Modular analytics e170

All specimens were analyzed with a Roche Modular Analytics E170 (Roche Diagnostics, Inc., Mannheim, Germany) to measure HBs Ag, anti-HBc, and anti-HBs. Specimens with ‘anti-HBc alone’ were stored at -70℃ for further HBV DNA analysis.

Measurements of biomarkers of bone resorption and baseline serum electrolyte, calcium and phosphorus, creatinine profile were retrospectively obtained from electronic medical records. All patients had collected 24-hour urine samples to estimate urine sodium, calcium, and creatinine excretion. Blood samples were obtained at their first visit at the outpatient clinic of Division of Endocrinology and Metabolism. Subjects were sampled in the morning after an overnight fast exceeding 8 hours. C-terminal crosslinking telopeptides of type I collagen (CTX-I), a marker of bone resorption, was measured in the serum by electrochemiluminescence immunoassay (ECLIA) (Roche Modular Analytics E170 [Roche Diagnostics, Mannheim, Germany]). The interassay coefficient of variation for CTX-I ranged between 2.6% and 2.9% depending on CTX-I level. All patients underwent baseline bone mineral density (BMD) using dual energy X-ray absorptiometry (DXA; GE Healthcare Lunar, Madison, WI, USA). In accordance with the WHO criteria, osteopenia was defined as a T-score for the lumbar spine, total hip of between -1 and -2.5, and osteoporosis was defined as T-score of less than -2.5.

The levels of TFF3 (Item ID: E-EL-H1108c) in serum were detected by ELISA kits, which were provided by Elabscience Biotechnology Co., Ltd. (Wuhan, China). The detection protocol was performed according to the manufacturer’s instructions. Briefly, 100 μL of standard, blank, or sample was added per well. Solutions were added to the bottom of the well and incubated for 90 min at 37 °C. The liquid was removed and 100 μL of biotinylated detection Ab working solution added to each well, followed by incubation for 1 h at 37 °C. The liquid was aspirated and the wells washed three times. Then, any remaining wash buffer was removed, and 100 μL of HRP conjugate working solution was added, followed by incubation for 30 min at 37 °C. The wash process was repeated five times, and then 90 μL of substrate solution was added. Incubation was performed for 15 min at 37 °C followed by the addition of 50 μL of stop solution to each well. The optical density (OD value) of each well at 450 nm was measured by a Bio-Rad iMark Microplate Absorbance Reader (Bio-Rad Laboratories Inc.). The level of TFF3 was calculated according to the standard curve. The coefficient of variation of all kits was less than 10%. The levels of CEA in serum were detected by a Roche Modular Analytics E 170 instrument (Roche Diagnostics, Mannheim, Germany). The detection assays were provided by Roche Diagnostics, United States.

Serum TSH values were assessed from frozen blood samples with the Roche Modular Analytics E170 electro-chemiluminescence immunoassay (Roche Diagnostic). The working range for this method was 0.005–100 mIU/L. The functional sensitivity amounts to 0.014 mIU/L. TSH values were used as a continuous variable except for some of the analysis, for which, the main analysis population was categorised into euthyroid, hypothyroid and hyperthyroid participants. The reference range of 0.27–4.20 mIU/L by the manufacturer was adopted to define participants as euthyroid. Hyperthyroid status was defined as TSH values <0.27 mIU/L, while hypothyroid status was defined as TSH values >4.20 mIU/L.