The base-quenched probe method for detecting single nucleotide polymorphisms (SNPs) relies on real-time PCR and melting-curve analysis, which might require only one pair of primers and one probe. At present, it has been successfully applied to detect SNPs of multiple genes. However, the mechanism of the base-quenched probe method remains unclear. Therefore, we investigated the possible mechanism of fluorescence quenching by DNA bases in aqueous solution using spectroscopic techniques. It showed that the possible mechanism might be photo-induced electron transfer. We next analyzed electron transfer or transmission between DNA bases and fluorophores. The data suggested that in single-stranded DNA, the electrons of the fluorophore are transferred to the orbital of pyrimidine bases (thymine (T) and cytosine (C)), or that the electron orbitals of the fluorophore are occupied by electrons from purine bases (guanine (G) and adenine (A)), which lead to fluorescence quenching. In addition, the electrons of a fluorophore excited by light can be transmitted along double-stranded DNA, which gives rise to stronger fluorescence quenching. Furthermore, we demonstrated that the quenching efficiency of bases is in the order of G > C ≥ A ≥ T and the capability of electron transmission of base-pairs in double-stranded DNA is in the order of CG[combining low line] ≥ GC[combining low line] > TA[combining low line] ≥ AT[combining low line] (letters representing bases on the complementary strand of the probe are bold and underlined), and the most common commercial fluorophores including FAM, HEX, TET, JOE, and TAMRA could be influenced by bases and are in line with this mechanism and regularity.

BackgroundRecently, many studies have shown that B7-H4 exhibits altered expression in various cancers. We performed a meta-analysis to evaluate the prognostic role of B7-H4 expression in solid tumors.ResultsData from 18 observational studies and 2467 patients were summarized. An elevated baseline B7-H4 was significantly associated with worse OS (pooled HR = 1.79; 95% CI = 1.56–2.06). Differences across subgroups of tumor type, patients' ethnicity, analysis type, HR obtain method and cut-off value were not significant (PD = 0.313, PD = 0.716, PD = 0.896, PD = 0.290 and PD = 0.153, respectively). Furthermore, patients with high B7-H4 had a significantly shorter DFS (pooled HR = 2.12; 95%CI = 1.45–3.09).Materials and MethodsWe searched PubMed, Embase and the Cochrane Library (last update by November 26, 2015) to identify studies assessing the effect of B7-H4 on survival of cancer patients. Pooled hazard ratios (HRs) for overall survival (OS) and disease-free survival (DFS) were estimated using fixed-effects models and random-effects models respectively.ConclusionsThis meta-analysis clarified that high B7-H4 expression in tissue was significantly associated with poor survival in patients with solid tumors. Future clinical studies are warranted to determine whether B7-H4 blockade has a favorable effect on disease recurrence and mortality.

Objective: This study aimed at exploring the correlation of microRNA (miR)-497/ fibroblast growth factor-23 (FGF-23) axis with major adverse cardiac and cerebral event (MACCE) occurrence in end-stage renal disease (ESRD) patients who underwent continuous ambulatory peritoneal dialysis (CAPD).Methods: Totally, 360 ESRD patients who underwent CAPD were enrolled. Their plasma samples were collected to detect miR-497 expression by real-time quantitative polymerase chain reaction, and FGF-23 level by enzyme-linked immunosorbent assay. All patients were followed up for 36 months, and the occurrence of MACCE during the follow-up was documented.Results: MiR-497 expression negatively correlated with FGF-23 level in ESRD patients who underwent CAPD (P < .001). The MACCE occurrence rate at 1, 2, and 3-year was 5.6%, 11.9%, and 15.0%, respectively. Furthermore, miR-497/FGF-23 axis high level (P < .001) and miR-497 high expression (P = .034) correlated with reduced accumulating MACCE occurrence, whereas FGF-23 high level (P = .008) correlated with increased accumulating MACCE occurrence. Forward stepwise multivariate Cox's regression disclosed that miR-497/FGF-23 axis high level (P = .008) was an independent predictive factor for lower accumulating MACCE occurrence, whereas age (≥55 years) (P < .001), body mass index (≥21.7 kg/m 2 ) (P = .006), peritoneal dialysis duration (≥61.0 months) (P < .001), C-reactive protein (≥4.7 mg/L) (P = .001), serum uric acid (≥409.4 μmol/L) (P = .009), β-fibrinogen (≥5.8 mmol/L) (P < .001), and lowdensity lipoprotein cholesterol (≥2.7 mmol/L) (P = .003) were independent factors for predicting higher accumulating MACCE occurrence. Conclusion:MiR-497/FGF-23 axis holds clinical significance for predicting attenuated MACCE risk in ESRD patients who underwent CAPD.