PIP5k1β is crucial to the generation of phosphotidylinosotol (4, 5)P₂. PIP5k1β participates in numerous cellular activities, such as B cell and platelet activation, cell phagocytosis and endocytosis, cell apoptosis, and cytoskeletal organization. In the present work, we aimed to examine the function of PIP5k1β in osteoclastogenesis and osteogenesis to provide promising strategies for osteoporosis prevention and treatment. We discovered that PIP5k1β deletion in mice resulted in obvious bone loss and that PIP5k1β was highly expressed during both osteoclast and osteoblast differentiation. Deletion of the gene was found to enhance the proliferation and migration of bone marrow-derived macrophage-like cells to promote osteoclast differentiation. PIP5k1β^−/− osteoclasts exhibited normal cytoskeleton architecture but stronger resorption activity. PIP5k1β deficiency also promoted activation of mitogen-activated kinase and Akt signaling, enhanced TRAF6 and c-Fos expression, facilitated the expression and nuclear translocation of NFATC1, and upregulated Grb2 expression, thereby accelerating osteoclast differentiation and function. Finally, PIP5k1β enhanced osteoblast differentiation by upregulating master gene expression through triggering smad1/5/8 signaling. Therefore, PIP5k1β modulates bone homeostasis and remodeling.

Objective: To develop a novel non-invasive CRISPR/Cas12a-RCA assay for the primary screening of human echinococcosis via detection of circulating Echinococcus cell-free DNA (cfDNA) in peripheral blood. Methods: Plasma cfDNA from 20 AE patients was analyzed via high-throughput sequencing to identify conserved repetitive Echinococcus fragments.A one-pot RCA system coupled with CRISPR/Cas12a was optimized for Echinococcus -cfDNA detection. The limit of detection (LOD) was determined using serially diluted synthetic standards, while specificity was validated through mismatch probes and cross-reactivity testing. Clinical validation included 50 AE cases, 22 cystic echinococcosis (CE) cases, 43 non- Echinococcus hepatic disease (HD) cases, and 53 healthy controls (CON). Results: A conserved repetitive 28S rDNA fragment (pan- Echinococcus -28S) was identified as a biomarker. The CRISPR/Cas12a-RCA assay achieved amplification within 30 minutes at 37 °C, with a linear range of 1 aM to 100 pM and an LOD of 1.41 aM. Visual detection limits were 10 aM (UV light) and 1 aM (blue light). The assay demonstrated high sensitivity (87.5%) and specificity (96.9%, AUC = 0.933) in distinguishing Echinococcus infection (AE/CE) from HD and CON. Conclusion: The optimized one-pot CRISPR/Cas12a-RCA system enables rapid and ultrasensitive detection of pan- Echinococcus cfDNA, providing a cost-effective and highly accurate solution for the primary screening of echinococcosis.

Black goji berry ( Lycium ruthenicum Murr.) has been recognized for its immune-regulating and anti-cancer potentials, but its key active components and underlying mechanisms remain unclear. Here, we applied LC–MS/MS to characterize the metabolite profile of L. ruthenicum and performed in vitro assays to evaluate its anti-inflammatory and anti-cancer activities. A total of 149 metabolites were identified, with organic acids, carbohydrates, and amino acid derivatives as the dominant components. Isorhamnetin was identified as a representative flavonoid. In LPS-stimulated Calu3 cells, isorhamnetin reduced the secretion and mRNA expression of pro-inflammatory cytokines (IL-6, TNF-alpha, IL-1beta). In cancer cells, isorhamnetin induces ROS accumulation, DNA damage, mitochondrial dysfunction, and caspase-3-dependent apoptosis, with modulation of the AKT signaling pathway. These findings suggest that isorhamnetin may contribute to the anti-inflammatory and anti-cancer effects of L. ruthenicum observed in vitro , providing preliminary experimental evidence for its potential as a functional food ingredient. However, this study is limited by the use of a single cell line (Calu3) and the absence of in vivo validation and pharmacokinetic evaluation; further studies are needed to confirm these observations.

Introduction:Age-related bone loss is closely linked to inflammaging in the bone marrow microenvironment, but the molecular factors connecting immune remodeling to bone metabolic imbalance remain incompletely defined. This study aimed to identify aging-associated genes related to inflammatory remodeling in the bone marrow and to provide preliminary cellular evidence for a prioritized candidate.Methods:Public bulk RNA-seq (GSE132040) and single-cell RNA-seq (GSE132042) datasets from physiologically aging mice were analyzed using an integrated workflow. Differentially expressed genes (DEGs) were identified from bulk RNA-seq data, followed by Gene Ontology (GO) and Gene Set Enrichment Analysis (GSEA). Candidate genes were prioritized using least absolute shrinkage and selection operator (LASSO) regression and support vector machine-recursive feature elimination (SVM-RFE). Single-cell RNA-seq was used to examine the cellular distribution of the candidate gene within bone marrow cell populations. For cellular validation, Ifi27l2a was knocked down in RAW264.7 cells using lentiviral shRNA, followed by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and transcriptomic analysis. Primary bone marrow-derived macrophages (BMMs) were further used for TRAP staining-based assessment under M-CSF/RANKL-induced osteoclastogenic conditions.Results:A total of 313 DEGs were identified between young and aged bone marrow samples, with enrichment mainly in antiviral, host-defense, and immune-related processes. Integrated LASSO and SVM-RFE screening prioritized Ifi27l2a for further analysis. Whole-bone-marrow bulk RNA-seq analysis showed an age-associated decline in Ifi27l2a expression, whereas single-cell analysis showed a monocyte-enriched distribution. In RAW264.7 cells, Ifi27l2a knockdown reduced Il6 and Tnf expression and was accompanied by broader transcriptomic remodeling involving inflammatory programs and osteoclast-related transcriptional changes. Pathway analysis showed significant positive enrichment of the rheumatoid arthritis pathway and suggestive enrichment of the osteoclast differentiation pathway. In primary BMMs, TRAP staining showed that Ifi27l2a knockdown was associated with altered osteoclast-like morphology under M-CSF/RANKL stimulation.Conclusion:These findings identify Ifi27l2a as a bone marrow aging-associated candidate gene with monocyte-enriched expression and a potential link to inflammatory and osteoclast-lineage remodeling. The current study supports an association between Ifi27l2a reduction, macrophage-lineage transcriptional remodeling, and osteoclast-like morphological changes, rather than definitive causality. Further in vivo validation of bone phenotypes and pathway-level studies are needed to clarify its role in age-related bone remodeling.

PIP5K1β is crucial to generation of phosphotidylinosotol (4, 5) P ₂ . PIP5K1β participates in numerous cellular activities, such as B cell and platelet activation, cell phagocytosis and endocytosis, cell apoptosis, and cytoskeletal organization. In the present work, we aimed to make insight into the function of PIP5K1β in osteoclastogenesis and osteogenesis to provide promising strategies for osteoporosis prevention and treatment. We discovered that PIP5k1β deletion in mice resulted in obvious bone loss and PIP5K1β was highly expressed both during osteoclast and osteoblast differentiation, besides, PIP5K1β deletion enhanced the proliferation and migration of BMMs to promote osteoclast differentiation. PIP5k1β ^−/− osteoclast exhibited normal cytoskeleton architecture but stronger resorption activity. PIP5k1β deficiency also promoted activation of MAPK and Akt signaling, enhanced TRAF6 and c-Fos expression, facilitated the expression and nuclear translocation of NFATC1 and upregulated Grb2 expression, thereby accelerating osteoclast differentiation and function. Finally, PIP5K1β enhanced osteoblast differentiation by upregulating master genes expression through triggering smad1/5/8 signaling. Thereby, PIP5K1β modulate bone homeostasis and remodeling.