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As the therapeutic landscape for peptides continues to evolve, improving the chemical and metabolic stability of disulfide-containing peptides has emerged as a critical priority in peptide drug development ( 2 The function and limitations of native disulfides in bioactive peptides A defining feature of many bioactive peptides is the formation of disulfide bond, a covalent linkage formed between the thiol groups of two cysteine residues (Cys) (Figure 1a) ( FIGURE 1 Disulfide bonds play a central role in stabilizing the three-dimensional structures by constraining conformational flexibility, thereby preserving bioactive peptide conformations essential for molecular recognition, receptor binding, and enzymatic regulation ( Despite their structural and functional advantages, disulfide-containing peptides face notable limitations that restrict their broader therapeutic applications ( in vivo circulation, can generate heterogeneous mixtures of isomers with variable or undesired biological activity

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Firstly, it is based on the NHANES database, which employs a multistage complex probability sampling design and weighted adjustments, enhancing the representativeness and reliability of the study resultsa crucial factor for improving the generalizability of our findings within the American context

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Secondary antibodies: Fluorescein donkey anti-rat IgG (Jackson ImmunoResearch, Cat#712-095-150, 1:5000), Fluorescein donkey anti-mouse IgG (Jackson ImmunoResearch, Cat#715-095-150, 1:5000), Anti-mouse IgG, HRP-linked (Cell Signaling Technology, Cat#7076, 1:5000), Anti-Rabbit IgG, HRP-linked (Cell Signaling Technology, 7074, 1:5000), Normal mouse IgG (Santa Cruz Biotechnology, Cat#sc-2025, 2g), Normal rabbit IgG (Cell Signaling Technology, Cat#2729S, 2g), Fluorescein (FITC)-AffiniPure Donkey Anti-Rabbit (Jackson Immuno, cat# 711-095-152, 1:5000), Cy3 Goat Anti-Rabbit (Jackson Immuno, cat# 111-165-003, 1:5000), Cy3-AffiniPure Donkey Anti-Mouse (Jackson Immuno, cat# 715-165-150, 1:5000)

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