Observing the clinical efficacy of combined serum microneedle therapy for moderate to severe androgenetic alopecia in scalp repair.

OBJECTIVE: In this study, the safety and efficacy of scalp repair serum microneedles combined with oral drug administration and topical medication were investigated for the treatment of moderate to severe androgenetic alopecia. METHODS: Twenty patients, consisting of 4 males and 16 females, who sought treatment for moderate to severe androgenetic alopecia at our hair medicine research center alopecia specialty clinic between August and December 2022 were randomly selected for the study. Male patients underwent oral administration of finasteride topical application of 5% minoxidil, and biweekly scalp repair serum microneedle therapy. Female patients were administered spironolactone or Diane-35 orally and applied 2% minoxidil topically, paired with biweekly scalp repair serum microneedle therapy sessions. After seven treatments, the scalp repair serum microneedle was discontinued, but oral administration and topical applications were continued, followed by a 1-month follow-up. Using a hair dermoscopy, hair follicles in a fixed region on the top of the head were manually counted per unit area to evaluate the hair restoration status of the patients quantitatively. RESULTS: All 20 patients completed 3 months of combined therapy and a 1-month follow-up. On average, the patients experienced an increase of 42.6 hairs, with an efficiency rate of 100%. Significant differences were observed in hair count between any two of the first seven treatments (p < 0.001). A significant negative correlation was discovered between the initial pre-treatment hair count and the total improvement of hair (p < 0.001), indicating that the greater the degree of hair loss before treatment, the more pronounced the improvement. CONCLUSION: Scalp repair serum microneedle combined therapy in moderate to severe androgenetic alopecia significantly reduces the number of microneedle treatments required, enhances treatment efficacy, and improves therapeutic outcomes.

Cloning and Direct Evolution of a Novel 7-O-Glycosyltransferase from Cucurbita moschata and Its Application in the Efficient Biocatalytic Synthesis of Luteolin-7-O-glucoside.

Luteolin-7-O-glucoside(L7G), a glycosylation product of luteolin, is present in a variety of foods, vegetables, and medicinal herbs and is commonly used in dietary supplements due to its health benefits. Meanwhile, luteolin-7-O-glucoside is an indicator component for the quality control of honeysuckle in the pharmacopoeia. However, its low content in plants has hindered its use in animal pharmacological studies and clinical practice. In this study, a novel 7-O-glycosyltransferase CmGT from Cucurbita moschata was cloned, which could efficiently convert luteolin into luteolin-7-O-glucoside under optimal conditions (40 °C and pH 8.5). To further improve the catalytic efficiency of CmGT, a 3D structure of CmGT was constructed, and directed evolution was performed. The mutant CmGT-S16A-T80W was obtained by using alanine scanning and iterative saturation mutagenesis. This mutant exhibited a kcat/Km value of 772 s-1·M-1, which was 3.16-fold of the wild-type enzyme CmGT. Finally, by introducing a soluble tag and UDPG synthesis pathway, the strain BXC was able to convert 1.25 g/L of luteolin into 1.91 g/L of luteolin-7-O-glucoside under optimal conditions, achieving a molar conversion rate of 96% and a space-time yield of 27.08 mg/L/h. This study provides an efficient method for the biosynthesis of luteolin-7-O-glucoside, which holds broad application prospects in the food and pharmaceutical industry.

Efficient and green production of flavone-5-O-glycosides by glycosyltransferases in Escherichia coli.

O-Glycosylflavonoids exhibit diverse biological activities but their low content in plants is difficult to extract and isolate, and chemical synthesis steps are cumbersome, which are harmful to the environment. Therefore, the biosynthesis of O-glycosylflavonoids represents a green and sustainable alternative strategy, with glycosyltransferases playing a crucial role in this process. However, there are few studies on flavone 5-O-glycosyltransferases, which limits the synthesis of rare flavone 5-O glycosides by microorganisms. In this study, we characterized a highly regioselectivity flavone 5-O glycosyltransferase from Panicum hallii. Site-directed mutagenesis at residue P141 switches glucosylation to xylosylation. Using a combinatorial strategy of metabolic engineering, we generated a series of Escherichia coli recombinant strains to biocatalyze glycosylation of the typical flavone apigenin. Ultimately, further optimization of transformation conditions, apigenin-5-O-glucoside and apigenin-5-O-xyloside were biosynthesized for the first time so far, and the yields were 1490 mg/L and 1210 mg/L, respectively. This study provides a biotechnological component for the biosynthesis of flavone-5-O-glycosides, and established a green and sustainable approach for the industrial production of high-value O-glycosylflavones by engineering, which lays a foundation for their further development and application in food and pharmaceutical fields.