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Circular RNAs (circRNAs) are natural outputs of eukaryotic transcription and RNA processing and have emerged as critical regulators in physiology and diseases. Although multiple cis-elements and trans-factors are reported to modulate the backsplicing of circRNA biogenesis, most of these regulations play roles in flanking introns of circRNAs. Here, using a genome-wide CRISPR knockout screen, we have identified an evolutionarily conserved RNA-binding protein ZC3H14 in regulating circRNA biogenesis. ZC3H14 binds to 3' and 5' exon-intron boundaries and 3' UTRs of cognate mRNAs to promote circRNA biogenesis through dimerization and the association with spliceosome. Yeast knockout of the ZC3H14 ortholog Nab2 has significantly lower levels of circRNAs. Zc3h14 -/- mice exhibit disrupted spermatogenesis and reduced testicular circRNA levels. Additionally, expression levels of human ZC3H14 are associated with non-obstructive azoospermia. Our findings reveal a conserved requirement for ZC3H14 in the modulation of backsplicing and link ZC3H14 and circRNA biogenesis to male fertility., Competing Interests: Declaration of interests G.S., X.W., and Q.L. have an ownership interest in a patent related to this research., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Introduction: Progression to Alzheimer's disease (AD) dementia from normal cognition (NC) can follow different trajectories, with most progressing through a recognizable mild cognitive impairment stage (NC-MCI-AD), while some individuals transition quickly from NC to AD dementia (NC-AD)., Methods: We compared demographic characteristics, health factors, and cognitive and functional assessments across three time points: the first NC visit, the last NC visit, and the first AD dementia visit., Results: The NC-MCI-AD group showed greater impairment in cognitive and functional scores at AD dementia diagnosis, despite maintaining better cognitive function during the NC stage. Analysis of yearly changes revealed negligible differences during NC. However, the yearly change during the AD dementia stage suggested potentially more rapid functional decline in the NC-AD group., Discussion: These findings highlight the heterogeneity in AD disease progression and emphasize the importance of considering diverse progression patterns in AD research and clinical practice., Highlights: We investigated the disease progression difference between patients who converted to Alzheimer's disease (AD) dementia from normal cognition (NC) directly or through the mild cognitive impairment (MCI) stage. We found that the NC-MCI-AD group showed greater impairment in cognitive and functional scores at AD dementia diagnosis. We discovered that the NC-AD group had rapid functional decline once patients were confirmed with AD onset., (© 2024 The Author(s). Alzheimer's & Dementia published by Wiley Periodicals LLC on behalf of Alzheimer's Association.)

Photodynamic therapy is an efficient approach to promote cytotoxic T lymphocyte tumor infiltration to convert immunologically cold tumors into hot tumors through the induction of immunogenic cell death . However, tumors usually overexpress immune checkpoints such as PD-L1 to suppress T lymphocyte antitumor activity and evade immune surveillance. Therefore, the design of efficient photosensitizers to overcome checkpoint-mediated immune evasion is highly necessary. In this work, we report the design of BRD-PS , a BRD4-targeting photosensitizer, as a new class of immunomodulatory photosensitizer termed an immune checkpoint-modulating photosensitizer, to solve this issue. On one hand, BRD-PS induces immunogenic pyroptosis and ferroptosis to promote the activation and tumor infiltration of cytotoxic T cells. On the other hand, BRD-PS suppresses the expression of PD-L1 to avoid immune evasion. This work demonstrated the feasibility of utilizing a single photosensitizer to simultaneously induce immunogenic cell death and PD-L1 downregulation for synergistic cancer photoimmunotherapy.

Simultaneous separation of compounds with multiple chiral centers and highly similar structures presents significant challenges. This study developed a novel supercritical fluid chromatography (SFC) method with reduced organic solvent consumption and robust separation capabilities to address these challenges. The method was applied to simultaneously achieve enantioselective, diastereoselective, and achiral separation of palonosetron hydrochloride and its six impurities. The effects of the polysaccharide-based chiral stationary phase (CSP), modifier, additive, and column temperature on retention and separation were comprehensively evaluated. It was found that a combination of a polysaccharide-based CSP and a single modifier or a mixture of protonic modifiers could not achieve complete separation due to high structural similarity. However, an ADH column and a ternary solvent mixture containing acetonitrile (methanol: acetonitrile: diethylamine, 60:40:0.2, v/v/v) provided satisfying separation, particularly for the enantiomer and diastereomers of palonosetron. Using the optimized method, the enantioselective, diastereoselective, and achiral separation of palonosetron hydrochloride and its six impurities can be accomplished in 18 min under gradient elution. Thermodynamic results indicated that the separation process was entropy driven. A molecular docking study revealed that the separation was mainly achieved through the differences in hydrogen bond and π - π interactions between the analytes and CSP. This study lays the foundation for SFC analysis of palonosetron hydrochloride and provides a reference for the simultaneous SFC separation of the enantiomers, diastereoisomers and structurally similar compounds., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The epidermal growth factor receptor (EGFR) is a recognized target in tumor treatment. While there is significant focus on inhibiting membrane EGFR and its downstream signaling activation, the ectopic accumulation of EGFR, particularly nuclear EGFR (nEGFR), has been implicated in tumor-associated activities and associated with poor prognosis. Within the nucleus, nEGFR functions as a transcriptional regulator to modulate transcriptional landscape and exerts tyrosine kinase activity to phosphorylate nuclear proteins and subsequently influences DNA repair, cell cycle, proliferation, and resistance to radiotherapy and chemotherapy. The nuclear localization of EGFR involves the internalization, subcellular trafficking, and nuclear envelope shuttling of membrane EGFR. Given the challenges of delivering drugs to the nucleus for targeting nEGFR, understanding the molecules affecting the translocation process is crucial for novel insights. This review initially explores the association between nEGFR expression and clinical outcomes and then elucidates how nEGFR fulfills its regulatory role within the nucleus. Subsequently, the mechanisms governing EGFR nuclear translocation and potential therapeutic targets during this process are summarized, highlighting avenues to target nEGFR as an innovative strategy in tumor treatment., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Aims: Circular RNAs (circRNAs) are considered important regulators of biological processes, but their impact on atherosclerosis development, a key factor in coronary artery disease (CAD), has not been fully elucidated. We aimed to investigate their potential use in patients with CAD and the pathogenesis of atherosclerosis., Methods and Results: Patients with stable angina (SA) or acute coronary syndrome (ACS) and controls were selected for transcriptomic screening and quantification of circRNAs in blood cells. We stained carotid plaque samples for circRNAs and performed gain- and loss-of-function studies in vitro. Western blots, protein interaction analysis, and molecular approaches were used to perform the mechanistic study. ApoE-/- mouse models were employed in functional studies with adeno-associated virus-mediated genetic intervention. We demonstrated elevated circARCN1 expression in peripheral blood mononuclear cells from patients with SA or ACS, especially in those with ACS. Furthermore, higher circARCN1 levels were associated with a higher risk of developing SA and ACS. We also observed elevated expression of circARCN1 in carotid artery plaques. Further analysis indicated that circARCN1 was mainly expressed in monocytes and macrophages, which was also confirmed in atherosclerotic plaques. Our in vitro studies provided evidence that circARCN1 affected the interaction between HuR and ubiquitin-specific peptidase 31 (USP31) mRNA, resulting in attenuated USP31-mediated NF-κB activation. Interestingly, macrophage accumulation and inflammation in atherosclerotic plaques were markedly decreased when circARCN1 was knocked down with adeno-associated virus in macrophages of ApoE-/- mice, while circARCN1 overexpression in the model exacerbated atherosclerotic lesions., Conclusions: Our findings provide solid evidence macrophagic-expressed circARCN1 plays a role in atherosclerosis development by regulating HuR-mediated USP31 mRNA stability and NF-κB activation, suggesting that circARCN1 may serve as a factor for atherosclerotic lesion formation., Competing Interests: Conflict of interest: none declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of the European Society of Cardiology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. The authors declare no competing interests related to the publication of this manuscript. Financial support for this study was provided by the Western Australian Future Health Research and Innovation Fund (Grant ID WANMA/EL2023-24/2). The funder had no role in the design, analysis, or interpretation of the study, nor in the decision to publish the results.

Background: Gongju is recognized as one of the four traditional Chinese medicinal herbs, and its main constituents are chlorogenic acid (CGA) and its derivative material. CGA content in autooctoploid Gongju are considerably elevated compared with those in parental tetraploid Gongju at different flowering stages. However, the underlying molecular mechanisms governing the regulation CGA content remain poorly understood., Methods: Therefore, we conducted integrated transcriptome and metabolome analyses of different flowering stages in autooctoploid and tetraploid Gongju to elucidate the underlying molecular mechanisms governing CGA biosynthesis., Results: Transcriptome analysis showed that the number of differentially expressed genes in the budding stage (BS), early flowering stage (EF), and full flowering stage (FF) of tetraploid and octoploid Gongju were 3859, 11,211, and 6837, respectively. A total of 563, 466, and 394 differential accumulated metabolites were respectively identified between the bud stages of tetraploid and octoploid Gongju (4BS vs. 8BS), between the early flowering stages of tetraploid and octoploid Gongju (4EF vs. 8EF), and the full flowering stages of tetraploid and octoploid Gongju (4FF vs. 8FF) groups. The integrated analysis of transcriptomics and metabolomics revealed that the expression of pma6460 and mws0178, which are key enzymes involved in the CGA synthesis pathway, increased during the flowering stages of octoploid Gongju relative to that of tetraploid Gongju. The expression levels of CmHQT and CmC3H genes associated with CGA synthesis were higher in octoploid plants than in tetraploid plants at various flowering stages. To investigate the potential regulation of transcription factors involved in CGA synthesis, we analyzed the coexpression of CmC3H and CmHQT with CmMYBs and CmbHLHs . Results indicated that transcription factors, such as CmMYB12 (Cluster-30519.0), CmMYB26 (Cluster-75874.0), CmMYB5 (Cluster-94106.0), CmMYB1 (Cluster-71968.7), CmbHLH62 (Cluster-32024.1), CmbHLH75 (Cluster-62341.0), CmbHLH62 (Cluster-32024.8), CmbHLH75 (Cluster-60210.0), and CmbHLH16 (Cluster-90665.1) play a pivotal role in CGA synthesis regulation. The present study provides novel insights into the molecular mechanisms underlying CGA accumulation in autopolyploid Gongju., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhao, Cao, Shan, Zhou, Li, Liu, Wang, An and Gao.)

KRAS is among the most commonly mutated oncogenes in human malignancies. Although the advent of sotorasib and adagrasib, has lifted the "undruggable" stigma of KRAS, the resistance to KRAS inhibitors quickly becomes a major issue. Here, we reported that aldehyde dehydrogenase 1 family member A1 (ALDH1A1), an enzyme in retinoic acid biosynthesis and redox balance, increases in response to KRAS inhibitors and confers resistance in a range of cancer types. KRAS inhibitors' efficacy is significantly improved in sensitive or drug-resistant cells, patient-derived organoids (PDO), and xenograft models by ALDH1A1 knockout, loss of enzyme function, or inhibitor. Furthermore, we discovered that ALDH1A1 suppresses the efficacy of KRAS inhibitors by counteracting ferroptosis. ALDH1A1 detoxicates deleterious aldehydes, boosts the synthesis of NADH and retinoic acid (RA), and improves RARA function. ALDH1A1 also activates the CREB1/GPX4 pathway, stimulates the production of lipid droplets in a pH-dependent manner, and subsequently prevents ferroptosis induced by KRAS inhibitors. Meanwhile, we established that GTF2I is dephosphorylated at S784 via ERK by KRAS inhibitors, which hinders its nuclear translocation and mediates ALDH1A1's upregulation in response to KRAS inhibitors. In summary, the results offer valuable insights into targeting ALDH1A1 to enhance the effectiveness of KRAS-targeted therapy through ferroptosis in cancer treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Inhibiting the shuttle of polysulfides is of great significance for promoting the practical application of lithium-sulfur batteries (LSBs). Here, an imine-linked covalent organic framework@carbon nanotube (COF@CNT) interlayer composed of triazine and boroxine rings is constructed between the sulfur cathode and the separator for polysulfides reception and reutilization. The introduction of CNT imparts the conductor characteristic to the interlayer attributed to electron tunneling in thin COF shell, and creates a hierarchical porous architecture for accommodating polysulfides. The uniform distribution of amphiphilic adsorption sites in COF microporous structure not only enables efficient entrapment of polysulfides while allowing the penetration of Li + ions, but also provides a stable electrocatalytic channel for bidirectional conversion of active sulfur to achieve the substantially improved capacity and stability. The interlayer-incorporated LSBs deliver an ultrahigh capacity of 1446 mA g -1 at 0.1C and an ultralow capacity decay rate of 0.019% at 1C over 1500 cycles. Even at an electrolyte/sulfur ratio of 6 µL mg -1 , an outstanding capacity of 995 mAh g -1 and capacity retention of 74.1% over 200 cycles at 0.2C are obtained. This work offers a compelling polysulfides entrapment and reactivation strategy for stimulating the study on ultra-stable LSBs., (© 2024 Wiley‐VCH GmbH.)

Ferroptosis is a regulated cell death driven by iron-dependent lipid peroxidation and associated with drug resistance in lung adenocarcinoma (LUAD). It's found that aldehyde dehydrogenase 2 (ALDH2), which is highly mutated in East Asian populations, is correlated with response to chemotherapy in LUAD patients. The rs671 variant knock-in, downregulation, and pharmacological inhibition of ALDH2 render LUAD cells more vulnerable to ferroptosis inducers and platinum-based chemotherapy. ALDH2 inhibits ferroptosis through the detoxification of 4-hydroxynonenal and malondialdehyde, the product of lipid peroxidation, as well as the production of NADH at the same time. Besides, ALDH2 deficiency leads to elevated intracellular pH (pHi), thus inhibiting the ERK/CREB1/GPX4 axis. Interestingly, ALDH2 is also regulated by CREB1, and the ALDH2 enzyme activity was decreased with elevated pHi. What's more, the elevated pHi caused by impaired ALDH2 activity promotes the biosynthesis of lipid droplets to counteract ferroptosis. At last, the effect of ALDH2 on ferroptosis and chemosensitivity is confirmed in patient-derived organoids and xenograft models. Collectively, this study demonstrates that ALDH2 deficiency confers sensitivity to platinum through ferroptosis in LUAD, and targeting ALDH2 is a promising new strategy to enhance the sensitivity of platinum-based chemotherapy for the treatment of LUAD patients., Competing Interests: Declaration of competing interest The authors declare no conflict of interest in this work., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Lay Abstract: Self-determination encompasses various components, including decision-making and independence, making it a complex process. While the importance of self-determination for individuals with autism spectrum disorder has been explored in previous studies, there is limited research focusing on individuals with moderate to severe autism spectrum disorder. Evidence-based practices such as visual activity schedules and video modeling have shown effectiveness in promoting independence among individuals with autism spectrum disorder. To address the need for independence and choice-making among individuals with moderate to severe autism spectrum disorder, this study developed a visual support package incorporating visual activity schedules, video modeling, preference assessments, and prompt procedures. By investigating the intervention's effectiveness in three participants, this study contributes to the existing literature on the use of a visual activity schedule and video modeling in enhancing choice-making and independent leisure engagement. Following the intervention, all participants were able to select three leisure activities, develop their own visual schedules, and complete them. Notably, this study conducted preference assessments to determine participants' preferred leisure activities and did not provide additional reinforcement. Practical implications of this research include incorporating video prompting as needed and adjusting activity engagement time. Future research should explore the long-term effectiveness of the visual support package and its application in developing novel skills or vocational activities for individuals with moderate to severe autism spectrum disorder. This study fills a critical gap in the literature, providing important insights for practices and research in the field of autism spectrum disorder interventions., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Introduction: Radiotherapy (RT) is the main treatment for patients with nasopharyngeal carcinoma (NPC). NPC patients at different stages have varying levels of damage to normal brain tissue after RT. No study has yet thoroughly analyzed the variations in radiation dosages in the brain for different stages of NPC patients treated with RT. This study aims to examine these variations., Methods: 1446 NPC patients' CT and RTdose data were retrospectively reviewed. Analysis of the radiation dosage was executed on these 803 patients. The RTdose images for several patient groups were averaged after registering each patient's RTdose data to the CT brain template created in our earlier study. The voxel-based (VB) analysis was used to examine the dose variations in the brains of three groups of NPC patients: the early-stage group, the stage III group, and the stage IV group., Results: As the disease progresses from early to advanced stages, the intensity and volume of radiation in the brain increase. The normal brain tissue accepted a substantially larger dosage in more advanced NPC patients. Differences in brain regions between stage III and early-stage patients were minimal compared to any other two groups. Brain regions exhibited substantial variations between the stage IV group and all other patient groups were broadly distributed., Conclusion: Our findings highlight the critical role of NPC staging in the therapeutic strategy, emphasizing the heterogeneity of radiation-induced tissue damage across disease stages and implying the need to develop stage-specific RT plans., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Blastocyst morphology has been demonstrated to be associated with ploidy status. Existing artificial intelligence models use manual grading or 2D images as the input for euploidy prediction, which suffer from subjectivity from observers and information loss due to incomplete features from 2D images. Here we aim to predict euploidy in human blastocysts using quantitative morphological parameters obtained by 3D morphology measurement., Methods: Multi-view images of 226 blastocysts on Day 6 were captured by manually rotating blastocysts during the preparation stage of trophectoderm biopsy. Quantitative morphological parameters were obtained by 3D morphology measurement. Six machine learning models were trained using 3D morphological parameters as the input and PGT-A results as the ground truth outcome. Model performance, including sensitivity, specificity, precision, accuracy and AUC, was evaluated on an additional test dataset. Model interpretation was conducted on the best-performing model., Results: All the 3D morphological parameters were significantly different between euploid and non-euploid blastocysts. Multivariate analysis revealed that three of the five parameters including trophectoderm cell number, trophectoderm cell size variance and inner cell mass area maintained statistical significance (P < 0.001, aOR = 1.054, 95% CI 1.034-1.073; P = 0.003, aOR = 0.994, 95% CI 0.991-0.998; P = 0.010, aOR = 1.003, 95% CI 1.001-1.006). The accuracy of euploidy prediction by the six machine learning models ranged from 80 to 95.6%, and the AUCs ranged from 0.881 to 0.984. Particularly, the decision tree model achieved the highest accuracy of 95.6% (95% CI 84.9-99.5%) with the AUC of 0.978 (95% CI 0.882-0.999), and the extreme gradient boosting model achieved the highest AUC of 0.984 (95% CI 0.892-1.000) with the accuracy of 93.3% (95% CI 81.7-98.6%). No significant difference was found between different age groups using either decision tree or extreme gradient boosting to predict euploid blastocysts. The quantitative criteria extracted from the decision tree imply that euploid blastocysts have a higher number of trophectoderm cells, larger inner cell mass area, and smaller trophectoderm cell size variance compared to non-euploid blastocysts., Conclusions: Using quantitative morphological parameters obtained by 3D morphology measurement, the decision tree-based machine learning model achieved an accuracy of 95.6% and AUC of 0.978 for predicting euploidy in Day 6 human blastocysts., Trial Registration: N/A., (© 2024. The Author(s).)

Arrhythmogenic cardiomyopathy (ACM) is a leading cause of sudden cardiac death among young adults. Aberrant gap junction remodeling has been linked to disease-causative mutations in plakophilin-2 ( PKP2 ). Although gap junctions are a key therapeutic target, measurement of gap junction function in preclinical disease models is technically challenging. To quantify gap junction function with high precision and high consistency, we developed a robotic cell manipulation system with visual feedback from digital holographic microscopy for three-dimensional and label-free imaging of human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). The robotic system can accurately determine the dynamic height changes in the cells' contraction and resting phases, microinject drug-treated healthy and diseased iPSC-CMs in their resting phase with constant injection depth across all cells, and deposit a membrane-impermeable dye that solely diffuses between cells through gap junctions for measuring the gap junction diffusion function. The robotic system was applied toward a targeted drug screen to identify gap junction modulators and potential therapeutics for ACM. Five compounds were found to dose-dependently enhance gap junction permeability in cardiomyocytes with PKP2 knockdown. In addition, PCO 400 (pinacidil) reduced beating irregularity in a mouse model of ACM expressing mutant PKP2 (R735X). These results highlight the utility of the robotic cell manipulation system to efficiently assess gap junction function in a relevant preclinical disease model, thus providing a technique to advance drug discovery for ACM and other gap junction-mediated diseases.

Oxidative stress is a critical pathogenic factor for age-related macular degeneration (AMD). Autophagy serves as a mechanism to counteract oxidative stress. LAMTOR1 regulates mTORC1 activity by recruiting or disassembling it on the lysosome under the addition or deprivation of amino acids. This regulation inhibits or enhances autophagy. Our study investigates whether oxidative stress impacts LAMTOR1, thereby adapting to oxidative conditions. We employed oxidative stressors, menadione (VK3) and 4-hydroxynonenal (4-HNE), and observed a reduction of LAMTOR1 in both human and mouse retinal pigment epithelium (RPE) following short-term (1h) and prolonged exposures (24h). Nrf2 overexpression increased both lamtor1 mRNA and LAMTOR1 protein in the RPE. To determine if Nrf2 regulates lamtor1 transcription, we cloned the deletion mutants of the lamtor1 promoter into a luciferase reporter. Although the promoter contained antioxidant response elements, transcriptional activity depended on the interaction between Nrf2 and the constructs containing the transcriptional start site. Moreover, Nrf2-driven transcription was significantly reduced by an inhibitor of histone acetyltransferase, p300. Correspondingly, Nrf2 overexpression increased levels of acetylated histone 3 and p300. The reduction in LAMTOR1 by 4-HNE was reversed by pepstatin A and NH 4 Cl which block lysosomal degradation. 4-HNE increased TFEB nuclear translocation which was reversed by LAMTOR1 overexpression. In vivo, LAMTOR1 levels decreased in the photoreceptor and RPE layers of NaIO 3 -injected mice, compared to PBS-injected controls. In conclusion, oxidative injury reduces LAMTOR1, predominantly through lysosomal degradation although Nrf2-mediated histone acetylation enhances lamtor1 transcription. This study reveals a previously unrecognized regulatory mechanism of lamtor1 by oxidative stress, suggesting a novel role for LAMTOR1 in the pathogenesis of AMD., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Tumor genomic profiling has a significant impact on the selection of targeted therapy. Circulating tumor DNA (ctDNA) has emerged as a noninvasive, and reproducible assay compared with tissue biopsy. We aimed to evaluate its utility in identifying mutations and guiding targeted therapy for lung cancer., Methods: A total of 173 lung cancer patients underwent next-generation sequencing (NGS) using a targeted enrichment panel covering 20 lung cancer-related genes. The performance of the ctDNA NGS assay in identifying genetic mutations or alterations was compared with tissue biopsy and droplet digital PCR (ddPCR). The treatment response to epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) therapies based on the ctDNA assay results was also assessed., Results: The ctDNA was detected in 61.85% of patients. Tissue mutations were detected in paired ctDNA in 38.57% of cases, while ctDNA mutations were detected in paired tissues in 89.1% of cases. The ctDNA increased the number of advanced non-small cell lung cancer (NSCLC) patients who received NCCN-recommended genetic testing by 12%. The concordance between ddPCR and ctDNA was relatively high reaching 99.43%. EGFR T790M/C797S c.G2390C and EGFR T790M/C797S c.T2389A were detected in tissue and ctDNA, respectively, in patient 01015. Moreover, ctDNA assay identified the EGFR T790M mutation, which was missed by tissue biopsy in patient 01149, who developed drug resistance after 1 year of EGFR-TKI therapy. Of the 17 patients who received EGFR-TKI targeted therapies based on the ctDNA NGS results, 12 patients achieved a partial response and two patients had stable disease., Conclusions: The results demonstrated that the ctDNA assay could partially overcome tumor heterogeneity in detecting mutations and provide complementary information on tumor genomic profiles. Moreover, the presence of EGFR mutations in ctDNA could offer valuable guidance for selecting appropriate EGFR-TKI treatment for advanced lung cancer patients. However, it is important to note that the ctDNA NGS assay has certain limitations in fully identifying all genomic alterations present in the tumor., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2024.)

The co-occurrence of glyphosate (GPS), a commonly used organophosphorus herbicide, and cadmium (Cd), a neurotoxic metal, in agricultural environments prompts concerns about their combined toxic effects on ecosystems. This study explores the combined effects of GPS and Cd on the model organism Caenorhabditis elegans (C. elegans), to understand their cumulative effects in organismal living environments. We investigated the interaction between GPS and Cd over 24 hours using a comprehensive approach that included a variety of toxicity endpoints as well as the novel Automated Recognition and Statistics Tool (NCLE) for body bend measurement. Our data show a concentration-dependent interplay in which antagonistic effects at lower concentrations reduce phenotypic damage while synergistic effects emerge at higher concentrations, particularly at GPS's LC50. Transcriptome analysis under antagonistic conditions revealed significant downregulation of Cd toxicity-related genes and identified Y22D7AL.16, which has a C2H2-type zinc finger domain, as a novel gene involved in metal stress response, implying an alternative Cd-resilience mechanism. The expression profile of this gene shows that it plays a larger role in both development and metal stress adaption. These findings highlight the complexities of compound pollutant interactions, emphasizing the importance of including such dynamics in environmental risk assessments and control techniques., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The escalating concern of perfluoroalkyl and polyfluoroalkyl substances (PFAS), particularly at contaminated sites, has prompted extensive investigations. In this study, samples of multimedia including air, rhizosphere soil, and tissues of wheat at various growing stages were collected near a mega fluorochemical industrial park in China. Perfluorooctanoic acid (PFOA) was predominant in both air and soil with a strong correlation, highlighting air deposition as an important source in the terrestrial system. PFAS concentrations in wheat decreased in the stem and ear but increased in the leaves as wheat matured. Specifically, perfluorobutanoic acid (PFBA) dominated in the aboveground tissues in the full-life-cycle, except that PFOA surpassed and became predominant in leaves during the filling and maturing stages, hinting at an airborne source. For all PFAS, both bioaccumulation factors and translocation factors (TFs) were inversely correlated with the carbon chain length during the full-life-cycle. The obtained TF values were considerably higher than those obtained from ambient sites reported previously, further suggesting an unneglectable foliar uptake from air, which was estimated to be 25% for PFOA. Moreover, spray irrigation remarkably enhanced the absorption of PFAS in wheat via foliar uptake relative to flood irrigation. The estimated daily intake of PFBA via wheat consumption and air inhalation was 0.50 μg/kg/day for local residents, at least one magnitude higher than the corresponding threshold, suggesting an alarmingly high exposure risk.

Background: Studies about the combined effects of gaseous air pollutants and particulate matters are still rare., Objectives: This study was performed based on baseline survey of the Diverse Life-Course Cohort in the Beijing-Tianjin-Hebei (BTH) Region of North China to evaluate the association of long-term air pollutants with blood pressure and the combined effect of the air pollutants mixture among 32821 natural han population aged 20 years or above., Methods: Three-year average exposure to air pollutants (PM 10 , PM 2.5 , PM 1 , O 3 , SO 2 , NO 2 , and CO) and PM 2.5 components [black carbon (BC), ammonium (NH 4 + ), nitrate (NO 3 ), and organic matter (OM)] of residential areas were calculated based on well-validated models. Generalized linear mixed models (GLMMs) were used to estimate the associations of air pollutants exposure with the systolic blood pressure (SBP), diastolic blood pressure (DBP), Mean arterial pressure (MAP), pulse pressure (PP) and prevalent hypertension. Quantile g-Computation and Bayesian Kernel Machine Regression (BKMR) were employed to assess the combined effect of the air pollutant mixture.- ), sulfate (SO 4 2- ), and organic matter (OM)] of residential areas were calculated based on well-validated models. Generalized linear mixed models (GLMMs) were used to estimate the associations of air pollutants exposure with the systolic blood pressure (SBP), diastolic blood pressure (DBP), Mean arterial pressure (MAP), pulse pressure (PP) and prevalent hypertension. Quantile g-Computation and Bayesian Kernel Machine Regression (BKMR) were employed to assess the combined effect of the air pollutant mixture., Results: We found that long-term exposures of O 3 , PM 2.5 , and PM 2.5 components were stably and strongly associated with elevated SBP, DBP, and MAP and prevalent hypertension. O 3 increased SBP, DBP, and MAP at a similar extent, but with greater effects; while, PM 2.5 and PM 2.5 components had a greater impact on SBP than DBP, which increased PP simultaneously. In multi-pollutant models, the combined effects of the air pollutant mixture on blood pressure and prevalent hypertension was predominantly influenced by O 3 , PM 2.5 , and O 3 , OM in different models, respectively. For example, O 3 , PM 2.5 contributed 57.25 %, 39.22 % of the positive combined effect of the air pollutant mixture on SBP; and O 3 , OM positively contributed 70.00 %, 30.00 % on prevalent hypertension, respectively. There were interactions between O 3 , CO, SO 2 and PM 2.5 components on hbp, SBP and PP., Conclusions: The results showed positive associations of air pollutant mixtures with blood pressure, where O 3 and PM 2.5 (especially OM) might be primary contributors. There were interactions between gaseous air pollutants and PM 2.5 components on blood pressure and prevalent hypertension., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: There have been few large-scale studies on the outcomes of cardiomyopathy-associated heart failure (HF) in infants aged <1 year. This study aimed to assess longitudinal echocardiographic outcomes of infants with HF secondary to cardiomyopathy who survived for >1 year., Methods and Results: A prospective observational study following 327 infant patients up to 5 years in 2 large pediatric heart centers in Northern China between January 2010 and December 2018. A total of 236 (72.2%) patients had reduced left ventricular ejection fraction (LVEF) (HF with reduced ejection fraction group; LVEF <40%), 91 (27.8%) patients had midrange LVEF (HF with midrange ejection fraction group; LVEF ≥40% but <55%). LVEF improved significantly within the first year and remained stable in years 2 through 5 for both groups. The HF with midrange ejection fraction group had a higher rate of LVEF normalization (hazard ratio, 1.65; P <0.001). Baseline LVEF ≥40%, baseline left ventricular end-diastolic diameter Z score <7.8, the absence of left bundle-branch block, and the absence of β-blocker use were 4 independent favorable predictors for future LVEF normalization. A total of 62.4% of enrolled patients were diagnosed with left ventricular noncompaction. No significant difference in LVEF normalization was found among the different types of cardiomyopathy studied., Conclusions: A significant number of infants with cardiomyopathy who survived >1 year were found to improve with medical therapies during the first year of diagnosis. Poorer outcomes were associated with decreased LVEF and increased heart size at diagnosis baseline, the presence of left bundle-branch block and use of β blockers. The Northern Chinese pediatric population may have a high proportion of left ventricular noncompaction.

Background: Esophageal squamous cell carcinoma (ESCC) is one of the most common malignant tumors, with high incidence and poor prognosis. Revealing mechanisms of ESCC progression and developing new therapeutic targets remains crucial. The aim of this study was to elucidate the molecular mechanism of miR-30c-5p in regulating the malignant progression of ESCC., Methods: TCGA, GEO, and other datasets were used to analyze the differential expression of miR-30c-5p in ESCC and adjacent tissues, and its impact on prognosis. Then the effects of miR-30c-5p on the proliferation, migration, and invasion of TE-1 and Eca9706 cells were investigated through proliferation experiments, transwell and wounding healing assays. The regulatory mechanism of miR-30c-5p on the PI3K/AKT signaling pathway and its interaction in cancer progression were investigated through Western blots, dual-luciferase reporter assay, and rescue experiments., Results: miR-30c-5p was significantly downregulated in ESCC tissue and represented a poor prognosis. miR-30c-5p mimic significantly inhibited the proliferation, migration, and invasion ability of ESCC, while miR-30c-5p inhibitor significantly promoted tumor cell progression. Through bioinformatic analysis and experimental results, miR-30c-5p interacted directly with PIK3CA mRNA and inhibited subsequent signaling pathway activation. PIK3CA activator could eliminate the inhibitory effects of miR-30c-5p mimic on the progression of ESCC, while PIK3CA inhibitors could rescue the promoting effect of miR-30c-5p inhibitor group cells., Conclusions: In summary, we found that miR-30c-5p inhibited the proliferation, invasion and migration of ESCC by inhibiting PI3K/AKT signaling pathway for the first time, and this study is expected to provide a novel insight and potential therapeutic target for managing ESCC., (© 2024 The Author(s). Thoracic Cancer published by John Wiley & Sons Australia, Ltd.)

Instance segmentation of biological cells is important in medical image analysis for identifying and segmenting individual cells, and quantitative measurement of subcellular structures requires further cell-level subcellular part segmentation. Subcellular structure measurements are critical for cell phenotyping and quality analysis. For these purposes, instance-aware part segmentation network is first introduced to distinguish individual cells and segment subcellular structures for each detected cell. This approach is demonstrated on human sperm cells since the World Health Organization has established quantitative standards for sperm quality assessment. Specifically, a novel Cell Parsing Net (CP-Net) is proposed for accurate instance-level cell parsing. An attention-based feature fusion module is designed to alleviate contour misalignments for cells with an irregular shape by using instance masks as spatial cues instead of as strict constraints to differentiate various instances. A coarse-to-fine segmentation module is developed to effectively segment tiny subcellular structures within a cell through hierarchical segmentation from whole to part instead of directly segmenting each cell part. Moreover, a sperm parsing dataset is built including 320 annotated sperm images with five semantic subcellular part labels. Extensive experiments on the collected dataset demonstrate that the proposed CP-Net outperforms state-of-the-art instance-aware part segmentation networks., Competing Interests: Declaration of competing interest There are no competing interest for all authors., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Rationale: Spirometry reference equations that are derived from a large, nationally representative general population are warranted in China, and the impact of using prebronchodilator (pre-BD) and post-BD spirometry reference values has yet to be assessed in Chinese populations. Objectives: To present the pre-BD and post-BD spirometry reference values for Chinese adults using the China Pulmonary Health (CPH) Study. Methods: A reference population of 17,969 healthy, nonsmoking participants in the CPH Study was used to calculate the pre- and post-BD reference values for FEV 1 , FVC, and FEV 1 /FVC ratio. Pre- and post-BD reference values were applied to the entire CPH population ( N  = 50,991) to illustrate the divergence between the use of different references in determining disease prevalence and severity grading. Measurements and Main Results: The prevalences of airflow limitation were 5.36% using the pre-BD reference and 8.02% using the post-BD reference. Individuals who had a post-BD FEV 1 /FVC ratio lower than the post-BD reference value but higher than the pre-BD reference value were found to have significantly higher rates of self-reported respiratory symptoms and significantly lower values on spirometry indicators than those whose post-BD FEV 1 /FVC ratio was greater than the post-BD reference value. An additional 3.51% of participants were identified as having grade II-IV chronic obstructive pulmonary disease using the post-BD FEV 1 predicted values. Conclusions: This study generated and applied pre- and post-BD spirometry reference values in a nationally representative Chinese adult population. Post-BD reference values may serve as an additional criterion in identifying individuals at risk for obstructive pulmonary diseases, and their diagnostic and prognostic values should be further investigated.

The past decade has witnessed the superior power of deep neural networks (DNNs) in applications across various domains. However, training a high-quality DNN remains a non-trivial task due to its massive number of parameters. Visualization has shown great potential in addressing this situation, as evidenced by numerous recent visualization works that aid in DNN training and interpretation. These works commonly employ a strategy of logging training-related data and conducting post-hoc analysis. Based on the results of offline analysis, the model can be further trained or fine-tuned. This strategy, however, does not cope with the increasing complexity of DNNs, because (1) the time-series data collected over the training are usually too large to be stored entirely; (2) the huge I/O overhead significantly impacts the training efficiency; (3) post-hoc analysis does not allow rapid human-interventions (e.g., stop training with improper hyper-parameter settings to save computational resources). To address these challenges, we propose an in-situ visualization and analysis framework for the training of DNNs. Specifically, we employ feature extraction algorithms to reduce the size of training-related data in-situ and use the reduced data for real-time visual analytics. The states of model training are disclosed to model designers in real-time, enabling human interventions on demand to steer the training. Through concrete case studies, we demonstrate how our in-situ framework helps deep learning experts optimize DNNs and improve their analysis efficiency.

Purpose: To investigate the epidemiological and clinical characteristics of pertussis in children and close contacts., Methods: Nasopharyngeal swabs and blood samples of clinically suspected children with pertussis and their close contacts from 2018 to 2022 were collected for pathogen detection of Bordetella pertussis. Questionnaires were designed to investigate the basic information and infection status of pertussis children cases and their close contacts. Descriptive epidemiological analysis was performed on the results., Results: 1229 confirmed children cases of pertussis were collected and infants < 1 year old were the most affected (77.7%). Etiological data were collected from 587 close contacts of 269 confirmed cases and the infection rate was high (24.4%). The positive detection rate of parents, especially mothers, was significantly higher than that of other groups (32.2% vs. 18.4%, P < 0.001); The rates of misdiagnosis or missed diagnosis in pertussis children (92.2%) and close contacts (99.8%) were very high, and the distribution of symptoms between pertussis children and their close contacts was different (χ 2  = 535.328, P < 0.001); The vast majority of pertussis children (84.0%) were diagnosed with upper respiratory tract infection or trachea/bronchitis while 91.0% of close contacts did not seek medical attention (χ 2  = 685.373, P < 0.001)., Conclusion: Infants < 1 year old are at high risk in pertussis. Pertussis infection in close contacts of confirmed children is underestimated. Caregivers who are positive for pertussis but missed or misdiagnosed seriously may be a main source of pertussis infections in children. Adjusting the current pertussis immunization strategy in China is necessary., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

As a salt-type compound, mosapride citrate's metabolism and side effects are correlated with its salt-forming ratio. Several techniques were developed in this work to compare various quantitative nuclear magnetic resonance (qNMR) methodologies and to quantitatively examine the content of raw materials. Among the qNMR techniques, methods for 1 H NMR and 19 F NMR were developed. Appropriate solvents were chosen, and temperature, number of scans, acquisition time, and relaxation delay parameter settings were optimized. Maleic acid was chosen as the internal standard in 1 H NMR, and the respective characteristic signals of mosapride and citrate were selected as quantitative peaks. The internal standard in 19 F NMR analysis was 4,4'-difluoro diphenylmethanone, and the distinctive signal peak at -116.15 ppm was utilized to quantify mosapride citrate. The precision, repeatability, linearity, stability, accuracy, and robustness of the qNMR methods were all validated according to the ICH guidelines. By contrasting the outcomes with those from high-performance liquid chromatography (HPLC), the accuracy of qNMR was assessed. As a result, we created a quicker and easier qNMR approach to measure the amount of mosapride citrate and evaluated several qNMR techniques to establish a foundation for choosing quantitative peaks for the qNMR method. Concurrently, it is anticipated that various selections of distinct quantitative objects will yield the mosapride citrate salt-forming ratio.

ESCC presents a significant global health challenge due to its high mortality rates and varying responses to treatment. This underscores the critical need for novel diagnostic and predictive biomarkers to improve treatment outcomes. Initially, we conducted single-cell transcriptome sequencing on a total of 128,688 cells obtained from 10 patients as part of our research. Utilizing machine learning and cross-validation techniques, we developed a model incorporating 12 genes that distinguish malignant cells from non-malignant ones. In vitro, we explored the effects of IGFBP2 knockdown on the proliferation, invasion, and migration of ESCC cells. The clinical relevance of IGFBP2 was confirmed through IHC and Kaplan-Meier survival analyses. Furthermore, using bioinformatics tools such as GSVA and xCell on public databases, we discovered that high expression of IGFBP2 is associated with an immunosuppressive tumor microenvironment in ESCC, characterized by reduced CD8 + T cell infiltration. This was validated then through IHC. In summary, our study integrates single-cell sequencing and sophisticated computational techniques to highlight IGFBP2 as a promising biomarker and therapeutic target in ESCC., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Background: The fat-to-muscle mass ratio (FMR), integrating the antagonistic effects of fat and muscle mass, has been suggested as a valuable indicator to assess cardiometabolic health independent of overall adiposity. However, the specific associations of total and regional FMR with cardiometabolic risk are poorly understood. We aimed to examine sex-specific associations of total and regional FMR with single and clustered cardiometabolic risk factors (CRFs)., Methods: 13,505 participants aged 20 years and above were included in the cross-sectional study. Fat mass and muscle mass were assessed using a bioelectrical impedance analysis device. FMR was estimated as fat mass divided by muscle mass in corresponding body parts (whole body, arm, leg, and trunk). Clustered CRFs was defined as the presence of two or more risk factors, including hypertension, elevated blood glucose, dyslipidemia, insulin resistance (IR), and hyperuricemia. IR was assessed by the triglyceride glucose (TyG) index. Multivariable logistic regression models were applied to explore the associations of FMR in the whole body and body parts with single and clustered CRFs., Results: The odds ratios (ORs) increased significantly for all single and clustered CRFs with the per quartile increase of total and regional FMR in both sexes (P for trend < 0.001), following adjustment for confounders. Among the regional parts, FMRs of the legs presented the strongest associations for clustered CRFs in both men and women, with adjusted OR of 8.54 (95% confidence interval (CI): 7.12-10.24) and 4.92 (95% CI: 4.24-5.71), respectively. Significant interactions (P for interaction < 0.05) were identified between age and FMRs across different body parts, as well as between BMI status and FMRs in different regions for clustered CRFs. Restricted cubic splines revealed significant non-linear relationships between FMRs of different body parts and clustered CRFs in both sexes (P for nonlinear < 0.05)., Conclusions: FMRs in the whole body and different regions were significantly associated with single and clustered CRFs in the general Chinese population. The association between FMR and clustered CRFs was more pronounced in youngers than in the elderly., (© 2024. The Author(s).)

Numerical simulation serves as a cornerstone in scientific modeling, yet the process of fine-tuning simulation parameters poses significant challenges. Conventionally, parameter adjustment relies on extensive numerical simulations, data analysis, and expert insights, resulting in substantial computational costs and low efficiency. The emergence of deep learning in recent years has provided promising avenues for more efficient exploration of parameter spaces. However, existing approaches often lack intuitive methods for precise parameter adjustment and optimization. To tackle these challenges, we introduce ParamsDrag, a model that facilitates parameter space exploration through direct interaction with visualizations. Inspired by DragGAN, our ParamsDrag model operates in three steps. First, the generative component of ParamsDrag generates visualizations based on the input simulation parameters. Second, by directly dragging structure-related features in the visualizations, users can intuitively understand the controlling effect of different parameters. Third, with the understanding from the earlier step, users can steer ParamsDrag to produce dynamic visual outcomes. Through experiments conducted on real-world simulations and comparisons with state-of-the-art deep learning-based approaches, we demonstrate the efficacy of our solution.

Study Objectives: To investigate the engagement and health outcomes of community-based intervention for obstructive sleep apnea (OSA) in the general population., Methods: We conducted a 3-month randomized controlled trial in two communities in southern China. We initially screened the general population for high-risk OSA and further diagnosis using home sleep testing. Eligible participants were randomly (1:1) assigned to either a control or continuous positive airway pressure-based integrated intervention group. The primary outcomes were multimodal indicators reflecting health outcomes, including health-related quality of life (Short Form-36 [SF-36]), sleep-related symptoms, and cardiometabolic risk., Results: Of the 2484 participants screened, 1423 identified as having high-risk OSA were considered for telephone invitations to participate in the trial. Of these, 401 participants responded positively (28.2%), 279 were diagnosed with OSA, and 212 were randomized. The intervention significantly improved several domains of SF-36, including physical functioning (intergroup difference, 2.8; p = .003), vitality (2.3; p = .031), and reported health transition (6.8; p = .005). Sleep-related symptoms, including Epworth Sleepiness Scale (-0.7; p = .017), Fatigue Severity Scale (-3.0; p = .022), Insomnia Severity Index (-1.8; p < .001), and Pittsburgh Sleep Quality Index (-0.7; p = .032), also showed significant improvements. Although the intervention did not significantly alter glycolipid metabolism, ventricular function, or cardiac structural remodeling, it achieved a significant reduction in systolic (-4.5 mmHg; p = .004) and diastolic blood pressure (BP; -3.7 mmHg; p < .001)., Conclusions: Community-based intervention for previously undiagnosed OSA in the general population yielded improvements in health-related quality of life, sleep-related symptoms, and BP. However, engagement in the intervention program was low., Clinical Trial Registration: "Community Intervention Trial for OSA";URL: https://www.chictr.org.cn/showprojEN.html?proj=144927;Trial identification number: ChiCTR2100054800., (© The Author(s) 2024. Published by Oxford University Press on behalf of Sleep Research Society. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Objective: This study aimed to investigate the heritability of various obesity indices and their shared genetic factors with cardiometabolic traits in the Chinese nuclear family., Methods: A total of 1270 individuals from 538 nuclear families were included in this cross-sectional study. Different indices were used to quantify fat mass and distribution, including body index mass (BMI), visceral fat index (VFI), and body fat percent (BFP). Heritability and genetic correlations for all quantitative traits were estimated using variance component models. The susceptibility-threshold model was utilized to estimate the heritability for binary traits., Results: Heritability estimates for obesity indices were highest for BMI (59%), followed by BFP (49%), and VFI (40%). Heritability estimates for continuous cardiometabolic traits varied from 24% to 50%. All obesity measures exhibited consistently significant positive genetic correlations with blood pressure, fasting blood glucose, and uric acid (r G range: 0.26-0.57). However, diverse genetic correlations between various obesity indices and lipid profiles were observed. Significant genetic correlations were limited to specific pairs: BFP and total cholesterol (r G  = 0.24), BFP and low-density lipoprotein cholesterol (r G  = 0.25), and VFI and triglyceride (r G  = 0.33)., Conclusion: The genetic overlap between various obesity indices and cardiometabolic traits underscores the importance of pleiotropic genes. Further studies are warranted to investigate specific shared genetic and environmental factors between obesity and cardiometabolic diseases., (© 2024 University College London (UCL) and John Wiley & Sons Ltd.)

Aberrant expression of circular RNAs (circRNAs) is frequently linked to colorectal cancer (CRC). Here, we identified circZFR as a promising biomarker for CRC diagnosis and prognosis. CircZFR was upregulated in CRC tissues and serum exosomes and its level was linked to cancer incidence, advanced-stages, and metastasis. In both in vitro and in vivo settings, circZFR promoted the growth and spread while suppressing apoptosis of CRC. Exosomes with circZFR overexpression promoted the proliferation and migration of cocultured CRC cells. Mechanistically, epithelial splicing regulatory protein 1 (ESRP1) in CRC cells may enhance the production of circZFR. BCL2-associated transcription factor 1 (BCLAF1) bound to circZFR, which prevented its ubiquitinated degradation. Additionally, circZFR sponged miR-3127-5p to boost rhotekin 2 (RTKN2) expression. Our TCP1-CD-QDs nanocarrier was able to carry and deliver circZFR siRNA (si-circZFR) to the vasculature of CRC tissues and cells, which inhibited the growth of tumors in patient-derived xenograft (PDX) models. Taken together, our results show that circZFR is an oncogenic circRNA, which promotes the development and spread of CRC in a BCLAF1 and miR-3127-5p-dependent manner. CircZFR is a possible serum biopsy marker for the diagnosis and a desirable target for further treatment of CRC., (© 2024. Science China Press.)

Wet distillers grains, as a waste biomass with a large annual output, pose a threat to the environment and food industry. Herein, artificial humic acid (AHA) was first produced from wet distillers grains in a dual-stage microwave-assisted hydrothermal process. The influence of temperature on AHA's characteristics was investigated and compared with natural humic acid (NHA) and standard humic acid (SHA). A high yield of AHA at 20.6% was obtained at 200 °C with a total reaction time of 1 h, which is 1.8-3.1 times that obtained in traditional single-stage hydrothermal process. Increasing the reaction temperature induced the formation of phenolic hydroxyl in AHA. AHA was rich in aromaticity and carboxylic acid structure, showing similar spectral characteristics to NHA. The distribution of molecular weight of AHA was mostly 5797 Da, which decreased by 15% compared to SHA. The optimal concentration of AHA to promote seedling growth was 0.2 g/L, and the root length was 2.0 times that of the control. The microwave hydrothermal process is a facile and efficient approach to preparing AHA from waste biomass with high moisture content., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

It was common to see that older adults were reluctant to be vaccinated for coronavirus disease 2019 (COVID-19) in China. There is a lack of practical prediction models to guide COVID-19 vaccination program. A nationwide, self-reported, cross-sectional survey was conducted from September 2022 to November 2022, including people aged 60 years or older. Stratified random sampling was used to divide the dataset into derivation, validation, and test datasets at a ratio of 6:2:2. Least absolute shrinkage and selection operator and multivariable logistic regression were used for variable screening and model construction. Discrimination and calibration were assessed primarily by area under the receiver operating characteristic curve (AUC) and calibration curve. A total of 35057 samples (53.65% males and mean age of 69.64 ± 7.24 years) were finally selected, which constitutes 93.73% of the valid samples. From 33 potential predictors, 19 variables were screened and included in the multivariable logistic regression model. The mean AUC in the validation dataset was 0.802, with sensitivity, specificity, and accuracy of 0.732, 0.718 and 0.729 respectively, which were similar to the parameters in the test dataset of 0.755, 0.715 and 0.720, respectively, and the mean AUC in the test dataset was 0.815. There were no significant differences between the model predicted values and the actual observed values for calibration in these groups. The prediction model based on self-reported characteristics of older adults was developed that could be useful for predicting the willingness for COVID-19 vaccines, as well as providing recommendations in improving vaccine acceptance.

Introduction: The investigation of brainwave changes during the recovery process of unilateral spatial neglect (USN) has garnered considerable attention in recent years. This paper presents an updated overview of the evolving brainwave patterns during USN rehabilitation, aiming to predict clinical outcomes and guide the selection of effective recovery strategies., Methods: A systematic review was conducted, encompassing English literature published up to June 2024. Databases including PubMed, Web of Science, and clinical trials were utilized. The included studies assessed brainwaves using electroencephalography (EEG) in at least one group with USN. However, the diverse nature of these studies posed challenges for a quantitative synthesis., Results: The final quantitative synthesis comprised 36 studies, incorporating a total of 4517 data points. The analysis revealed abnormalities in alpha, beta, and gamma brainwave activity, along with alterations in the functional monitoring of the alpha band during USN rehabilitation. Additionally, reductions were observed in specific brainwave components such as P1, N1, P2, P300, early directing attention negativity (EDAN), late directing attention positivity (LDAP), and contingent negative variation (CNV). However, findings regarding measures of synchrony, connectivity, and evoked responses across different frequency bands exhibited variability., Conclusions: Various indicators of brainwave activity displayed changes at different stages of post-stroke neglect rehabilitation, highlighting the significance of neural network dysfunction in this process. Nonetheless, due to the diversity of the studies, further investigation is necessary to achieve a more comprehensive understanding in future research endeavors., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Environmental stimuli not only alter gene expression profiles but also induce structural changes in cells. How distinct nuclear bodies respond to cellular stress is poorly understood. Here, we identify a subnuclear organelle named the nucleolar stress body (NoSB), the formation of which is induced by the inhibition of rRNA transcription or inactivation of rRNA processing and maturation in C. elegans. NoSB does not colocalize with other previously described subnuclear organelles. We conduct forward genetic screening and identify a bZIP transcription factor, named nucleolar stress response-1 (NOSR-1), that is required for NoSB formation. The inhibition of rRNA transcription or inactivation of rRNA processing and maturation increases nosr-1 expression. By using transcriptome analysis of wild-type animals subjected to different nucleolar stress conditions and nosr-1 mutants, we identify that the SR-like protein NUMR-1 (nuclear localized metal responsive) is the target of NOSR-1. Interestingly, NUMR-1 is a component of NoSB and itself per se is required for the formation of NoSB. We conclude that the NOSR-1/NUMR-1 axis likely responds to nucleolar stress and mediates downstream stress-responsive transcription programs and subnuclear morphology alterations in C. elegans., (© 2024. The Author(s).)

Background: The tumor microenvironment (TME) in lung adenocarcinoma (LUAD) influences tumor progression and immunosuppressive phenotypes through cell communication. We aimed to decipher cellular communication and molecular patterns in LUAD., Methods: We analyzed scRNA-seq data from LUAD patients in multiple cohorts, revealing complex cell communication networks within the TME. Using cell chat analysis and COSmap technology, we inferred LUAD's spatial organization. Employing the NMF algorithm and survival screening, we identified a cell communication interactions (CCIs) model and validated it across various datasets., Results: We uncovered intricate cell communication interactions within the TME, identifying three LUAD patient subtypes with distinct prognosis, clinical characteristics, mutation status, expression patterns, and immune infiltration. Our CCI model exhibited robust performance in prognosis and immunotherapy response prediction. Several potential therapeutic targets and agents for high CCI score patients with immunosuppressive profiles were identified. Machine learning algorithms pinpointed the novel candidate gene ITGB1 and validated its role in LUAD tumor phenotype in vitro., Conclusion: Our study elucidates molecular patterns and cell communication interactions in LUAD as effective biomarkers and predictors of immunotherapy response. Targeting cell communication interactions offers novel avenues for LUAD immunotherapy and prognostic evaluations, with ITGB1 emerging as a promising therapeutic target., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Zhencong Chen reports financial support was provided by Shanghai Sailing Program (NO. 22YF1407500). If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

Purpose: This study aimed to investigate the prevalence of refractive error (RE) and risk factors for myopia among older adults in the Han and various minority ethnic groups across seven provinces in China., Methods: This cross-sectional study forms a part of the ophthalmic dataset of the China National Health Survey (CNHS). Face-to-face interviews and ophthalmic examinations were conducted in seven provinces located in western and northern China. The age- and sex-adjusted prevalence of RE among Han and seven other ethnic groups aged 50-80 years were compared. A mixed-effects model was used to identify the risk factors associated with RE., Results: A total of 12,902 participants, including 8800 Han and 4102 from ethnic minorities, were included in the study. The age- and sex-adjusted prevalence of myopia, high myopia, hyperopia, and astigmatism ranged from 15.3 % (Manchu) to 22.9 % (Han), 0.2 % (Yugur) to 2.8 % (Han), 21.6 % (Tibetan) to 48.9 % (Uyghur), and 38.7 % (Yi) to 57.5 % (Manchu) across different ethnicities, respectively. Compared to the Han population, the Mongolian (odds ratios (OR) 0.62, 95 % confidence interval (CI) 0.46-0.84, p = 0.002), Tibetan (OR 0.66, 95 % CI 0.52-0.85, p = 0.001), Uyghur (OR 0.63, 95 % CI 0.49-0.80, p < 0.001), Yi (OR 0.65, 95 % CI 0.46-0.92, p = 0.014), and Yugur (OR 0.65, 95 % CI 0.50-0.85, p = 0.001) ethnicities were less likely to have myopia. There was no significant difference in the prevalence of myopia between the Manchu, Korean, and Han ethnic groups. Factors associated with a lower prevalence of myopia included rural residence (p < 0.001), a body mass index (BMI) > 18.5 kg/m 2 (all p < 0.001), residence in higher latitude areas (p = 0.020), and a history of smoking (p = 0.002 in the past smoking group, p = 0.031 in the current smoking group). The Mongolian (p = 0.006) and Yugur (p = 0.007) populations, participants living in rural areas (p = 0.012), and those with a BMI >24 kg/m 2 (p = 0.038 in the >24.0 ≤ 27.0 kg/m 2 group or p = 0.041 in the >27.0 kg/m 2 group) were less likely to have high myopia. Factors associated with a higher prevalence of hyperopia included older age (all p < 0.001), rural residence (p = 0.039), higher latitude areas (p = 0.031), smoking history (p = 0.040), and Mongolian (p = 0.001), Uyghur (p < 0.001), Yi (p < 0.001), and Yugur (p = 0.002) ethnicities. Conversely, the Manchu population (p = 0.004) and individuals with higher education levels than illiteracy (p = 0.024 or p < 0.001) were less likely to have hyperopia., Conclusions: Myopia affected more than one-fifth of the older adults in the Han population in this survey. Significant differences in the prevalence of RE were observed between minority ethnicities and Han individuals, except for the Manchu and Korean groups., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Background: Acute post-subarachnoid hemorrhage (SAH) headaches are common and severe. Management strategies for post-SAH headaches are limited, with heavy reliance on opioids, and pain control is overall poor. Pterygopalatine fossa (PPF) nerve blocks have shown promising results in treatment of acute headache, including our preliminary and published experience with PPF-blocks for refractory post-SAH headache during hospitalization. The BLOCK-SAH trial was designed to assess the efficacy and safety of bilateral PPF-blocks in awake patients with severe headaches from aneurysmal SAH who require opioids for pain control and are able to verbalize pain scores., Methods: BLOCK-SAH is a phase II, multicenter, randomized, double-blinded, placebo-controlled clinical trial using the sequential parallel comparison design (SPCD), followed by an open-label phase., Results: Across 12 sites in the United States, 195 eligible study participants will be randomized into three groups to receive bilateral active or placebo PPF-injections for 2 consecutive days with periprocedural monitoring of intracranial arterial mean flow velocities with transcranial Doppler, according to SPCD (group 1: active block followed by placebo; group 2: placebo followed by active block; group 3: placebo followed by placebo). PPF-injections will be delivered under ultrasound guidance and will comprise 5-mL injectates of 20 mg of ropivacaine plus 4 mg of dexamethasone (active PPF-block) or saline solution (placebo PPF-injection)., Conclusions: The trial has a primary efficacy end point (oral morphine equivalent/day use within 24 h after each PPF-injection), a primary safety end point (incidence of radiographic vasospasm at 48 h from first PPF-injection), and a primary tolerability end point (rate of acceptance of second PPF-injection following the first PPF-injection). BLOCK-SAH will inform the design of a phase III trial to establish the efficacy of PPF-block, accounting for different headache phenotypes., (© 2024. The Author(s).)

Sulfonate esters, one class of genotoxic impurities (GTIs), have gained significant attention in recent years due to their potential to cause genetic mutations and cancer. In the current study, we employed the dummy template molecular imprinting technology with a dummy template molecule replacing the target molecule to establish a pretreatment method for samples containing p-toluene sulfonate esters. Through computer simulation and ultraviolet-visible spectroscopy analysis, the optimal functional monomer acrylamide and polymerization solvent chloroform were selected. Subsequently, a dummy template molecularly imprinted polymer (DMIP) was prepared by the precipitation polymerization method, and the polymer was characterized in morphology, particle size, and composition. The results of the adsorption and enrichment study demonstrated that the DMIP has high adsorption capability (Q = 7.88 mg/g) and favorable imprinting effects (IF = 1.37); Further, it could simultaneously adsorb three p-toluene sulfonate esters. The optimal adsorption conditions were obtained by conditional optimization of solid-phase extraction (SPE). A pH 7 solution was selected as the loading condition, the methanol/1 % phosphoric acid solution (20:80, v/v) was selected as the washing solution, and acetonitrile containing 10 % acetic acid in 6 mL was selected as the elution solvent. Finally, we determined methyl p-toluene sulfonate alkyl esters, ethyl p-toluene sulfonate alkyl esters, and isopropyl p-toluene sulfonate alkyl esters in tosufloxacin toluene sulfonate and capecitabine at the 10 ppm level (relative to 1 mg/mL active pharmaceutical ingredient (API) samples) by using DMIP-based SPE coupled with HPLC. This approach facilitated the selective enrichment of p-toluene sulfonate esters GTIs from complex API samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Purpose: Determination of the appropriate ablative parameters is the key to the success and safety of microwave ablation (MWA) of lung tumors. The purpose of this study was to provide guidelines and recommendations for the optimal time and power for lung tumor MWA., Material and Methods: MWA using a 2450-MHz system was evaluated in a porcine lung. The independent variables were power (30, 40, 50, 60, 70, and 80 W) and time (2, 4, 6, 8, 10, and 12 min), and the outcome variable was the volume of ablation. Lung tissues were procured after MWA for measurement and histological evaluation. Analysis of variance was used for statistical analysis, followed by least significant difference (LSD) t-tests where appropriate. A P value of <0.05 was considered statistically significant., Results: The outcome variable (ablative volume) was significantly affected by time, power, and time/power interaction (P < 0.05). When the total output energy was kept constant, the combination of higher power and shorter time obtained a larger ablative volume, especially in the low- and medium-energy groups (P < 0.01)., Conclusions: We propose guidelines for ablative volume based on different time and power variables to provide a reference for clinical applications., (Copyright © 2024 Copyright: © 2024 Journal of Cancer Research and Therapeutics.)

Introduction: Estimating treatment effects as time savings in disease progression may be more easily interpretable than assessing the absolute difference or a percentage reduction. In this study, we investigate the statistical considerations of the existing method for estimating time savings and propose alternative complementary methods., Methods: We propose five alternative methods to estimate the time savings from different perspectives. These methods are applied to simulated clinical trial data that mimic or modify the Clinical Dementia Rating Sum of Boxes progression trajectories observed in the Clarity AD lecanemab trial., Results: Our study demonstrates that the proposed methods can generate more precise estimates by considering two crucial factors: (1) the absolute difference between treatment arms, and (2) the observed progression rate in the treatment arm., Discussion: Quantifying treatment effects as time savings in disease progression offers distinct advantages. To provide comprehensive estimations, it is important to use various methods., Highlights: We explore the statistical considerations of the current method for estimating time savings. We proposed alternative methods that provide time savings estimations based on the observed absolute differences. By using various methods, a more comprehensive estimation of time savings can be achieved., (© 2024 The Author(s). Alzheimer's & Dementia published by Wiley Periodicals LLC on behalf of Alzheimer's Association.)

The nitrogen loss in composting is primarily driven by the transformation of organic nitrogen, yet the mechanisms underlying the degradation process remain incompletely understood. This study employed protein family domains (Pfams) analysis based on metagenomic sequencing to investigate the functional characteristics, key microorganisms, and environmental parameters influencing organic nitrogen degradation in chicken manure and pig manure composting. 154 Pfams associated with ammonification function were identified. Predominant Pfams: proteolytic peptidase, followed by chitin/cell wall degraders, least involved in nucleic acid degradation. Ammonifying microbial diversity was basically consistent among compost types, particularly in the thermophilic stage with the peak of abundance of dominant ammonifying microorganisms. Viruses played an important role in ammonification process, especially Uroviricota. 26 key ammonifying genera were identified by the microbial network. pH dominated the metabolic activity of ammonifying microorganisms in various manure compost types, primarily consisting of protein-degrading bacteria with stable community structures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In recent years, the utilization of aerogel templates in oleogels to replace animal fats has garnered considerable attention due to health concerns. This study employed a "fiber-particle core-shell nanostructure model" to combine sodium carboxymethylcellulose (CMCNa) and soy protein isolate (SPI) or SPI hydrolysate (SPIH), and freeze-dried to form aerogel template, which was then dipped into oil to induce oleogels. The results showed that adding SPIH significantly improved the physicochemical properties of oleogels. The results of ζ-potential, FTIR, and rheology demonstrated a stronger binding of SPIH to CMC-Na compared to SPI. The CMC-Na-SPIH aerogels exhibited a coarser surface and denser network structure in contrast to CMC-Na-SPI aerogels, with an oil holding capacity (OHC) of up to 84.6 % and oil absorption capacity (OAC) of 47.4 g/g. The mechanical strength of oleogels was further enhanced through chemical crosslinking. Both CMC-Na-SPI and CMC-Na-SPIH oleogels displayed excellent elasticity and reversible compressibility, with CMC-Na-SPIH oleogels demonstrating superior mechanical strength. Additionally, CMC-Na-SPIH oleogels exhibited enhanced slow release of antimicrobial substances and antioxidant properties. Increasing the content of SPI/SPIH significantly improved the mechanical strength, antioxidant capacity, and OHC of the oleogels. This research presents a straightforward and promising approach to enhance the performance of aerogel template oleogels., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to be declared., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Hydrogels have emerged as promising candidates for anticounterfeiting materials, owing to their unique stimulus-responsive capabilities. To improve the security of encrypted information, efforts are devoted to constructing transient anticounterfeiting hydrogels with a dynamic information display. However, current studies to design such hydrogel materials inevitably include sophisticated chemistry, complex preparation processes, and particular experimental setups. Herein, a facile strategy is proposed to realize the transient anticounterfeiting by constructing bivalent metal (M 2+ )-coordination complexes in poly(acrylic acid) gels, where the cloud temperature (T c ) of the gels can be feasibly tuned by M 2+ concentration. Therefore, the multi-T c parts in the gel can be locally programmed by leveraging the spatially selective diffusion of M 2+ with different concentrations. With the increase of temperature or the addition of a complexing agent, the transparency of the multi-T c parts in the gel spontaneously evolves in natural light, enabling the transient information anticounterfeiting process. This work has provided a new strategy and mechanism to fabricate advanced anticounterfeiting hydrogel materials., (© 2024 Wiley‐VCH GmbH.)

Aims: The evidence for joint and independent associations of low muscle mass and low muscle strength with diabetes is limited and mixed. The study aimed to determine the associations of muscle parameters (muscle mass, strength, quality, and sarcopenia) and sarcopenia obesity with diabetes, and the previously unstudied mediating effect of inflammation., Materials and Methods: A total of 13,420 adults from the 2023 China National Health Survey (CNHS) and 5,380 adults from the 2011-2014 National Health and Nutrition Examination Survey (NHANES) were included in this study. Muscle mass was determined using bioelectrical impedance analysis (BIA) in the CNHS, and whole-body dual X-ray absorptiometry (DXA) in the NHANES. Muscle strength was assessed using digital hand dynamometer. Multivariate logistic regression models were used to evaluate the associations of muscle parameters and sarcopenia obesity with diabetes. Inflammatory status was assessed using blood cell counts and two systemic inflammation indices (platelet-to-lymphocyte ratio (PLR) and system inflammation response index (SIRI)). Mediation analysis was conducted to examine inflammation's role in these associations., Results: Low muscle mass and strength were independently related to diabetes. Low muscle quality was associated with elevated diabetes risk. Sarcopenia has a stronger association with diabetes compared to low muscle strength alone or mass alone (CNHS, odds ratio (OR) = 1.93, 95 % confidence interval (CI):1.64-2.27; NHANES, OR = 3.80, 95 %CI:2.58-5.58). Participants with sarcopenia obesity exhibit a higher risk of diabetes than those with obesity or sarcopenia alone (CNHS, OR = 2.21, 95 %CI:1.72-2.84; NHANES, OR = 6.06, 95 %CI:3.64-10.08). Associations between muscle parameters and diabetes were partially mediated by inflammation (mediation proportion: 1.99 %-36.64 %, P < 0.05)., Conclusion: Low muscle mass and muscle strength are independently or jointly associated with diabetes, and inflammation might be a potential mechanism underlying this association. Furthermore, the synergistic effects of sarcopenia and obesity could significantly increase diabetes risk., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sensitive detection and effective inactivation of bacteria are essential in preventing foodborne bacterial infection that poses a significant threat to human health. Herein, a near-infrared (NIR)-driven multifunctional photoelectrochemical (PEC) biosensor was constructed for detection and inactivation of S. aureus. Based on the covalent bonding between amine and carboxyl groups, carboxyl-functionalized SA31 aptamer was immobilized on the PDA/MnO 2 photoelectrode. In the presence of S. aureus, SA31 aptamer can specifically capture S. aureus, causing the decrease of photocurrent signal owing to steric hindrance effect. Leveraging photocurrent-off signal, there existed a satisfied linear relationship between the photocurrent variation and the logarithm of S. aureus concentration, achieving a wide linear range from 10 to 10 7  CFU/mL with a low detection limit of 2.0 CFU/mL. Notably, PDA/MnO 2 with peroxidase-like activity facilitated the catalytic oxidation of S. aureus with assistance of hydrogen peroxide (H 2 O 2 ) to cause the inactivation of S. aureus. Desorption of inactivated S. aureus from the photoelectrode led to a recovery of photocurrent signal, enabling a "signal on" switch. Simultaneously, the excellent photothermal performance of the PDA/MnO 2 converted light energy into heat energy under the irradiation of NIR light (808 nm, 1.5 W/cm 2 ), triggering the synergistic antibacterial effect against S. aureus (97.36%). This work provides a novel strategy for fabricating the detection and inactivation of bacteria in practical applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The purpose of this study is to develop an electronic portal imaging device-based multi-leaf collimator calibration procedure using log files. Picket fence fields with 2-14 mm nominal strip widths were performed and normalized by open field. Normalized pixel intensity profiles along the direction of leaf motion for each leaf pair were taken. Three independent algorithms and an integration method derived from them were developed according to the valley value, valley area, full-width half-maximum (FWHM) of the profile, and the abutment width of the leaf pairs obtained from the log files. Three data processing schemes (Scheme A, Scheme B, and Scheme C) were performed based on different data processing methods. To test the usefulness and robustness of the algorithm, the known leaf position errors along the direction of perpendicular leaf motion via the treatment planning system were introduced in the picket fence field with nominal 5, 8, and 11 mm. Algorithm tests were performed every 2 weeks over 4 months. According to the log files, about 17.628% and 1.060% of the leaves had position errors beyond ± 0.1 and ± 0.2 mm, respectively. The absolute position errors of the algorithm tests for different data schemes were 0.062 ± 0.067 (Scheme A), 0.041 ± 0.045 (Scheme B), and 0.037 ± 0.043 (Scheme C). The absolute position errors of the algorithms developed by Scheme C were 0.054 ± 0.063 (valley depth method), 0.040 ± 0.038 (valley area method), 0.031 ± 0.031 (FWHM method), and 0.021 ± 0.024 (integrated method). For the efficiency and robustness test of the algorithm, the absolute position errors of the integration method of Scheme C were 0.020 ± 0.024 (5 mm), 0.024 ± 0.026 (8 mm), and 0.018 ± 0.024 (11 mm). Different data processing schemes could affect the accuracy of the developed algorithms. The integration method could integrate the benefits of each algorithm, which improved the level of robustness and accuracy of the algorithm. The integration method can perform multi-leaf collimator (MLC) quality assurance with an accuracy of 0.1 mm. This method is simple, effective, robust, quantitative, and can detect a wide range of MLC leaf position errors., (© 2024 The Author(s). Journal of Applied Clinical Medical Physics published by Wiley Periodicals LLC on behalf of American Association of Physicists in Medicine.)