Fibromyalgia syndrome (FMS) is a complex illness to diagnose and treat, which presents symptoms that may be part of, or overlap with other diseases or syndromes.The most widely used diagnostic criteria are those of the American College of Rheumatology [1]; no laboratory test has been validated for FMS diagnosis which remains primarily clinically based.Oxidative stress has been proposed as a relevant event in the pathogenesis of FMS with an increase of lipid peroxidation (LPO) [2] and a decrease in vitamin A and E concentrations [3].Although the etiology of FMS remains unclear, genetic predisposition is likely to be an important factor and transmission is thought to be polygenic [4,5]. If FMS is a multi-genetic disease then it could be hypothesized that differences exist in the type of proteins or protein expression levels in sera of FMS patients compared with healthy controls.Proteomic analysis is a powerful tool for the global evaluation of protein expression and plays a central role in clinical diagnosis and monitoring. Some studies [6] reported interesting proteomic analysis data with an overexpression of transaldolase and phosphoglicerate mutase I in salivary fluid of FMS patients and focused on the role of oxidative stress in the pathogenesis of the disease.The aim of this preliminary study was to evaluate, using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), changes in protein profiles of FMS patients with respect to control subjects to identify potential serum biomarkers useful for disease diagnosis and management.Sixteen females (52 ± 12 years) affected by FMS and 12 healthy females (48 ± 13 years) were enrolled; all patients fulfilled new ACR diagnostic criteria [1] ( Table 1). The presence of a major clinical condition other than fibromyalgia was excluded by physical examination and routine blood and urine screening. No patients suffered of thyroid dysfunction. Psychiatric examination data of patients were reported in a previous study [7].Study was approved from our Local Ethical Committee and informed consent was obtained from all subjects.Serum protein concentrations were quantified using the PlusOne 2D Quant Kit (GE Healthcare); first dimension isoelectric focusing (IEF) was carried out with 18 cm immobilized pH gradient strips of pH 3-10. The IPG strips were applied onto 10% acrylamide sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) slab gels (25.5 cm × 20.5 cm × 1.0 mm) and overlaid with a solution of 0.5% agarose with a trace of bromophenol blue. Gels were fixed, stained with Coomassie Brilliant Blue G-250 colloidal for 24 h and stored at room temperature. At least three sample replicates were performed.2D data were processed with Progenesis SameSpots software, which computed multiplication fold, false discovery rate (FDR) q-value, p-values of all spots using one-way ANOVA analysis. A p-value < 0.05 was considered statistically significant; no difference has been found in protein expression according to patient therapy.After normalization, volume calcul...