Histopathology of type C liver disease for determining hepatocellular carcinoma risk factors

Table 1 Comparison of clinical background factors at the time of liver biopsy in sustained virological response patients, between F0 + 1 + 2 and F3 + 4 groups n (%)

Parameter		F0 + 1 + 2	F3 + 4	P value
Number	232	182	50
Gender (male)	136 (58.6)	104 (57.1)	32 (64.0)	0.3832
Age (yr)	47.9 ± 12.2	45.9 ± 12.4	55.2 ± 8.3	0.0001
BMI	22.6 ± 3.3	22.7 ± 3.3	22.2 ± 3.6	0.5466
AST (U/L)	55.2 ± 45.9	49.8 ± 46.6	74.5 ± 37.6	0.0007
ALT (U/L)	75.5 ± 62.7	70.5 ± 63.0	93.5 ± 58.9	0.0216
γ-GT (U/L)	62.3 ± 71.8	61.4 ± 76.7	65.8 ± 50.4	0.7048
Platelet count (× 104/μL)	18.8 ± 6.2	20.2 ± 6.0	13.9 ± 3.9	0.0001
Genotype type 1/2	108 (46.5)/ 124 (53.5)	86 (47.2)/ 96 (52.8)	22 (44.0)/ 28 (56.0)	0.6830
AFP (ng/mL)	7.7 ± 13.3	5.4 ± 8.6	15.5 ± 21.4	0.0008
Observation period (yr)	7.5 ± 4.9	7.4 ± 4.7	8.0 ± 5.5	0.4350

These 232 patients were divided into two groups according to the F stage: stages 0, 1, and 2 (F0 + 1 + 2 group, n = 182) and stages 3-4 (F3 + 4 group, n = 50). Comparison of clinical background factors at the time of liver biopsy in SVR patients according to the F stage (F0 + 1 + 2 and F3 + 4) was performed, and the data are shown. The clinical background factors demonstrated that there was a higher level of deterioration of the liver in the F3+4 group as compared to the F0 + 1 + 2 group; Age, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and α-fetoprotein (AFP) was higher, and platelet count was lower. The parameters are presented as mean ± SD. AST: Upper limit of the normal range is 38 U/L; ALT: Upper limit of the normal range is 44 U/L; γ-glutamyltransferase (GT): Upper limit of normal is 73 U/L; AFP: Upper limit of normal is 20.0 ng/mL; Platelet count: reference value is 15.0 × 10⁴-35.0 × 10⁴/μL. BMI: Body mass index.

Table 2 Comparison of liver histological findings at the time of liver biopsy in sustained virological response patients, between F0 + 1 + 2 and F3 + 4 groups

Parameter	Score					F0+1+2	F3+4	P value
	0	1	2	3	4
Irregular regeneration
Dysplastic change	109	50	49	21	3	0.769 ± 0.998	1.660 ± 1.081	0.0001
Oncocytes	211	16	4	1	0	0.066 ± 0.290	0.300 ± 0.647	0.0003
Map-like distribution	160	37	22	13	0	0.396 ± 0.756	0.960 ± 1.142	0.0001
Nodular arrangement	216	13	1	2	0	0.027 ± 0.164	0.320 ± 0.713	0.0001
Bulging	206	18	6	2	0	0.104 ± 0.400	0.340 ± 0.688	0.0022
Atypical hepatocytes	227	5	0	0	0	0.005 ± 0.074	0.080 ± 0.274	0.0012
Inflammatory cell infiltration
Peri-portal	0	65	127	35	5	1.819 ± 0.652	2.260 ± 0.828	0.0001
Parenchymal	0	37	149	44	2	1.978 ± 0.585	2.300 ± 0.678	0.0010
Portal lymphocyte	0	5	103	121	3	2.456 ± 0.562	2.780 ± 0.507	0.0003
Portal lymphoid reaction	0	6	47	78	101	3.132 ± 0.850	3.360 ± 0.802	0.0904
Fibrosis
F stage	5	128	49	34	16
Portal sclerosis	113	81	32	6	0	0.615 ± 0.783	1.020 ± 0.795	0.0014
Pericellular fibrosis	89	90	41	11	1	0.802 ± 0.870	1.260 ± 0.853	0.0011
Perivenular fibrosis	63	66	65	25	0	1.236 ± 1.016	1.243 ± 0.895	0.9691
Bridging necrosis Y (presence)						0/228	4/228 (1.7%)	0.0020
Bile duct damage	123	71	26	9	2	0.597 ± 0.861	1.000 ± 0.904	0.0041
Steatosis	96	46	68	18	1	0.944 ± 0.990	1.429 ± 1.118	0.0035

Comparison of liver histological findings in sustained virological response patients according to the F stage (F0 + 1 + 2 and F3 + 4) was performed, and the data are shown. The histological findings demonstrated that there was a higher level of deterioration of the liver in the F3 + 4 group as compared to the F0 + 1 + 2 group; liver fibrosis was more progressive and inflammatory reaction was stronger. The number of patients for each score was compared, then the presence of bridging necrosis (Y/N) was compared and the remaining parameters are presented as mean ± SD. Scoring: score 0, none; score 1, minimal (observed in less than one third of the field); score 2, mild (observed in one third to less than two thirds of the field); score 3, moderate (observed in two thirds or more of the field) and score 4, severe (diffusely in all fields).

Table 3 Factors associated with the development of hepatocellular carcinoma, identified by comparing patients with sustained virological response by univariate analysis using Cox proportional hazard regression method (n = 232)

Parameter	F0 + 1 + 2			F3 + 4
	RR	95%CI	P value	RR	95%CI	P value
Clinical background factors1
Gender (male)	1.200	0.109-13.252	0.8815	-	-	-
Age (yr)	1.308	0.985-1.737	0.0635	1.123	0.959-1.314	0.1494
BMI	1.121	0.693-1.814	0.6415	0.987	0.549-1.772	0.9644
AST (U/L)	0.996	0.964-1.028	0.7903	1.003	0.971-1.037	0.8540
ALT (U/L)	0.992	0.965-1.020	0.5678	0.996	0.972-1.020	0.7494
γ-GT (U/L)	0.996	0.975-1.018	0.7169	1.001	0.977-1.025	0.9436
Platelet count (× 104/μL)	1.004	0.837-1.204	0.9634	1.088	0.855-1.384	0.4936
Genotype type 1	-	-	-	0.285	0.030-2.752	0.2781
AFP (ng/mL)	0.989	0.867-1.129	0.8698	0.995	0.919-1.077	0.9046
Histological findings
Irregular regeneration2
Dysplastic change	1.920	0.720-5.124	0.1926	1.330	0.543-3.256	0.5322
Oncocytes	-	-	-	1.788	0.510-6.265	0.3639
Map-like distribution	-	-	-	3.082	1.066-8.913	0.0378
Nodularity	-	-	-	-	-	-
Bulging	-	-	-	-	-	-
Atypical hepatocytes	-	-	-	42.055	4.303-411.029	0.0013
Infiltration
Peri-portal	3.248	0.537-19.655	0.1995	0.477	0.110-2.067	0.3225
Parenchymal	3.028	0.389-23.604	0.2902	0.420	0.092-1.921	0.2634
Portal lymphocyte	2.085	0.217-20.014	0.5241	1.213	0.177-8.333	0.8443
Portal lymphoid reaction	2.362	0.345-16.168	0.3812	3.364	0.484-23.407	0.2202
Fibrosis
F stage
Portal sclerosis	0.551	0.078-3.884	0.5500	1.293	0.444-3769	0.6373
Pericellular fibrosis	0.963	0.243-3.822	0.9576	0.534	0.117-2.440	0.4184
Perivenular fibrosis	0.627	0.168-2.344	0.4878	0.429	0.076-2.412	0.3370
Bridging necrosis Y	-	-	-	-	-	-
Bile duct damage	0.678	0.113-4.061	0.6707	1.305	0.374-4.555	0.6766
Steatosis	1.064	0.343-3.306	0.9142	1.660	0.623-4.421	0.3106

We compared the clinical background factors and histological findings at liver biopsy of the patients with sustained virological response in the F0 + 1 + 2 and F3 + 4 groups, data of which are shown. Univariate analysis using the Cox proportional hazard regression method was applied to these parameters to determine the clinical background factors that were associated with the development of hepatocellular carcinoma (HCC).

¹None of the clinical background factors showed any significant associations with HCC. Blank in the table signifies that the bias was applied, but could not be statistically processed;

²Significant correlations were found for map-like distribution (RR = 3.082, 95%CI: 1.066-8.913, P = 0.0378) and atypical hepatocytes (RR = 42.055, 95%CI: 4.303-411.0, P = 0.0013) in the F3 + 4 patients. None of the liver histopathological findings showed any significant associations with HCC in the F0 + 1 + 2 patients. Blank values signify that the bias was applied, but could not be statistically processed. AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; AFP: α-fetoprotein; BMI: Body mass index; GT: glutamyltransferase.

Table 4 Factors associated with the development of hepatocellular carcinoma, identified by comparing patients with sustained virological response by multivariate analysis using Cox proportional hazard regression method

F3 + 4 parameter	RR	95%CI	P value
Map-like distribution	1.687	0.404-7.053	0.4734
Atypical hepatocytes	20.748	1.335-322.530	0.0303

Multivariate analysis using the Cox proportional hazard regression method was applied to the parameters in each of the F stage groups. In the F0 + 1 + 2 group, none of the factors showed any significant correlation with the development of hepatocellular carcinoma (HCC). In the F3 + 4 group, map-like distribution (RR = 1.687, 95%CI: 0.404-7.053, P = 0.4734) was not found to have any correlation while the presence of atypical hepatocytes (RR = 20.748, 95%CI: 1.335-322.5, P = 0.0303) was shown to be significantly associated with HCC development.