Table_1_Proximity to major roads and the incidence of osteoporotic fractures in elderly women: The BONE study in Beijing.DOCX

BackgroundThere is growing evidence to suggest that living near major roads (and suffering from the air pollution of urban streets) can have an adverse effect on bone health. However, little is known about its relationship to fractures caused by osteoporosis.ObjectiveThis study was designed to investigate the relationship between residents living near major roads and the incidence of osteoporotic fractures.MethodsA retrospective cohort of 529 subjects was established based on community populations in older women aged 65–91. All participants lived in Beijing between September 27, 2007 and September 26, 2017. The distance between the residential sites of the subjects and the main roads was determined by the authors. Osteoporotic fracture diagnosis was based on medical histories and imaging examinations (DXA and X-rays). The Cox proportional hazard model was used to assess the association between traffic proximity and osteoporotic fractures, with suitable adjustments for individual and background factors.ResultsThe age range of all participants was 65–91 years, with an average age of 75.8 years (and a standard deviation 6.8 years). Of these, 19 (3.59%) suffered from diabetes, and 48 (9%) had hypertension; 85 (14%) families had annual incomes below US $30,000 and 402 (76%) had received a secondary school education or higher. Nearly 25% of people lived within 50 m of a main road, while 50% lived within 300 m. Between 2007 and 2017, a total of 96 osteoporotic fractures were observed. For people living ConclusionIn this community-based cohort, living near a major road was associated with a higher incidence of osteoporotic fractures.</p

Additional file 1 of Genetic insights into the ‘sandwich fusion’ subtype of Klippel–Feil syndrome: novel FGFR2 mutations identified by 21 cases of whole-exome sequencing

Additional file 1: Fig. S1. Bioinformatics analysis workflow (the filtration strategy). Samples were initially mapped to the human reference genome GRCh37 (hg19) using BWA, followed by the removal of low-quality reads (< 80 bps) using CutAdapt and elimination of duplicate reads with Picard. Subsequently, SNP and INDEL variants were detected using GATK. In the third step, various software tools, including ANNOVAR, 1000Genome, ESP6500, dbSNP, ExAC, HGMD, ClinVar, were employed for sample annotation and further exclusion of deep intron variants. Comparative analysis with databases such as 1000Genome, ESP6500, and ExAC was performed to eliminate common SNPs. Pathogenicity prediction scores were determined using SIFT, Polyphen-2, MutationTaster, and Gerp++. GLUSTAL W and UGENE were utilized for cross-species comparison to ascertain sequence conservation, while ESE Finder 3.0 was employed to predict potential changes in protein structure. Finally, all suspicious variants underwent Sanger sequencing for further validation. Fig. S2. The animated diagram of FGFR2 (RCSB PDB number: 3B2T) and the arrowhead indicated the M584 site. A Ball-and-stick model of FGFR2; the sequence from 582 to 596 is shown in a dotted line. B Molecular surface model of FGFR2; the pink bulky region is P582. Mutation of M584V might result in a steric hindrance effect in the structural region of the protein and affect the folding of FGFR2 and reduce its catalytic capability. Table S1. Quality control of sequencing data

Comparison between immature ossification (imOS) and mature ossification (mOS) of TOLF.

(A) Micro-CT images of immature ossification (imOS) and mature ossification (mOS) of TOLF; (B) morphological parameters; * P< 0.05 compared with imOS; (C) Volcano plots showing the DEGs among imOS and OS; (D) Differential GO terms distribution.</p

DataSheet_1_Deciphering Obesity-Related Gene Clusters Unearths SOCS3 Immune Infiltrates and 5mC/m6A Modifiers in Ossification of Ligamentum Flavum Pathogenesis.docx

BackgroundOssification of ligamentum flavum (OLF) is an insidious and debilitating heterotopic ossifying disease with etiological heterogeneity and undefined pathogenesis. Obese individuals predispose to OLF, whereas the underlying connections between obesity phenotype and OLF pathomechanism are not fully understood. Therefore, this study aims to explore distinct obesity-related genes and their functional signatures in OLF.MethodsThe transcriptome sequencing data related to OLF were downloaded from the GSE106253 in the Gene Expression Omnibus (GEO) database. The obesity-related differentially expressed genes (ORDEGs) in OLF were screened, and functional and pathway enrichment analysis were applied for these genes. Furthermore, protein-protein interactions (PPI), module analysis, transcription factor enrichment analysis (TFEA), and experiment validation were used to identify hub ORDEGs. The immune infiltration landscape in OLF was depicted, and correlation analysis between core gene SOCS3 and OLF-related infiltrating immune cells (OIICs) as well as 5mC/m6A modifiers in OLF was constructed.ResultsNinety-nine ORDEGs were preliminarily identified, and functional annotations showed these genes were mainly involved in metabolism, inflammation, and immune-related biological functions and pathways. Integrative bioinformatic algorithms determined a crucial gene cluster associated with inflammatory/immune responses, such as TNF signaling pathway, JAK-STAT signaling pathway, and regulation of interferon-gamma-mediated signaling. Eight hub ORDEGs were validated, including 6 down-regulated genes (SOCS3, PPARG, ICAM-1, CCL2, MYC, and NT5E) and 2 up-regulated genes (PTGS2 and VEGFA). Furthermore, 14 differential OIICs were identified by ssGSEA and xCell, and SOCS3 was overlapped to be the core gene, which was associated with multiple immune infiltrates (dendritic cells, macrophage, and T cells) and six m6A modifiers as well as four 5mC regulators in OLF. Reduced SOCS3 and FTO expression and up-regulated DNMT1 level in OLF were validated by Western blotting.ConclusionThis study deciphered immune/inflammatory signatures of obesity-related gene clusters for the first time, and defined SOCS3 as one core gene. The crosstalk between 5mC/m6A methylation may be a key mediator of SOCS3 expression and immune infiltration. These findings will provide more insights into molecular mechanisms and therapeutic targets of obesity-related OLF.</p

Effects of ANGTP2 knockdown on notch signaling and osteogenic differentiation of ligamentum flavum cells.

(A) Effect of siANGPT2 on Notch1 and Notch2 expressions. ANGPT2, Notch1 and Notch2 expressions were assessed via qRT-PCR in mOS ligamentum flavum cells afterANGPT2 siRNA transfection (siANGPT2); * PPP< 0.05 compared with NC; (G) Effect of siNotch2 on ANGPT2 protein level.</p

ANGTP2 stimulation enhances notch signaling and osteogenic differentiation of ligamentum flavum cells.

(A) Effect of ANGPT2 on Notch1 and Notch2 expressions. Notch1 and Notch2 expressions were assessed via qRT-PCR in imOS ligamentum flavum cells with ANGPT2 stimulation (St.); * PP< 0.05 compared with Non-St; (D) Effect of ANGPT2 on protein levels of osteogenic markers; (E) ALP staining following ANGPT2 stimulation; scale bar represents 200 μm.</p

Gene activity comparison between immature ossification (imOS) and mature ossification (mOS) of TOLF.

(A) Expression level of genes in primary thoracic ligamentum flavum cells. Expressions of ANGPT2, Notch1 and Notch2 were assessed via qRT-PCR.* PP< 0.05 compared with imOS; (D) Osteogenic marker protein levels in primary thoracic ligamentum flavum cells via Western blot; (E) ALP staining of ligamentum flavum cells from imOS and mOS patterns; scale bar represents 200 μm.</p

Einstein Probe discovery of EP240408a: A peculiar X-ray transient with an intermediate timescale

We report the discovery of a peculiar X-ray transient, EP240408a, by Einstein Probe (EP) and follow-up studies made with EP, Swift, NICER, GROND, ATCA and other ground-based multiwavelength telescopes. The new transient was first detected with Wide-field X-ray Telescope (WXT) on board EP on April 8th, 2024, manifested in an intense yet brief X-ray flare lasting for 12 s. The flare reached a peak flux of 3.9 × 10−9 erg cm−2 s−1 in 0.5–4 keV, ∼300 times brighter than the underlying X-ray emission detected throughout the observation. Rapid and more precise follow-up observations by EP/FXT, Swift and NICER confirmed the finding of this new transient. Its X-ray spectrum is non-thermal in 0.5–10 keV, with apower-law photon index varying within 1.8–2.5. The X-ray light curve shows a plateau lasting for ∼4 d, followed by a steep decay till becoming undetectable ∼10 d after the initial detection. Based on its temporal property and constraints from previous EP observations, an unusual timescale in the range of 7–23 d is found for EP240408a, which is intermediate between the commonly found fast and long-term transients. No counterparts have been found in optical and near-infrared, with the earliest observation at 17 h after the initial X-ray detection, suggestive of intrinsically weak emission in these bands. We demonstrate that the remarkable properties of EP240408a are inconsistent with any of the transient types known so far, by comparison with, in particular, jetted tidal disruption events, gamma-ray bursts, X-ray binaries and fast blue optical transients. The nature of EP240408a thus remains an enigma. We suggest that EP240408a may represent a new type of transients with intermediate timescales of the order of ∼10 d. The detection and follow-ups of more of such objects are essential for revealing their origin.</p