Long-term Outcomes of Anti-viral Therapies in Patients With Chronic Viral Hepatitis B: A Multicenter, Real-world Study
Conditions: Chronic Hepatitis b
Sponsor: Huashan Hospital

Protocol ID：KY2020-911 
Page 1 of 19 
Version: 03 3 04/12/2023 
1 
 
                                                                     
 
 
 
 
Long-term Outcomes of Anti-viral Therapies  
in Patients with Chronic Viral Hepatitis B : 
A Multicenter, Real-world Study (OASIS) 
 
Principal Investigator: Wen-Hong Zhang  
Department: Department of Infectious Diseases  
Sponsor: None 
Protocol Number: KY2020-911 
NCT Number: NCT04896255 
Version: 03 3 04/12/2023 
 
 
 
 
 
 
 
 
 

Protocol ID：KY2020-911 
Page 2 of 19 
Version: 03 3 04/12/2023 
2 
 
Contents 
1 
Scientific Background ............................................................................................... 3 
2 
Aims .......................................................................................................................... 5 
3 
Study design .............................................................................................................. 5 
4 
Eligibility Criteria, Exclusion Criteria, and Withdrawal Criteria .............................. 7 
4.1 
Inclusion Criteria ............................................................................................... 7 
4.2 
Exclusion Criteria .............................................................................................. 7 
4.3 
Withdrawal Criteria ........................................................................................... 7 
4.4 
Lost to Follow-up Criteria ................................................................................. 7 
5 
Sample Size Estimation ............................................................................................. 8 
6 
Study Procedures ....................................................................................................... 8 
6.1 
Clinical Information to be Collected ................................................................. 8 
6.1.1 
The collection information and time point of PS Cohort .......................... 8 
6.1.2 
The collection information of RPS Cohort .............................................. 11 
6.1.3 
The collection information of RS Cohort ................................................ 11 
6.2 
Follow-up Management and Measure to Ensure Patient Compliance ............. 11 
7 
Study Termination Criteria ...................................................................................... 11 
8 
Efficacy Evaluation ................................................................................................. 12 
8.1 
Effectiveness.................................................................................................... 12 
8.1.1 
Primary Objectives: ................................................................................. 12 
8.1.2 
Secondary Objectives: ............................................................................. 12 
8.1.3 
Exploratory Objectives: ........................................................................... 12 
8.2 
Safety Objectives: ............................................................................................ 13 
9 
Adverse Event Recording ........................................................................................ 13 
10 
Statistical Analysis Plan ...................................................................................... 14 
11 
Data Management and Confidentiality ................................................................ 14 
12 
Quality Control and Assurance............................................................................ 15 
13 
Ethical Considerations ......................................................................................... 15 
14 
Recruitment and Informed Consent Process........................................................ 15 
15 
Responsibilities of Participating Parties and Other Provisions ........................... 16 
16 
Reference ............................................................................................................. 16 

Protocol ID：KY2020-911 
Page 3 of 19 
Version: 03 3 04/12/2023 
3 
 
1 
Scientific Background 
Chronic hepatitis B (CHB) is an infectious disease caused by hepatitis B virus (HBV), which can 
last longer than six months, and characterized primarily by liver injury. Epidemiological data 
estimate that over 70 million people in China are currently infected with HBV, of whom more 
than 20 million have developed CHB. Since 2012, the annual incidence of CHB in China has 
exceeded 1.05 million cases [1,2]. In the liver of CHB patients, direct effects of viral replication 
and transcription activity, together with virus-triggered host antiviral immune responses, lead to 
persistent inflammatory cell infiltration in the liver, ongoing hepatocellular injury and necrosis, 
and activation of collagen fiber proliferation and deposition. This results in abnormal liver 
function, progressive hepatic fibrosis, and eventually cirrhosis or hepatocellular carcinoma (HCC) 
[3,4]. CHB imposes a heavy disease burden and remains a major public health challenge in China. 
 
After HBV infects the human body, its partially double-stranded relaxed circular DNA (rcDNA) 
together with HBV DNA polymerase enters into the nucleus. With the assistance of host DNA 
repair mechanisms, rcDNA is converted into covalently closed circular DNA (cccDNA), which 
serves as the template for viral replication. cccDNA binds to host histones and non-histone 
proteins to form a highly stable viral minichromosome [5,6]. Due to the long half-life of cccDNA, 
progeny rcDNA generated through transcription and reverse transcription can replenish the 
nuclear cccDNA pool without requiring new viral entry, it maintains a persistent transcriptional 
template. When cccDNA continues to exist, some progeny rcDNA replenishes the nuclear 
cccDNA pool, and others acquire an envelope through the endoplasmic reticulum3Golgi complex 
to form viral particles that are secreted from hepatocytes into the bloodstream [5,7]. 
 
Because of this unique and complex replication mechanism, HBV cannot be completely 
eradicated. The persistence of cccDNA in hepatocyte nuclei means that even patients who achieve 
apparent recovery, as indicated by serological markers, remain at lifelong risk of viral reactivation 
[5,8]. 
 
Currently, the main antiviral drugs used in CHB include interferons (IFNs) and nucleos(t)ide 
analogues (NAs) [2,9311]. NAs mainly inhibit viral replication by blocking the reverse 
transcription process. They are administered orally, easy to use, and generally well tolerated 
[5,18,19]. However, because NAs do not directly inhibit cccDNA transcriptional activity, they fail 
to effectively suppress the production of viral proteins such as HBsAg expression. As a result, 
HBsAg declines slowly during treatment, and the annual clearance rate of HBsAg remains only 03

Protocol ID：KY2020-911 
Page 4 of 19 
Version: 03 3 04/12/2023 
4 
 
1.3% under the long-term NAs therapy, and this means CHB patients require indefinite treatment 
[5,12,13]. IFNs exert a dual effect by enhancing host immune responses and directly inhibiting 
viral replication at multiple stages [14316]. In addition to suppressing viral replication, IFNs also 
inhibit cccDNA transcription, promote cccDNA degradation, and facilitate the clearance of HBV-
infected cells, thus favoring viral elimination. In some patients, HBsAg declines rapidly during 
IFN therapy. Previous studies, varying by selected populations, reported HBsAg seroclearance 
rates of 2.837% after 48 weeks of IFN monotherapy, which are higher than those observed with 
NAs monotherapy. And its benefits may continue even after IFN discontinuation [17319]. 
However, IFNs require subcutaneous injection and are associated with adverse effects such as 
bone marrow suppression and flu-like symptoms, leading to relatively poor tolerability. 
 
Pegylated interferon (PegIFN) in combination with NAs has become a recent focus of clinical 
practice and research. As these drugs target different aspects of the viral life cycle, their 
combination may produce additive or synergistic effects, potentially improving efficacy [19,20]. 
Combination therapy with PegIFN and NAs can be applied either as initial therapy or sequential 
therapy. In the former, PegIFN and NAs are administered together as first-line treatment in 
treatment-naïve CHB patients. In the latter, PegIFN is added to or substituted for NAs in patients 
who have already achieved a virological response with NAs. Whether PegIFN plus NAs provides 
clear additional benefit compared with PegIFN or NAs alone, or whether it merely increases 
patient burden with limited additional benefit, remains controversial. Details regarding treatment 
indications, timing of initiation, and duration of combination therapy remain debated. Regarding 
this issue, existing studies are often limited by sample size, geographical region, or study 
population, provide inconsistent results. Most studies include fewer than 1,000 patients, with 
follow-up typically limited to 132 years. Due to limited sample size and duration, evaluation of 
long-term outcomes such as HCC incidence or decompensated cirrhosis is difficult. Consequently, 
most studies use HBsAg loss as the primary endpoint. 
 
For example, retrospective analyses of previous studies show that although PegIFN plus NAs as 
initial combination therapy yields significantly higher HBsAg clearance rates than NA 
monotherapy, comparisons with PegIFN monotherapy yield inconsistent results. Sequential 
PegIFN therapy after NAs also improves HBsAg clearance compared with continued NA 
monotherapy, but few studies compare it directly with PegIFN monotherapy [12,13,17,20327]. 
Improvements in fibrosis observed with PegIFN sequential therapy suggest additional benefit over 
NAs alone, but comparative data remain limited. Patients treated with PegIFN show a lower 

Protocol ID：KY2020-911 
Page 5 of 19 
Version: 03 3 04/12/2023 
5 
 
incidence of HCC compared with NA monotherapy, but comparative data among PegIFN 
monotherapy, sequential therapy, and initial combination therapy are lacking. Regarding treatment 
duration, whether extending PegIFN therapy beyond the currently recommended 48 weeks 
provides additional benefit remains uncertain. Some studies report that extending therapy to 72 or 
96 weeks in HBeAg-negative patients yields higher HBsAg clearance rates than 48 weeks, though 
without statistical significance due to small sample sizes [28,29]. Other studies indicate that 
extending sequential PegIFN therapy to 96 weeks after NA-induced HBeAg clearance does not 
significantly improve HBsAg clearance compared with 48 weeks [24]. 
 
Therefore, the advantages and disadvantages of different antiviral strategies for CHB4
particularly their effects on long-term outcomes such as HCC and decompensated cirrhosis4
remains insufficient. Large-scale, long-term, high-quality clinical studies are urgently needed to 
provide robust evidence for optimizing antiviral regimens. 
 
This study aims to establish a nationwide, multicenter, real-world cohort to compare long-term 
outcomes under different treatment regimens. The goal is to generate high-quality evidence to 
guide treatment and follow-up of chronic viral hepatitis, support optimization of therapeutic 
strategies, and accelerate progress toward a functional cure. 
 
2 
Aims 
To explore the long-term outcomes of antiviral therapy for CHB with the incidence of HCC as the 
primary endpoint, in order to identify optimal clinical decision-making strategies for the 
management of CHB. 
 
3 
Study design 
This is an observational, multicenter, real-world study led by Huashan Hospital. Approximately 
98 sub-centers are expected to participate. Randomization and blinding are not involved in this 
study. It will only observe and collect data related to the changes of clinical indicators during 
routine treatment. The physicians will make clinical decisions based on the applicable guidelines 
and evidence at the time of enrollment, combined with clinical experience and patient-specific 
conditions. No additional interventions will be imposed, and clinical decisions will not be 
influenced by this study. 
 
The study consists of three cohorts (Figure 1): 

Protocol ID：KY2020-911 
Page 6 of 19 
Version: 03 3 04/12/2023 
6 
 
Prospective cohort (PS cohort): Patients enrolled before the implementation of Protocol Version 
3.0 who were planned or currently undergoing antiviral therapy. Clinical data will be 
prospectively collected for five years after informed consent has been obtained. The total follow-
up period will last five years. 
 
Retrospective-prospective cohort (RPS cohort): Patients received antiviral therapy whose initial 
clinical visit occurred before Protocol Version 3.0 but signed informed consent after its 
implementation. The study will retrospectively collect treatment and examination records from 
baseline (defined as the date patient first visited the study center for CHB starting from September 
2020, with concurrent HBV DNA, liver function, and abdominal ultrasound results) and will 
prospectively collect subsequent clinical data up to five years from baseline. The total follow-up 
period will last five years. 
 
Retrospective cohort (RS cohort): Patients receiving antiviral therapy whose clinical records 
from September 2020 until the implementation of Protocol Version 3.0 will be retrospectively 
collected. 
Figure 1 Schematic diagram of the study design 
 
Patients enrolled at sites before Version 3.0 are assigned to the PS cohort, those enrolled before 
Version 3.0 belong to the RS cohort, and whether to belong to the RPS cohort is depending on 
September 
2020 
Collect clinical data according to the visit form requirements 
Prospective cohort 
Retrospective- 
prospective cohort 
Retrospective cohort 
Implementation of 
Version 3.0 
October 
2027 
October 
2022 
Collect clinical data according  
to the visit form requirements 
Retrospective collection of  
clinical data 
Retrospective collection of  
clinical data 
Enrollment period 
Enrollment & follow-up period 
follow-up period 

Protocol ID：KY2020-911 
Page 7 of 19 
Version: 03 3 04/12/2023 
7 
 
patients’ willingness. Patients at newly added sites after Version 3.0 are included in the RS cohort, 
and may, depending on patients’ willingness, be included in the RPS cohort. 
 
4 
Eligibility Criteria, Exclusion Criteria, and Withdrawal Criteria 
4.1 Inclusion Criteria 
1) 
Chronic HBV infection (HBsAg positive for ≥6 months, or <6 months with liver histology 
consistent with CHB and other liver diseases excluded); 
2) 
Age ≥18 years; 
3) 
Planned or currently receiving potent low-resistance NAs [entecavir (ETV), tenofovir 
disoproxil fumarate (TDF), tenofovir alafenamide fumarate (TAF), or tenofovir 
amibufenamide (TMF)], or planned to receive PegIFNα-2b, either treated or treatment-
naïve; 
4) 
Written informed consent. 
 
4.2 Exclusion Criteria 
1) 
Hepatocellular carcinoma (diagnosed or planned for treatment) or liver failure at 
baseline; 
2) 
Concurrently participating in other interventional clinical trials; 
3) 
Any other conditions deemed unsuitable by investigators or preventing compliance with 
study requirements. 
 
4.3 Withdrawal Criteria 
1) 
Request to withdraw at any time; 
2) 
Investigators determine continuation is not in the patient’s best interest; 
3) 
Occurrence of clinical Objectives, including HCC, liver transplantation, or death; 
4) 
Lost to follow-up. 
 
4.4 Lost to Follow-up Criteria 
Patients in the PS or RPS cohort who fail to return within 13 months and cannot be contacted after 
at least two phone calls (each at least 24 hours apart) or other attempts via registered mail, courier, 
or email will be classified as lost to follow-up. 
 
Exception: If patients notify investigators in advance of inability to attend a scheduled visit or 
examinations due to special reasons, this is not considered lost to follow-up. In such cases, 

Protocol ID：KY2020-911 
Page 8 of 19 
Version: 03 3 04/12/2023 
8 
 
investigators should encourage patients to undergo examinations at other medical institutions and 
provide results through appropriate forms. 
 
5 
Sample Size Estimation 
As an observational real-world study, sufficient sample size is needed to reflect real-world 
conditions and balance patient heterogeneity. Based on the number of participating centers and 
patient flow, approximately 33,000 patients are expected to be enrolled for analysis. 
 
6 
Study Procedures 
6.1 Clinical Information to be Collected 
6.1.1 The collection information and time point of PS Cohort 
Starting from September 2020, outpatient or inpatient patients with CHB who meet inclusion 
criteria and have been screened through the exclusion criteria will be enrolled after informed 
consent. Baseline data and scheduled follow-up assessments will be collected at fixed time points 
after treatment initiation. Recommendations are based on the guidelines and clinical evidence. 
Actual schedules may be adjusted by treating physicians. No additional visits will be imposed by 
the study [2]. 

Protocol ID：KY2020-911 
Page 9 of 19 
Version: 03 3 04/12/2023 
9 
 
A recommended follow-up schedule and assessments are summarized in Table 1. 
Table 1 Study visit table 
 
Baseline 
V1 
V2 
V3 
V4 
V5 
V6 
V7 
V8 
V9 
V10 
V11 
V12 
V13 
Study week 
0 
4±1 
12±2 
24±4 
36±4 
48±4 
72±4 
96±4 
120±4 
144±4 
168±8 
192±8 
216±8 
240±8 
Time point 
0 
1m 
3m 
6m 
9m 
1y 
1.5y 
2y 
2.5y 
3y 
3.5y 
4y 
4.5y 
5y 
Demographics 
★ 
 
 
 
 
 
 
 
 
 
 
 
 
 
Height & weight 
★ 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical history 
★ 
 
 
 
 
 
 
 
 
 
 
 
 
 
Family 
history       
★ 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical assessment 
Symptoms and signs 
★ 
 
 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
 
★ 
 
★ 
Treatment regimen 
★ 
 
 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
 
★ 
 
★ 
Clinical eventsa 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
Laboratory tests 
Quantitative HBsAg 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
HBeAg, anti-HBe, 
anti-HBc, anti-HBs 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
HBV DNA 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
Complete blood count 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
Hepatic panel 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
★ 
Creatinine 
★ 
 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 

Protocol ID：KY2020-911 
Page 10 of 19 
Version: 03 3 04/12/2023 
10 
 
a Clinical events include decompensated cirrhosis, HCC, death, or liver transplantation 
b If abdominal ultrasound is unavailable, MRI or CT may be used as an alternative and will be treated as required completion items 
c The study does not mandate MRI/CT/liver histology, but when these tests are performed in routine care, their results will be collected to enrich observational data 
AFP & CEA 
★ 
 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
INR & PT 
★ 
 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
TC, TG, HDL-c, 
LDL-c 
★ 
 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
Glucose 
★ 
 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
HbA1c 
As clinically indicated c 
Creatine kinase 
ElectrolytesÿK，P，
CaĀ 
Urinalysis 
Imagingb and liver pathology 
Transient 
elastography 
★ 
 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
Abdominal 
ultrasound 
★ 
 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
 
★ 
Abdominal MRI 
As clinically indicated b,c 
Abdominal CT 
As clinically indicated b,c 
Liver biopsy 
As indicated for clinical events a or by clinical need 

Protocol ID：KY2020-911 
Page 11 of 19 
Version: 03 3 04/12/2023 
11 
 
6.1.2 The collection information of RPS Cohort 
6.1.2.1 Retrospective data collection 
Clinical records from baseline up to the date of informed consent will be collected retrospectively, 
according to the study visit table (Table 1). Any items not performed in routine care will be 
recorded as missing data. 
6.1.2.2 Prospective data collection 
Subsequent visits will follow the windows specified in the study visit table (Table 1). The initial 
prospective visit will depend on the interval between baseline date and the consent date. 
Example: If baseline was October 2020 and consent was signed in December 2023 (an interval of 
3 years), prospective follow-up would begin at V9 (corresponding to 3 years). 
6.1.3  The collection information of RS Cohort 
Clinical data generated between September 2020 and the implementation of Protocol Version 3.0 
will be retrospectively collected (items based on the study visit table, Table 1, with time points 
recorded according to when they actually occurred). Any items not collected in routine practice 
will be recorded as missing data. 
 
6.2 Follow-up Management and Measure to Ensure Patient Compliance 
The study aims to achieve a follow-up rate of more than 80%. As this is a non-interventional 
study, measures will be taken to maximize patient compliance. At both initial and follow-up visits, 
patients will receive education and reminders to attend subsequent scheduled follow-ups on time. 
The study database system includes functions that regularly notify site investigators of 
recommended follow-up time points, prompting physicians to conduct visits and complete data 
collection and entry as needed. In addition, patient education booklets containing follow-up record 
forms will be distributed, encouraging patients to adhere to regular follow-up, consistently 
document their test results, and gain a better understanding of their disease status and treatment 
outcomes. 
 
7 
Study Termination Criteria 
The study will be terminated if: 
1) 
Major flaws or serious deviations occur during implementation, making evaluation 
infeasible. 
2) 
Adverse events or serious adverse events related to treatment occur during the study, and 
investigators deem it unsafe to continue. 
3) 
Regulatory authorities, government departments, or ethics committees request suspension 

Protocol ID：KY2020-911 
Page 12 of 19 
Version: 03 3 04/12/2023 
12 
 
or termination of the study. 
4) 
Other circumstances arise that investigators consider make continuation inappropriate or 
impractical. 
 
8 
Efficacy Evaluation 
8.1 Effectiveness 
8.1.1 Primary Objectives: 
To compare the 5-year incidence of HCC under different treatment regimens. 
8.1.2 Secondary Objectives: 
1) 
Compare the 1-year, 2-year, 3-year, 4-year, and 5-year rate of HBsAg seroclearance and 
HBsAg seroconversion under different treatment regimens. 
2) 
Compare the 5-year incidence of decompensated cirrhosis and its complications under 
different treatment regimens 
3) 
Compare the 5-year incidence of cirrhosis under different treatment regimens 
4) 
Compare proportion of patients with improvement or progression of liver fibrosis under 
different treatment regimens 
5) 
Compare the 5-year incidence of liver transplantation under different treatment regimens 
6) 
Compare the 1-year, 2-year, 3-year, 4-year, and 5-year HBsAg levels, the decline changes 
of HBsAg from baseline, the rate of HBeAg seroconversion, the decline changes of HBV 
DNA from baseline, and the proportion of HBV DNA undetectable (<20 IU/mL) under 
different treatment regimens 
7) 
Compare the relapse rate after treatment cessation under different treatment regimens 
8.1.3 Exploratory Objectives: 
Describe the demographic characteristics of patients in the real-world clinical setting. 
8.1.4 Methods for Assessing Efficacy Endpoints: 
8.1.4.1  HCC: According to the 2019 National Health Commission diagnostic criteria for 
hepatocellular carcinoma, a diagnosis of HCC will be considered confirmed when either 
pathological or clinical criteria are met. Pathological confirmation requires diagnosis by two or 
more pathologists as hepatocellular carcinoma. Clinical diagnosis requires at least one typical 
imaging features about nodules and liver cancer on dynamic contrast-enhanced MRI, dynamic 
contrast-enhanced CT, contrast-enhanced ultrasound, or hepatocyte-specific contrast (Gd-EOB-
DTPA) MRI; a diagnosis is established if either of the following is satisfied: 1) any nodule size >2 
cm with one typical imaging feature; 2) all nodules sizes ≤2 cm with two typical imaging 
features. During follow-up, any HCC event must include recording of pathology reports (including 

Protocol ID：KY2020-911 
Page 13 of 19 
Version: 03 3 04/12/2023 
13 
 
subtype, differentiation, and presence/distribution of intravascular tumor clusters) or imaging 
descriptions (including imaging modality that showed typical HCC features and nodule size), and 
scanned copies of original reports must be provided for centralized verification by the study team. 
8.1.4.2  Decompensated cirrhosis: According to 2019 Chinese Society of Hepatology 
guidelines on the management of liver cirrhosis, a diagnosis of cirrhosis will be considered 
confirmed when either pathological or clinical criteria are met, and when clinical manifestations 
suggest decompensated will be considered to be decompensated cirrhosis [31]. Pathological 
confirmation requires diagnosis by two or more pathologists as decompensated cirrhosis. Clinical 
criteria require that endoscopic evidence of esophagogastric varices, ultrasound or CT findings 
suggesting cirrhosis or portal hypertension, or liver stiffness measurement [LSM] values reaching 
corresponding thresholds. Clinical features of decompensation include ascites, variceal 
hemorrhage, spontaneous bacterial peritonitis/sepsis, hepatic encephalopathy, or hepatorenal 
syndrome, confirmed by laboratory tests, imaging, or clinical scoring systems. On occurrence of 
decompensated cirrhosis during follow-up, investigators must document pathology (including 
number of pseudo-lobules, inflammation and fibrosis staging), or supporting ancillary test results 
(e.g., presence of varices, imaging features suggesting cirrhosis/portal hypertension, specific LSM 
values, ascites, variceal bleeding), and laboratory or clinical records indicating decompensation 
(e.g., temperature, cognitive assessment scales, inflammatory markers, blood ammonia, liver and 
renal function). Original reports and scans must be provided for verification by the study team. 
8.1.4.3  Changes in HBsAg, HBeAg, and HBV DNA will be analyzed by comparing 
baseline and follow-up values. 
8.2 Safety Objectives: 
Observe the drug safety under different regimens 
 
9 
Adverse Event Recording 
Adverse event (AE) is any unfavorable medical occurrence in a research participant, not 
necessarily having a causal relationship with the study. Serious adverse event (SAE) or reaction is 
any untoward medical occurrence that results in death, life-threatening, requiring or prolonging 
hospitalization, persistent or significant disability or incapacity, congenital anomaly or birth 
defect, or an important medical event in the investigator’s judgment. 
Expected AEs: As a observational real-world study that only collects clinical data and medical 
history without imposing additional procedures, visits, or costs, no extra risks are anticipated 
theoretically. However, obtaining consent and taking medical histories may cause psychological 
discomfort; investigators should therefore be careful with phrasing and patient communications.  

Protocol ID：KY2020-911 
Page 14 of 19 
Version: 03 3 04/12/2023 
14 
 
Recording and management of AEs: If a patient experiences psychological discomfort during 
history taking, study staff should provide verbal reassurance and record the incident within 2 
hours, notifying the designated study personnel. For patients showing persistent or severe 
psychological symptoms (e.g., depression, mania), psychiatric consultation and psychological 
support should be arranged within 12 hours, and the event recorded by designated personnel. For 
unexpected injuries or harms related to the study, appropriate treatment and compensation will be 
provided. All AEs observed during the study must be documented in detail. If SAEs occur, 
designated personnel will manage and record the event promptly and report to the ethics 
committee and relevant authorities within required timelines. Serious or unexpected adverse 
reactions not consistent with drug labeling will be reported to the hospital’s 
pharmacovigilance/ADR monitoring department per relevant requirements.  
 
10 Statistical Analysis Plan 
Details of statistical methods and definitions of analysis datasets are provided in the separate 
Statistical Analysis Plan (SAP). 
 
11 Data Management and Confidentiality 
(1) Data Entry and Modification 
Data entry and corrections are performed by investigators. All entered data must originate from 
source documents (e.g., original medical records, laboratory reports) and be consistent with 
original records. All observations and test results should be entered promptly, accurately, 
completely, clearly, and truthfully into case report forms and the electronic database. Data 
managers will review and manage entered data. For data queries, data managers will send queries 
to investigators, who should respond promptly; data managers may re-query if necessary. 
(2) Confidentiality of Participant Information 
All study participant information must be strictly confidential. Participant records and data will be 
identified by study ID numbers rather than names. Identifiable information will not be disclosed 
outside the study team without participant consent. All study staff are required to maintain 
confidentiality. Participant files will be stored in locked cabinets accessible only to authorized 
study personnel. To ensure regulatory compliance, government regulators or ethics committee 
members may inspect personal records at the study site per regulations. Any publications will not 
disclose participants’ personal identifiers. 
(3) Confidentiality of Study Data 

Protocol ID：KY2020-911 
Page 15 of 19 
Version: 03 3 04/12/2023 
15 
 
Study data are confidential. Study staff must not disclose data to individuals outside the project 
without the principal investigator’s authorization, nor transfer data to external institutions without 
hospital authorization. Transfer of human genetic resources or related data to foreign entities or 
domestic entities with foreign capital requires approval by the National Human Genetic Resources 
Administration; normal academic result publication that complies with regulations is permitted. 
 
12 Quality Control and Assurance 
The principal investigator will organize relevant training to ensure the study is conducted 
according to standards, and that case report forms and other documents comply with GCP 
principles and the protocol. All data and materials must be verifiable. Quality control will be 
applied at every study stage to ensure data reliability and correct execution of procedures. The 
principal investigator must ensure investigators adhere to the protocol, confirm data accuracy, and 
ensure completeness of reports and informed consent documentation prior to study initiation. Any 
protocol deviations should be reported to the ethics committee in a timely manner. 
When necessary, the study team will develop standard operating procedures (SOPs) and 
implement quality control procedures throughout study conduct and data handling to ensure 
standardization and reliability. 
 
13 Ethical Considerations 
This observational study collects medical history and routine laboratory/imaging results and does 
not impose additional burden or costs on participants; therefore, it does not present additional 
theoretical risks. The study will be conducted after approval by the Huashan Hospital Ethics 
Committee and other relevant local ethics committees; participating sites must obtain approval 
from their own ethics committees prior to commencement. All eligible patients will sign informed 
consent forms. Participant information will be kept strictly confidential and recorded using study 
ID numbers rather than names. The study will be conducted in compliance with applicable 
national regulations (e.g., Measures for Ethical Review of Biomedical Research Involving 
Humans) and international ethical guidelines such as the Declaration of Helsinki. The Huashan 
Hospital Ethics Committee will perform continuing oversight of the study. 
 
14 Recruitment and Informed Consent Process 
Participants will be recruited from outpatient clinics and inpatient wards at participating centers 
when they are diagnosed with CHB. The study team will inform clinicians across relevant 
departments about recruitment criteria. Clinicians who identify eligible patients during routine 

Protocol ID：KY2020-911 
Page 16 of 19 
Version: 03 3 04/12/2023 
16 
 
care will notify the study team. Study staff will then visit the patient, fully inform them of the 
study’s risks and benefits, and obtain written informed consent. 
 
15 Responsibilities of Participating Parties and Other Provisions 
Responsibilities of lead site (Huashan Hospital): 
1. 
Establish a standard operating procedure for the multicenter registry within the study, and 
define standards for each process (including enrollment criteria, database audit standards, and 
data statistical workflow); appoint database managers and quality control personnel to conduct 
regular checks on data entry quality and study progress at each site; 
2. 
Coordinate data aggregation and analysis; 
3. 
Assign personnel responsible for enrollment, follow-up, and data collection/recording at 
Huashan Hospital. 
Sub-centers responsibilities: 
Appoint personnel responsible for patient enrollment, follow-up, data collection, and record-
keeping at each sub-center. 
 
16 Reference 
[1] Schweitzer A, Horn J, Mikolajczyk R T, et al. Estimations of worldwide prevalence of chronic 
hepatitis B virus infection: a systematic review of data published between 1965 and 2013[J]. 
Lancet, 2015, 386(10003): 1546-55.  
[2] Chinese Society of Infectious Diseases, Chinese Medical Association; Chinese Society of 
Hepatology, Chinese Medical Association. [The guidelines of prevention and treatment for 
chronic hepatitis B (2019 version)]. Zhonghua Gan Zang Bing Za Zhi, 2019, 20;27(12):938-961.  
[3] Jung M C, Pape G R. Immunology of hepatitis B infection[J]. Lancet Infectious Diseases, 
2002, 2(1): 43-50.  
[4] Shin E-C, Sung P S, Park S-H. Immune responses and immunopathology in acute and chronic 
viral hepatitis[J]. Nature reviews. Immunology, 2016, 16(8): 509-523.  
[5] Nassal M. HBV cccDNA: viral persistence reservoir and key obstacle for a cure of chronic 
hepatitis B[J]. Gut, 2015, 64(12): 1972-1984.  

Protocol ID：KY2020-911 
Page 17 of 19 
Version: 03 3 04/12/2023 
17 
 
[6] Pollicino T, Belloni L, Raffa G, et al. Hepatitis B virus replication is regulated by the 
acetylation status of hepatitis B virus cccDNA-bound H3 and H4 histones[J]. Gastroenterology, 
2006, 130(3): 823-837.  
[7] Ko C, Chakraborty A, Chou W-M, et al. Hepatitis B virus genome recycling and de novo 
secondary infection events maintain stable cccDNA levels[J]. Journal of hepatology, 2018, 69(6): 
1231-1241.  
[8] Tang L S Y, Covert E, Wilson E, et al. Chronic Hepatitis B Infection: A Review[J]. JAMA, 
2018, 319(17): 1802-1813.  
[9] Terrault N A, Lok A S F, Mcmahon B J, et al. Update on prevention, diagnosis, and treatment 
of chronic hepatitis B: AASLD 2018 hepatitis B guidance[J]. Hepatology (Baltimore, Md.), 2018, 
67(4): 1560-1599.  
[10] E.A.S.L. EASL 2017 Clinical Practice Guidelines on the management of hepatitis B virus 
infection[J]. Journal of hepatology, 2017, 67(2): 370-398.  
[11] Omata M, Cheng A-L, Kokudo N, et al. Asia-Pacific clinical practice guidelines on the 
management of hepatocellular carcinoma: a 2017 update[J]. Hepatology international, 2017, 
11(4): 317-370.  
[12] Kim G-A, Lim Y-S, An J, et al. HBsAg seroclearance after nucleoside analogue therapy in 
patients with chronic hepatitis B: clinical outcomes and durability[J]. Gut, 2014, 63(8): 1325-
1332.  
[13] Lim T H, Gane E, Moyes C, et al. HBsAg loss in a New Zealand community study with 28-
year follow-up: rates, predictors and long-term outcomes[J]. Hepatology international, 2016, 
10(5): 829-837. 
[14] Wang Y-X, Niklasch M, Liu T, et al. Interferon-inducible MX2 is a host restriction factor of 
hepatitis B virus replication[J]. Journal of hepatology, 2019.  
[15] Tan G, Yi Z, Song H, et al. Type-I-IFN-Stimulated Gene TRIM5γ Inhibits HBV Replication 
by Promoting HBx Degradation[J]. Cell reports, 2019, 29(11).  
[16] Sadler A J, Williams B R G. Interferon-inducible antiviral effectors[J]. Nature reviews. 
Immunology, 2008, 8(7): 559-568.  

Protocol ID：KY2020-911 
Page 18 of 19 
Version: 03 3 04/12/2023 
18 
 
[17] Marcellin P, Ahn S H, Ma X, et al. Combination of Tenofovir Disoproxil Fumarate and 
Peginterferon α-2a Increases Loss of Hepatitis B Surface Antigen in Patients With Chronic 
Hepatitis B[J]. Gastroenterology, 2016, 150(1).  
[18] Reijnders J G P, Rijckborst V, Sonneveld M J, et al. Kinetics of hepatitis B surface antigen 
differ between treatment with peginterferon and entecavir[J]. Journal of hepatology, 2011, 54(3): 
449-454.  
[19] Wursthorn K, Lutgehetmann M, Dandri M, et al. Peginterferon alpha-2b plus adefovir induce 
strong cccDNA decline and HBsAg reduction in patients with chronic hepatitis B[J]. Hepatology 
(Baltimore, Md.), 2006, 44(3): 675-684.  
[20] Chineses Society of Infectious Disease, Chinese Society of Hepatology, Chinese Medical 
Association. [The expert consensus on clinical cure (functional cure) of chronic hepatitis B][J]. J 
Clin Hepatol, 2019, 35(8): 1693-1701.  
[21] Marcellin P, Lau G K K, Bonino F, et al. Peginterferon alfa-2a alone, lamivudine alone, and 
the two in combination in patients with HBeAg-negative chronic hepatitis B[J]. The New England 
journal of medicine, 2004, 351(12): 1206-1217.  
[22] Lau G K K, Piratvisuth T, Luo K X, et al. Peginterferon Alfa-2a, lamivudine, and the 
combination for HBeAg-positive chronic hepatitis B[J]. The New England journal of medicine, 
2005, 352(26): 2682-2695.  
[23] Tamaki N, Kurosaki M, Kusakabe A, et al. Hepatitis B surface antigen reduction by 
switching from long-term nucleoside/nucleotide analogue administration to pegylated 
interferon[J]. Journal of viral hepatitis, 2017, 24(8): 672-678.  
[24] Peng H, Jia S, Zhang W, et al. HBsAg Loss with Peg-interferon Alfa-2a in Hepatitis B 
Patients with Partial Response to Nucleos(t)ide Analog: New Switch Study[J]. Journal of Clinical 
& Translational Hepatology, 2018, 6(1): 25-34.  
[25] Ning Q, Han M, Sun Y, et al. Switching from entecavir to PegIFN alfa-2a in patients with 
HBeAg-positive chronic hepatitis B: a randomised open-label trial (OSST trial)[J]. Journal of 
hepatology, 2014, 61(4): 777-784.  
[26] Chang T T, Lai C L, Kew Yoon S, et al. Entecavir treatment for up to 5 years in patients with 
hepatitis B e antigen-positive chronic hepatitis B[J]. Hepatology, 2010, 51(2): 422-30.  

Protocol ID：KY2020-911 
Page 19 of 19 
Version: 03 3 04/12/2023 
19 
 
[27] Cheng J, Wang Y, Hou J, et al. Peginterferon alfa-2b in the treatment of Chinese patients 
with HBeAg-positive chronic hepatitis B: a randomized trial[J]. J Clin Virol, 2014, 61(4): 509-16.  
[28] Chen X, Chen X, Chen W, et al. Extended peginterferon alfa-2a (Pegasys) therapy in Chinese 
patients with HBeAg-negative chronic hepatitis B[J]. J Med Virol, 2014, 86(10): 1705-13.  
[29] Lampertico P, Viganò M, Di Costanzo G G, et al. Randomised study comparing 48 and 96 
weeks peginterferon α-2a therapy in genotype D HBeAg-negative chronic hepatitis B[J]. Gut, 
2013, 62(2): 290-8.  
[30] Bureau of Medical Administration, National Health Commission of the People’s Republic of 
China. [Guidelines for diagnosis and treatment of primary liver cancer in China (2019 edition)][J]. 
J Clin Hepatol, 2020, 36(2): 277-292.  
[31] Chinese Society of Hepatology, Chinese Medical Association. [Chinese guidelines on the 
management of liver cirrhosis][J] Zhonghua Gan Zang Bing Za Zhi, 2019, 20;27(11):846-865.