Serum N-glycan markers for diagnosing liver fibrosis induced by hepatitis B virus

Table 1 Characteristics of 432 hepatitis B virus-infected patients in different fibrosis stages diagnosed by liver biopsy (mean ± SD)

Variables	Fibrosis stage				P value
	F0-F1	F2	F3	F4
Cases, n	95	126	103	108	/
Male, n (%)	62 (65.26)	88 (69.84)	82 (79.61)	89 (82.41)	/
Age (yr)	37.91 ± 10.71	36.52 ± 11.11	41.71 ± 10.01	46.63 ± 9.95	< 0.001c
HBV DNA (IU/mL)	1.26 × 108 ± 3.64 ×108	1.33 × 108 ± 4.01 × 108	6.39 × 107 ± 1.62 × 108	5.63 × 107 ± 1.56 × 108	0.229
AFP (ng/mL)	3.20 ± 3.01	19.37 ± 100.27	12.56 ± 23.25	40.98 ± 160.29	0.118
ALT (IU/L)	42.58 ± 61.49	125.21 ± 211.29	263.82 ± 421.57	115.84 ± 165.90	< 0.001c
AST (IU/L)	30.45 ± 39.36	85.65 ± 171.17	185.61 ± 292.67	92.18 ± 127.81	< 0.001c
GGT (IU/L)	31.59 ± 26.74	42.10 ± 50.37	78.13 ± 107.06	64.47 ± 68.93	< 0.001c
ALP (IU/L)	75.63 ± 21.27	81.56 ± 31.47	109.30 ± 120.10	103.82 ± 41.55	0.002b
PLT (109/L)	206.91 ± 51.14	198.36 ± 55.72	185.36 ± 58.29	134.95 ± 46.41	< 0.001c
Total bilirubin (μmol/L)	13.09 ± 5.30	18.34 ± 16.99	29.93 ± 31.44	22.67 ± 12.60	< 0.001c
Indirect bilirubin (μmol /L)	7.78 ± 4.31	8.40 ± 9.40	13.46 ± 17.05	10.01 ± 7.94	0.003b
Total protein (g/L)	73.46 ± 4.78	73.61 ± 10.53	72.48 ± 11.14	74.44 ± 7.66	0.635
Albumin (g/L)	45.80 ± 3.30	45.44 ± 4.40	45.33 ± 6.51	44.39 ± 5.39	0.320

^bP < 0.01.

^cP < 0.001. Reference values: AST: 8-38 IU/L; ALT: 4-44 IU/L; GGT: 8-64 IU/L; ALP: 40-129 IU/L; AFP: 0-20 μg/L; PLT: 100-300 × 10⁹/L; TB: 5.1-18.8 μmol/L; IB: 0-12 μmol/L; TP: 60-87 g/L; ALB: 34-55 g/L. HBV: Hepatitis B virus; AFP: Alpha-fetoprotein; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; PLT: Platelet; GGT: γ-glutamyl-transpeptidase; ALP: Alkaline phosphatase.

Table 2 Inflammation grade and fibrosis stage of 432 hepatitis B virus-infected patients

Fibrosis stage	Inflammation grade
	G0	G1	G2	G3	G4	Total
F0-F1	8	71	16	0	0	95
F2	0	31	83	12	0	126
F3	0	12	40	47	4	103
F4	0	11	38	55	4	108
Total	8	125	177	114	8	432

Table 3 N-glycome profiling results in hepatitis B virus-infected patients with liver fibrosis F0-F4 and differences between different fibrosis stages in the modeling group (mean ± SD)

Peak	F0-F1	F2	F3	F4	F0-F1 vs F2-F4	F0-F2 vs F3-F4	F0-F3 vs F4
1	5.92 ± 1.46	7.62 ± 2.57	7.51 ± 2.09	8.68 ± 3.60	< 0.001c	< 0.001c	< 0.001c
2	1.95 ± 0.67	1.78 ± 0.76	1.60 ± 0.57	1.85 ± 0.76	0.161	0.235	0.312
3	4.57 ± 1.16	5.96 ± 1.43	6.04 ± 1.17	6.21 ± 1.66	< 0.001c	< 0.001c	0.003b
4	7.34 ± 1.37	5.96 ± 1.15	5.54 ± 0.81	5.43 ± 0.81	< 0.001c	< 0.001c	< 0.001c
5	47.90 ± 3.77	46.93 ± 4.64	46.75 ± 4.59	46.27 ± 6.23	0.097	0.163	0.197
6	16.89 ± 2.92	17.66 ± 3.01	18.13 ± 2.89	17.30 ± 2.58	0.086	0.323	0.413
7	4.48 ± 1.08	4.53 ± 0.95	4.86 ± 1.22	4.90 ± 1.16	0.089	0.004b	0.058
8	8.07 ± 2.07	6.91 ± 1.90	6.74 ± 1.70	6.22 ± 2.16	< 0.001c	< 0.001c	0.001b
9	2.87 ± 1.27	2.65 ± 1.01	2.82 ± 1.12	3.12 ± 1.33	0.684	0.136	0.035a

^aP < 0.05.

^bP < 0.01.

^cP < 0.001.

Table 4 Predictive value of fibrosis diagnostic models in the modeling group

Fibrosis stage	Model	Cutoff value	Sen	Spec	PPV	NPV	AUROC
F0-F1 vs F2-F4	Model A	0.8032	0.871	0.839	0.951	0.642	0.890
	GlycoFibroTest	-0.4404	0.280	0.903	0.913	0.257	0.538
	GlycoCirrhoTest	-0.2946	0.778	0.452	0.837	0.359	0.644
	APRI	0.4871	0.626	0.839	0.933	0.385	0.790
	FIB-4	0.4720	0.401	0.855	0.908	0.285	0.599
	S index	0.0543	0.742	0.596	0.871	0.386	0.701
	Model A + ALT + PLT	0.7779	0.923	0.806	0.945	0.746	0.912
F0-F2 vs F3-F4	Model B	0.3418	0.986	0.514	0.662	0.974	0.752
	GlycoFibroTest	-0.2295	0.128	1.000	1.000	0.543	0.524
	GlycoCirrhoTest	0.0033	0.270	0.911	0.745	0.564	0.618
	APRI	0.3964	0.819	0.562	0.638	0.766	0.740
	FIB-4	0.3930	0.609	0.726	0.677	0.663	0.699
	S index	0.0890	0.664	0.711	0.690	0.686	0.738
	Model B + ALT + PLT	0.4404	0.855	0.685	0.720	0.833	0.829
F0-F3 vs F4	Model C	0.2258	0.808	0.598	0.407	0.901	0.747
	GlycoFibroTest	-0.3200	0.274	0.939	0.606	0.791	0.571
	GlycoCirrhoTest	-0.0287	0.356	0.827	0.413	0.790	0.607
	APRI	0.4206	0.859	0.488	0.359	0.912	0.666
	FIB-4	0.3762	0.831	0.657	0.447	0.921	0.795
	S index	0.0817	0.765	0.571	0.380	0.876	0.717
	Model C + ALT + PLT	0.3010	0.789	0.845	0.629	0.923	0.885

F¹ APRI = [AST (IU/L) / 40] × 100/PLT (10⁹/L). F² FIB-4 = [age (yr) × AST (IU/L)] / [PLT (10⁹ /L) × ALT^1/2 (IU/L)]. F³ S index = 1000 × GGT (IU/L) / [PLT (10⁹/L) × ALB² (g/L)]. Sen: Sensitivity; Spec: Specificity; PPV: Positive predictive value; NPV: Negative predictive value; ALT: Alanine aminotransferase; GGT: γ-glutamyl-transpeptidase; PLT: Platelet; AUROC: Area under receiver operating characteristic curves.