Clinical assessment
A detailed clinical assessment was performed in each patient.
Information on medical history, past and current drugs and alcohol
consumption were collected. In this study, we analyzed alcohol
consumption by asking the patient about the type and amount of alcohol
consumption per day during the prior year, then estimating the mean
alcohol intake per day. Anthropometric assessments were performed by
well-trained nurses and included body mass index (BMI) and waist
circumference (WC). Obesity was defined in patients that had BMI ≥ 30
kg/m2, and central obesity as WC >94 cm
for male and >80 cm for female. Hypertension was defined if
there was positive medical history or if the patient was taking
anti-hypertensive drugs or if the average blood pressure after three
repeated measures was ≥140/90 mmHg. Dyslipidemia was defined as positive
medical history, using of lipid-lowering drugs, or if the serum total
cholesterol level was ≥5.2 mmol/L, serum triglyceride (TG) level ≥1.7
mmol/L, serum high-density lipoprotein (HDL) cholesterol level
<1.0 mmol/L for male or <1.3 mmol/L for female, or
serum low-density lipoprotein (LDL) cholesterol level ≥3.4 mmol/L. The
MetS was defined according to the International Diabetes Federation
criteria that are including central obesity (waist circumference
>80 cm for women and >94 cm for men) plus at
least two of the following risk abnormalities: blood pressure ≥130/85
mmHg or drug treatment; previously diagnosed type 2 diabetes, or drug
treatment or a fasting plasma glucose level ≥5.6 mmol/L; triglyceride
levels >1.7 mmol/L and/or HDL-cholesterol <1.03
mmol/L for men and <1.29 mmol/L for women or drug treatment.
An extensive laboratory characteristic was analyzed for each patient
including and all necessary laboratory tests for exclusion of other CLD.
Fasting blood samples for analyzing laboratory parameters were taken in
the morning hours after overnight fasting at the day of TE and US
examinations. Sleeping duration,
coffee consumption and smoking were recorded using quantitate
questionnaire. Sleeping duration was categorized in three groups: short
(S) (<6h), moderate (M) (6-8h) and long (L) (>8h)
sleep duration. Each patient was asked if he had drunk coffee in the
previous year, and how many cups per day. Therefore, coffee drinking was
categorized into no (0), moderate (1–2) and frequent (≥3) consumption
(in cups/day, one cup equals 150 g). Smoking was categorized as yes vs.
no. Those patients who reported as
smoking at least sporadically during the last two year were defined as
smokers. On the other hand, those who reported as having stopped smoking
at least two year before their enrolment in our study and those who had
not smoked during their life were defined as non-smokers.
Transient elastography and liver ultrasound
LSM and CAP were obtained by using Fibroscan® 502 (Echosens, Paris,
France) by a single operator who had performed more than 2.000
examinations before the start of this study. In each patient a minimum
of 10 seral measurements were done using the M- or the XL probe
according to the instructions. The examination was defined as successful
if there was ≥10 valid measurements with interquartile range
(IQR)-to-median ratio of LSM was ≤0.3. Liver steatosis was defined by
the mean CAP values of ≥238 dB/m
(19). According to earlier
published data, significant liver fibrosis (≥F2) was defined as an LSM
≥7kPa and advanced fibrosis (≥F3) if LSM was ≥9.6 kPa using the M probe
or ≥9.3 kPa using the XL probe. On the other hand, cirrhosis was
considered if the LSM was ≥ 11.5 kPa using the M probe or ≥ 11.0 kPa
using the XL probe (20,21). Abdominal US was carried out by a certified
and experienced gastroenterologist on Philips Affiniti (PC Best,
Nederland). Diagnosis of steatosis was determined dichotomously
according to the protocol of Hamaguchi et al (22), as presence or
absence of a hyper-echogenic liver parenchyma.