Longgang Central Hospital

Periodontitis can impair the osteogenic function of periodontal ligament stem cells (PDLSCs), thereby compromising their capacity for periodontal tissue regeneration. In this study, we explored the impact of a synthetic small molecule, DS96432529 (DS), on the osteogenic differentiation potential of PDLSCs and its underlying mechanism. The viability of DS was assessed by cell proliferation assays and apoptosis analysis. Osteogenic potential was evaluated through alkaline phosphatase (ALP) activity staining and Alizarin Red S (ARS) staining for mineralized nodule formation. Inflammatory injury was induced using recombinant tumor necrosis factor-alpha (TNF-α). RNA sequencing analyzed signaling pathways involved in DS-enhanced osteogenic differentiation. Western blotting quantified key pathway protein expression. Specific small molecule inhibitors and agonists modulated relevant signaling pathways. Therapeutic efficacy was evaluated in a ligature-induced rat periodontitis model. DS inhibited cell proliferation at lower concentrations but did not induce significant apoptosis at concentrations up to 250 nM. Across tested concentrations, DS significantly enhanced ALP activity and accelerated mineralized nodule formation in PDLSCs. DS upregulated mitophagy-related protein expression under both inflammatory and non-inflammatory conditions. Additionally, DS restored TNF-α-inhibited ALP activity and attenuated TNF-α-induced activation of the RIG-I-like receptor (RLR) signaling pathway. The RIG-I activator Poly(I: C) counteracted DS-mediated repair of inflammatory injury during osteogenesis. Mitophagy inhibition diminished DS's beneficial effects on osteogenic differentiation under inflammation and reduced its suppression of RIG-I expression. DS alleviated ligation-induced alveolar bone loss in rats with periodontitis. DS enhances the osteogenic potential of PDLSCs in association with the activation of mitophagy-related processes. It mitigates inflammation-impaired osteogenesis, potentially via modulation of the RIG-I-mediated RLR signaling pathway, in association with increased mitophagy-related activity. DS represents a potent therapeutic small molecule for ameliorating periodontitis-induced bone loss.

Artificial intelligence has transformed vision and audition, but olfaction remains difficult to formalize because smell lacks a consensus perceptual coordinate system and similar scent targets can be reached through multiple molecular routes. Here we present DeepClaw, an automated workflow system built on the OpenClaw agent substrate that converts receptor-grounded odor digitization into an end-to-end pipeline for perfume design, physical synthesis, and expert evaluation. Each molecule is represented as a 409-dimensional vector of receptor-odorant interaction proxy scores derived from AlphaFold 3-supported structural scoring across 409 human olfactory receptors, capturing receptor engagement rather than statistical labels. DeepClaw coordinates three task-specific agent roles-Formulator, Receptor modeler, and Experimenterthrough a shared artifact workspace linked to a precomputed knowledge base of 4,000 annotated odorants and 1.636 million interaction proxy scores, on-demand structural computation, and automated synthesis equipment. In a proof-of-concept study, DeepClaw formulated and synthesized a kapok-character perfume that was confirmed by blind evaluation from ten trained perfumers. These results show that a generalpurpose agent substrate can be extended into a domain-specific closed-loop system for receptor-grounded fragrance research.

Despite the use of lipid-lowering and anti-inflammatory treatments, the progression of atherosclerosis is relentless in most patients. This suggests the presence of in situ pathological factors that continuously exacerbate lesions. We hypothesized that extracellular vesicles (EVs) within the atherosclerotic microenvironment might act as in situ stimulatory factors on vascular smooth muscle cells (VSMCs), thereby exacerbating atherosclerosis. A local atherosclerosis model was induced using Ldlr knockout (Ldlr KO) rats fed a high-cholesterol diet and subjected to partial carotid ligation. Immunofluorescence, Western blot (WB), and single-cell sequencing confirmed the phenotypic switching of VSMCs in atherosclerotic plaques from both rats and humans. The phenotypic switching of VSMCs in atherosclerotic rats was characterized by reduced expression of VSMC contraction markers and increased expression of LGALS3, PDGFRB, and SCA1. GW4869 inhibited the phenotypic switching of VSMCs in atherosclerotic plaques in a rat model. EVs were extracted from atherosclerotic carotid tissues using differential centrifugation. Chitosan thermosensitive hydrogels were used for in situ delivery of EVs into the arterial wall of the carotid artery. Immunofluorescence staining revealed that atherosclerotic plaque-derived EVs (AS-EVs) promoted VSMC phenotypic switching and downregulated the expression of VSMC contractile markers in vitro. miRNA analysis of EVs derived from atherosclerotic plaques of rats identified miR-23a-3p and its target gene Myl12b. To investigate the underlying mechanisms, engineered EVs loaded with miR-23a-3p were used to mimic AS-EVs. AS-EVs potentially regulate MRTFA nuclear translocation via miR-23a-3p/Myl12b, inhibit contractile marker expression, promote VSMC phenotypic switching, and further promote carotid artery remodelling. We conclude that AS-EVs promote VSMC phenotypic switching and exacerbate atherosclerosis.

BackgroundDysbiosis of gut microbiota and increased intestinal permeability are associated with various diseases, and their relationship with cognitive dysfunction such as cerebral small vessel disease (CSVD) and Alzheimer's disease remains to be elucidated.ObjectiveThis study investigates the role of gut microbiota dysbiosis and its relationship with intestinal permeability in patients with cognitive impairment associated with CSVD (CSVD-CI).MethodsIntestinal permeability was detected in 21 patients with CSVD-CI and 20 healthy controls by testing urine lactulose/mannitol, and correlation analysis was performed between the test results and cognitive function assessment. 16S rRNA sequencing was used to analyze the different combination of gut microbiota. Feces of the patients or controls were gavaged into C57 mice, and gut barrier function, behavior, and metabolites were assessed.ResultsPatients with CSVD-CI have a higher incidence of hypertension, higher homocysteine levels, higher scores for white matter hyperintensities, and worse cognitive function. Their urinary mannitol recovery rate is higher, which is correlated with lower scores of cognitive function assessment. Alterations in the gut microbiota involve a reduction in Prevotella-9 alongside increases in Proteobacteria and Fusobacteria. Fecal microbiota transplantation (FMT) from patients with CSVD-CI increases intestinal permeability in mice, but does not change their cognitive function; meanwhile, fecal metabolomics analysis has identified alterations in bile acids and vitamins, which are associated with shifts in the gut microbiota.ConclusionsPatients with CSVD-CI have gut microbiota imbalance and increased intestinal permeability, which are associated with cognitive decline. FMT from these patients can cause intestinal leakage and the production of harmful metabolites in mice.