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4.5.3.2 Principal factors influencing biomarker application Impact of renal functional status : when glomerular filtration rate declines, multiple metabolic intermediates accumulate systemically, including branched-chain amino acids and aromatic amino acid catabolites, rendering differentiation between authentic pathogenic markers and consequences of diminished renal clearance capacity difficult ( Temporal dependence of nutritional intake : short-term dietary composition adjustments can rapidly alter amino acid profiles in serum and urine ( Inter-individual microecological variations : gut microbiota compositional architecture exhibits 2–3-fold fluctuations across different timepoints within individual subjects and among different individuals ( Compound effects of pharmacological therapy : standard treatment regimens for DKD patients incorporate sodium-glucose cotransporter 2 inhibitors and renin-angiotensin system inhibitors, which exert influence through multiple pathways including intestinal pH environment modification, membrane transporter expression regulation, and metabolomic landscape remodeling (156, 157)