3.1 Baseline characteristics
Based on the inclusion criteria established for the enrolled patients, a total of 625 patients diagnosed with CHB were selected for statistical analysis, comprising 278 individuals in the uncured group and 347 individuals in the cured group (Figure 1). The baseline characteristics of the patients are presented in Table 1. Prior to matching, the average age of patients with CHB in the untreated group was notably higher (43.3 years vs. 39.1 years, P < 0.001). After matching, there were no statistically significant differences in the average age or sex ratio between the CHB patients in the uncured group and those in the cured group (P = 0.479, P = 0.054). Additionally, there were no statistically significant differences in the treatment regimen or baseline HBsAg level between the two groups (P = 0.513, P = 0.492). Moreover, there was no statistically significant difference in baseline renal function between the two groups of CHB patients, as indicated by the estimated glomerular filtration rate (eGFR) and the inorganic phosphorus (IPHOS) and cystatin C (CysC) levels (all P > 0.05).
Table1. Baseline characteristics of participants before and after PS matching.
Baseline Characteristics
|
Before PSM(N=625)
|
P Value
|
After PSM(N=250)
|
P
Value
|
Uncured Cured
(N=278) (N=347)
|
Uncured Cured
(N=125) (N=125)
|
Age (years)
|
|
43.35±8.66
|
39.15±8.52
|
<0.001
|
41.38±9.33
|
41.75±8.65
|
0.479
|
|
Sex
|
|
|
|
|
|
|
|
|
|
male
|
|
23
|
57
|
|
<0.001
|
113
|
110
|
0.054
|
|
female
Treatment
The types of NAs in combination therapy
ETV
TDF
TAF
Other NAs
Peg-IFN (n)
Blood routine
|
|
255
135 (49%)
110 (40%)
20 (7%)
13 (4%)
53 (48-60)
|
290
162 (47%)
149 (43%)
15 (4%)
21 (6%)
45 (48-52)
|
|
0.341
|
12
60 (48%)
110 (40%)
8 (6%)
5 (4%)
49 (37-60)
|
15
57 (46%)
149 (43%)
14 (10%)
7 (6%)
48 (3-60)
|
0.513
|
|
WBC (×109/L)
|
|
6.13±1.48
|
5.88±1.56
|
|
0.058
|
6.07±1.33
|
5.87±1.56
|
0.291
|
|
HGB (g/L)
|
|
153.43±17.82
|
152.52±16.91
|
|
0.513
|
153.45±18.20
|
152.33±15.06
|
0.597
|
|
PLT (×109/L)
|
|
206.74±52.77
|
208.80±58.96
|
|
0.649
|
209.08±49.65
|
206.28±58.26
|
0.291
|
|
LYM (×109/L)
|
|
2.04±0.58
|
1.98±0.58
|
|
0.260
|
1.98±0.56
|
2.00±0.57
|
0.807
|
|
NEUT (×109/L)
|
|
3.48±1.19
|
3.27±1.23
|
|
0.026
|
3.47±1.07
|
3.25±1.21
|
0.122
|
|
HBsAg (iu/ml)
|
|
751.01±643.79
|
243.58±283.47
|
|
<0.001
|
454.42±288.32
|
426.81±343.72
|
0.492
|
|
Blood biochemistry data
|
|
|
|
|
|
|
|
|
|
AST (U/L)
|
|
31.52±18.67
|
31.07±25.92
|
|
0.811
|
29.82±16.17
|
32.54±31.26
|
0.388
|
|
ALT (U/L)
|
|
36.15±25.84
|
36.80±37.17
|
|
0.805
|
34.65±24.62
|
39.49±48.62
|
0.322
|
|
TBIL (µmol/L)
|
|
12.21±6.73
|
11.65±5.04
|
|
0.235
|
12.96±7.58
|
11.96±5.73
|
0.243
|
|
DBIL (µmol/L)
|
|
3.65±3.76
|
3.24±1.66
|
|
0.072
|
4.41±5.29
|
3.33±1.79
|
0.032
|
|
ALB (g/L)
|
|
47.58±3.18
|
48.20±2.74
|
|
0.010
|
47.84±3.20
|
48.24±2.66
|
0.280
|
|
eGFR(ml/min/1.73 m²)
|
|
101.64±14.70
|
104.64±16.11
|
|
0.016
|
103.07±14.18
|
102.70±16.10
|
0.850
|
|
IPHOS (mmol/L)
|
|
1.03±0.16
|
1.06±0.29
|
|
0.119
|
1.04±0.14
|
1.08±0.34
|
0.308
|
|
Cysc (mg/L)
|
|
0.90±0.14
|
0.87±0.14
|
|
0.005
|
0.89±0.12
|
0.88±0.13
|
0.564
|
|
Note: Continuous variables are expressed as the mean ± standard deviation (SD), and categorical variables are expressed as numbers and percentages (%).
Abbreviations: WBC, white blood cell; HGB, haemoglobin; PLT, platelet count; Lym, lymphocyte count; Neut, neutrophil neutrophil count; HBsAg, hepatitis B surface antigen; Ast, aspartate aminotransferase; Alt, alanine aminotransferase; TBIL, total bilirubin serum total bilirubin; DBIL, direct bilirubin; ALB, albumin, albumin; eGFR, estimated glomerular filtration rate; IPHOS, iphos, serum inorganic phosphorus; CysC, Cystatin C, serum cystatin.
a1: One covariate in the matched model included age, total number of interferon needles, baseline HBsAg, baseline ALT, and baseline eGFR.
b2:Data conforming to a normal distribution were analysed for differences (Wilcoxon rank sum test and chi square test) using the two independent samples t test.
3.2 Renal function
The eGFR of the study population, stratified by cure status, sex, and age, decreased at 12 weeks of treatment among women and patients under 40 years of age in the cured group. After 12 weeks of treatment, the eGFR in the uncured group decreased, whereas the eGFR in the cured group continuously increased until the 24-week mark(Figure2). A statistically significant decrease was subsequently detected in both groups at the 48 weeks (P < 0.05, Figure2). During the 48-week treatment period, women exhibited a greater eGFR than men did. Specifically, women exhibited an increase in the eGFR at 36 weeks, while men continued to experience a decrease. The difference between the two groups was found to be statistically significant at 48 weeks (P < 0.001, Figure2). After initially decreasing from 0 to 12 weeks in the cured state, the IPHOS levels increased according to sex and age group. CysC levels increased up to 36 weeks and then decreased in both groups, with no significant difference between them. CysC levels were greater in males older than 40 years than in the other groups at 48 weeks (P < 0.05,Figure2 ).
Renal function between different healing, sex and age groups. (A) Changes in the eGFR between different healing states. (B) Changes in eGFR between different sexes. (C) Changes in the eGFR at different ages (< 40 years, ≥ 40 years). (D) Changes in the IPHOS between different healing states. (E) Changes in IPHOS between different genders. (F) Changes in the IPHOS at different ages (< 40 years old, ≥ 40 years old). (G) Changes in CysC between different cure states. (H) Changes in CysC between different sexes. (I) Changes in CysC at different ages (< 40 years old, ≥ 40 years old). nsp>0.05, *p<0.05, **p<0.001.
3.3 Renal function after various treatment regimens
There were no statistically significant differences in the age or sex distributions among patients receiving pegIFN in combination with ETV, pegIFN in combination with TDF, or pegIFN monotherapy. The analysis of eGFR levels indicated that the three treatment regimens exhibited similar patterns of change over the 48 weeks treatment period, with no statistically significant differences observed (all P > 0.05,Table2). Similarly, the levels of IPHOS demonstrated consistent trends among the three treatment groups throughout the 48 weeks treatment duration, with no statistically significant differences at 0, 12, 24, or 36 weeks (all P > 0.05,Table2). However, a statistically significant difference was noted at the 48 weeks mark (P = 0.013,Table2,). The levels of CysC indicated that the three treatment regimens exhibited a consistent pattern of change over a 48 weeks treatment period, with no statistically significant differences observed at 0, 12, 24, or 48 weeks (all P > 0.05,Table2). However, a significant difference was noted at 36 weeks (P = 0.029,Table2).
Table2. Participant baseline characteristics after propensity score matching.
Baseline Characteristics
|
pegIFN-ETV
(N=208)
|
pegIFN-TDF
(N=208)
|
pegIFN alone
(N=104)
|
P value
|
Age(years)
|
36.1±9.1
|
35.3±9.2
|
36.6±8.8
|
0.462
|
Sex
|
|
|
|
0.955
|
male
|
176
|
178
|
89
|
|
female
|
32
|
30
|
15
|
|
Peg-IFN
|
47(36-60)
|
47(36-59)
|
51(38-60)
|
0.261
|
HBsAg(iu/ml)
|
|
|
|
|
0 w
|
418.33±366.09
|
444.98±477.61
|
344.93±295.55
|
0.117
|
12 w
|
278.93±374.78
|
329.81±469.90
|
245.42±332.25
|
0.188
|
24 w
|
170.05±300.37
|
207.46±360.38
|
192.31±345.52
|
0.520
|
36 w
|
134.71±260.41
|
168.98±330.66
|
288.64±1515.15
|
0.205
|
48 w
|
131.54±238.79
|
174.70±495.13
|
122.08±225.05
|
0.353
|
Biochemistry data(renal function)
|
|
|
|
|
eGFR(ml/min/1.73 m²)
|
|
|
|
|
0 w
|
103.44±14.74
|
104.34±16.03
|
106.97±15.06
|
0.156
|
12 w
|
107.54±13.57
|
109.59±14.01
|
111.29±14.52
|
0.067
|
24 w
|
107.98±12.87
|
110.06±14.28
|
108.99±13.79
|
0.297
|
36 w
|
106.17±13.20
|
108.96±12.84
|
107.16±15.51
|
0.107
|
48 w
|
105.06±14.08
|
106.51±13.92
|
105.17±15.02
|
0.537
|
IPHOS(mmol/L)
|
|
|
|
|
0 w
|
1.04±2.67
|
1.05±0.18
|
1.08±0.33
|
0.384
|
12 w
|
1.00±0.26
|
1.05±0.39
|
1.00±0.23
|
0.258
|
24 w
|
1.02±0.20
|
1.05±0.33
|
1.06±0.30
|
0.466
|
36 w
|
1.07±0.20
|
1.08±0.16
|
1.09±0.35
|
0.641
|
48 w
|
1.09±0.15
|
1.11±0.15
|
1.05±0.18
|
0.013
|
Cysc(mg/L)
|
|
|
|
|
0 w
|
0.87±0.13
|
0.88±0.14
|
0.85±0.16
|
0.304
|
12 w
|
0.97±0.16
|
0.95±0.16
|
0.94±0.16
|
0.191
|
24 w
|
1.01±0.16
|
1.01±0.16
|
1.00±0.19
|
0.875
|
36 w
|
1.03±0.17
|
1.00±0.15
|
0.98±0.20
|
0.029
|
48 w
|
1.01±0.78
|
0.99±0.16
|
1.00±0.17
|
0.373
|
Abbreviations: pegIFN-ETV, pegylated interferon enteric cavity, pegylated interferon combined with entecavir; pegIFN-TDF, pegylated interferon tenofovir disoproxil, pegylated interferon combined with tenofovir fumarate; pegIFN, pegylated interferon, pegylated interferon
3.4 Baseline characteristics of patients with various stages of renal dysfunction
Patients were categorized into a stage 1 group and a stage 2 group based on their eGFR at 48 weeks of treatment. The present study revealed significant differences in age between the two groups, with the stage 1 group being significantly older than the stage 2 group (P < 0.001,Table3). There was no significant difference in the use of pegylated interferon (P = 0.452,Table3).
Baseline levels of eGFR, IPHOS, and CysC showed significant differences (all P < 0.05,Table3). eGFR and CysC levels showed significant differences over the 48 weeks of treatment (P < 0.001,Table3), while IPHOS levels did not differ significantly between the two groups at 12, 24, 36, and 48 weeks (all P > 0.05,Table3).Meanwhile, we observed patients in both phases separately according to gender subgroups and found that among male patients, there was a significant difference in baseline renal function and 48 weeks changes between the two stages (P < 0.001,TableS1).The same results were seen in female patients, with significant differences in baseline and 48 weeks renal function between the two stages(all P < 0.05,TableS2).
Baseline Characteristics
|
Renal function(stage1)
(N=73)
|
Renal function(stage2)
(N=447)
|
P value
|
Age
|
41.60±8.80
|
34.90±8.80
|
<0.001
|
HBsAg(iu/ml)
|
|
|
|
0 w
|
362.63±365.66
|
422.75±409.54
|
0.234
|
12 w
|
276.23±371.15
|
295.25±414.89
|
0.842
|
24 w
|
213.59±366.87
|
185.53±328.96
|
0.236
|
36 w
|
176.57±341.80
|
179.63±341.80
|
0.257
|
48 w
|
159.59±316.37
|
144.84±369.29
|
0.477
|
Biochemistry data
|
|
|
|
AST(U/L)
|
|
|
|
0 w
|
27.55±9.12
|
28.08±14.64
|
0.185
|
12 w
|
55.68±41.35
|
58.46±45.35
|
0.444
|
24 w
|
52.48±48.36
|
52.80±33.64
|
0.327
|
36 w
|
41.24±19.85
|
43.61±26.16
|
0.844
|
48 w
|
34.72±17.16
|
41.84±80.71
|
0.913
|
ALT(U/L)
|
|
|
|
0 w
|
30.29±14.09
|
31.62±20.42
|
0.471
|
12 w
|
63.62±62.01
|
68.44±45.81
|
0.084
|
24 w
|
54.25±65.32
|
60.93±58.19
|
0.016
|
36 w
|
42.73±19.47
|
47.34±38.32
|
0.558
|
48 w
|
45.33±65.76
|
44.68±60.80
|
0.560
|
eGFR(ml/min/1.73 m²)
|
|
|
|
0 w
|
87.48±12.92
|
107.29±13.87
|
<0.001
|
12 w
|
92.06±11.52
|
111.90±12.28
|
<0.001
|
24 w
|
92.10±11.49
|
111.78±11.86
|
<0.001
|
36 w
|
89.57±12.08
|
110.41±11.41
|
<0.001
|
48 w
|
81.01±7.67
|
109.68±10.46
|
<0.001
|
IPHOS(mmol/L)
|
|
|
|
0 w
|
1.03±0.25
|
1.06±0.25
|
0.009
|
12 w
|
1.01±0.20
|
1.02±0.25
|
0.104
|
24 w
|
1.00±0.20
|
1.04±0.29
|
0.103
|
36 w
|
1.09±0.29
|
1.08±0.21
|
0.256
|
48 w
|
1.09±0.15
|
1.09±0.16
|
0.947
|
Cysc(mg/L)
|
|
|
|
0 w
|
0.94±0.14
|
0.86±0.14
|
<0.001
|
12 w
|
1.08±0.18
|
0.94±0.15
|
<0.001
|
24 w
|
1.11±0.19
|
0.99±0.16
|
<0.001
|
36 w
|
1.13±0.19
|
0.99±0.16
|
<0.001
|
48 w
|
1.12±0.17
|
0.98±0.16
|
<0.001
|
Table 3. Characteristics of participants with different renal function stages.
3.5 Variation tendencies at the different stages of renal function.
Participants were divided into stage 1 and stage 2 groups based on their baseline renal function. The renal function in both groups increased and then decreased at 48 weeks, with higher eGFR in the pegIFN-TDF group than in the other two groups. There was no significant difference in renal function between the stage 1 and stage 2 groups; however, there was a significant difference in the eGFR at 12, 24, and 36 weeks in the stage 2 group (P < 0.05,Figure3).In addition, we also analyzed the changes in renal function among the three treatment regimens for the two staged patients, and the results showed that there was no significant difference in the changes in renal function among the three treatment regimens at 12, 24, 36 and 48 weeks(FigureS3).
3.6 Efficacy test of predictors for renal outcome
Four factors were chosen for predictive testing based on LASSO regression and multivariate analysis(FigureS4). The first two factors, baseline eGFR and age, were selected using the AUC. Logistic regression was used to predict the eGFR at 48 weeks in patients. The area under the receiver operating characteristic curve (AUC) of the prediction model was 0.851 (95% CI: 0.807, 0.895,Figure4 and Table4). At a prediction probability threshold of 0.853, the Youden index was maximized for patients with normal renal function at 48 weeks of treatment, yielding a sensitivity and specificity of 0.781 each. We created nomograms and analysed a calibration curve to evaluate the impact of sex, age, cure status, and eGFR on renal function after 48 weeks of treatment. The calibration curve had a C index of 0.729, indicating moderate accuracy.
Table4. Age and Baseline Renal Function Indicators Predict 48-Week Renal Outcomes
Variables
|
AUC
|
P value
|
95%Confidence interval
|
Age
|
0.698
|
<0.001
|
0.630-0.765
|
eGFR0W
|
0.849
|
<0.001
|
0.807-0.891
|
IPHOS0W
|
0.596
|
0.009
|
0.525-0.666
|
Cysc0W
Age-eGFR0W
|
0.659
0.851
|
<0.001
<0.001
|
0.591-0.727
0.807-0.895
|