Research on the relationship between work-family conflict and burnout among civil aviation pilots after the COVID-19 pandemic

by Weiheng Chen, Shouxi Zhu, Kequan Shao, Kejia Liang

To examine the impact of work-family conflict on occupational burnout among pilots following the COVID-19 pandemic, this study employed the Maslach Burnout Inventory-General Survey and the Zhao Xinyuan Bi-directional Scale of Work-Family Conflict as research instruments. The data were analyzed using correlation analysis and Structural Equation Modeling (SEM) to explore the relationship between work-family conflict and burnout. The results indicate a significant correlation between work-family conflict and burnout, as well as the three dimensions of burnout. Specifically, the correlation coefficients between work interfering with family (WIF) and family interfering with work (FIW) with occupational burnout were found to be 0.737 and 0.496, respectively. In the path analysis of the SEM, the estimated effects of WIF and FIW on burnout were both 0.49. Mediation analysis revealed that WIF indirectly influences burnout through FIW, with a mediation effect value of 0.117. Additionally, while pilot-related factors did not significantly affect burnout, they were found to have a notable impact on work-family conflict. These findings underscore the significant role of work-family conflict in contributing to burnout and provide a theoretical foundation for targeted interventions aimed at mitigating burnout among pilots.

A non-invasive urinary diagnostic signature for diabetic kidney disease revealed by machine learning and single-cell analysis

by Yonggang Chen, Jintai Luo, Yingying Zheng, Xiaomei Jiang, Zixiang Yang, Xiaobing Liu

Background

Diabetic kidney disease (DKD) poses a significant health burden with inadequate diagnostic sensitivity. This study develops non-invasive biomarkers by integrating urinary and renal single-cell sequencing with machine learning.

Methods

This study analyzed DKD single-cell and bulk transcriptomic data from public repositories. We established a computational pipeline to distinguish kidney-originating cells in urinary sediments, enabling the identification of injury-associated gene signatures. These signatures were refined using machine learning to develop a diagnostic model, which was validated in independent cohorts. The biomarkers were further verified in DKD renal tissues at single-cell resolution and across multiple nephropathies. Functional and spatial analyses confirmed biological relevance using transcriptomic and histological validation.

Results

Single-cell analysis of 2,089 urine-derived cells identified eight renal cell types, including injured proximal tubule cells (Inj-PTC) showing upregulated injury markers (HAVCR1, VCAM1) and enriched apoptotic/TGF-β pathways. A machine learning-selected biomarker panel (PDK4, RHCG, FBP1) demonstrated strong diagnostic value (area under the curve, AUC > 0.9), with consistent downregulation across multiple chronic kidney diseases. PDK4 and FBP1 were specifically suppressed in DKD renal Inj-PTC (p  Conclusions

This study identifies a three-gene biomarker panel (PDK4, RHCG, FBP1) as a promising non-invasive diagnostic tool for DKD. While demonstrating excellent diagnostic performance. It represents a tubular injury-associated gene signature that is detectable in urinary cells and shows strong association with DKD in transcriptomic datasets, presenting a promising candidate for a non-invasive diagnostic assay.

Correlation study of brain function changes after chemotherapy in breast cancer patients by automated fiber quantification based on diffusion tensor imaging

by Yun Feng, Wei Chen, Yang Lu, Haifei Zhou

Purpose

The aim of this study is to evaluate the changes of white matter microstructure in breast cancer patients before and after chemotherapy based on automated fiber quantification (AFQ),as well as determine if these dispersion indexes are significantly correlated with clinical data.

Materials and Methods

Twenty-four breast cancer patients scheduled for chemotherapy were enrolled. Diffusion tensor imaging (DTI), neuropsychological tests and self-report measures, and hematological tests were conducted before chemotherapy(time 0,T0) and within one week after chemotherapy(time 1,T1). AFQ was used to track 20 fiber tracts in the brain. The correlation between average abnormal tracts and changes in neuropsychological tests and self-report measures and blood indicators was analyzed.

Results

Compared to T0, subjects at T1 showed decreased scores on the verbal fluency test; increased scores on the self-rating anxiety scale(SAS)and self-rating depression scale (SDS). Estrogen concentration was lower while luteinizing hormone(LH), follicle-stimulating hormone, and triglyceride levels were higher. Mean fractional anisotropy (FA) value decreased in the right cingulum cingulate(CGC)while mean radial diffusivity (RD) increased in the right CGC; mean axial diffusivity (AD) value decreased in callosum forceps major and callosum forceps minor. Changes in FA with in the right CGC were positively correlated with changes in SDS and LH, while changes in RD with in the right CGC were negatively correlated with changes in SDS and LH.

Conclusion

Early changes observed in brain white matter fiber tracts, along with persistent hormone and triglyceride metabolism disorders, could potentially serve as neurobiological markers for monitoring chemotherapy-induced cognitive impairment.

Real-world optimization of tunnel lengths in tunneled peripherally inserted central catheters for cancer patients: A multi-center retrospective cohort study

by Yinyin Wu, Wei Ding, Yuying Liu, Qianhong Deng, Fengqin Tao, Hanbin Chen, Chang Chen, Meng Xiao, Bilong Feng

Background

Standardized guidelines for optimal tunnel length in tunneled peripherally inserted central catheters (PICCs) are lacking.

Objectives

The objective of this study was to evaluate the real-world impact of tunnel length on clinical outcomes.

Methods

This retrospective cohort study included 207 cancer patients who received tunneled PICCs, categorized into a control group (tunnel length > 4 cm, n = 134) and an observation group (tunnel length ≤ 4 cm, n = 73). Propensity score matching (PSM) was used to address baseline heterogeneity. Cox regression analyses were used to assess the risk of complication during a 120-day follow-up.

Results

Compared to the control group (tunnel length > 4 cm), the observation group (tunnel length ≤ 4 cm) had a significantly higher adjusted overall complication risk (HR = 2.92, 95% CI: 1.07–7.94, P = 0.036) and unplanned catheter removal rate (4.4% vs. 0.0%, P = 0.027), confirming the safety of longer tunnels despite comparable comfort levels between groups. After PSM, Cox regression analysis showed results consistent with those from the unmatched cohort. Subgroup analyses revealed a reduced risk of complications with longer tunnels in patients with BMI ≤ 25 kg/m² (HR = 0.29, 95% CI: 0.11–0.82), without hypertension (HR = 0.36, 95% CI: 0.13–1.00), without diabetes (HR = 0.38, 95% CI: 0.15–0.97), and with solid tumors (HR = 0.31, 95% CI: 0.11–0.85).

Conclusion

The results show that tunnel lengths > 4 cm reduce overall complications and prolong catheter retention, supporting the implementation of standardized protocols while advocating for personalized adjustments based on BMI, comorbidities, and cancer type.

Multi-omics analysis reveals diagnostic and therapeutic biomarkers for aging phenotypes in ulcerative colitis

by Lei Guo, Jun Ge, Li Cheng, Xinyi Zhang, Zhengzheng Wu, Meili Liu, Hanmei Jiang, Wei Gong, Yi Liu

Background

The incidence of ulcerative colitis (UC) remains high, with an increasing prevalence among elderly patients. Cellular senescence has been widely recognized as a contributor to UC susceptibility; however, the underlying molecular mechanisms remain incompletely understood. This study aimed to identify senescence-associated biomarkers in UC to provide new insight for diagnosis and treatment.

Methods

By integrating transcriptomic data from UC patients with established aging-related databases, we identified aging-associated differentially expressed genes (DEGs). Using weighted gene co-expression network analysis (WGCNA) and Cytoscape, we pinpointed the core genes involved. A diagnostic model for UC was then developed based on these core genes, and their expression patterns were characterized at single-cell resolution. The roles of these genes were ultimately validated through in vitro and animal experiments.

Results

We identified 24 aging-related DEGs in UC, which were primarily implicated in inflammatory responses and cytokine-receptor interactions. Further analyses pinpointed three core genes (CXCL1, MMP9, and STAT1) that were predominantly expressed in macrophages. A diagnostic model constructed using these genes exhibited robust predictive performance. Experimental validation confirmed that the expression levels of all three core genes were significantly upregulated in both a UC mouse model and in macrophages compared to controls. Additionally, pathway analyses revealed elevated levels of CXCL12 and VEGFA in the enriched pathways.

Discussion

Our findings underscore the pivotal roles of CXCL1, MMP9, and STAT1 in UC-associated cellular senescence. The analysis positions these molecules as promising macrophage-mediated diagnostic biomarkers and therapeutic targets. Collectively, this work provides novel insights into UC pathogenesis and lays a foundation for developing precision medicine strategies that target senescence pathways.

Concentration-dependent effects of fermented spent coffee grounds and contrasting effects of earthworms on growth and phytochemicals in medicinal plant <i>Glechoma longituba</i>

by Bing-Nan Zhao, Zi-Yang Xie, Jia-Ning Liu, Xiao-Ran Chen, Xin-Xin Wang, Jia-Yi Li, Rui Zhang, Chao Si

Fermented spent coffee grounds (FSCG) serve as a valuable soil amendment to improve soil structure and fertility, while earthworms play a well-established role in enhancing soil processes and plant growth. However, their combined effects on bioactive compound accumulation in medicinal plants remain unclear. This study investigated the individual and interactive effects of FSCG (0%, 10%, and 20%, v/v) and earthworms (with and without Pheretima guillelmi) on the growth and phytochemical content of Glechoma longituba, a common medicinal herb, under greenhouse conditions. Results showed that 10% FSCG generally promoted plant growth, whereas 20% FSCG generally enhanced the accumulation of total flavonoids, chlorogenic acid, and soluble protein. Earthworms enhanced aboveground biomass and node number but significantly reduced chlorogenic acid content. These findings highlight the potential of FSCG as a sustainable soil amendment in medicinal plant cultivation and underscore the need to consider earthworm activity when optimizing both plant biomass and phytochemical quality.

GV effects of diabetes mellitus on clinical outcomes of patients with acute heart failure: A systematic review and meta-analysis

by Linna Zhao, Juanjuan Zhang, Weizhe Liu, Cheng Dai, Aiying Li

Diabetes mellitus (DM) is identified as a potential modifier of clinical outcomes in acute heart failure (AHF), yet its prognostic impact is not fully determined. This systematic review and meta-analysis aimed to assess the prognostic impact of DM on survival outcomes in AHF patients by synthesizing evidence from 26 studies involving 326,928 subjects collected from Cochrane Library, PubMed, Web of Science, and Embase databases up to 1 June 2024. Both prospective/retrospective cohort and case-control studies published since 2000 were included, with outcomes evaluated through multivariate, univariate, and binary analyses using the Newcastle-Ottawa Scale for quality assessment. Multivariate analysis indicated that DM significantly increased the risk of all-cause mortality in AHF patients (cohort studies: HR = 1.21, 95%CI (1.13, 1.29), OR=1.15, 95%CI (1.05, 1.26); case-control studies: HR = 1.39, 95%CI (1.26, 1.53), OR=1.43, 95%CI (1.10, 1.84)]. Univariate analysis confirmed this finding in case-control studies [HR = 1.30, 95%CI (1.01, 1.67)], but not in cohort studies. In both cohort [RR = 1.27, 95%CI (1.12, 1.43)] and case-control [OR=1.21, 95%CI (1.08, 1.35)] studies, DM increased the risk of all-cause mortality. AHF patients with DM had a higher risk of cardiovascular mortality [cohort studies: HR = 1.85, 95%CI (1.46, 2.33); case-control: OR=1.70, 95%CI (1.17, 2.47)]. While multivariate analysis showed no association between DM and in-hospital mortality, case-control studies indicated an increased risk [OR=1.21, 95%CI (1.03, 1.42)]. DM also increased the risk of readmission [cohort studies: HR = 1.32, 95%CI (1.14, 1.53); case-control studies: HR = 1.44, 95%CI (1.23, 1.69); binary data: OR=1.19, 95%CI (1.07, 1.31)].This updated meta-analysis demonstrates that DM imposes significant adverse effects on all-cause mortality, cardiovascular-related mortality, and readmission risk in AHF patients. However, no significant connection was found between diabetes and survival outcomes with respect to the co-endpoint of death or readmission and the endpoint of in-hospital mortality. These findings underscore the necessity for implementing targeted diabetes management within AHF care protocols to enhance clinical outcomes, an essential consideration for future practice.

Exosomes from bone marrow mesenchymal stem cells protect melanocytes under vitiligo-related conditions through induction of NRF2/HO1 expression

by Xuecheng Sun, Bo Huang, Gaobo Ruan, Aie Xu

Background

Vitiligo, a chronic autoimmune disease linked to excess oxidative stress, can be temporarily improved. Bone marrow mesenchymal stem cells (BMSCs)-derived exosomes (BMSCs-Exos) have recently emerged as a promising novel therapeutic means for vitiligo.

Methods

Exosomes were isolated and characterized from BMSCs-conditioned medium. PIG3V cells and those transfected with NRF2 siRNA or negative control were cultured under normal conditions or exposure to hydrogen peroxide (H₂O₂) to induce oxidative stress, with addition of BMSCs-conditioned medium, conditioned medium from BMSCs pretreated with GW4869 (referred to as BMSCs-GW4869), or BMSCs-Exos. Cell viability, apoptosis, and oxidative stress parameters, including cellular glutathione (GSH)/oxidized glutathione (GSSG) ratio, superoxide dismutase (SOD), reactive oxygen species (ROS), and malondialdehyde (MDA), were assessed. The expression of Ki67, NRF2, HO1, BAC, and Bcl-2 was measured.

Results

BMSC-Exos significantly enhanced cell viability and reduced apoptosis and oxidative stress in H₂O₂-treated PIG3V cells. Simultaneously, BMSCs-Exos reversed H₂O₂-induced downregulation of Ki67, NRF2, HO1, and Bcl-2, and upregulation of BAX in PIG3V cells. Silencing NRF2 by siRNA in PIG3V cells prior to H₂O₂ treatment abolished the protective effect of BMSCs-Exos and decreased the HO1 expression.

Conclusions

BMSCs-Exos protect melanocytes from vitiliog-related oxidative stress by mitigating oxidative damage through induction of NRF2/HO1 expression.

Visit-to-visit glycemic variability is associated with lung function variables and lung function impairment in individuals with type 2 diabetes

by Yi-Hua Wu, Chia-Ing Li, Chiu-Shong Liu, Chih-Hsueh Lin, Shing-Yu Yang, Cheng-Chieh Lin, Tsai-Chung Li

Glycemic variability (GV) is an emerging biomarker of glycemic control and may be a predictor for lung function impairment in persons with type 2 diabetes mellitus (T2DM). However, the associations between GV and lung function variables and lung function impairment have not been fully evaluated. The objective of this study was to assess the associations of glycemic variability (GV) with lung function impairment in persons with T2DM. A follow-up study was conducted on the data of 3,108 subjects collected from 2001 to 2020 using the diabetes care management program database in Taiwan. GV in fasting plasma glucose (FPG) was calculated using standard deviation (SD), average real variability (ARV), coefficient of variation (CV), variability independent of the mean (VIM), and slope of 1-year repeated measurements. A ratio of forced expiratory volume in 1 s (FEV1) to forced vital capacity (FVC) less than 0.70 was used to define lung function impairment. Multivariable linear and logistic regression models were applied to explore the relationships of GV with lung function variables and lung function impairment. A total of 359 (11.6%) subjects were defined as having lung function impairment. After multivariable adjustment, FPG‐SD, FPG-CV, FPG-AVR, FPG-VIM and were found to be negatively linked with FEV1, % predicted FEV1, and FVC but not FEV1/FVC. Relative to those for the first tertile, the odds ratios (ORs) of lung function impairment for the second and third tertiles were 1.37 (95% confidence interval [CI]: 1.01, 1.87) and 1.51 (1.10, 2.08) for FPG-CV, respectively; 1.59 (1.16, 2.17) and 1.73 (1.24, 2.40) for FPG‐SD, respectively; and 1.57 (1.15, 2.13) and 1.69 (1.22, 2.33) for FPG-AVR, respectively. GV, measured by CV, SD, VIM, and VIM, is linked with lung function impairment and all lung function variables, except for FEV1/FVC ratio. GV may serve as a useful biomarker for assessing lung function impairment in persons with T2DM.

Dysregulated serum chloride and clinical outcomes in critically ill adults: A systematic review and meta-analysis

by Xiaoliang Wan, Feiyao Deng, Xue Bai, Chenxi Xiang, Chuan Xu, Linxiao Qiu

Dysregulated serum chloride levels are prevalent in critically ill patients. However, their clinical impact remains unclear. This first systematic review and meta-analysis quantified the prevalence of hypochloremia and hyperchloremia, and their associations with mortality and acute kidney injury (AKI) in critically ill populations. We searched PubMed, Embase, Web of Science, and the Cochrane Library for studies reporting hyperchloremia prevalence or outcomes in adult ICU patients until August 2025. Statistical analyses were conducted using Stata v16.0, and study quality was assessed using the Newcastle-Ottawa Scale. 34 studies (n = 175,021 patients) were included. The aggregated prevalence of hyperchloremia was 34% (95% CI [26%−43%]) and hypochloremia was 14% (95% CI [1%−28%]). Meta-analysis demonstrated that both hyperchloremia and hypochloremia were significantly associated with increased mortality, conferring a 28% (OR = 1.28, 95% CI [1.08–1.52]) and 55% (OR = 1.55%, 95% CI [1.33–1.81]) elevated risk for mortality, respectively. Crucially, a dose-response analysis revealed a non-linear relationship between serum chloride levels and mortality, confirming that the risk is independently elevated at both extremes. Furthermore, hyperchloremia was linked to an increased risk of AKI (OR = 1.40, 95% CI [1.07–1.85]). These findings establish dysregulated serum chloride as a common and clinically significant biomarker, underscoring the necessity of monitoring and managing both high and low chloride levels in critically ill patients. Future large-scale studies are warranted to validate these results and elucidate the mechanistic pathways linking chloride dysregulation to such adverse outcomes.

Aerial small target detection algorithm based on cross-scale separated attention

by Ju Liang, Fan Wang, Jia Chen, Hai-Yan Huang, Zu-Fan Dou

In UAV aerial photography scenarios, targets exhibit characteristics such as multi-scale distribution, a high proportion of small targets, complex occlusions, and strong background interference. These characteristics impose high demands on detection algorithms in terms of fine-grained feature extraction, cross-scale fusion capability, and occlusion resistance.The YOLOv11s model has significant limitations in practical applications: its feature extraction module has a single semantic representation, the traditional feature pyramid network has limited capability to detect multi-scale targets, and it lacks an effective feature compensation mechanism when targets are occluded.To address these issues, we propose a UAV aerial small target detection algorithm named UAS-YOLO (Universal Inverted Bottleneck with Adaptive BiFPN and Separated and Enhancement Attention module YOLO), which incorporates three key optimizations. First, an Adaptive Bidirectional Feature Pyramid Network (ABiFPN) is designed as the Neck structure. Through cross-scale connections and dynamic weighted fusion, ABiFPN adjusts weight allocation based on target scale characteristics, focusing on enhancing feature integration for scales related to small targets and improving multi-scale feature representation capability. Second, a Separated and Enhancement Attention Module (SEAM) is introduced to replace the original SPPF module. This module focuses on key target regions, enhances effective feature responses in unoccluded areas, and specifically compensates for information loss in occluded regions, thereby improving the detection stability of occluded small targets. Third, a Universal Inverted Bottleneck (UIB) structure is proposed, which is fused with the C3K2 module to form the C3K2_UIB module. By leveraging dynamic channel attention and spatial feature recalibration, C3K2_UIB suppresses background noise; although this increases parameters by 34%, it achieves improved detection accuracy through efficient feature selection, striking a balance between accuracy and complexity.Experimental results show that on the VisDrone2019 dataset and the TinyPerson dataset from Kaggle, the mean Average Precision (mAP) of the algorithm is increased by 4.9 and 2.1 percentage points, respectively. Moreover, it demonstrates greater advantages compared to existing advanced algorithms, effectively addressing the challenge of small target detection in complex UAV scenarios.

Disruption of <i>yqhG</i> attenuates virulence in <i>methicillin-resistant Staphylococcus aureus</i> by compromising membrane stability and oxidative stress resistance

by Jianhua Liao, Jun Cheng, Baoqing Liu, Yuzhi Shao, Chunyan Meng

The growing prevalence of methicillin-resistant Staphylococcus aureus (MRSA) infections, coupled with the increasing resistance to existing antibiotics, underscores the critical need for novel therapeutic approaches to combat this pathogen. In this study, the role of yqhG, a conserved gene encoding a periplasmic protein, in MRSA virulence and stress adaptation was investigated. yqhG deletion in MRSA significantly attenuated virulence in a murine infection model, leading to reduced bacterial burden in infected organs and improved host survival. In vitro, the yqhG mutant exhibited impaired membrane integrity, reduced motility, and increased sensitivity to oxidative stress, but did not affect biofilm formation. These defects were fully restored upon genetic complementation. These findings highlight the critical role of yqhG in maintaining MRSA’s ability to withstand host-imposed stresses, suggesting that yqhG is a key determinant of MRSA pathogenesis. The study provides new insights into the stress-defense mechanisms employed by MRSA and underscores yqhG as a potential target for therapeutic strategies aimed at combating MRSA infections.

Safety and efficacy of radiotherapy combined with immunotherapy in limited-stage small cell lung cancer a single-arm meta-analysis and systematic review

by Li Yu, Xinlin Yu, Cheng Ma, Xialin Zhang, Ran Cui

Background

Limited-stage small cell lung cancer (LS-SCLC) has a poor prognosis despite being potentially curable with standard concurrent chemoradiotherapy. The success of immune checkpoint inhibitors (ICIs) in extensive-stage SCLC has prompted investigation into combining immunotherapy with radiotherapy for LS-SCLC. This systematic review and single-arm meta-analysis aims to synthesize the evidence on this combined modality, providing pooled estimates of efficacy and safety to inform clinical practice and future trials.

Methods

Following PRISMA guidelines, we systematically searched PubMed, Embase, Cochrane Library, and Web of Science through July 2025 for studies evaluating radiotherapy combined with immunotherapy in patients with LS-SCLC. The primary outcomes analyzed included pooled objective response rate (ORR), median progression-free survival (mPFS), and median overall survival (mOS).

Results

Six studies, encompassing 487 patients, met the inclusion criteria. The pooled analysis demonstrated an ORR of 57.7% (95% CI: 24.9–90.5%), a weighted mPFS of 13.6 months (95% CI: 11.3–15.9 months), and a pooled mOS of 33.7 months (95% CI: 26.7–40.7 months). Grade 3−4 treatment-related adverse events occurred in 42.2% of patients. Subgroup analyses revealed that a concurrent treatment sequence yielded a significantly higher ORR compared to sequential approaches (77.6% vs. 65.2% for immunotherapy followed by radiation vs. 25.8% for radiation followed by immunotherapy). Radiation dose was also identified as a critical determinant of efficacy. Anti-PD-L1 agents showed a numerically higher ORR than anti-PD-1 agents (96.0% vs. 65.0%).

Conclusion

The combination of radiotherapy and immunotherapy is a promising therapeutic strategy for LS-SCLC, demonstrating encouraging efficacy outcomes that appear favorable compared to historical benchmarks for chemoradiotherapy alone. Optimizing treatment sequencing, particularly favoring a concurrent approach, is crucial for maximizing clinical benefit. These findings support further investigation in randomized controlled trials to confirm the value of this combined modality and to identify predictive biomarkers for patient selection.

Adjuvant Icotinib in EGFR-mutated stage IB non-small cell lung cancer with high-risk factors: A retrospective case series

by Mengzhi Cheng, Jianbin Zhang, Lili Jin, Caihua Yu, Zhonghai Xie, Dong Li, Qinhua Gu, Qibin Shen

Primary results of the CORIN trial indicated that, compared with chemotherapy, icotinib significantly improved 3-year disease-free survival (DFS) in patients with Epidermal Growth Factor Receptor (EGFR)-mutated stage IB non-small cell lung cancer (NSCLC). However, evidence regarding the outcomes of adjuvant icotinib in patients with high-risk factors remains limited. This retrospective study evaluated the efficacy and safety of adjuvant icotinib in patients with EGFR-mutated high-risk stage IB NSCLC. We enrolled 37 patients with completely resected EGFR-mutated high-risk stage IB NSCLC. The median follow-up time was 31 months, and the 3-year DFS rate was 91.4%. Two patients experienced disease recurrence and were successfully switched to osimertinib upon identification of an EGFR (T790M) mutation. Although overall survival (OS) and central nervous system (CNS)-DFS data were not mature, no deaths or central nervous system metastases were observed by the end of follow-up. 29 (78.4%) patients experienced grade 1–2 adverse events (AEs), no grade 3 or higher AEs occurred. This study suggests a potential DFS benefit and well-tolerated profile of adjuvant icotinib in patients with EGFR-mutated high-risk stage IB NSCLC. However, longer-term follow-up is necessary to assess the long-term outcomes.

Generational mutation patterns in a honey bee Deformed wing virus via infectious clones

by Anthony Nearman, Alriana Buller-Jarrett, Dawn Boncristiani, Eugene Ryabov, Yanping Chen, Jay D. Evans

Efforts to improve honey bee colony health continue due to persistent high loss rates. A major focus in this area is Deformed wing virus (DWV), a key driver of colony loss. The application of modern molecular techniques has characterized the DWV genome and its high mutational rate that enables the formation of diverse quasi-species populations capable of evading host immune responses, while other work has led to the development of DWV clones suitable for sequence-specific tracking of viral dynamics. In this work we combine knowledge of these efforts to track the mutational progression in a DWV clone surrounding an area of low nucleotide diversity and compare it to its wild-type source. We achieve this through amplicon sequencing of the structural viral protein, VP2, after incubation across three generations and multiple host genetic sources. Inocula were injected into pupae, allowed to replicate, then extracted for a further two generations of injections. For the final injection generation, recipient pupae were injected with preparations from either the same genetic source or cross-fostered from other colonies. Overall, we compared the mean number and type of mutations, their proportional abundance in the read pool, and specific locations across strains. Sequencing results indicate a limited number of mutational hotspots, which were driven by silent mutations in the final injection generation of the wild-type strains. No significant differences were found among other mutation types, cross-fostering status, or interactions with host genetics. This work is an initial attempt at examining viral dynamics in a cloned system across multiple generations and treatment groups. The results provide valuable insights, which may further enhance our understanding of viral dynamics and potentially improve future honey bee therapeutics.

Temporal dynamics of emotional face processing in social anxiety

by Ya-Chun Feng, Bo-Cheng Kuo, Wen-Yau Hsu

Previous studies have demonstrated that emotional facial expressions influence attention and perception in individuals with social anxiety. However, the relative influence of positive versus negative expressions on distinct subprocesses of attention and perception remains unclear. This event-related potential (ERP) study investigates the temporal dynamics of electrophysiological responses to emotional faces in high (HSA; N = 56) or low (LSA; N = 47) social anxiety individuals using a dot-probe task. Four face pairs (angry-neutral, happy-neutral, angry-happy, and neutral-neutral) were presented to probe the influence of positive and negative expressions. While behavioural results showed no significant group differences in attention bias, ERP results showed a reduced N170 amplitude for the HSA vs. LSA group in angry-neutral, happy-neutral, and angry-happy face pairs. Furthermore, enhanced N2pc effects to emotional expressions were found only in the HSA group when angry-neutral and happy-neutral face pairs were presented. No N2pc effect emerged when both positive and negative expressions were presented simultaneously. Finally, no significant P1 effect was found. Together, both positive and negative expressions influenced attentional deployment and face-specific processing in relation to social anxiety. Socially anxious individuals perceived less emotional facial information, yet their attention was biased by both negative and positive expressions.

Identification of pathogenic variants for the development of ultra-long axial length in myopic children

by YanYing Zhu, XueYan Li, YueXin Chen, HaiYan Xie, YuKun Liu, XiaoChen Xu, Jing Wang

Purpose

Axial elongation is a key factor in myopia progression, yet its genetic basis remains incompletely understood. This study aims to identify pathogenic genetic variants associated with excessively elongated axial length in children.

Methods

This study included 56 children with axial lengths exceeding the normal range for their age group, and whole-exome sequencing (WES) was performed on their oral mucosal samples. Clinical evaluations included axial length measurement, refraction testing, and fundus photography to assess the degree of myopia and retinal changes. Co-segregation analysis was conducted in selected families (F#1, F#2, F#5) to validate the familial inheritance patterns of the variants.

Results

Fifteen children carried variants in genes including BBS2, OPN1LW, P4HA2, FBN1, LOXL3, FZD4, USH2A, COL2A1, and BFSP2, with five novel variants identified: BBS2 (c.700C > T), P4HA2 (c.1382C > G), FBN1 (c.7130T > C), LOXL3 (c.1580delC), and FZD4 (c.1315G > A). Notably, a rare compound heterozygous BBS2 variant (c.700C > T/c.534 + 1G > T) was found in a non-syndromic child, and the P4HA2 (c.419A > G) variant in family F#5 exhibited a phenotype distinct from previous studies.

Conclusions

This study identified five novel variants sites and discovered two cases with phenotypes distinct from previous studies, thereby expanding the genetic variant spectrum associated with myopia and providing new targets for genetic screening and intervention.

Analytical validation of a homologous recombination deficiency signature (HRDsig) in pan-tumor tissue samples

by Wenshu Li, Jeffrey A. Leibowitz, Shuoguo Wang, Louisa Walker, Chang Xu, Kuei-Ting Chen, Alexa B. Schrock, Jason Hughes, Nimesh Patel, Julia A. Elvin, Lauren L. Ritterhouse, Ethan Sokol, Garrett Frampton, Lucas Dennis, Bahar Yilmazel, Brennan Decker

Homologous recombination repair (HRR) is a cellular pathway for high-fidelity double strand DNA break repair that uses the sister chromatid as a guide to ensure chromosomal integrity and cell viability. Deficiency in the HRR pathway (HRD) can sensitize tumors to poly (ADP-ribose) polymerase inhibitors (PARPi) and platinum-based chemotherapy, offering an avenue to identify patients who may benefit from targeted therapies. HRD signature (HRDsig) is a pan-solid-tumor biomarker on the FoundationOne®CDx (F1CDx®) assay that employs a DNA scar-based approach to calculate a score based on copy number features (e.g., segment size, oscillation patterns, and breakpoints per chromosome arm) and does not rely on HRR gene alterations, enabling detection of genomic and epigenetic mechanisms of HRD. After finalizing the HRDsig algorithm, analytical validation was conducted in a CAP-accredited, CLIA-certified laboratory on 278 solid tumor and normal tissue specimens. HRDsig results were compared with an independent HRD biomarker, defined by the presence of a reversion mutation restoring HRR gene function. In this evaluation, 100 HRD-positive and 126 HRD-negative samples showed a positive percent agreement of 90.00% and a negative percent agreement of 94.44%. The limit of detection (LoD) was estimated at 23.04% tumor purity, with the limit of blank (LoB) confirmed as zero in 60 normal tissue replicates. Reproducibility testing on 11 positive and 11 negative samples across multiple labs, reagent lots, and sequencers yielded agreement in 99.49% of positive and 99.73% of negative replicates. HRDsig status remained consistent in the presence of interfering substances, demonstrating 100% concordance in spiked samples. These validation results underscore the high analytical concordance, low false-positive rate, and overall robustness of HRDsig for reliable assessment of homologous recombination deficiency.

Computational identification of novel SIRT4 inhibitors for diabetic nephropathy using pharmacophore modeling, molecular simulations, and DFT calculations

by Wenxiang He, Jianwu Chen

Sirtuin 4 (SIRT4) plays a critical role in regulating oxidative stress, apoptosis, and mitochondrial dysfunction in diabetic nephropathy (DN). This study employed a multi-step in silico strategy to identify novel SIRT4 modulators with potential therapeutic relevance for DN. A ligand-based pharmacophore model was developed using UBCS182, followed by virtual screening of 3,285 compounds from major chemical libraries. Molecular docking revealed strong binding affinities (−9.46 to −8.41 kcal/mol), with CSC057320968, PubChem-162316407, and ChemDiv-V013-1548 emerging as top candidates. ADMET analysis confirmed their favorable pharmacokinetic and toxicity profiles. Subsequent 200 ns molecular dynamics simulations demonstrated the stability of protein–ligand complexes, with CSC057320968 exhibiting the most stable interaction profile based on RMSD, RMSF, Rg, and contact frequency analyses. Principal component analysis and free energy landscapes indicated conformational rigidity and energetic favorability for CSC057320968. Density Functional Theory (DFT) analysis further validated its reactivity and chemical softness, supporting its potential as a lead scaffold. This integrated computational pipeline provides novel insights into SIRT4 modulation and offers a rational framework for targeting mitochondrial dysfunction in DN.

Prevalence of extended spectrum β-lactamase and carbapenemase-producing <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> in raw bulk cow milk from dairy cooperatives, Northwest Amhara, Ethiopia

by Chalachew Genet, Wendemagegn Enbiale, Anna Rommerskirchen, Rajiha Abubeker, Wudu Tafere, Tsehaynesh Gebre-Eyesus, Michael Getie, Alem Tsega, Muluken Acham, Addisu Melese, Tewachew Awoke, Wondemagegn Mulu, Degu Ashagrie, Tadele Amsalu, Achenef Motbainor, Endalew Gebeyehu, Mulugeta Kibret, Bayeh Abera, Endalkachew Nibret, Abaineh Munshea

Introduction

Extended spectrum β-lactamase (ESBL) and carbapenemase-producing Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) emanating from raw cow milk are among the leading contributors to the spread of antimicrobial resistance (AMR). Due to the misuse and overuse of antibiotics in dairy farms, cow’s milk has become a reservoir of ESBL- and carbapenemase-producing E. coli and K. pneumoniae posing a growing public health threat, especially in areas where the consumption of raw milk is common. However, compared to the clinical sector, the prevalence of ESBL- and carbapenemase-producing E. coli and K. pneumoniae in the food sector is under-studied.

Objective

This study aimed to determine the prevalence of ESBL and carbapenemase-producing E. coli and K. pneumoniae in raw bulk cow milk from Dairy Cooperatives in Northwest Amhara, Ethiopia.

Methods

A cross-sectional study was conducted from January to April, 2025 among 257 dairy cooperative member farms. Sociodemographic and related data were collected using a structured questionnaire. Five milliliters of raw bulk cow milk were collected aseptically from each farm in four Dairy Cooperatives (DCs) (DC-A to D). 10 microliters of milk sample were directly inoculated into MacConkey agar. Escherichia coli and K. pneumoniae were identified using standard microbiological techniques. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method. ESBL and carbapenemase production were confirmed phenotypically via combination disk tests and modified carbapenem inactivation methods, respectively.

Results

The prevalence of E. coli and/or K. pneumoniae in raw cow milk was 21% (95% CI, 16.5–26.4%), with respective individual prevalence of 8.2% and 14.8%. ESBL-producing E. coli and K. pneumoniae accounted for 23.8% and 15.8% of isolates, respectively, while 2.6% of isolates (only K. pneumoniae) were carbapenemase producers. Resistance to ampicillin and amoxicillin-clavulanic acid exceeded 70%. All E. coli and 94.7% of K. pneumoniae isolates remained susceptible to carbapenems. Nearly half of all isolates (45.8%) were multidrug resistant (MDR), and 51.9% of MDR isolates were co-resistant to at least six antibiotics. Having additional non-farming occupations (AOR: 4.17, 95% CI: 1.49–11.67), large herd size (AOR: 3.21, 95% CI: 1.26–8.18), having pet animals (AOR: 6.53, 95% CI: 1.39–30.7), and use of calabash milk pail (AOR: 7.37, 95% CI: 1.45–37.49) were significantly associated with milk culture positive result for E. coli and/or K. pneumoniae.

Conclusion

Raw milk in Northwest Amhara harbors ESBL and carbapenemase-producing E. coli and K. pneumoniae posing a substantial public health risk coupled with MDR and resistance to critically important antimicrobials. Strengthened AMR surveillance, improved farm hygiene, restricted antibiotic use, and public education on milk safety are urgently needed.