Objective  To investigate the serum expression levels of Notch homolog 1 (Notch1), vitamin K2 (VK2), and fibroblast growth factor receptor 1 (FGFR1) in patients undergoing maintenance hemodialysis (MHD), and to analyze their relationship with the development of vascular calcification.  Methods This cross-sectional study enrolled MHD patients treated at Hebei PetroChina Central Hospital from January 2022 to June 2023. Patients were divided into a calcification group and a non-calcification group based on abdominal lateral X-ray plain films. Serum levels of Notch1 and FGFR1 were measured using enzyme-linked immunosorbent assay (ELISA), and serum VK2 levels were determined by high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS). Clinical data of both groups were collected. Multivariate logistic regression analysis was used to identify independent risk factors for vascular calcification, and receiver operating characteristic (ROC) curves were employed to evaluate the predictive value of Notch1, FGFR1, and VK2.  Results  A total of 120 patients were included in this study, comprising 78 patients in the calcification group and 42 in the non-calcification group. Serum levels of Notch1 [(95.67±1.16) ng/L vs. (91.21±1.20) ng/L, t=81.189, P<0.001] and FGFR1 [(12.62±0.28) μg/L vs. (11.09±0.38) μg/L, t=2.600, P=0.011] were significantly higher in the calcification group than in the non-calcification group, while serum VK2 levels were significantly lower [(1.31±0.03)ng/L vs. (1.48±0.04) ng/L, t=40.391, P<0.001]. Multivariate logistic regression analysis identified prolonged dialysis vintage (OR=1.132, 95% CI: 1.022～1.253, P=0.017), elevated serum phosphorus (OR=3.228, 95% CI: 1.290～8.079, P=0.012), elevated intact parathyroid hormone (iPTH) (OR=1.030, 95% CI: 1.012～1.056, P=0.003), elevated Notch1 (OR=1.147, 95% CI: 1.002～1.313, P=0.047), low VK2 (OR=0.503, 95% CI: 0.339～0.746, P=0.001), and elevated high-sensitivity C-reactive protein (hs-CRP) (OR=5.447, 95% CI: 1.768～16.778, P=0.003) as independent risk factors for vascular calcification in MHD patients. ROC analysis demonstrated that the area under the curve (AUC) values for Notch1, VK2, and FGFR1 alone were 0.639, 0.700, and 0.656, respectively, whereas the combined detection of the three markers yielded an improved AUC of 0.785. The sensitivities of Notch1, VK2, and FGFR1 alone were 33.33%, 80.95%, and 78.57%, with specificities of 100.00%, 59.52%, and 47.62%, respectively; the combined detection achieved a sensitivity of 76.19% and a specificity of 73.81%. Conclusion  Serum Notch1 and FGFR1 expressions are up-regulated, while VK2 expression is down-regulated in MHD patients. Notch1 and VK2 are independent influencing factors for VC. The combined detection of these markers demonstrates high predictive value for VC, potentially providing a basis for early clinical screening and intervention.