Evaluation of Antiviral Indications and Therapeutic Effect by Liver Biopsy Combined With Clinical Trial Parameters on Chronic HBV Infection With Different Transaminase Levels
Conditions: Chronic Hepatitis b, ALT
Sponsor: First Affiliated Hospital Xi'an Jiaotong University

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Evaluation of antiviral indications and therapeutic 
effect by liver biopsy combined with clinical trial 
parameters on chronic HBV infection with different 
transaminase levels 
 
 
 
 
 
 
Study Protocol with Statistical Analysis Plan (SAP) 
and Informed Consent Form (ICF) 
 
 
 
 
 
October 25, 2018 
 
 
 
 

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Evaluation of antiviral indications and therapeutic 
effect by liver biopsy combined with clinical trial 
parameters on chronic HBV infection with different 
transaminase levels 
 
 
 
 
 
 
 
Study Protocol 
(Pages 2-5) 
 
 
 
 
 
October 25, 2018 
 
 
 

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Study Protocol 
Study titled ‘Evaluation of antiviral indications and therapeutic effect by liver biopsy 
combined with clinical trial parameters on chronic HBV infection with different 
transaminase levels’was approved by the First Affiliated Hospital of Xi’an Jiaotong 
University of Medicine Clinical Researches Ethics Committee meeting on 10/19/2018 
with approval number: XJTU1AF2018LSK-146. 
The patients or legal representatives of the children who participated in the study 
were informed about the aim and the programs to be applied throughout the 
study.Volunteer Information Form has been signed and approved in accordance with 
the standards deemed appropriate by the First Affiliated Hospital of Xi’an Jiaotong 
University of Medicine Clinical Researches Ethics Committee (This form can be seen 
on Informed Consent Form (ICF) section). The study was conducted in accordance 
with the Declaration of Helsinki. 
Hypothesis of the Study 
H0:Initiating treatment is not recommend for the patients with ALT <2ULN, high 
viral load,because some studies considered that the potential harms of long-term 
therapy,including cost,low response rates,drug adverse effects and development of 
resistance,outweigh its benefits. 
H1:Initiating treatment is recommend for the patients with ALT <2ULN,high 
viral load,because some studies considered that high levels of serum HBV DNA 
(>104copies/ml)are associated with a high risk of hepatocellular carcinoma (HCC) and 
the histological activity and HBV-specific immune responses do also occur in this 

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phase. 
Inclusion Criteria 
   (1)Patients with a history of chronic HBV infections,defined as those in whom 
presence of serum hepatitis B surface(HBsAg) for more than 6 months. 
(2)All patients had underwent a liver biopsy. 
(3)All patients signed the informed consent. 
(4)No history of interferon or Nucleoside analogue treatment. 
Exclusion Criteria 
(1)Co-infection 
with 
human 
immunodeficiency 
virus(HIV),hepatitis 
A 
virus(HAV),hepatitis C virus(HCV),hepatitis D virus(HDV) and/or hepatitis E 
virus(HEV). 
(2)Decompensated cirrhosis. 
(3)History of liver transplantation. 
(4)History of hepatocellular carcinoma(HCC). 
(5)Having received antiviral treatment. 
(6)Taking immunoregulation drugs such as cytotoxic agents and hormones. 
(7)Use of hepatotoxic drugs or regular consumption of alcohol. 
(8)Autoimmune disease or antinuclear antibody titers higher than 1:160. 
(9)And other chronic liver disease. 
(10)Incomplete data. 
Applied Evaluations 
Response indexes as the follows: virological response(HBV DNA below the 

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detection 
limit);HBsAg 
response(HBsAg 
decline≥0.5log10 
IU/ml);HBeAg 
seroconversion(HBeAg loss and HBeAb appearance);biochemical response(ALT 
normalization);LSM response(LSM decrease as at least a 1-kPa). 
Research programme 
 
HBV-infected received liver biopsy in this study between October 2013 and July 2018
Liver biopsy
Biochemical tests +HBV markers+HBV DNA+HBV genotypes+Liver 
stiffness Measurement(LSM)
Baseline characteristics of 
different ALT groups 
Changes of indexes in treated and 
untreated groups  
Identify factors associated with 
achieving antiviral responses
1.Make a thorough inquiry of diagnostic bounds for parameters related to 
antiviral timing for patients with ALT<2ULN.
2.Response indexes of clinical results such as virological,serological, 
biochemical and histological responses
 
Research group 
 
 
 

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Evaluation of antiviral indications and therapeutic 
effect by liver biopsy combined with clinical trial 
parameters on chronic HBV infection with different 
transaminase levels 
 
 
 
 
 
Statistical Analysis Plan (SAP) 
(Pages 6-7) 
 
 
 
 
 
 
 
October 25, 2018 
 
 
 

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Statistical Analysis Plan (SAP) 
The statistical program Statistical Package for Social Sciences (SPSS) Version 
20.0(SPSS Inc., Chicago, IL, USA) was used in the data analysis of the study.The p 
value of p<0.05 was considered to be statistically significant in the data analysis. 
Continuous quantitative variables were expressed as mean±standard deviation (SD) 
for normal distributions or median and interquartile range (IQR)(25th-75th percentile) 
for abnormal distributions,which were assessed with an independent samples T test or 
the Mann-Whitney U test as comparison of two sets of samples while comparison of 
multiple samples were assessed with ANOVAs or Kruskal-Wallis tests.Repeated 
measures date of general linear model were used to calculate variation trend between 
treated and untreated follow-up patients. Categorical variables were expressed 
aspercentages that were analyzed using the χ2 test, such as gender, HBV 
genotype,ALT, HBsAg, HBeAg, HBeAb, HBcAb and LSM.  
The receiver operating characteristic (ROC) curve was plotted in order to find 
the most appropriate HBV DNA ,HBsAg, HBeAg cutoff value for diagnosis of 
patients with ALT<2ULN who are chronic HBV infection or chronic hepatitis B. 
Univariate logistic regression and multivariate forward stepwise logistic 
regression were used to assess the incidences of antiviral responses, such as HBV 
DNA response, serological responses, biochemical response 
 
 
 

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Evaluation of antiviral indications and therapeutic 
effect by liver biopsy combined with clinical trial 
parameters on chronic HBV infection with different 
transaminase levels 
 
 
 
 
 
Informed Consent Form (ICF) 
(Pages 8-11)  
 
 
 
 
 
 
 
October 25, 2018 
 
 
 

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Informed Consent Form (ICF) 
Description of the Physiotherapist 
This study is a retrospective observational cohort study,some baseline demographics 
and clinical data were obtained from the electronic integrated clinical records 
system.In consideration of achieving complete follow-up information, if you accept 
yourself or your child to be participated,it is a scientific research and it is titled as 
‘Evaluation of antiviral indications and therapeutic effect by liver biopsy combined 
with clinical trial parameters on chronic HBV infection with different transaminase 
levels’,which aimed to explore the effect of antiviral therapy in CHB patients with 
these characteristics.All the patients are planed to achieve follow-up information at 
least 12 months and follow-up assessments such as HBV DNA,HBV Markers,liver 
function,LSM were performed at months 12, 24, 36 and 48.This research will be 
carried out by the department of the Infectious Disease,The Affiliated Hospital of 
Yan’an University and the First Affiliated Hospital of Xi’an Jiaotong 
University,Xi'an,China.  
Benefits of participation in this study 
Chronic HBV infection is a lifelong disease and high levels of serum HBV DNA 
(>104copies/ml)are associated with a high risk of liver cirrhosis and hepatocellular 
carcinoma (HCC).Therefore,the goal of antiviral therapy is to inhibit HBV replication 
in a sustained manner and to improve the quality of life and prolong the survival 
time.But the patients with ALT<2ULN are recommended follow-up and no need for 
antiviral therapy.Comprehensive assessment of the degree of liver inflammation and 

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fibrosis is conducive to the timely diagnosis of the patients and to determine whether 
the active anti-viral treatment.Through longitudinal follow-up of the clinical outcome 
of treated and untreated patients,further lay a theoretical foundation for whether such 
patients should be treated with antiviral therapy,so as to alleviate the psychological 
stress they have to endure because of the long follow-up waiting.It also avoids the 
waste of medical resources caused by improper treatment, and helps to alleviate the 
economic burden of patients, families and society.Therefore,it is important for these 
patients to particate in this study achieve individualized management scheme. 
See the risks or inconveniences of this study and the compensatory measures 
This subject is a retrospective study,so there is no direct conflict of security 
interests between the researcher and the participators. 
Confidentiality of research 
Your personal information collected in this study is confidential and only used 
for research and scientific analysis.By signing this informed consent,you can allow 
people with legitimate reasons to collect and view your personal data. 
Your rights 
The participation in the study is entirely voluntary, and you can withdraw at any 
stage of the study without any reasons,which will not affect your relationship with the 
medical staff and future treatment. 
    Finally,thank you for your great support for this study,and for your contribution 
to the exploration of the diagnosis and treatment of the disease. 
Agree to declare 

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I have understood the purpose,process,potential benefits and possible adverse 
reactions of this study,volunteered to participate in the study and follow the research 
process as far as possible. 
 
 
 
 
 
 
 
Name and the Surname of the Participant and Legal Representative: 
Date:                           Signature: 
Name and Surname of the Witness: 
Date:                           Signature: