Strategies that concurrently reactivate latent reservoirs and enhance immune-mediated clearance hold significant promise for achieving an HIV cure. Here, we developed hyperIL-15×sCD4-Fc (15×sCD4-Fc), a fusion protein that integrates latency reactivation, targeted immune engagement, and effector-mediated killing into a single molecule. This agent not only potently reverses HIV-1 latency in CD4+ T cells from people living with HIV-1 (PLWH) through coordinated IL-15 receptor signaling and sCD4-mediated HIV-1 envelope (Env) engagement, but also enhances antigen-specific CD8+ T cell response in PBMCs derived from PLWH. Furthermore, 15×sCD4-Fc enables Env-specific elimination of reactivated latently infected cells by NK cells while preventing off-target cytotoxicity. In PBMCs from PLWH, 15×sCD4-Fc reduced replication-competent HIV-1 DNA by 93.8%. In antiretroviral treated HIV-1-infected humanized mice, the molecule demonstrated both safety and efficacy in diminishing viral reservoir in lymphoid organs. This spatiotemporally coupled approach to reservoir exposure and immune recognition establishes a clinically viable strategy for clearing the HIV-1 reservoir.
This study evaluated the efficacy and safety of a sedation protocol through the nares combining dexmedetomidine with midazolam and butorphanol in cockatiels. Six adults were enrolled in a randomized, 4 treatment, 4 period crossover trial comparing a control regimen (midazolam 3 mg/kg and butorphanol 3 mg/kg) with three combinations including dexmedetomidine at 0.04, 0.06, or 0.08 mg/kg, administered through the nares. The volume of all treatments was standardized to the volume of the highest dose of dexmedetomidine administered (2 mL/kg) with sterile saline solution. Sedation scores, respiratory rate, heart rate, and cloacal temperature were recorded for 10 min after drug administration and during 15 min of manual restraint, followed by flumazenil and atipamezole via the nares for reversal. All birds completed all treatments; mild sneezing occurred but no clinically relevant respiratory complications were observed. Medium and high dose dexmedetomidine produced higher sedation scores and lower respiratory rate, heart rate, and body temperature during restraint than the control regimen, whereas the control group showed increased respiratory rate, suggesting insufficient suppression of restraint induced stress. In all groups, sedation and physiological variables returned to near baseline values within 15 min after reversal, and no serious adverse events occurred. These findings indicate that administration through the nares dexmedetomidine at 0.06 or 0.08 mg/kg combined with midazolam and butorphanol provides deeper, more stable, and reversible sedation and improved control of stress responses in cockatiels and may represent a clinically useful protocol in this species.
Outcome assessment after claw hand correction is inconsistent, with prior reports relying on heterogeneous combinations of joint-angle measurements, photographs, grip testing, and qualitative functional grading. We developed the Contralateral Hand Function Score (CHFS), a simple contralateral hand-referenced composite measure, and evaluated its reproducibility and construct validity. We performed a retrospective study of 51 patients who underwent surgical correction of claw hand between 2009 and 2025. CHFS was calculated as the mean of three equally weighted domains-motion, strength, and function-each expressed as a percentage of the contralateral normal hand. A separate modifier system (CHFS-M) recorded claw-specific contextual determinants of residual impairment but was not included in score calculation. Validation included interobserver reproducibility, interdomain coherence, and predefined known-group comparisons. Cohort characteristics, outcome distribution, and validation metrics were summarized from the study dataset. Mean age was 41.8 ± 12.6 years and mean follow-up was 18.6 ± 7.2 months. Mean CHFS was 66.2 ± 18.4%, with 24% excellent, 55% good, and 21% poor outcomes. Interobserver reproducibility was excellent for the final CHFS (intraclass correlation coefficient, 0.994) and for all component domains (>0.97). Cronbach's alpha was 0.984 for the three domains and 0.932 for the function subitems. CHFS differed significantly by claw type, contracture status, and surgical technique. The most frequent contextual determinants were diminished workspace (42%), arch reversal (38%), thumb pinch deficit (34%), and flexor digitorum profundus weakness (30%). CHFS is a simple, reproducible, and clinically interpretable outcome measure for claw hand correction. It provides a practical contralateral-referenced summary of recovery, whereas CHFS-M improves clinical interpretation by documenting residual contextual deficits not captured by the composite score alone. Therapeutic Level IV.
The relative age effect (RAE) is well-documented in traditional sport, but its applicability to professional esports remains largely unexplored. The two existing studies examined only the entry question, without controlling for birth seasonality, testing causal mechanisms, or analyzing post-entry career trajectories. This study examined (1) whether RAE exists in professional esports after adjusting for birth seasonality; (2) whether academic cutoff dates causally influence esports career outcomes; (3) whether birth quarter affects career longevity and earnings after professional entry; and (4) whether birth quarter operates indirectly through debut age. The sample comprised 15,065 professional esports players with confirmed birthdates from the Esports Earnings database, spanning 177 countries and 609 game titles. Methods included chi-square tests with population-adjusted expected frequencies, a cross-national quasi-experiment exploiting cutoff variation across 20 countries, Kaplan-Meier survival analysis and Cox regression, propensity score matching, and Baron-Kenny mediation analysis with the Sobel test. After adjusting for birth seasonality, no aggregate RAE was detected [χ2(3) = 1.38, p = 0.711]. A cohort reversal emerged: players born after 2006 exhibited a traditional RAE (OR = 1.92, p < 0.001), while those born in 1991-2000 showed an inverse RAE (OR = 0.81-0.85, p < 0.05). Cross-national analyses revealed no causal effect of cutoff dates on career outcomes. Post-entry, birth quarter had no direct effect on career duration (log-rank p = 0.593) or earnings (propensity-score-matched p = 0.487). Birth quarter operated indirectly through debut age: late-born players debuted 0.16 years earlier, mediating 48.5% of the total association with earnings (Sobel z = 2.57, p = 0.010). At the aggregate level, professional esports shows no relative age effect after adjusting for birth seasonality, and cross-national evidence is consistent with academic cutoff dates having no causal effect on career outcomes. Birth quarter is associated with career earnings only indirectly, through the timing of debut. The traditional RAE pattern emerging in the youngest cohort (born after 2006) may warrant attention as esports adopts more structured youth development systems.
Acquired tonsillar descent with syringomyelia secondary to intracranial space-occupying lesions is rare, and supratentorial causes are particularly uncommon. These cases are of particular pathophysiological interest because they suggest that tonsillar descent and syrinx formation may arise as downstream consequences of altered intracranial compliance, compartmental pressure transmission, and impaired craniocervical cerebrospinal fluid buffering, rather than from a primary hindbrain malformation. We report a previously healthy 39-year-old woman who presented with progressive headache, papilledema, and left homonymous hemianopia. Imaging demonstrated a giant posterior tentorial dural-based mass confined to the supratentorial compartment, with marked mass effect, inferior tentorial deformation, 7-mm caudal tonsillar descent below McRae's line, and a small C1 syrinx. The patient underwent preoperative embolization and near-total microsurgical resection, followed 6 weeks later by single-fraction LINAC-based stereotactic radiosurgery to a small residual remnant. Follow-up MRI showed reduction of the remnant, resolution of the midline shift, restoration of normal tonsillar morphology and position, and complete resolution of the syrinx, accompanied by substantial visual recovery. The observed reversal after treatment of the causative lesion supports a model in which the supratentorial mass secondarily disturbed craniocervical hydrodynamics and produced a reversible acquired Chiari-like state with upper cervical syringomyelia. This case highlights the need for careful cranial imaging before posterior fossa decompression is considered in adults with newly identified tonsillar descent and syringomyelia.
Intranasal (IN) naloxone achieves rapid systemic exposure necessary for effective opioid overdose reversal; yet establishing bioequivalence (BE) for fast-acting drug-device combination products remains challenging due to the interplay of formulation attributes, device performance, and nasal physiology. This study developed an in vitro permeation test (IVPT) approach designed to quantitatively relate the nasal permeation behavior of naloxone to its clinical pharmacokinetic (PK) performance through an in vitro-in vivo relationship (IVIVR). Naloxone hydrochloride (Narcan®, 4 mg/0.1 mL) was deposited onto artificial membranes and EpiAirway™ mucociliary tissues using a controlled aerosol-deposition system (VITROCELL® Cloud Alpha 12). Naloxone permeation was assessed under sink conditions using a validated LC-MS/MS method. Cumulative permeation at 20 and 120 min (F₂₀ and F₁₂₀, respectively) was correlated with clinical maximum plasma concentration (Cmax) and area under the curve from time zero to infinity (AUC₀-∞) to construct IVIVR models, supplemented by exploratory point-to-point in vitro-in vivo extrapolation (IVIVE) using Wagner-Nelson deconvolution. Permeation profiles differed by substrate, with the hydrophilic membranes showing higher dissolution rates and EpiAirway™ tissues demonstrating dose-proportional transport despite lower deposited mass. The tissue-based IVIVR models showed strong linearity (R2 > 0.98) and mean prediction errors within accepted limits (≤ 10%), while artificial membranes consistently overpredicted the systemic exposure. IVIVE analysis further supported close temporal agreement with clinical absorption patterns. These findings indicate that a tissue-based IVPT-IVIVR framework may provide a translational tool for relating in vitro permeation behavior to systemic exposure, supporting its utility in formulation development and BE risk assessment of rapidly acting IN naloxone products.
Sleep-disordered breathing (SDB) is common in childhood and is associated with attentional and behavioral impairments despite largely preserved sleep macrostructure and minimal abnormalities in conventional electroencephalographic measures. This discrepancy has contributed to the perception that sleep is relatively preserved in pediatric SDB and has limited understanding of the physiological mechanisms underlying morbidity. To determine whether pediatric SDB is associated with disruption of the regional organization and homeostatic dynamics of slow-wave activity (SWA), a key physiological marker of sleep- dependent neural recovery and development. Cross-sectional study of 62 children aged 4 to 12 years who underwent overnight polysomnography with high-density electroencephalography in a laboratory setting. Participants were recruited from clinical referrals and the community, spanning the full spectrum of SDB severity. SDB severity indexed by hypopnea index (HI), apnea-hypopnea index (AHI), and obstructive apnea index (OAI). Regional electroencephalogram-derived SWA (0.5-4 Hz) topography and exponential decay parameters derived from frontal and posterior cortical regions. The frontal-to-posterior decay- rate ratio was evaluated as a summary measure of regional sleep homeostasis. In children with lower hypopnea index, SWA demonstrated the expected developmental pattern, with posterior predominance in younger children and a progressive shift toward a more balanced anterior-posterior distribution with age. Increasing HI was associated with attenuation or reversal of this spatial organization. Global SWA showed no meaningful association with SDB severity. In contrast, regional frontal and posterior decay parameters were strongly associated with HI (adjusted R² = 0.53; p < 1 × 10⁻⁶) but not OAI (adjusted R² = 0.05; p = .95). The frontal- to-posterior decay-rate ratio showed the strongest association with HI β = 4.15; 95% CI, 3.17- 5.13; p < 1 × 10⁻¹⁰; adjusted R² = 0.55. Pediatric SDB was associated with regional disruption of slow-wave sleep homeostasis rather than global loss of deep sleep. These alterations affected both the spatial organization and temporal dynamics of SWA during a period of active cortical maturation and were not captured by conventional sleep metrics. Regional SWA dynamics may provide a developmentally sensitive marker of physiological disease burden in children with SDB. Question: Does pediatric sleep-disordered breathing disrupt the regional organization and homeostatic dynamics of slow-wave activity during development in ways that are not captured by conventional sleep metrics?Findings: In this cross-sectional study of 62 children across the spectrum of sleep-disordered breathing, hypopnea burden was associated with altered regional organization and overnight dissipation of NREM slow-wave activity (SWA) despite preserved global SWA. A frontal-to- posterior SWA decay-rate ratio was strongly associated with hypopnea severity, whereas global SWA was not.Meaning: Pediatric sleep-disordered breathing may disrupt sleep physiology in a regional, developmentally meaningful manner not captured by conventional polysomnography, suggesting a potential physiological marker of disease burden beyond event counts.
Existing models of thermoset deconstruction facilitated by incorporating cleavable comonomers rely on a mean-field reverse gel point paradigm, which predicts network dissolution once cleavable bonds reach a critical stoichiometric threshold, but does not account for where those bonds reside within the network architecture. Using reactive coarse-grained molecular dynamics simulations coupled with graph-theoretic analysis, we extend this stoichiometric picture to show that deconstructability is governed by the curing-imprinted network topology rather than stoichiometry alone. This topological organization is hierarchical: at the local scale, the elastic effectiveness of cross-link junctions determines which cross-links constitute the load-bearing scaffold; at the mesoscale, the cross-linking rate kinetically templates that scaffold into topologically modular communitiesdensely cross-linked clusters connected by sparse bridging strands that sustain network connectivity. Using betweenness centrality to identify nodes that disproportionately lie on intercommunity shortest paths, we demonstrate that effective deconstruction of the network into macromolecular fragments requires cleavable comonomers to intercept these high-centrality bridging strands. We further find that under uniform, disassortative comonomer incorporation, this topological requirement provides a mechanistic basis for extending the reverse gel point to incorporate network topology. We also show that modularity imposes a fundamental limit on fragment uniformity that persists even when the centrality requirement is met. Finally, we demonstrate that chain stiffness provides a nearly independent lever to suppress mechanically redundant cross-links and raise the glass transition temperature without significantly altering the deconstruction outcome. Together, these findings reframe the thermoset design space around network topology and provide actionable guidelines for engineering thermoset copolymers with predictable deconstructability and targeted thermomechanical performance.
Osteoarthritis (OA) involves cartilage degradation and subchondral bone alterations, yet the mechanisms of chondro-endothelial crosstalk remain unclear. Stress-sensitive Piezo1 and SP1/Angptl7 signaling may play key roles in this process. This research aimed to investigate whether Platelet-derived growth factor (PDGF)-BB regulates chondro-endothelial crosstalk via Piezo1-mediated SP1/Angptl7 inhibition. Single- and multiple-cell-component organoids (chondrocytes SW1353 and endothelial HMEC-1) were constructed using high-throughput 3D culture. Organoids were treated with MIA to induce OA-like changes, followed by PDGF-BB with or without Yoda1 (Piezo1 activator) or Angptl7. In vivo, OA was induced in rats by intraarticular MIA injection, and PDGF-BB or sodium hyaluronate (SH)-PDGF was administered. Histology and immunofluorescence were used to assess F-actin formation, Piezo1 activation, SP1 phosphorylation, Angptl7 and VEGF/Notch/DLL4 expression. PDGF-BB inhibited F-actin formation and reduced Piezo1 activation in OA chondrocytes. It suppressed SP1 phosphorylation and Angptl7 expression, downregulating VEGF/Notch/DLL4 signaling and reducing endothelial invasion in organoids. These effects were partially reversed by Yoda1 and fully reversed by Angptl7. In vivo, PDGF-BB and SH-PDGF attenuated cartilage degeneration and reduced Piezo1 activation, SP1 phosphorylation, and Angptl7 expression. PDGF-BB alleviates OA by inhibiting SP1/Angptl7-mediated chondro-endothelial crosstalk, partially via stress-sensitive Piezo1 regulation through cytoskeletal remodeling. Multiple-cell-component organoids provide a valuable in vitro model for studying cartilage pathophysiology.
Oral frailty is a potentially reversible state associated with systemic frailty and adverse outcomes, but evidence on non-pharmacological programmes remains scattered. To evaluate non-pharmacological interventions for oral frailty and related oral functional decline in older adults and to identify domain-specific patterns of effect across different intervention models. We searched PubMed, Embase, Web of Science, Cochrane Library, CNKI, Weipu, and CBM from inception to 22 November 2025 for randomised controlled trials and quasi-experimental studies in adults aged ≥ 65 years. Owing to heterogeneity, we conducted a synthesis without meta-analysis (SWiM) using vote counting by direction of effect and rated certainty using GRADE. Interventions were classified as oral exercise alone, oral exercise plus education or self-management, or multicomponent programmes adding physical activity and/or nutrition. Twenty-three studies (18 randomised; 5 quasi-experimental) involving 2 412 participants were included, predominantly from East Asia. Overall, interventions tended to show favourable directions in several oral-function outcomes, with domain-specific patterns. Oral exercise alone showed the most consistent improvement direction for oral motor performance, such as tongue pressure and oral diadochokinesis. Adding education or self-management more often showed improvement directions in salivary-related outcomes and xerostomia-related symptoms. Multicomponent programmes suggested favourable directions across several dimensions, including composite oral health status, although the evidence was limited by few trials. The overall certainty of evidence ranged from very low to low, reflecting heterogeneity in intervention dose and follow-up, outcome measurement and incomplete reporting. Non-pharmacological interventions were generally associated with favourable directions in several oral-function outcomes related to oral frailty and related oral functional decline, with programme design corresponding to different outcome domains; however, the certainty of evidence was very low to low. Future trials should standardise core outcomes, report dose and adherence clearly, and assess longer-term effects.
A coherent sense of self depends not only on how individuals evaluate themselves, but also on how they believe they are evaluated by others. These internal and socially inferred self-evaluations-self-appraisal and reflected appraisal-are central to identity regulation and emotional wellbeing, yet their momentary interplay in everyday life remains poorly understood. In particular, little is known about how self- and reflected appraisal are associated with one another across moments, how social context shapes their coupling, and whether discrepancies between them carry affective consequences. Using ecological momentary assessment, student participants reported their self-appraisal, reflected appraisal, social context, and mood eight times daily over a 10-day period. Self-appraisal and reflected appraisal were strongly associated within the same moment, with concurrent associations substantially larger than lagged effects. Social context selectively moderated the momentary association from self-appraisal to reflected appraisal, such that self-appraisal was more strongly associated with reflected appraisal when individuals were with others compared to when they were alone, whereas the reverse association was not moderated by social context. Greater discrepancy between self-appraisal and reflected appraisal was initially associated with lower concurrent mood but was not associated with subsequent mood. Moreover, the concurrent discrepancy association was no longer significant after accounting for self-appraisal and reflected appraisal directly. Additional analyses suggested that mood was associated more strongly with self-appraisals than with discrepancies between self-appraisal and reflected appraisal. Together, the findings indicate that self- and reflected appraisal are closely coupled yet distinguishable processes that show substantial momentary covariation in daily life. Social context was associated with stronger coupling between self-appraisal and reflected appraisal, although additional decomposition analyses suggested that this effect was not driven by within-person fluctuations in social engagement. Strong associations between self-appraisal and reflected appraisal were evident even when individuals were alone, consistent with the notion that representations of others' evaluations remain psychologically relevant outside of direct social interaction. At the same time, the findings provide limited support for discrepancy as an independent correlate of mood, suggesting that momentary affect may be associated more strongly with the positivity or negativity of self-appraisals and reflected appraisals than with discrepancies between them. These results contribute to a more nuanced understanding of how internal and socially inferred self-evaluations are integrated in everyday life.
Insulin-like growth factor-1 (IGF-1) receptor (IGF-1R) inhibitors have changed the treatment landscape for moderate-to-severe thyroid eye disease (TED), but the longitudinal behavior of circulating IGF-1 during and after therapy remains insufficiently characterized. This study aimed to describe serum IGF-1 dynamics in patients with TED treated with IGF-1R inhibitor teprotumumab N01 and to explore their relationship with glycemic changes. In this retrospective cohort study, 92 patients with moderate-to-severe TED treated with teprotumumab N01 who had longitudinal IGF-1 measurements were included. IGF-1 dynamics were characterized at infusion-based visits and during post-treatment follow-up. Glycemic changes were assessed using fasting blood glucose (FBG), hemoglobin A1c (HbA1c), and glycated albumin (GA). Patients were classified by baseline glycemic status. Associations between IGF-1 metrics and glycemic changes were evaluated using multivariable linear regression for continuous glycemic outcomes and logistic regression for threshold-defined glycemic events, with sequential adjustment for age, sex, baseline glycemic markers, and baseline IGF-1 when applicable. Additional analyses were performed according to baseline glycemic status and baseline HbA1c quartiles. Baseline serum IGF-1 was 151.0 ng/mL (IQR 116.8-187.3). IGF-1 increased markedly after treatment initiation, with a median early-treatment peak fold change of 3.5 (IQR 3.0-4.3) and a median on-treatment peak concentration of 649.5 ng/mL (IQR 520.8-753.8), corresponding to a 4.2-fold increase from baseline (IQR 3.5-5.0). IGF-1 remained elevated during the first 3 months after the last infusion and then declined progressively, generally approaching baseline by 6-9 months or later. Glycemic markers showed modest increases, with median peak increases from baseline of 0.40% (IQR 0.20-0.77) for HbA1c, 1.63% (IQR 0.95-2.39) for GA, and 0.65 mmol/L (IQR 0.30-1.14) for FBG. Glycemic deterioration was most pronounced in patients with baseline dysglycemia. In contrast, neither baseline IGF-1 nor IGF-1 dynamic metrics were consistently identified as independent correlates of glycemic changes after adjustment for age, sex, and baseline glycemic markers and IGF-1. Similar findings were observed in subgroup and HbA1c quartile analyses. Teprotumumab N01 treatment was associated with a substantial, early, and broadly reversible increase in circulating IGF-1 in patients with moderate-to-severe TED, but IGF-1 dynamics did not serve as an independent indicator of glycemic deterioration.
Muscle atrophy and fatty infiltration of the shoulder muscles are known to influence clinical outcomes after reverse total shoulder arthroplasty (rTSA). Although quantitative evaluation of rotator cuff muscles has been increasingly reported, most previous studies have focused on intrinsic muscles and relied on two-dimensional or manual assessment methods. The purpose of this study was to evaluate shoulder muscle volume and composition using an artificial intelligence-based automated segmentation method on pre-operative computed tomography (CT) images and to investigate their associations with post-operative range of motion (ROM) after rTSA. This retrospective study included 13 patients (14 shoulders) who underwent rTSA for cuff tear arthropathy and related conditions with a minimum post-operative follow-up of 1 year. Pre-operative CT scans with 1-mm slice thickness were obtained within four weeks before surgery. Shoulder muscles, including intrinsic and extrinsic muscles, were automatically segmented using a deep learning-based model. Muscle volume and composition-functional muscle (FM), low-attenuation muscle, and adipose tissue (AT)-were quantified and normalized to scapular bone volume. Associations between muscle parameters and post-operative active ROM were analyzed using Spearman correlation coefficients. The supraspinatus (SSP) exhibited a higher proportion of AT, possibly reflecting fatty infiltration, and was the only muscle showing a relatively balanced distribution of FM, low-attenuation muscle, and AT. The proportion of FM in the SSP was positively associated with active anterior elevation (AE), whereas the proportion of AT in the SSP and subscapularis (SSc) was negatively associated with active AE. In addition, total deltoid muscle volume and FM volume of the SSP were positively associated with post-operative AE. The AT volume of the SSc were negatively associated with active AE. No significant associations were identified between muscle composition or volume and post-operative external rotation. Artificial intelligence-based automated segmentation on pre-operative CT enabled detailed quantitative assessment of shoulder muscle volume and composition. Post-operative ROM tended to be associated with the deltoid, SSP, and SSc. Although these findings should be interpreted with caution due to limited statistical power, this exploratory feasibility study provides preliminary insights and may serve as a basis for future investigations.
In the post-pandemic era, co-circulation of multiple respiratory RNA viruses has increased the need for timely diagnosis and reliable recognition of mixed infections. Although reverse transcription quantitative polymerase chain reaction (RT-qPCR) remains the clinical standard for respiratory virus detection, its target-restricted design limits the detection of unexpected or coinfecting pathogens. Conventional metagenomic next-generation sequencing (mNGS) provides hypothesis-free pathogen detection, but routine clinical use is still limited by long turnaround times and complex library preparation. Therefore, a sequencing-based strategy that preserves broad, unbiased detection while offering a simplified workflow and clinically acceptable turnaround time is needed. We optimized and clinically validated CATCH, a rapid RNA/DNA hybrid tagmentation-based mNGS workflow, for respiratory RNA virus detection. Analytical performance was assessed using standardized reference materials, including SARS-CoV-2 and influenza A virus, with evaluations of sensitivity, reproducibility, short-term stability, and host-background interference. Clinical validation was performed in retrospective and prospective respiratory infection cohorts, and assay performance was benchmarked against RT-qPCR and multiplex PCR. The same sequencing data were further examined for semiquantitative viral assessment, coinfection detection, and exploratory respiratory microbial profiling. The optimized CATCH workflow shortened library preparation to approximately 3 h, with about 35 min of hands-on time, enabling same-day sequencing-based diagnostics. Broad detection was achieved across seven clinically relevant respiratory RNA viruses. Sequencing-derived viral abundance showed a significant overall correlation with viral input concentration, supporting semiquantitative interpretation, although virus- and subtype-specific variability highlighted biological constraints on absolute quantification. Using SARS-CoV-2 and influenza A virus as representative targets, CATCH achieved clinically actionable limits of detection with high reproducibility and stability. In clinical cohorts, CATCH showed high concordance with routine molecular assays and identified mixed respiratory infections missed by targeted testing. Exploratory analyses also demonstrated the feasibility of respiratory microbial community profiling from the same sequencing dataset. CATCH is a rapid and clinically deployable RNA virus mNGS workflow that helps bridge targeted molecular diagnostics and conventional metagenomic sequencing. By combining broad pathogen detection, coinfection identification, and semiquantitative assessment within a streamlined workflow, CATCH provides a practical framework for comprehensive respiratory RNA virus diagnosis and syndromic surveillance.
Alzheimer's disease (AD) is a neurodegenerative disorder causing progressive neuronal damage. Incidence rises with age, and early diagnosis is difficult. This study examined DLX6-AS1 clinical relevance and regulatory mechanism in AD, and its interaction with miR-204-5p in AD pathology. It offers new insights into early diagnosis and treatment. A total of 133 AD patients and 105 healthy controls were selected. Their serum levels of DLX6-AS1 and miR-204-5p were analyzed using quantitative polymerase chain reaction. β-amyloid (Aβ)1-42-induced SH-SY5Y neuronal injury and okadaic acid (OA)-induced Neuro-2a tau abnormal phosphorylation models were constructed. The role/function of DLX6-AS1/miR-204-5p axis was then investigated using cell counting kit-8, flow cytometry, western blotting, enzyme-linked immunosorbent assay and a dual-luciferase reporter gene assay. AD patients had higher serum DLX6-AS1 and lower miR-204-5p levels. DLX6-AS1 showed an AUC of 0.838 for AD diagnosis. DLX6-AS1 levels were negatively associated with cognitive function, brain structural integrity, and benign pathology-and positively associated with disease severity, functional impairment, and pathological markers. In AD cell models, DLX6-AS1 was upregulated. Silencing it promoted cell proliferation, reduced apoptosis and oxidative stress, improved mitochondrial and synaptic function, decreased tau phosphorylation, and enhanced microtubule stability and axonal transport. Dual-luciferase assays confirmed direct binding between DLX6-AS1 and miR-204-5p. Co-inhibition reversed the protective effects of DLX6-AS1 silencing. Serum DLX6-AS1 is a potential biomarker for early diagnosis and assessment of AD. It regulates Aβ-induced neuronal damage and tau phosphorylation by targeting miR-204-5p, offering a new mechanism target for AD molecular therapy.
Alcohol use is a major global health issue, causing about 3.3 million deaths each year, or roughly 5.9% of all deaths worldwide. Alcohol-related liver disease develops in stages: steatosis, steatohepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma. If alcohol consumption is discontinued at early stages, alcohol-related fatty liver disease can be reversed; however, continued exposure leads to progressive liver injury and increased mortality risk. Current diagnostic tools lack sufficient sensitivity and specificity to detect early-stage disease or accurately assess disease progression. This highlights the need for reliable and mechanistically relevant biomarkers for early diagnosis and staging. This review examines alterations in gut microbiota across different stages of alcohol-associated liver disease and evaluates gut microbiota-associated biomarkers, including microbial metabolites, in the context of their potential diagnostic and prognostic utility. In addition, the review discusses the limitations of existing biomarkers and highlights the emerging role of microbiome-derived signals in reflecting disease mechanisms. These findings suggest that gut microbiota-related biomarkers may provide a promising but still evolving approach for improving early detection and understanding disease progression in alcohol-associated liver disease.
The human cochlea is a fragile, spiral-shaped sensory organ for hearing that is deeply embedded within the dense otic capsule bone, making accurate post-mortem anatomical studies difficult when using conventional histological approaches requiring sectioning. X-ray microscopy (XRM) is a non-destructive radiological imaging modality that combines geometric and optical magnification to achieve high-resolution 3D visualization. Here, we evaluated the ability of XRM to resolve the entire inner ear, including delineating soft tissue and bony structures, in trimmed human temporal bones ex vivo following lumen perfusion with a reversible iodine-based contrast-enhancing solution. After optimizing contrast incubation conditions, XRM datasets were acquired, reconstructed, and used to generate 3D volume renderings of the inner ear. XRM enabled visualization of the entire cochlea and vestibular system, from millimeter-scale labyrinthine anatomy to submillimeter and micrometer-scale cochlear structures, including spiral ganglion nerve fiber bundles and sensory cell rows. Quantitative analysis in a representative case demonstrated an increasing width-to-height ratio of the organ of Corti from the basal to the apical cochlear turns, consistent with histology images and prior anatomical studies. These findings demonstrate that XRM provides high-resolution, sectioning-free ex vivo 3D visualization of human cochlear anatomy and may enable quicker and more faithful correlation of cochlear pathology with otologic disorders than traditional histology.
Sjögren's disease (SjD) is a chronic autoimmune disease characterized by lymphocytic infiltration of the exocrine glands and progressive secretory dysfunction. Although the major molecular abnormalities in SjD, including prominent type I interferon activation and abnormal B-cell responses, are now better defined, these findings have not consistently translated into effective disease-modifying therapy. A persistent gap therefore remains between biological target engagement and clinical benefit. This review argues that this gap may partly reflect a mismatch between current pathological models of SjD and the way clinical trials are designed. Conventional frameworks often treat SjD as a relatively homogeneous inflammatory disease and rely heavily on systemic disease-activity measures. Such approaches may underrepresent the spatiotemporal evolution of the salivary gland microenvironment and the heterogeneity of treatment responses across tissue states. Drawing on recent evidence, including single-cell RNA sequencing, spatial transcriptomics, and longitudinal histological studies, we propose a tissue-centered interpretive framework for SjD. This framework highlights three partially overlapping dimensions: epithelial activation states linked to nucleic-acid-sensing and interferon-related programs; lymphoid organization and B-cell-supportive niches associated with IFN-BAFF and Tfh/Tph-related activity; and later structural remodeling associated with fibrosis, impaired regeneration, and reduced functional reversibility. These dimensions are used here as interpretive constructs rather than as validated clinical staging categories. On this basis, precision medicine in SjD may need to move beyond a uniform anti-inflammatory strategy toward mechanism-based stratification anchored in tissue state and disease stage. Biomarkers such as salivary gland ultrasonography, TLS-related features, and microenvironment-linked molecular readouts may help refine patient stratification, endpoint selection, and clinical trial design, although these applications still require prospective validation.
The tumour microenvironment (TME) of relapsed/refractory (R/R) diffuse large B-cell lymphoma (DLBCL) patients is associated with resistance of DLBCL cells to CD19 CAR-T cells. How to improve TME in DLBCL and improve the efficacy of CAR-T cell therapy remains to be further explored. We observed the sensitivity of HBL-1/U2932 cells pretreated with BTK inhibitors (BTKi) to CAR-T cells with flow cytometry (FCM). The effect of pretreatment of BTKi on the polarisation state of alternative activated M2 macrophages was observed with FCM, real-time PCR and Western blot method. The effect of Notch1 agonist on expressions of Arg-1 protein, iNOS protein, Notch1 protein and RBP-J protein in alternative activated M2 macrophages was observed by Western blot method. Then the expression consistency of Notch-1 and RBP-J in activated M2 macrophages was observed by siRNA transfection of Notch-1. The cytotoxicity of CD19 CAR-T cells on HBL-1/U2932 cells pretreated with ibrutinib/orelabrutinib was higher than that of HBL-1/U2932 cells unpretreated with ibrutinib/orelabrutinib. Cytotoxicity of CAR-T cells to HBL-1 cells in coculture system with alternative activated M2 macrophages was very low. This drug resistance could be reversed by replacing the M2 macrophages (M2 macrophages after 48 h pretreatment with BTKi) in coculture system. Pretreatment with BTKi could down-regulate the expression of CD206 and IL-10 in activated M2 macrophages. And pretreatment with BTKi down-regulated the expression of Arg-1 and upregulated the expression of iNOS in activated M2 macrophages. The upregulation polarisation of M2 macrophages by Notch1 agonist could be reversed by BTKi. Expression of RBP-J protein decreased in alternative activated M2 macrophages by siRNA silencing Notch 1. Pretreatment with BTKi could down-regulate the polarisation of M2 macrophages and reverse the resistance of DLBCL cells which were cocultured with alternative activated M2 macrophages to CAR-T cells. This effect might be achieved by downregulating the Notch-RBP-J pathway.
Metabolic syndrome (MetS) is a multifaceted disorder influenced by genetic and environmental factors. MetS is associated with obesity, dyslipidemia, hypertension, and hyperglycemia. Recently, attention has turned to gut microbiota, a diverse microbial community in the gastrointestinal tract implicated in metabolic diseases, including MetS. Berry cactus (Myrtillocactus geometrizans) contains polyphenols, pectins, sterols, and betalains with reported hypoglycemic, hypolipemic, anti-inflammatory, and antiproliferative properties. This study evaluated the impact of berry cactus juice concentrate (BJC) on metabolic markers, gut microbiota composition and predicted microbial functions in high-fat diet-induced MetS rat model. Metabolic markers related to MetS and comprehensive analyses of microbial 16 S rRNA gene were obtained after 140 days of treatment. HFD feeding induced a MetS-like phenotype in rats; however, BJC supplementation did not significantly reverse the main metabolic alterations under the experimental conditions evaluated. Notably, BJC treatment was associated with changes in gut microbiota composition, including alterations in dominant phyla and relevant genera such as Parabacteroides. Predicted functional analyses suggested associations with pathways involved in fatty acid metabolism and promotion of the availability of berry cactus bioactive molecules. In addition, the presence of betalains and flavonoid derivatives was identified in BJC, which may contribute to its microbiota-modulating effects. These findings suggest that BJC may act primarily as a dietary modulator of microbiota in the context of HFD-induced metabolic disturbances. Although further studies are needed to determine whether these microbial changes translate into significant metabolic benefits.