Efficacy of a Chinese herbal formula on hepatitis B e antigen-positive chronic hepatitis B patients

Table 1 Hospitals or medical centers that participated in this study

Name	Location (city and province)
Shanghai Shuguang hospital, Shanghai University of Traditional Chinese Medicine	Shanghai
The Second Hospital Affiliated with Zhejiang University of Traditional Chinese Medicine	Hangzhou, Zhejiang
Xiamen Hospital of Traditional Chinese Medicine	Xiamen, Fujian
Shenzhen Hospital Affiliated with Guangzhou University of Chinese Medicine	Shenzhen, Guangdong
Foshan Hospital of Traditional Chinese Medicine	Foshan, Guangdong
The Third People’s Hospital of Shenzhen	Shenzhen, Guangdong
Guangdong Hospital of Traditional Chinese Medicine	Guangzhou, Guangdong
The Third Affiliated Hospital of Sun Yat-sen University	Guangzhou, Guangdong
Ruikang Hospital of Guangxi College of Traditional Chinese Medicine	Nanning, Guangxi
The First Affiliated Hospital of Guangxi College of Traditional Chinese Medicine	Nanning, Guangxi
Attached Hospital of Chengdu University of Traditional Chinese Medicine	Chengdu, Sichuan
West China Hospital, West China School of Medicine, Sichuan University	Chengdu, Sichuan
Beijing Ditan Hospital, Capital Medical University	Beijing
Xiyuan Hospital, China Academy of Traditional Chinese Medicine	Beijing
302 Military Hospital of China	Beijing
Tianjin Infectious Disease Hospital	Tianjin
Beijing Youan Hospital, Capital Medical University	Beijing
Hubei Provincial Hospital of TCM	Wuhan, Hubei
People's Hospital of Wuhan University	Wuhan, Hubei
The First Hospital of Hunan University of Chinese Medicine	Changsha, Hunan

Table 2 Inclusion and exclusion criteria of patients

Inclusion criteria	Exclusion criteria
Conform with the diagnostic criteria of HBeAg (+) chronic hepatitis B	Inactive HBsAg (+) carriers
Conform with the pathogenesis and syndromes of kidney deficiency	Serum a-fetoprotein abnormal
Age 30-65 yr	Pregnancy or breast feeding
ALT ≤ 40 IU/L	Coinfection with HIV, HCV, HDV
HBsAg > 10 IU/mL and < 105 IU/mL HBV DNA (105-109 IU/mL)	Histologic evidence of cirrhosis; Evidence of any other chronic liver disease
Liver biopsy: Liver histology showed Knodell HAI > 4, Ishak fibrosis score > 3 were also included	Mental illness or any other serious systemic illness
Voluntary	Interferon-γ within 6 mo; Antivirus treatment with nucleoside
	Abuse alcohol or illegal drugs; Allergic to the drug ingredients

HBeAg: Hepatitis B e antigen; HBsAg: Hepatitis B surface antigen; ALT: Alanine aminotransferase; HBV: Hepatitis B virus; HIV: Human immunodeficiency virus; HCV: Hepatitis C virus; HDV: Hepatitis D virus; HAI: Histological activity index.

Table 3 The list of raw herbs composing the Chinese herbal formula

Chinese name	Latin name	Parts of plant used	Dose of dryplant (grams)	Dose afterextraction (grams)
Ye xia zhu	Phyllanthus urinaria Linn	Whole plant	30	12.00
Ci wu jia	Radix et caulis acanthopanacis senticosi	Root and rhizome	10	0.50
Xian ling pi	Herba Epimedii	Overground part	30	1.50
Nv zhen zi	Fructus ligustri lucidi	Mature fruit	15	1.50
Han lian cao	Herba ecliptae	Overground part	15	1.50
Chai hu	Radix bupleuri	Root	10	1.67
Bai shao	Radix paeoniae alba	Root	10	1.00
Zhi shi	Fructus aurantii immaturus	Fruitlet	10	1.67
Tao ren	Semen persicae	Nuts	10	0.50
Gan cao	Radix glycyrrhizae	Root and rhizome	5	0.83
Hu zhang	Rhizoma polygoni cuspidati	Root and rhizome	15	1.00
Xi huang cao	Herba rabdosiae serrae	Whole plant	30	9.00
Total			190	32.67

Table 4 Baseline characteristics of all study patients

Variable	Treatment (ICE) group (n = 199)	Control group(n = 196)	χ2/t/ Z	P value
Age (mean ± SD), yr	38.51 ± 7.63	38.90 ± 7.54	-0.904	0.366a
Range	30-65	30-63
Sex, n (%)			0.013	0.910b
Male	128 (64.3)	125 (63.8)
Female	71 (35.7)	71 (36.2)
Regions (N)			2.796	0.593b
Eastern	30 (14.42)	28 (14.97)
Western	22 (10.58)	18 (9.63)
Southern	89 (42.79)	69 (36.90)
Northern	40 (19.23)	48 (25.67)
Central	27 (12.98)	24 (12.83)
Smoking			0.491	0.484b
Yes	57 (28.64)	50 (25.51)
No	142 (71.36)	146 (74.49)
Alcohol consumption			3.626	0.057b
Yes	18 (9.05)	30 (15.31)
No	181 (90.95)	166 (84.69)
Genotype			0.522	0.770b
B	99 (49.75)	93 (47.45)
C	90 (45.23)	95 (48.47)
D	10 (5.02)	8 (4.08)
Genealogy of hepatocellular carcinoma	2 (1.00)	3 (1.53)	0.000	0.986b
Clinical course (mean ± SD) week	90.21 ± 22.40	86.69 ± 27.20	-1.607	0.108a
Liver function (mean ± SD)
ALT, IU/L	29.29 ± 8.19	30.12 ± 6.32	1.126	0.261c
AST, IU/L	24.86 ± 7.53	25.79 ± 6.19	1.340	0.181c
TB, μmol/L	14.34 ± 3.25	13.98 ± 4.15	0.961	0.337c
HBV DNA baseline level, (%)			0.154	0.695b
2 to < 5 log10 IU/mL	0 (0)	0 (0)
5 to < 7 log10 IU/mL	14 (2.01)	12 (1.02)
7to < 9 log10 IU/mL	185 (97.99)	184 (98.98)
HBsAg (mean ± SD), log10 IU/mL	3.86 ± 0.52	3.89 ± 0.42	-0.758	0.449a
HBeAg (mean ± SD), SCO/mL	1138.18 ± 423.99	1158.40 ± 401.86	-0.393	0.695a
HBeAb (mean ± SD), SCO/mL	38.43 ± 14.28	40.01 ± 12.15	-0.904	0.366a
Histological scores
Knodell (HAI), n (%)	168 (84.40)	156 (79.60)	0.010	0.922b
≥ 4	60 (35.71)	55(35.26)
< 4	108 (64.29)	101 (64.74)
Ishak (FIB), n (%)			0.035	0.851b
≥ 2	67 (39.90)	60 (38.46)
< 2	101 (60.10)	96 (61.54)

^a: Mann-Whitney U test;

^b: Chi-square test;

^c: t-test. ICE: Invigorating kidney and clearing away the heat and expelling superficial evils; SD: Standard deviation; ALT: Alanine aminotransferase; AST: Aspartate transaminase; TB: Total bilirubin; HBV: Hepatitis B virus; HBsAg: Hepatitis B virus surface antigen; HBeAg: Hepatitis B virus e antigen; HBeAb: Hepatitis B virus e antibody.

Table 5 Virologic response and change in serum hepatitis B virus DNA level after treatment

Treatment response	Treatment (ICE) group (n = 199)	Control group ( n = 196)	χ2/Z	P value
48 wk
Patients with HBV DNA level decline > 2 log10 IU/mL, n (%)	30 (15.08)	13 (6.63)	7.255	0.007
Patients with HBV DNA level ≤ 4 log10 IU/mL, n (%)	20 (10.05)	5 (2.55)	9.367	0.002
Patients with undetectable HBV DNA (≤ 20 IU/mL), n (%)	2 (1.01)	0 (0.00)	1.980	0.159
96 wk
Patients with HBV DNA level decline > 2 log10 IU/mL, n (%)	60 (30.15)	12 (6.12)	38.249	0.000
Patients with HBV DNA level ≤ 4 log10 IU/mL, n (%)	37 (18.59)	6 (3.06)	24.555	0.000
Patients with undetectable HBV DNA (≤ 20 IU/mL), n (%)	4 (2.01)	0 (0.00)	2.227	0.136

Chi-square test. ICE: Invigorating kidney and clearing away the heat and expelling superficial evils; HBV: hepatitis B virus.

Table 6 Virologic response and change in serum hepatitis B surface antigen and hepatitis B virus e antigen levels after treatment

Treatment response	Treatment (ICE) group (n = 199)	Control group (n = 196)	χ2/Z	P value
48 wk
Patients with HBsAg level decline ≥ 0.5 log10 IU/mL, n (%)	49 (24.62)	19 (9.69)	15.443	0.000
Patients with HBsAg level decline ≥ 1 log10 IU/mL, n (%)	29 (14.57)	11 (5.61)	8.712	0.003
Patients with HBsAg level decline ≥ 2 log10 IU/mL, n (%)	7 (3.52)	2 (1.02)	1.758	0.185
Patients with undetectable HBsAg (≤ 0.05 IU/mL), n (%)	0 (0.00)	0 (0.00)	0.000	1
Patients with HBeAg level decline ≥ 1 log10 S/CO), n (%)	45 (22.61)	5 (2.55)	35.947	0.000
Patients with undetectable HBeAg (≤ 1.00 S/CO), n (%)	17 (8.54)	5 (2.55)	6.740	0.009
Seroconversion rates of HBeAga, n (%)	16 (8.04)	4 (2.04)	7.394	0.007
96 wk
Patients with HBsAg level decline ≥ 0.5 log10 IU/mL, n (%)	83 (41.71)	41 (20.92)	19.817	0.000
Patients with HBsAg level decline ≥ 1 log10 IU/mL, n (%)	63 (31.66)	22 (11.22)	24.413	0.000
Patients with HBsAg level decline ≥ 2 log10 IU/mL, n (%)	17 (8.54)	1 (0.51)	7.129	0.008
Patients with undetectable HBsAg (≤ 0.05 IU/mL), n (%)	0 (0)	0 (0)	0.000	1
Patients with HBeAg level decline ≥ 1 log10 S/CO), n (%)	51 (25.63)	9 (4.59)	33.919	0.000
Patients with undetectable HBeAg (≤ 1.00 S/CO), n (%)	32 (16.08)	11 (5.61)	11.154	0.001
Seroconversion rates of HBeAga, n (%)	29 (14.57)	9 (4.46)	11.962	0.001

^aPrevious HBeAg-positive patient HBeAg ≤ 1.00 S/CO and HBeAb > 1.0 S/CO. Chi-square test. ICE: Invigorating kidney and clearing away the heat and expelling superficial evils; HBsAg: Hepatitis B surface antigen; HBeAg: Hepatitis B e antigen.

Table 7 Baseline variables and change in hepatitis B virus e antigen and hepatitis B surface antigen from week 12 to week 48 associated with HBeAg clearance

Variables	Univariable analysis			Multivariable analysis
	OR	95%CI	P	OR	95%CI	P
Sex	0.788	(0.367-1.986)	0.453
Age	0.947	(0.886-1.198)	0.715
HBV DNA	0.633	(0.574-1.393)	0.214
IFN-γ	0.915	(0.837-2.131)	0.656
IL-2	0.773	(0.512-1.318)	0.375
Baseline HBeAg	1.653	(1.332-2.257)	0.030	1.027	(1.145-1. 908)	0.047
Baseline HBsAg	2.431	(1.236-3.915)	0.004	1.339	(1.131-1.862)	0.009
Week 24 HBeAg change from baseline	2.762	(1.562-4.256)	< 0.001	2.338	(1.636-4.863)	< 0.001
Week 36 HBeAg change from baseline	3.411	(1.976-4.526)	< 0.001	3.185	(1.977-5.466)	< 0.001
Week 24 HBsAg change from baseline	4.458	(2.153-10.198)	< 0.001	3.273	(1.375-5.216)	< 0.001
Week 36 HBsAg change from baseline	5.371	(3.239-6.392)	< 0.001	5.788	(2.726-10.612)	< 0.001
Week 12 ALT elevation	2.676	(1.133-5.432)	0.016	2.049	(1.363-9.198)	0.006
Week 24 ALT elevation	3.373	(2.637-7.568)	0.003	3.788	(2.728-7.687)	0.003
Week 48 IFN-γ elevation	2.735	(1.317-6.682)	0.002	2.171	(1.163-2.961)	0.007
Week 48 IL-2 elevation	2.133	(1.171-8.616)	0.008	1.882	(1.026-2.613)	0.020

CI: Confidence interval; ALT: Alanine aminotransferase; HBV: Hepatitis B virus; HBsAg: Hepatitis B virus surface antigen; HBeAg: Hepatitis B virus e antigen; IL-2: Interleukin-2; IFN-γ: Interferon-γ.