Background/Aims: The diagnostic criteria for nonerosive reflux disease (NERD) and functional heartburn (FH) have been changed. We investigated demographic, clinical, and psychological characteristics of the heartburn groups classified using the Rome III criteria and factors associated with the responsiveness to proton pump inhibitors (PPIs) in the gastroesophageal reflux disease (GERD) group. Methods: Ninety-five patients with heartburn underwent endoscopy, 24-hour esophageal pH-metry and then a PPI test. NERD was diagnosed when % time with pH <4 was >4%, a symptom index (SI) ≥50% or a positive PPI test in patients without erosive esophagitis. Patients without such findings were classified as FH. Results: Thirty-six patientshad erosive reflux disease (ERD), 36 had NERD, and 23 had FH. The proportion of males was significantly higher in ERD than in FH. Atypical symptoms and IBS were more prevalent in FH than in ERD. Anxiety was more prevalent in FH than in NERD. The prevalence of pathologic acid reflux, a positive SI and a positive PPI test was similar between ERD and NERD patients. In the ERD and NERD groups, depression was independently associated with nonresponsiveness to PPIs. Conclusions: FH is a different entity from ERD or NERD, particularly in terms of gender, acid reflux patterns, psychological profiles, and the responsiveness to PPIs.

Reverse transcription-quantitative (RT-q) PCR is the most feasible and useful technique for identifying and evaluating cancer biomarkers; however, the method requires suitable reference genes for gene expression analysis. The aim of the present study was to identify the most suitable reference gene for the normalization of relative gene expression in human hepatocellular carcinoma (HCC) tissue and blood samples. First, 14 candidate reference genes were selected through a systematic literature search. The expression levels of these genes ( ACTB, B2M, GAPDH, GUSB, HMBS, HPRT1, PGK1, PPIA, RPLP0, RPL13A, SDHA, TBP, TFRC and YWHAZ ) were evaluated using human multistage HCC transcriptome data (dataset GSE114564), which included normal liver (n=15), chronic hepatitis (n=20), liver cirrhosis (n=10), and early (n=18) and advanced HCC (n=45). From the 14 selected genes, five genes, whose expression levels were stable in all liver disease statuses ( ACTB, GAPDH, HMBS, PPIA and RPLP0 ), were further assessed using RT-qPCR in 40 tissues (20 paired healthy tissues and 20 tissues from patients with HCC) and 40 blood samples (20 healthy controls and 20 samples from patients with HCC). BestKeeper statistical algorithms were used to identify the most stable reference genes, of which HMBS was found to be the most stable in both HCC tissues and blood samples. Therefore, the results of the present study suggest HMBS as a promising reference gene for the normalization of relative RT-qPCR techniques in HCC-related studies.

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