Zhuang-Gu-Fang promotes osteoblast differentiation via myoblasts and myoblast-derived exosomal miRNAs:miR-5100, miR-126a-3p, miR-450b-5p, and miR-669a-5p.

BACKGROUND: Senile osteoporosis (SOP) is an age-related systemic metabolic bone disorder. Previous studies have proved that Zhuang-Gu-Fang (ZGF) modulates myokines, stimulates osteogenic differentiation, and mitigates osteoporosis. OBJECTIVE: To elucidate the mechanism by which ZGF promotes osteogenic differentiation via myoblast and myoblast exosomal microRNAs (miRNAs) and investigate its potential implications in senile osteoporosis. METHODS: Characterization of ZGF and ZGF serum using UHPLC-MS/MS. An alkaline phosphatase (ALP) activity assay and staining techniques were employed to corroborate the impacts of ZGF on the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) via myoblasts. Subsequently, exosomes derived from myoblasts were isolated through ultracentrifugation. The effects of ZGF on the BMSCs' osteogenic differentiation were substantiated through ALP activity, alizarin red staining, and a quantitative real-time polymerase reaction system (qRT-PCR). Selected miRNAs were identified via high-throughput sequencing and subjected to differential expression analysis, and subsequently validated through qRT-PCR. The senescence-accelerated (SAMP6) mice were selected as the SOP models. qRT-PCR analyses were further conducted to confirm the expression levels of these selected miRNAs in the muscle and bone tissues of the SAMP6 mice, and the protein expression of osteogenesis-related transcription factors OCN and Osterix in its bone tissue was evaluated by immunofluorescence staining analysis (IF). RESULTS: ZGF may enhance the osteogenic differentiation of BMSCs through myoblasts and myoblast-derived exosomes. High-throughput sequencing, differential expression analysis, and subsequent qRT-PCR validation identified four miRNAs that stood out due to their significant differential expression: miR-5100, miR-142a-3p, miR-126a-3p, miR-450b-5p and miR-669a-5p. Moreover, the mice experiment corroborated these findings, which revealed that ZGF not only up-regulated the expression of miR-5100, miR-450b-5p and miR-126a-3p in muscle and bone tissues but also concurrently down-regulated the expression of miR-669a-5p in these tissues. IF staining analysis indicated that ZGF can significantly increase the protein expression of the osteogenic transcription factors OCN and Osterix in the bone tissue of mice with SOP. CONCLUSIONS: ZGF can promote osteogenic differentiation of osteoblasts, regulate bone metabolism, and thereby delay the process of SOP. Perhaps, its mechanism is to upregulate myoblast-derived exosomes miR-5100, miR-126a-3p, and miR-450b-5p or downregulate miR-669a-5p. This study reports for the first time that myoblast exosomes miR-669a-5p and miR-450b-5p are novel targets for the regulation of osteoblastic differentiation and the treatment of SOP.

Research on Single-Event Burnout Reinforcement Structure of SiC MOSFET.

In this paper, the single-event burnout (SEB) and reinforcement structure of 1200 V SiC MOSFET (SG-SBD-MOSFET) with split gate and Schottky barrier diode (SBD) embedded were studied. The device structure was established using Sentaurus TCAD, and the transient current changes of single-event effect (SEE), SEB threshold voltage, as well as the regularity of electric field peak distribution transfer were studied when heavy ions were incident from different regions of the device. Based on SEE analysis of the new structural device, two reinforcement structure designs for SEB resistance were studied, namely the expansion of the P+ body contact area and the design of a multi-layer N-type interval buffer layer. Firstly, two reinforcement schemes for SEB were analyzed separately, and then comprehensive design and analysis were carried out. The results showed that the SEB threshold voltage of heavy ions incident from the N+ source region was increased by 16% when using the P+ body contact area extension alone; when the device is reinforced with a multi-layer N-type interval buffer layer alone, the SEB threshold voltage increases by 29%; the comprehensive use of the P+ body contact area expansion and a multi-layer N-type interval buffer layer reinforcement increased the SEB threshold voltage by 33%. Overall, the breakdown voltage of the reinforced device decreased from 1632.935 V to 1403.135 V, which can be seen as reducing the remaining redundant voltage to 17%. The device's performance was not significantly affected.