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BACKGROUND AND AIMS: Acupuncture is a widely used therapeutic practice that targets specific points on or beneath the skin, known as acupoints. While acupoints are thought to exhibit low electrical resistance, their underlying biophysical and molecular characteristics remain incompletely understood. This study investigates the presence and properties of high-conductivity regions in surgically isolated human skin.METHODS: High-conductivity points (HCPs) were identified on human skin explants using a clinical conductivity-based acupoint detector. Histological, transcriptomic, and metabolomic analyses were performed on HCP and control regions, with and without needle stimulation. To probe mechanistic pathways, human keratinocytes and fibroblasts were cultured under biotin-deficient conditions, mimicking stimulation-induced metabolic changes.RESULTS: No significant structural differences were observed between HCPs and control skin. However, RNA sequencing revealed that needled HCPs activated gene programs resembling those of anatomically defined murine acupoints. Metabolomic profiling showed a stimulation-specific decrease in biotin levels at HCPs. In vitro, biotin deficiency altered acetyl-CoA carboxylase regulation and increased ATP production via mitochondrial respiration and glycolysis.CONCLUSION: These findings suggest that biotin-dependent metabolic reprogramming occurs at electrically distinct skin regions in response to physical stimulation. While the relationship between HCPs and classical acupoints remains to be fully established, this study provides novel insights into the local biochemical responses associated with acupuncture.