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. 2025 Jan 18;18:827–846. doi: 10.2147/JIR.S499343

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© 2025 Cai et al.

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Screened candidate diagnostic biomarkers of AD progression and infiltration analysis of biomarkers. (A) Candidate gene LASSO coefficient profiles. (B) A cross-validation study was conducted to determine which tuning parameter log (Lambda) is most effective. (C) Candidate gene profiles according to RF coefficients. (D) Random forest algorithm MeanDecreaseGini for hub genes. (E) Biomarker screening using the SVM-RFE algorithm. (F) Biomarker screening using the GBM algorithm. (G) Four machine learning algorithm screening results are shown in a Venn diagram, in which the overlap genes were defined as candidate biomarkers. (H) The expression of 5 candidate biomarkers (EGFR, HIF1A, JUN, MAPK3, and RELA) between control and AD groups in the GSE5281 test dataset. (I) The expression of 5 candidate biomarkers (EGFR, HIF1A, JUN, MAPK3, and RELA) between control and AD groups in the GSE122063 validation dataset. (J) The ROC curves of the 5 candidate biomarkers (EGFR, HIF1A, JUN, MAPK3, and RELA) are based on the GSE5281 test dataset. (K) The ROC curves of the 5 candidate biomarkers (EGFR, HIF1A, JUN, MAPK3, and RELA) in the GSE122063 validation dataset. (L) Nomograms of AD. (M) Calibration curves of the nomogram. (N) Western blot detection of EGFR and RELA (n = 3). (O) The protein levels were determined by Western blot, and β-actin was employed as internal references (n = 3). The serum levels of MDA (P) and GPx (Q) were detected using an ELISA assay (n = 6). (R) The protein levels were determined by Western blot, and β-actin was employed as internal references (n = 3). The serum levels of MDA (S) and GPx (T) were detected using an ELISA assay (n = 6). (U) The protein levels were determined by Western blot, and β-actin was employed as internal references (n = 3). The serum levels of MDA (V) and GPx (W) were detected using an ELISA assay (n = 6). *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; #, p < 0.05; ###, p < 0.001; ####, p < 0.0001.