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A 10 mL syringe preloaded with 2 mL of oocyte-washing medium (TCM199-HEPES buffer, Gibco, #12340030) containing 1 mg/mL heparin sodium (Sigma, #H3149), 10% (v/v) bovine calf serum (FBS, Gibco, #16030074), and 0.1% (v/v) penicillinstreptomycin was used to aspirate cumulusoocyte complexes (COCs) from 3 to 8 mm follicles on the ovarian surface

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21-23 These risk factors include: Sedentary lifestyle 21,22 Low educational attainment 21,22 Smoking 21,22 Obesity 21,22 Insulin resistance and type 2 diabetes 21,22,582 Chronic inflammation 582 Hypertension 21,22 High total and LDL cholesterol 664 Chronic kidney disease 21 Atrial fibrillation (a type of arrhythmia) 21 Cardiovascular disease 24 Cerebrovascular disease 512 Traumatic brain injury 58-60 Depression 21 Sleep disorders 25 Sleep apnea 26 Hearing loss 583 High homocysteine levels 27 Heavy metal toxicity 28 Excessive alcohol consumption 650 Air pollution 650 Anticholinergic medications block the effects of acetylcholine and are used to treat a wide variety of illnesses, including asthma, chronic obstructive pulmonary disease (COPD), Parkinson disease, depression, and allergies, among other conditions