*2.1. Subjects*

The study protocol was approved by the Medical Ethics and Institutional Review Board of Taoyuan General Hospital, Ministry of Health and Welfare (TYGH109009) (Taoyuan, Taiwan), and conducted as per the tenets of the Declaration of Helsinki. All study participants provided written informed consent before their enrollment, and the nature and possible consequences of the study were explained to them. Human AH samples from treatmentnaive patients with a single risk factor (*n* = 10) of DM (*n* = 5) or cigarette smoking (*n* = 5), double risk factors (*n* = 8) of DM combined with cigarette smoking, and aged-matched cataract controls with neither risk factor (*n* = 9) were collected while patients were undergoing cataract surgery at Taoyuan General Hospital. The diagnostic criterion for cataracts was defined with a slit lamp with no other ocular diseases, trauma, or previous intraocular operation history. The presence of type 2 diabetes was defined as any one or more of the following: (1) having had a diagnosis of type 2 diabetes that was confirmed by a physician (ICD10: E11); (2) self-report of a diabetes diagnosis and use of hypoglycemic medications; (3) a fasting glucose level of ≥126 mg/dL; (4) a 2 h post-challenge plasma glucose level of ≥200 mg/dL. All subjects were included as cases of type 2 diabetes within a follow-up time of five years. A cigarette smoking history was obtained from all patients. Their cigarette consumption varied with a mean duration of more than 20 years. Data on control eyes were collected from senior cataract patients who were free from other ocular or systemic

diseases. In these three groups, inclusion criteria were cataract patients aged older than 55 years. Exclusion criteria were a history of any systemic or ocular disorder or condition including ocular surgery, trauma, or disease. Best corrected visual acuity (BCVA) was measured as the logarithm of the minimum angle of resolution (logMAR).

#### *2.2. AH Sample Collection*

AH samples were obtained from patients during the implantation of phakic intraocular lenses. To avoid hemorrhaging and ocular surface contamination, a sample was collected using a 1 mL tuberculin syringe with a 30 gauge needle at the limbus before any other entry into the eye under a surgical microscope. Note that 50–100 μL of AH was collected from each patient by anterior chamber paracentesis. Undiluted AH samples were collected and stored at −80 ◦C within 24 h until preparation was initiated.
