Editor, Fibrosis is characterized by fibroblast dysfunction and excessive deposition of cell-matrix that affect the normal functioning of the original tissue or organ. 1 The pathogenesis of keloids is complex and remains elusive. Multiple studies have suggested that skin color and tension, 2 immunity, 3 hormones, inflammatory stimulation, 4 genes 5 and other factors play critical roles in the onset and development of keloids. In addition to fibroblasts, immune cells that include macrophages, 6 mast cells 7 and lymphocytes 8 are also important to keloid pathogenesis via cytokine secretion and the triggering of abnormal signaling pathways. 9,10 Therefore, performing single-cell RNA-sEquation (scRNA-seq) that can identify heterogenous cell populations and cellular developmental pathways is thus valuable, as it will provide insight into the key pathogenic cell-types and the molecular patterns.In this study, we performed scRNA-seq on six pairs of keloid samples to elucidate the pathogenesis of skin fibrosis. Detailed methods and single-cell sequencing analysis are described in the Supplemental materials.We obtained lesion and non-lesion skin biopsies from six keloid patients (Figure 1A, Figure S1A,B, Table S1) and performed hematoxylin and eosin (H&E) and Masson staining (Figure 1B). Then we applied single-cell sequencing (scRNA-seq) to 12 skin samples that included both lesion and non-lesion areas (Figure 1C). After stringent quality-control procedures, 60,732 high-quality cells were obtained for further analysis. We first visualized all subgroups by uniform manifold approximation and projection (UMAP) and then determined the different cell types that included fibroblasts, keratinocytes, endothelial cells, T cells, pericytes, myeloid cells, mast cells, B cells, adipocytes, melanocytes and salivary gland cells (Figure 1C,D). All cell types were identified in each sample, and we observed minimal batch effects in our study (Figure S1C--F). Top-marked signatures for each cell typeThis is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Axillary osmidrosis (AO) and primary hyperhidrosis (PH) are common diseases, but there are still difficulties in treatment. Microwave therapy may become a new method. In order to evaluate long‐time efficacy of patients with AO or PH treated by microwave and to discuss possible mechanism of microwave therapy by combining results of clinical and pathological, the study was carried out. Ten AO or PH patients with moderate or severe level were selected as subjects, and each subject received microwave treatment of bilateral armpits. The follow‐up period lasted 2 years, and the changes of perspiration and odor were evaluated in subjective and objective ways. Each subject took skin biopsy in the treatment area before and after treatment or each follow‐up. Hematoxylin‐eosin and immunohistochemical staining were performed. Both subjective and objective index reflected the significant improvement of AO and PH after treatment (p < 0.05). Dermatology life quality index score decreased by 10.4 ± 4.6 (p < 0.05). The number of apocrine glands decreased significantly after treatment, and most of them changed from secretory phase to quiescent phase. In conclusion, microwave therapy can destroy apocrine sweat glands, reduce number of functional glands, so as to improve symptoms of AO and PH and elevate quality of life, which is safe, effective, and stable.

Systemic sclerosis (SSc) and keloid are typical skin fibrotic diseases with unclear epigenetic mechanisms and clinical targets, this study aimed to assess the role of miR-3606-3p in skin fibrosis and the therapeutic potential. MiR-3606-3p was reduced in the skin tissues and fibroblasts from both SSc and keloid patients. RNA-seq analysis and in silico prediction indicated GRB2 associated binding protein 1 (GAB1) and integrin subunit alpha V (ITGAV) were potential targets of miR-3606-3p. We then found that miR-3606-3p downregulated both GAB1 and ITGAV by directly targeting their 3′-UTRs, and further reduced p-AKT and p-ERK activities to inhibit collagen synthesis and fibroblast inflammation. Furthermore, miR-3606-3p inhibited fibroblast migration in primary fibroblasts and keloid-bearing nude mice by wound healing scratch assay and in vivo imaging techniques respectively. In contrast, GAB1 and ITGAV were upregulated in SSc and keloid patients, and siRNA-mediated GAB1 or ITGAV knockdown replicated the phenotypes observed in miR-3606-3p-overexpressing fibroblasts, including inflammation, migration and fibrogenesis. Finally, in vivo experiments showed that miR-3606-3p treatment significantly alleviates skin fibrosis in keloid-bearing mice. Our results indicated miR-3606-3p inhibits ECM deposition, inflammation, and migration of fibroblasts by downregulating GAB1 and ITGAV. miR-3606-3p-enhancing strategies may have beneficial effects on skin fibrosis through lowing p-AKT/p-ERK activity.