Am J Transl Res 2016;8(2):1218-1227

www.ajtr.org /ISSN:1943-8141/AJTR0021601

Original Article
Prevalence of ABCB4 polymorphisms in gallstone
disease in han-Chinese population
Lei Zhan1, Yao-Zhen Pan1, Ling Chen1, Hao Zhang1, Hong Zhang1, Jian Song1, Chi-Meng Tzeng2, Cheng-Yi
Sun1
1
Department of Biliary-Hepatic Surgery, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou,
China; 2Translational Medicine Research Center, School of Pharmaceutical Sciences, Xiamen University, Xiamen,
Fujian, China

Received December 10, 2015; Accepted February 5, 2016; Epub February 15, 2016; Published February 29,
2016
Abstract: ATP Binding Cassette Transporter A4 (ABCB4) is a sterol export pump that regulates excretion of biliary
cholesterol. We tested association between ABCB4 polymorphisms and gallstone disease using meta-analysis. In
a cross-sectional study, 296 subjects were recruited from a hospital-based population. Total of 171 subjects were
diagnosed as gallstone disease by abdominal ultrasonography from three cohort studies. We evaluated prevalence
of ABCG8 rs11887534 (D19H) as a positive control, and the ABCB4 rs1202283 and rs2230028 polymorphisms on
Chinese population were screened by meta-analysis and genotyped using TaqMan SNP assay. Stata/SE 11.0 software and random-effects model were used in meta-analyzing 3 cohort between study heterogeneity. Four studies
including three cohorts were used for final meta-analysis. In allelic model, minor alleles of ABCB4 rs1202283 (OR =
0.41, 95% CI: 0.25-0.67, P<0.001) and of ABCB4 rs2230028 (OR = 0.12, 95% CI: 0.06-0.22, P = 0.001) were associated with an increased risk for gallstone disease in Europeans. Funnel plot and Eggers test suggested absence
of publication bias. Concentration of total cholesterol, low-density lipoprotein cholesterol (LDLC) (P = 0.015) and
high-density lipoprotein cholesterol (HDLC) (P = 0.028) were significantly higher in subjects with gallstones disease
than controls. ABCB4 rs1202283 (heterozygote AG) (P<0.0001), rs2230028 (heterozygote CT) (P = 0.023) and
ABCG8 rs11887534 (heterozygote CG) (P = 0.006) were significantly associated with gallstone disease in Chinese
population. Genetic risk associated with ABCB4 rs2230028 (homozygote GG) polymorphism was dominated in
asymptomatic gallstone disease (95% C.I.: 0.219-0.768; P = 0.005). In conclusion, carriers of ABCB4 rs1202283,
rs2230028 are at an increased risk for gallstone disease, while ABCB4 rs2230028 is associated with asymptomatic gallstone disease.
Keywords: Gallstone disease, single nucleotide polymorphism, ATP binding cassette transporter A4

Introduction
Gallstone disease is common in Western countries [1]. In China, the prevalence of gallstone
disease has increased to 10.8% in the general
population compared to 5% five years ago.
While an additional 2.97% gallstone disease
subjects have a previous history of cholecystectomy. The majority of the gallstone disease
is associated with high cholesterol. Recently,
studies have suggested that genetic factors, in
interaction with environmental factors, are the
predisposition to gallstone disease [2]. Katsika
et al. [3], suggested that genetic heredity contributes 25% of factors to gallstone formation
after an elegant analysis of data from Swedish
twins.

Since late 1980s, studies have attempted to

reveal susceptible genes associated with gallstone disease in different populations. The possible genes studies included apolipoprotein
(Apo)-E [4-7], Apo-B, [5, 6] and cholesterol
alpha-hydroxylase [5, 6]. In the last decade, the
ABCB4 [adenosine triphosphate (ATP)-binding
cassette, sub-family B (MDR/TAP), member 4]
transporter located in the hepatocyte canalicular membrane was identified as involving in the
transport of phosphatidylcholine into bile.
Mutation in the ABCB4 gene may lead to low
levels of biliary phosphatidylcholine, resulting in
enhanced cholesterol precipitation and formation of crystals and gallstones. The first evidence that human gallstone disease might be
caused by a single gene defect came [8, 9].

Three risk factors of gallstone disease in Chinese population

Table 1. List of references
References

Year

Ethnicity

SNPs

Methods

Oliver Rosmorduc et al.

Karl Esten Nakken et al.
Bronsky et al.
Milan Jirsa et al.

2003
2008
2010
2014

French
Norwegian
Czech
Czech

Exon 4, 5, 6, 8, 16
Exon 5, 6, 13, 14, 15, 16
Exon 6, 16
Exon 6, 16

PCR-RFLP
AD
PCR-RFLP
AD

Number
Case
Control
108
66
16
95
35
150
370
150

AD: allelic discrimination; PCR: polymerase chain reaction; RFLP: restricted fragment length polymorphism.

Table 2. Baseline data for 296 recruited subjects

IndexIndex
Sex ratio (M:F)
Age (years)
BMI (kg/m2)
AST (units/l)
ALT (units/l)
LDLC (mg/dl)
HDLC (mg/dl)

Gallstones
No stones
(n = 171)
(n = 125)
72:99
51:74
46.19.2
53.25.6
24.93.1
24.62.9
34.0717.8 30.520.4
33.219.3 29.221.6
130.735.1 113.328.9
46.810.5 51.812.0

P
0.905
0.267
0.889
0.668
0.867
0.015*
0.028*

Two-sample Students t test, except Pearson test. Values

are presented as mean s.d. BMI: body mass index; AST:
aspartate aminotransferase; ALT: alanine aminotransferase;
LDLC: low-density lipoprotein cholesterol; HDLC: high-density
lipoprotein cholesterol. *P represents the significant differences between the Gallstones group and No stones group. *:
P value <0.05.

These studies showed that ABCB4 gene mutations represented a genetic risk factor in a
symptomatic and recurring form of gallstone
disease in young adults. Furthermore, gallstone
formation is a consistent feature of ABCB4
knockout mice [10]. Accordingly, ABCB4 genotyping could be used to confirm the diagnosis
of LPAC in young adults who present with symptomatic gallstone disease and allow familial
screening [9]. With the understanding of ABCB4
as important in regulating biliary cholesterol
content and cholesterol absorption, studies on
association of ABCB4 polymorphisms and gallstone disease have been published [8, 11-18].
The most studied loci are ABCB4 exon 6
(Rs1202283) and exon 16 (Rs2230028). Due
to difference in allele frequency at each polymorphic locus between different ethnicities,
the associations between these SNPs and gallstone disease have not been consistent.
Furthermore, the functional roles of these polymorphic loci have not been fully clarified.
Additionally, no study has ever determined
whether any difference exists for these SNPs
are associated with gallstone disease risk in
Chinese people.

1219

In this study, we evaluated the association

between polymorphisms at ABCB4 Exon 6
(Rs1202283), Exon 16 (Rs2230028) and gallstone disease using meta-analysis. We evaluated two loci attaining a genome-wide significance level of a P value less than 0.01 in this
meta-analysis of four studies including 529
gallstone patients and 461 controls. Then we
screened a Chinese population with these two
polymorphisms and examined the risk stratification and their interaction with age, sex, body
mass index (BMI) status and symptoms in gallstone disease. According to the likely mode of
inheritance extracted from the meta-analysis
for each locus, we illustrated that these two loci
are associated with the gallstone disease risk
of Chinese people by TaqMan SNP genotyping
assay.
Materials and methods
Literature search
We searched public databases PubMed (http://
www.ncbi.nlm.nih.gov/pubmed/), Embase (http://www.embase.com), ISI Wed of Knowledge
(http://isiknowledge.com), Wanfang (http://
www.wanfangdata.com.cn) and China Biological
Medicine (CBM) (http://cbm.imicams.ac.cn)
with the last update as of May 2015. The keywords used for search were gallstone disease
and ATP Binding Cassette Transporter A4 or
ABCB4 combined with gene or variants or
polymorphism or alleles, all of which were
MeSH terms (Medical Subject Headings in the
US National Library of Medicine). Only studies
published in English or Chinese were identified.
Afterwards, the full texts of the retrieved articles were scrutinized to ensure that data of
interest was included. If two or more studies
shared the same studied populations, the study
with the small size was excluded. If more than
one geographical or ethnic population were

Am J Transl Res 2016;8(2):1218-1227

Three risk factors of gallstone disease in Chinese population

Subjects
For the validation study, all
the cases and controls were
recruited in the Department
of Biliary-Hepatic Surgery at
The Affiliated Hospital of
Guizhou Medical University,
the Gallstone Disease Biobank project. The study protocol was approved by
Ethical Committee at The
Affiliated Hospital of Guizhou Medical University and
written informed consent
was obtained from all participants. We examined a
Figure 1. Flow diagram of
sub-cohort that consisted of
search strategy and study
296 individuals who were
selection. Sixteen papers
randomly selected from the
potentially relevant for our
total cohort using a randstudy. Twelve papers were
excluded and four papers
om selection algorithm. The
including three cohorts were
whole blood samples were
included for the final analyobtained from these individsis.
uals. Demographic characteristic were described in
Table 2. Among these paincluded in one paper, each ethnical population
tients, whole blood sample was collected from
was considered separately.
171 patients with gallstone disease and 125
gallstone-free patients. All samples were stored
Study criteria
in -80C until analysis. Patients with hepatic,
renal or endocrine disorders were not included
Inclusion criteria were listed as the followings:
in the study.
1. evaluation of the association between
ABCB4 polymorphism and gallstone disease; 2.
SNP genotyping assay
case-control study using either hospital-based
or population-based designs; 3. genotype/
We used MagCore Genomic DNA Whole Blood
allele counts of ABCB4 polymorphisms between
Kit (RBC Bioscience, Taipei, Taiwan) for purificacases and controls for estimating binary varition of total DNA (including genomic and mitoable. Gallstone disease was diagnosed by ultrachondrial DNA) from whole blood by MagCore
sonography or operation. Studies were excludauto-extraction instrument. Analysis was pered if they were published in a non-English and
formed with the TaqMan SNP genotyping
non-Chinese language or published in conferassay (Applied Biosystems, Foster City, CA,
ence abstract form only.
USA). The assay was designed for human SNP
genotyping studies with the precision of
Extracted information
TaqMan reagent-based chemistry, ABCG8
gene
D19H (C_26135643_10 for rs11887534)
We abstracted the following information from
as
positive
control in Chinese [19], and the
qualified studies (Table 1), including first
ABCB4
gene
Exon 6 (C_8317490_30 for
authors last name, publication date, populars1202283)
and
Exon 16 (C_25472183_30
tion ethnicity, methods of diagnosis of gallfor
rs2230028)
were
evaluated. Detection of
stone, study design, methods of genotyping
the
above
gene
variants
was performed accordand the genotype in cases and controls.
ing to the SNP genotyping reaction protocol,
Information such as cases and controls age,
under the following conditions, 95C for 10
gender and BMI were also collected.
1220

Am J Transl Res 2016;8(2):1218-1227

Three risk factors of gallstone disease in Chinese population

Figure 2. Pooled random-effect-based odds ratio of gallstone disease for ABCB4 polymorphism. A. Minor alleles
of Exon 6 polymorphism (R = 0.41, 95% CI: 0.25-0.67, P<0.001) was related with an increased risk of gallstone
disease in European. B. Exon 16 polymorphism (OR = 0.12, 95% CI: 0.06-0.22, P = 0.001) was also related with an
increased risk of gallstone disease in Europeans.

min, then 40 cycles of 95C for 15 s alternating with 60C for 90 s. The Agilent Mx3000p
sequence detection system (Agilent Technologies, Santa Clara, CA, USA) was used to
detect gene variants using VIC and FAM
dyes.
Statistical analysis
The meta-analysis was performed in Stata/SE
11.0 software (StataCorp LP, College Station,
USA). We used the random-effects model with
the method of DerSimonian & Laird to pool
effect-size estimates from the individual stud-

1221

ies. Heterogeneity was estimated qualitatively

using the Mantel-Haenszel method [20], and
quantified using the I2 statistic, with the ranges
of 0 to 100% (I2 = 0-25%, no heterogeneity; I2 =
25-50%, moderate heterogeneity; I2 = 50-75%,
large heterogeneity; I2 = 75-100%, extreme heterogeneity) [21]. Publication bias was evaluated by using funnel plots and the Egger test
[22].
The biochemical and gene expression data
analysis were performed using SPSS 13.0 for
Windows (SPSS, Chicago, Illinois, USA). The
comparison among groups for categorical vari-

Am J Transl Res 2016;8(2):1218-1227

Three risk factors of gallstone disease in Chinese population

dent variable. P<0.05 was considered significant with a twotailed test.
Results
Association of rs1202283 and
rs2230028 at ABCB4 with
gallstone disease in European:
meta-analysis
We identified 16 studies potentially relevant for our analysis. 12
papers were excluded, leaving
four in the final analysis (Figure
1). The studies comprised of
French, Norwegian and Czech
cohorts. All studies had genotypic information for ABCB4
rs1202283, rs2230028. A summary of study population characteristics is presented in Table 1.
In allelic model, minor alleles of
rs1202283 (pooled OR = 0.41,
95% CI: 0.25-0.67, P<0.001,
Figure 2A) and rs2230028
(pooled OR = 0.12, 95% CI: 0.060.22, P = 0.001, Figure 2B) were
significantly associated with an
increased risk of gallstone disease in Europeans.
Figure 3. Beggs funnel plot of the Eggers test for publication bias of
ABCB4 polymorphism and gallstone disease. The horizontal line in the
funnel plot indicates the fixed-effects summary estimates, and the sloping lines indicate the expected 95% CI for a given standard error. A.
rs1202283. B. rs2230028.

ables was performed with Pearson tests or

Fishers exact tests. Continuous variables were
compared by two-sample Students t tests.
Allelic frequencies were estimated by direct
gene counting. A test or a Fishers exact test
was used for testing Hardy-Weinberg equilibrium and power calculations for the sample size
of each genotype. Haploview was used for estimating haploblocks, and haplotype analysis
used a Hap-clustering program [23]. Binary
logistic regression gave odds ratios of having
gallstones under the influence of genotypes
ABCB4 rs1202283 and rs2230028. For the
crude odds ratio, the presence or absence of
gallstones was the dependent variable, and
rs1202283 or rs2230028 was the indepen-

1222

Publication bias

We used Funnel plot and Eggers

test to assess for publication
bias. The resultant symmetrical
funnel plot was symmetrical, consistent with an absence of publication bias.
This was consistent with results of the Egger
test for Exon 6 (P = 0.461) and Exon 16 (P =
0.315), Figure 3A and 3B.
Association of rs1202283 and rs2230028 at
ABCB4 with gallstone disease in Chinese

Allelic frequencies of ABCB4 rs1202283 and

rs2230028 are shown in Table 3. The distributions of genotypes in the whole study group
were in Hardy-Weinberg equilibrium. The allelic
frequencies were different from those reported
previously in the European-American population. The homozygote AA allele of rs1202283
was quite rare (2.3%). Compared with Caucasians, the dominant allele distribution heterozy-

Am J Transl Res 2016;8(2):1218-1227

Three risk factors of gallstone disease in Chinese population

Table 3. Allelic frequencies of the polymorphisms in ABCB4
rs1202283 and rs2230028 in 296 subjects
Gene
ABCB4: Exon 6
ABCB4: Exon 16
ABCG8: D19H

Allele ratio
AA, AG, GG
CC, CT, TT
CC, CG, GG

NCBI SNP reference

Rs1202283
Rs2230028
Rs11887534

Frequency
23:194:79
0:288:8
0:286:10

ABCB4 rs1202283 GG genotype

as risk with asymptomatic gallstone
disease

For further analysis of the odds

ratios of minor genotypes with gallstone disease, carriers of homozyNCBI: National Center for Biotechnology Information; SNP: single nucleotide
gote AA, heterozygote AG and homopolymorphism.
zygote GG genotypes of Exon 6 and
those of homozygote CC, heterozygote CT and homozygote TT genotypes of Exon
Table 4. Association of genotype frequency
16 were combined. The risk stratification of
of ABCB4 rs1202283 and rs2230028 with
these minor genotypes for gallstone disease
gallstone development
was determined and adjusted by age, sex, BMI
Gallstones No stones
P*
and symptoms, which were well established as
(n = 171) (n = 125)
independent risk factors for gallstone disease.
ABCB4: Exon 6
The rs1202283 heterozygote AG and rs22Genotype
30028 homozygote TT variant contributed a
AG
90
104
3.5E-8
significant and independent risk of gallstone
AA + GG
81
21
formation. Epidemiological surveys demonABCB4: Exon 16
strated that the risk of asymptomatic gallstone
Genotype
disease increased with rs1202283 homozyCT
163
125
0.023
gote GG genotype at ABCB4. The gallstone risk
TT
8
0
associated with rs1202283 and rs2230028
ABCG8: D19H
polymorphisms was further stratified into different symptoms groups. The result clearly
Genotype
demonstrated a significant risk of asymptomCG
10
0
0.006
atic gallstone disease associated with rs12GG
161
125
02283 homozygote GG genotype than homozy*Pearson test.
gote AA, heterozygote AG genotype (95% C.I.:
0.219-0.768; P = 0.005) and a greater risk
gote CT of rs2230028 was high (97.6%) and the
than rs2230028 (95% C.I.: 0.264-4.506; P =
homozygote CC genotype was absent in the
0.906). On the basis of these two SNPs in
study population. ABCG8 rs11887534 (D19H)
ABCB4, a haplotype analysis study was conallele as a positive control exhibited the same
ducted. There was no significant difference in
results as previously reported in Chinese
the distribution of each haplotype between the
Taiwan population [19]. The association begallstone disease and control groups (Figure
tween the genotypes of ABCB4 rs1202283
4).
and rs2230028, and the presence of gallstones are shown in Table 4, with ABCG8
Discussion
rs11887534 (P = 0.006) as a positive control.
There was a significant correlation of gallstone
In this study, we pooled data from 4 studies
disease with the genotype distribution of
which comprised of French, Norwegian and
rs1202283 (P = 3.5E-8) and rs2230028 (P =
Czech cohorts. The meta-analysis of samples
0.023) polymorphisms. The minor alleles of
were selected from 529 gallstone patients and
rs1202283 and rs2230028 increased in
461 control samples, and the results showed
patients with gallstones compared with conthat ABCB4 rs1202283 and rs2230028 mutatrols. For example, in the rs1202283 polymortions were associated with an increased risk of
phism, the frequency of the heterozygote AG
gallstone disease in Europeans. An obvious difgenotype was 52.6% in the gallstone group
ference of gallstone prevalence between ethcompared with 83.2% in the control group. The
nicities was present.
frequency of the rs2230028 heterozygote CT
Gallstone disease is highly prevalent in
variant was 95.3% in the gallstone group comEuropean, Pima, Hispanic populations, and is
pared with 100% in the control group.

1223

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Three risk factors of gallstone disease in Chinese population

Figure 4. Overview of the physical and genetic structure of the ABCB4. The
physical position of the investigated SNPs and a schematic representation
of the gene structures are shown above. Haplotype blocks derived from the
HapMap genotypes from individuals of Han Chinese Beijing (HCB) are outlined in black. No significant difference in the distribution of each haplotype
between the gallstone disease and stone free groups.

relatively lower in Asian, and lowest in African

populations [1]. Gallstone disease is a common
clinical entity, with a prevalence of 10-15%
detected by ultrasonography in Western populations. The overall prevalence is lower in Asia,
ranging from 3-15% [24-26]. This difference
may relate to both dietary habits and genetic
factors. The relationship between diet and gallstones is still controversial [27]. Cholesterol
gallstones are responsible for 80-90% of all
gallstones in Western countries [24, 25, 28]. As
a result of westernization of dietary habits, the
composition of gallstones has also altered: up
to 77% in Chinese populations are now cholesterol gallstones [25]. Therefore, we divided the
population into Asian and Western.
1224

We evaluated the two loci

associated with gallstone disease attaining genome-wide
significance in our meta-analysis as predictors of gallstone
risk in 296 recruited subjects
including 171 gallstone samples and 125 no gallstone
controls. We found a significant correlation between the
distribution of rs1202283
(AG) and rs2230028 (CT) polymorphisms and gallstone disease in the Chinese population. These similar results
were present between ABCB4
polymorphism in relation with
gallstone disease in both populations. Furthermore, upon
dividing the population into
Chinese and non-Chinese, the
association of rs1202283
and rs2230028 at ABCB4
polymorphisms with gallstone
disease did not change. The
risk associated with Exon 6
(GG) was strong in asymptomatic gallstone disease. Additionally as a positive control,
all the clinical conditions of
ABCG8 D19H were similar to
the Chinese Taiwan population previously reported [19].
The study also found that serum levels of total cholesterol,
LDLC were significantly lower
and HDLC were higher in subjects with gallstones than in
those without gallstones.

Gallstone formation results from a complex

interaction between multiple predisposing
genes and environmental factors. Genetic susceptibility for gallstone formation is influenced
by well-known risk factors including age, sex,
obesity, parity, and medication [24, 25, 28, 29].
In this study population, polymorphisms of
ABCB4 and age, sex, obesity, parity, and medication were not associated with gallstone disease in univariable analysis. However, we found
GG allele of rs2230028 polymorphism at
ABCB4 gene was a common allele predicting
susceptibility to asymptomatic gallstone disease independent of ethnicities. Previous studies also showed that Mdr2 gene knockout mice,
the murine homolog of ABCB4, may develop
Am J Transl Res 2016;8(2):1218-1227

Three risk factors of gallstone disease in Chinese population

bile cholesterol crystals [30, 31]. The study by
Oliver Rosmorduc et al. strongly suggested that
ABCB4 mutations may lead to cholesterol gallstone when residual ABCB4 activity and subsequent biliary phospholipid secretion decreases
below a critical threshold, which may depend
on the type of mutation and other host or environmental factors [9]. Consistent with previous
studies [13], this finding supports the hypothesis that the Exon 6 at ABCB4 mutation may
affect the transporter function resulting in gallstone formation in earlier life. These polymorphisms may alter the conformation of ABCB4
transporters to modulate the intestinal absorption efficiency or the biliary secretion of
cholesterol.
Epidemiological studies have indicated an
association between low plasma HDLC levels
and gallstone prevalence [26]. Super-saturation
of bile with excess cholesterol is an essential
step in gallstone formation. Bile is the major
route for the elimination of excess cholesterol
from the liver. In human studies, most of the
cholesterol secreted into bile comes from circulating plasma lipoproteins, the free cholesterol
carried by HDL particles binding to the scavenger receptor class B type 1 (SRB1), the uptake
of cholesterol by LDL receptors or LDL receptor
related proteins, and the dietary cholesterol in
the chylomicron remnants. Newly synthesized
cholesterol contributes very little (5-20%) to
biliary cholesterol secretion [24, 26, 28, 32]. In
cholesterol homeostasis, plasma HDLC plays a
key role in reverse cholesterol transport to the
liver. The biliary secretion of free cholesterol
markedly increases in Srb1 transgenic mice
and transport of cholesterol into bile is mediated mainly by SRB1 [33]. More interestingly, the
present study demonstrated that serum levels
of total cholesterol, LDLC, and HDLC were significantly associated with abdominal pain in
pancreatitis and other digestive system diseases [34, 35]. As previously reported, polymorphism in ABCB4 was strongly associated (P =
9E-14) with the plasma LDLC concentrations on
the high-cholesterol, high-fat diet. They indicated that genetic variation in ABCB4, or closely
linked genes, was responsible for the dramatic
differences among opossums in their LDLC
response to an atherogenic diet [36, 37].
Polymorphisms of ABCB4 and ABCB4-related
genes may alter the transporter function, modify the intrahepatic cholesterol content and
1225

regulate the reverse cholesterol transport in

cholesterol homeostasis. It is possible that gallstone formation may be a by-product of excess
cholesterol excretion into bile to keep serum
levels of cholesterol-containing lipoproteins
lower and significantly associated with the risk
of gallstone disease without symptoms, such
as abdominal pain.
In summary, our results strongly support the
role of rs1202283 (AG) and rs2230028 (CT) at
ABCB4, and rs11887534 (CG) at ABCG8 were
associated with the risk of gallstone disease.
More notable is that the risk associated with
rs1202283 (GG) was strong in asymptomatic
gallstone disease. In the near future, ABCB4
genotyping should be used to confirm the diagnosis of asymptomatic gallstone disease and
allow familial screening. Depending on the
results, long-term curative or prophylactic ursodeoxycholic acid therapy could be initiated
early to prevent the occurrence or recurrence of
this disease and its severe complications.
Acknowledgements
This study was granted by the Brainstorm
Project on Social Development by Department
of Science and Technology of Guizhou Province
(Grant No. SY [2015] 3047).
Disclosure of conflict of interest
None.
Address correspondence to: Dr. Cheng-Yi Sun, Department of Biliary-Hepatic Surgery, The Affiliated
Hospital of Guizhou Medical University, 28 Guiyi
Street, Guiyang 550001, Guizhou Province, China.
Tel: +86-851-6771326; Fax: +86-851-6771326;
E-mail: [email protected]

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