Ningbo University Affiliated Hospital

Understanding biological processes requires spatiotemporal mapping of proliferative and transcriptional dynamics. Current spatial transcriptomics methods capture only protein-coding transcripts and static snapshots, obscuring non-coding RNAs (ncRNAs) and dynamic events. We developed SPTEdU-seq, integrating spatial total transcriptomics with 5-ethynyl-2'-deoxyuridine tracking to co-profile gene expression and proliferation dynamics. SPTEdU-seq demonstrates ultrahigh sensitivity for coding and non-coding transcripts and for splicing isoforms, with single-molecule probe design eliminating optical imaging. Applied to developing and adult mouse brains, it revealed spatial lncRNA patterns, reconstructed developmental trajectories, and enabled spatiotemporal lineage tracing. In murine ischemic stroke, it mapped regeneration dynamics and identified an Igfbp5+ astrocyte subtype within a pro-repair niche. In mouse and human renal tumors, it uncovered tumor-associated splicing and detected diagnostic 3p loss. By profiling newborn and resident cells in intact microenvironments, it unveiled previously inaccessible interaction networks. SPTEdU-seq thus establishes a powerful framework for investigating cell fate dynamics in regeneration, development, and cancer.

Allergic rhinitis (AR) is a prevalent chronic inflammatory disorder of the upper airways, driven by Th2-mediated immune dysregulation and compromised epithelial barrier function. Current therapies, such as intranasal corticosteroids, often exhibit limited efficacy and adverse effects upon prolonged use. Herein, we developed a novel dual-action nasal spray, termed Mefp, by combining mussel adhesive protein (MAP) and xylitol to simultaneously target epithelial barrier repair and inflammatory pathways. The Mefp formulation exhibited strong mucoadhesive properties, with nasal retention lasting up to 24h, facilitated by catechol-mediated adhesion of 3,4-dihydroxyphenylalanine (DOPA) residues in MAP. In vitro experiments using LPS-stimulated human nasal epithelial cells (HNEpCs) demonstrated that Mefp significantly restored tight junction proteins (ZO-1 and occludin), suppressed NF-κB activation, reduced oxidative stress, and attenuated the release of pro-inflammatory (TNF-α, IL-6) and Th2 cytokines (IL-4, IL-5, IL-13). In an ovalbumin-induced murine AR model, Mefp treatment markedly alleviated allergic symptoms, reduced eosinophil infiltration, decreased OVA-specific IgE levels, and enhanced antioxidant enzyme activities. Histological and immunofluorescence analyses confirmed the restoration of mucosal integrity and tight junction assembly. Notably, Mefp demonstrated comparable or superior efficacy to mometasone furoate (MF) in alleviating allergic symptoms and suppressing key inflammatory mediators, while showing a better safety profile regarding epithelial integrity in vitro. These findings indicate that Mefp effectively addresses both epithelial barrier dysfunction and type 2 inflammation, offering a promising and safe therapeutic strategy for AR.

Abstract As a pivotal aromatic precursor for phytocannabinoids, olivetolic acid (OLA) is traditionally sourced from resource-intensive plant extraction or complex chemical synthesis. In this study, we established a sustainable photosynthetic production platform by engineering the model cyanobacterium Synechocystis sp. PCC 6803. We successfully reconstituted the OLA biosynthetic pathway by heterologously expressing Cannabis sativa-derived tetraketide synthase (CsTKS), olivetolic acid cyclase (CsOAC), and acyl-activating enzyme (CsAAE1). To overcome metabolic bottlenecks, the pathway was optimized using the strong promoter P cpc560 and carbon flux redirection. Disruption of the glycogen storage pathway and reinforcement of the malonyl-CoA pool via maeB overexpression significantly enhanced precursor availability. Furthermore, increasing CO 2 supplementation achieved a maximum OLA titer of 9.41 mg/L. This work demonstrates the feasibility of utilizing photosynthetic chassis for the production of olivetolic acid, providing a robust foundation for the sustainable manufacturing of cannabinoid-related metabolites.

To meet the requirements for remote optical thermometry, Er3+- and Ho3+-doped Cs2NaYbCl6 double perovskites with intense upconversion (UC) emissions were designed. Excited by 980 nm, characteristic emissions of Er3+ and Ho3+ were detected, with optimal doping concentrations of 30 and 50 mol % in the Cs2NaYbCl6 host, respectively. The near-infrared light-triggered UC luminescence mechanism was investigated through the pump power-dependent UC emission spectra. Moreover, using the fluorescence intensity ratio (FIR) technique, temperature sensing performance was explored by analyzing the UC emission evolution from thermally coupled levels (TCLs) and non-TCLs of rare-earth ions. Based on the TCLs of Er3+, the maximum relative sensitivity (Sr) of the resulting compound reached 1.15% K–1, while it declined to 0.43% K–1 when the non-TCLs of Er3+ were employed, featuring an ultrabroad operating range of 303–813 K, demonstrating that the selection of a proper sensing strategy is an efficient route to regulate the thermometric properties of the studied samples. Furthermore, when the non-TCLs of Ho3+ were used, the target material exhibited a maximum Sr value of 1.12% K–1, which is dependent on the sensing mode, and its working temperature range was 303–543 K. The distinct Sr values confirmed that doping engineering effectively tailors the thermometric behavior. This work demonstrates that rare-earth-ion-doped Cs2NaYbCl6 double perovskites are promising for optical thermometry, with tunable thermometric capabilities via the dual engineering of doping and sensing strategies.

Lung adenocarcinoma (LUAD) is a major subtype of lung cancer that frequently develops brain metastases. Tumor heterogeneity substantially complicates disease management, and reliable biomarkers for metastatic progression remain lacking. LIM domain kinase 1 (LIMK1), a key regulator of cytoskeletal dynamics, has been implicated in tumor progression; however, its role in regulating brain metastasis in LUAD remains unclear. Single-cell RNA sequencing data from 11 primary LUAD tumors and 10 brain metastases were analyzed to identify malignant epithelial subclusters associated with metastatic dissemination. Tumor evolutionary trajectories and gene regulatory networks were explored using pseudotime trajectory analysis and weighted gene co-expression network analysis. LIMK1 expression was validated using The Cancer Genome Atlas dataset, as well as RT–qPCR and Western blotting in LUAD tissues and cell lines. Functional assays, including cell proliferation, colony formation, migration, and invasion, were performed following LIMK1 knockdown in LUAD cell lines. Six malignant epithelial subclusters were identified, among which clusters 0 and 4 were significantly enriched in brain metastases and associated with poor survival outcomes. LIMK1 was identified as a hub gene within these metastasis-associated clusters. LIMK1 expression was markedly upregulated in LUAD tissues and cell lines, and elevated LIMK1 levels predicted an unfavorable prognosis. Functional experiments demonstrated that LIMK1 depletion significantly inhibited tumor cell proliferation, migration, and invasion, while partially reversing epithelial–mesenchymal transition (EMT). Mechanistically, LIMK1 was found to promote LUAD progression through activation of the PI3K/AKT/mTOR signaling pathway. LIMK1 represents a novel prognostic biomarker and metastasis-associated regulator in LUAD. By driving EMT and activating PI3K/AKT/mTOR signaling, LIMK1 facilitates malignant progression, highlighting its potential as a therapeutic target for LUAD patients with brain metastases.

The concurrent use of serotonergic antidepressants (SSRIs, SNRIs) and opioid analgesics is frequent in the perioperative setting. While this combination carries a known risk of serotonin syndrome, large-scale epidemiological evidence remains scarce. We performed a multinational pharmacovigilance study using data from the US FDA Adverse Event Reporting System (FAERS) and the Japanese Adverse Drug Event Report database (JADER) from Q1 2004 to Q2 2025. Disproportionality analyses, including Reporting Odds Ratio (ROR) with a multi-algorithm framework, were employed to quantify signals for serotonin syndrome and other adverse events. We further characterized clinical profiles and conducted gender-stratified and time-to-onset analyses. Opioids (e.g., tramadol, fentanyl, oxycodone) were consistently among the top drugs associated with serotonin syndrome reports. Concomitant SSRI/SNRI-opioid use showed exceptionally strong signals for serotonin syndrome (SSRI-opioid ROR 95.94, 95% CI 88.9-103.53; SNRI-opioid ROR 57.68, 95% CI 52.32-63.59). SSRI-opioid combinations were uniquely associated with cardiac disorders (ROR 1.87, 95% CI 1.77-1.98), whereas SNRI-opioid use was linked to drug withdrawal syndrome. Gender-stratified analysis indicated a higher reporting proportion for serotonin syndrome in males prescribed SSRI-opioids. Notably, up to 30% of adverse events occurred more than 300 days after therapy initiation, indicating a persistent risk. This study provides robust multinational evidence that concomitant serotonergic antidepressant and opioid use is strongly associated with a high risk of serotonin syndrome and distinct adverse event profiles. These findings highlight the urgent need for systematic preoperative medication review, tailored perioperative monitoring, and sustained clinical vigilance.

Background: The value of radical prostatectomy (RP) in large volume and unresectable locally advanced prostate cancer (LAPC) or oligometastatic prostate cancer (OMPC) is largely unknown. This retrospective study aims to evaluate the conversion efficacy and safety profile of preoperative androgen deprivation therapy combined with docetaxel (A + D) in patients with large-volume (≥60 mL) and initially unresectable LAPC or OMPC. Methods: Patients with LAPC or OMPC from three large tertiary medical centers (between April 2017 and October 2024) were included. Eligibility criteria included prostate volume ≥60 mL, unresectability, and an expected survival of more than 5 years, as confirmed by two senior urologists and one radiologist. All patients received preoperative A + D therapy (≥3 cycles), followed by reassessment for RP feasibility with multimodal therapy discussion. The primary endpoint was surgical conversion rate. Secondary endpoints included prostate-specific antigen (PSA) progression-free survival (PSA-PFS), overall survival (OS), PSA response rate, adverse events, pathological response, perioperative complications, urinary continence, and sexual function. Results: Among 31 enrolled patients, the mean age was 67.71 ± 6.69 years, and 29 (93.55%) successfully underwent RP. The mean follow-up duration was 42.83 ± 23.58 months (up to October 2024). Median PSA-PFS was 49 months (95% CI: 24.20–73.79 months), and median OS was not reached. Hematologic toxicity was the most common adverse event during chemotherapy. After A + D therapy, median prostate tumor volume decreased from 67.50 mL to 25.81 mL, and median PSA declined from 168 to 0.49 ng/mL. After RP, 11 patients (37.93%) experienced a downgrade in Gleason score, and 14 patients (48.28%) experienced a downgrade in T-stage. Positive surgical margins were found in 11 patients (37.93%). Perioperative complications included blood transfusion (11.11%), rectal injury (3.70%), lymphorrhea (7.41%), and fever >38.5°C (3.70%). The 1-year urinary continence rate was 89.66%, while 100% of patients experienced sexual dysfunction. Conclusion: Preliminary data suggest that preoperative A + D therapy followed by RP may be integrated into multimodal management for large-volume, initially unresectable LAPC and OMPC, demonstrating promising conversion efficacy and clinical outcomes.