Tibet University

Wuweiganlu (WGL) is a traditional formulation widely applied in the treatment of rheumatoid arthritis (RA), yet the identity of its bioactive constituents remains inadequately defined. In this study, ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and untargeted serum metabolomics were employed to characterize the active components of WGL. Fraxin was identified as a principal compound from WGL. To investigate its therapeutic mechanism in RA, a series of in silico and experimental approaches were conducted. Network pharmacology analysis and RNA sequencing identified heat shock protein family member 8 (HSPA8) as a potential molecular target of Fraxin, which was further validated by molecular docking studies. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that Fraxin exerts its effects primarily by modulating cell apoptosis through the PI3K signaling pathway. In vitro experiments demonstrated that Fraxin significantly reduced inflammatory responses and downregulated HSPA8 expression in lipopolysaccharide (LPS)-stimulated fibroblast-like synoviocytes (FLs) and macrophages. In vivo, Fraxin administration markedly reduced paw swelling, alleviated bone deformities, and improved bone volume fraction (BV/TV) in male IL1RA-deficient mice exhibiting spontaneous arthritis. Histological analysis confirmed that Fraxin attenuated joint inflammation by modulating the inflammatory microenvironment. Additionally, Fraxin inhibited synovial hyperplasia by regulating mitochondrial membrane potential collapse in FLs. Functional assays revealed that this regulation occurred via the inhibition of HSPA8/PI3K/AKT signaling axis, thereby suppressing aberrant FLS proliferation and contributing to the attenuation of RA progression.

Grain-scale heterogeneity strongly influences hydraulic fracture initiation and trajectory in crystalline rocks, yet its contributions to non-planar growth and the interaction of multiple nearby cracks remain insufficiently quantified. To address this gap, we perform numerical experiments on a model containing two parallel pre-existing cracks using a hydro-mechanical phase-field framework, systematically quantifying how mineral distribution and axial compression govern non-planar hydraulic fracture growth and inter-fracture competition. The results demonstrate that mineral distribution is the primary driver of fracture complexity. Even within the same Voronoi tessellation, redistributing minerals alone yields markedly different trajectories, deflections, branching patterns, and final morphologies. Furthermore, non-planar growth follows a stepwise, energy-threshold-driven mechanism. When cracks penetrate strong grains or undergo large-angle deflections, propagation is impeded, and injection pressure builds up. Once a critical energy threshold is reached, accumulated energy is rapidly released along the path of minimum incremental energy, manifested as abrupt pressure drops and rapid crack advance. Additionally, the two nearby fractures exhibit strong mechanical competition. Despite negligible hydraulic interference in low-permeability granite, early growth of one fracture redistributes stresses and suppresses the driving force of the other, resulting in asymmetric development. Finally, axial compression primarily governs the overall propagation orientation and influences local failure modes but has a limited effect on peak pressure relative to mineral distribution.

Drying significantly influences the quality of micellar casein (MC) powder. This study investigated the effects of three pre-freezing temperatures (-20 °C, -80 °C, and liquid nitrogen) prior to freeze drying on the structure and rennet-induced gelation properties of MC powder. The results showed that as the pretreatment temperature decreased, the degree of disruption to the secondary and tertiary protein structures was reduced, while the particle size gradually increased. In terms of rennet-induced gel properties, the untreated raw MC consistently outperformed all MC powder samples. Among the MC powders, the sample pre-frozen at -80 °C and then freeze-dried (FD-80) exhibited the highest gel strength and a relatively shorter rennet coagulation time. The observed microstructures of the rennet-induced gel were consistent with the rheological results, showing that samples with smaller particle sizes formed more regular and compact gel networks. In conclusion, the MC powder prepared via pre-freezing at -80 °C and then freeze-drying better preserved protein structure and demonstrated superior rennet-induced gel properties, which were closely related to particle size. This study provides theoretical insights for the application of MC powder in products such as cheese, processed cheese, and protein-fortified foods.