By Fisher's exact test, Wilcoxon test, or Kruskal–Wallis test.

#### *2.4. Protein Identification*

Then, the acquired MS/MS raw data files were applied to search against a UniProt human protein database (containing 20,387 protein sequences; released on 9 April 2021; http://www.uniprot.org/ (accessed on 6 December 2021)) with PEAKS Studio 7.5 (Bioinformatic Solution, Ontario, CA, USA). The search settings of PEAKS Studio 7.5 combined with UniProt's protein database were as follows: enzyme set to trypsin; up to two missing cut sites; precursor and fragment mass tolerances of 20 ppm and 0.8 Da, respectively; false discovery rate (FDR) of <1%, obtained from a search of the decoy database. Furthermore, based on a label-free quantitative analysis, each identified protein had to contain at least one unique peptide and protein quantification method. Moreover, spectral counts were normalized to the total identification spectrum of each biological sample.

#### *2.5. Enzyme-Linked Immunosorbent Assay (ELISA)*

An alpha-2-Heremans–Schmid (HS)-glycoprotein ELISA assay was performed to measure concentrations of AH samples among the single-risk group, double-risk group, and the age-matched cataract controls with a Human Alpha-2-HS-glycoprotein ELISA Kit (EH310RB, ThermoFisher Scientific), as per the manufacturer's protocol.

#### *2.6. Statistical Analysis*

Clinical data were analyzed using Stata (vers. 16.1, StataCorp, College Station, TX, USA) to define the statistical significance between groups by a *t*-test or Chi-squared test, and *p* < 0.05 was considered to be statistically significant. Statistical analysis by Fisher's exact test, Wilcoxon test, or Kruskal–Wallis test was used to confirm that there were no statistically significant differences in age among the single-risk group, double-risk group, and the age-matched cataract control group (Table 1).

Note: Single risk, patients with the DM or smoking risk factor; double risk, patients with both the DM and smoking risk factors; control, cataract patients with neither of these cataract risk factors; VA, visual acuity; AXL, axial length.
