1 levels in patients with middle-stage hepatocellular carcinoma were significantly higher than those in patients with early-stage hepatocellular carcinoma (P < 0.05). As shown in Fig. 2C, serum ApoB/ApoA1 levels in patients with Class C liver cirrhosis [1.40(0.80,2.06)] were significantly higher than those in patients with Class A liver cirrhosis [0.69(0.53,0.95)] (P < 0.05), serum ApoB/ApoA1 levels in patients with Class B liver cirrhosis [0.82(0.68,1.31)] had no statistically significant difference with patients with Class A and Class C liver cirrhosis(P > 0.05).
As shown in Fig. 2D, further analysis of the diagnostic role of ApoB/ApoA1 in differentiating hepatocellular carcinoma in cirrhosis and in combination with AFP, a common diagnostic marker for hepatocellular carcinoma. The analysis showed that the area under the diagnostic curve (AUC) for ApoB/ApoA1 alone was 0.649 (95% CI: 0.565,0.733) and for AFP alone was 0.752 (95% CI༚0.714,0.849). After combining the two, the area under the curve for the combined index was significantly higher at 0.782 (95% CI༚0.749,0.862) compared with the AUC for ApoB/ApoA1 alone (P < 0.05). The diagnostic specificity of the combined index (71.96%) > that of ApoB/ApoA1 (67.29%) > that of AFP (63.55%). The cut-off values for ApoB/ApoA1 and AFP were 0.727 and 12.45 ng/mL, respectively. As shown in Table 2.
Table 2
The performance of ApoB/ApoA1 and AFP for diagnosing HCC
| AUC (95%CI) | Sensitivity (%) | Specificity (%) | Youden Index | Cut-off |
ApoB/ApoA1 | 0.649(0.565,0.733) | 59.38 | 67.29 | 0.267 | 0.727 |
AFP | 0.752(0.714,0.849) | 75.00 | 63.55 | 0.386 | 12.45 ng/mL |
Combination | 0.782(0.749,0.862) | 71.88 | 71.96 | 0.438 | 0.550 |
3 Logistic analysis of the prevalence of HCC in LC patients and prevalence of LC in CHB patients and ApoB/ApoA1 ratio
To further analyze the correlation between ApoB/ApoA1 ratio and HCC in the LC patients, and the correlation between ApoB/ApoA1 ratio and LC in the CHB patients, the data were divided into quartiles of ApoB/ApoA1 ratio, taking the first quartiles as the reference to calculate the odds ratio (OR) for HCC or LC, and the results were shown in Tables 3 and 4. The prevalence of HCC in LC patients and prevalence of LC in CHB patients presented a positive association with the ApoB/ApoA1 ratio in the Crude model (p < 0.01). After age and gender adjusted in Model 1, the results were similar to those of Crude model (p < 0.01).The difference also remained statistically significant after further control of HBV infection time, anti-HBV therapy, smoking, drinking, Fn, CHE, FFA, Urea, TCh, TG, HDL-C, LDL-C, sdLDL-C, T/HDL-C, LP(a), LDH, ALT, AST, ALP and GGT. The fully adjusted OR in Model 2 was 0.043(95%CI:0.009,0.201) for quartile 4 of circulating ApoB/ApoA1 in the LC patients (the highest) versus quartile 1 (the lowest), and was 0.196(95%CI:0.037,1.033) for quartile 4 of ApoB/ApoA1 in the CHB patients (the highest) versus quartile 1 (the lowest).
Table 3
Association of prevalence of HCC with serum ApoB/ApoA1 ratios in LC patients.
ApoB/ApoA1 quartile | n | Ratio range | OR (95%CI) |
Crude | Model 1 | Model 2 |
Quartile 1 (low) | 44 | ≤ 0.5147 | Reference | Reference | Reference |
Quartile 2 | 43 | 0.5147–0.699 | 0.480(0.183,1.259) | 0.432(0.160,1.165) | 0.182(0.046,0.710) |
Quartile 3 | 42 | 0.699–0.8977 | 0.343(0.132,0.890) | 0.333(0.124,0.893) | 0.194(0.053,0.717) |
Quartile 4 (high) | 42 | ≥ 0.8977 | 0.234(0.090,0.605) | 0.205(0.076,0.549) | 0.043(0.009,0.201) |
β | | | -0.459 | -0.489 | -0.891 |
SE | | | 0.149 | 0.153 | 0.232 |
P for trend | | | 0.002 | 0.001 | <0.001 |
Logistic regression was used to examine the associations between serum levels of ApoB/ApoA1 and HCC in LC patients. Serum ApoB/ApoA1 was divided into quartiles (quartile 4: ≥75th, quartile 3: 50–75th, quartile 2: 25–50th, quartile 1: < 25th percentile) |
Crude: no adjustment |
Model 1: adjusted for age, gender |
Model 2: adjusted for the same variables as Model 1 as well as HBV infection time, Anti-HBV therapy, smoking, drinking, Fn, CHE, FFA, Urea, TCh, TG, HDL-C, LDL-C, sdLDL-C, T/HDL-C, LP(a), LDH, ALT, AST, ALP and GGT |
Table 4
Association of prevalence of LC with serum ApoB/ApoA1 ratios in CHB patients.
ApoB/ApoA1 quartile | n | Ratio range | OR (95%CI) |
Crude | Model 1 | Model 2 |
Quartile 1 (low) | 31 | ≤ 0.6876 | Reference | Reference | Reference |
Quartile 2 | 33 | 0.6876–0.8626 | 0.491(0.174,1.382) | 0.562(0.196,1.614) | 0.603(0.122,2.970) |
Quartile 3 | 30 | 0.8626–1.1784 | 0.237(0.081,0.693) | 0.243(0.081,0.727) | 0.111(0.020,0.604) |
Quartile 4 (high) | 32 | ≥ 1.1784 | 0.280(0.098,0.799) | 0.278(0.096,0.806) | 0.196(0.037,1.033) |
β | | | -0.446 | -0.461 | -0.637 |
SE | | | 0.168 | 0.171 | 0.264 |
P for trend | | | 0.008 | 0.007 | 0.016 |
Logistic regression was used to examine the associations between serum levels of ApoB/ApoA1 and LC in CHB patients. Serum ApoB/ApoA1 was divided into quartiles (quartile 4: ≥75th, quartile 3: 50–75th, quartile 2: 25–50th, quartile 1: < 25th percentile) |
Crude: no adjustment |
Model 1: adjusted for age, gender |
Model 2: adjusted for the same variables as Model 1 as well as HBV infection time, Anti-HBV therapy, smoking, drinking, Fn, CHE, FFA, Urea, TCh, TG, HDL-C, LDL-C, sdLDL-C, T/HDL-C, LP(a), LDH, ALT, AST, ALP and GGT |