fig1

Figure 1. Overall experimental design for biomarkers of stroke prognosis. This is a single-center, observational, multi-stage study conducted between June 2020 and June 2022. Patients with IS were categorized into LE and OE prognosis groups according to ADL assessments using the Longshi Scale and BI at admission and after 3 months of rehabilitation. The discovery subset underwent LC-MS/MS proteomic analysis to clarify stroke-associated pathway enrichments. Surface protein profiling of individual EVs using PBA was conducted in the exploration subset to screen initial candidates. Finally, ELISA and ROC analyses in an expanded cohort were performed to validate the sensitivity and specificity of the targeted biomarkers in distinguishing between the LE and OE groups. LC-MS/MS: Liquid chromatography-tandem mass spectrometry; PBA: proximity barcoding assay; ELISA: enzyme-linked immunosorbent assay; ROC: receiver operating characteristic; HCs: healthy controls; IS: ischemic stroke; OE: obvious-effect recovery group; LE: little-effect recovery group; ADL: activities of daily living; MMP9: matrix metalloproteinase 9; CEACAM1: carcinoembryonic antigen-related cell adhesion molecule 1.