One-tube B7-H3 detection based on isothermal exponential amplification and dendritic hybridization chain reaction.

We have developed a one-tube fluorescence strategy for the detection of B7-H3 based on a proximity hybridization-mediated protein-to-DNA signal transducer, isothermal exponential amplification (EXPAR), and dendritic hybridization chain reaction (D-HCR). In this assay, a protein signal transducer was employed to convert the input protein to output single-stranded DNA with a nicking site. Antibody-conjugated DNA1 was first hybridized with the output DNA (DNA3). The binding of antibodies conjugated DNA1 and DNA2 to the same protein was able to increase the local concentrations, resulting in strand displacement between DNA3 and DNA2. DNA3 with a nicking endonuclease recognition sequence at the 5' end then hybridized with hairpin probe 1 to mediate EXPAR in the presence of nicking endonuclease and DNA polymerase. A large number of single-strand DNA were produced in the circle of nicking, polymerization, and strand displacement. The resulting ssDNA products were further amplified by D-HCR to produce many large-molecular concatemers. The resulting DNA products can be monitored in real-time fluorescence signaling. Our proposed assay can realize one-tube detection due to the same reaction temperature of the protein-to-DNA signal transducer, EXPAR, and DHCR. This assay has a linear range from 100 fg mL-1 to 1 µg mL-1 with a detection limit down to 100 fg mL-1. This work shows a good performance in clinical specimen detection.

JAML promotes the antitumor role of tumor-resident CD8+ T cells by facilitating their innate-like function in human lung cancer.

Tissue-resident memory CD8+T cells (CD8+TRMs) are thought to play a crucial role in cancer immunosurveillance. However, the characteristics of CD8+TRMs in the tumor microenvironment (TME) of human non-small cell lung cancer (NSCLC) remain unclear. Here, we report that CD8+TRMs accumulate explicitly and exhibit a unique gene expression profile in the TME of NSCLC. Interestingly, these tumor-associated CD8+TRMs uniquely exhibit an innate-like phenotype. Importantly, we found that junction adhesion molecule-like (JAML) provides an alternative costimulatory signal to activate tumor-associated CD8+TRMs via combination with cancer cell-derived CXADR (CXADR Ig-like cell adhesion molecule). Furthermore, we demonstrated that activating JAML could promote the expression of TLR1/2 on CD8+TRMs, inhibit tumor progression and prolong the survival of tumor-bearing mice. Finally, we found that higher CD8+TRMs and JAML expression in the TME could predict favorable clinical outcomes in NSCLC patients. Our study reveals an intrinsic bias of CD8+TRMs for receiving the tumor-derived costimulatory signal in the TME, which sustains their innate-like function and antitumor role. These findings will shed more light on the biology of CD8+TRMs and aid in the development of potential targeted treatment strategies for NSCLC.

ROCK1 inhibition improves wound healing in diabetes via RIPK4/AMPK pathway.

Refractory wounds are a severe complication of diabetes mellitus that often leads to amputation because of the lack of effective treatments and therapeutic targets. The pathogenesis of refractory wounds is complex, involving many types of cells. Rho-associated protein kinase-1 (ROCK1) phosphorylates a series of substrates that trigger downstream signaling pathways, affecting multiple cellular processes, including cell migration, communication, and proliferation. The present study investigated the role of ROCK1 in diabetic wound healing and molecular mechanisms. Our results showed that ROCK1 expression significantly increased in wound granulation tissues in diabetic patients, streptozotocin (STZ)-induced diabetic mice, and db/db diabetic mice. Wound healing and blood perfusion were dose-dependently improved by the ROCK1 inhibitor fasudil in diabetic mice. In endothelial cells, fasudil and ROCK1 siRNA significantly elevated the phosphorylation of adenosine monophosphate-activated protein kinase at Thr172 (pThr172-AMPKα), the activity of endothelial nitric oxide synthase (eNOS), and suppressed the levels of mitochondrial reactive oxygen species (mtROS) and nitrotyrosine formation. Experiments using integrated bioinformatics analysis and coimmunoprecipitation established that ROCK1 inhibited pThr172-AMPKα by binding to receptor-interacting serine/threonine kinase 4 (RIPK4). These results suggest that fasudil accelerated wound repair and improved angiogenesis at least partially through the ROCK1/RIPK4/AMPK pathway. Fasudil may be a potential treatment for refractory wounds in diabetic patients.

Wemics: A Single-Base Resolution Methylation Quantification Method for Enhanced Prediction of Epigenetic Regulation.

DNA methylation, an epigenetic mechanism that alters gene expression without changing DNA sequence, is essential for organism development and key biological processes like genomic imprinting and X-chromosome inactivation. Despite tremendous efforts in DNA methylation research, accurate quantification of cytosine methylation remains a challenge. Here, a single-base methylation quantification approach is introduced by weighting methylation of consecutive CpG sites (Wemics) in genomic regions. Wemics quantification of DNA methylation better predicts its regulatory impact on gene transcription and identifies differentially methylated regions (DMRs) with more biological relevance. Most Wemics-quantified DMRs in lung cancer are epigenetically conserved and recurrently occurred in other primary cancers from The Cancer Genome Atlas (TCGA), and their aberrant alterations can serve as promising pan-cancer diagnostic markers. It is further revealed that these detected DMRs are enriched in transcription factor (TF) binding motifs, and methylation of these TF binding motifs and TF expression synergistically regulate target gene expression. Using Wemics on epigenomic-transcriptomic data from the large lung cancer cohort, a dozen novel genes with oncogenic potential are discovered that are upregulated by hypomethylation but overlooked by other quantification methods. These findings increase the understanding of the epigenetic mechanism by which DNA methylation regulates gene expression.

Microbial metabolite enhances immunotherapy efficacy by modulating T cell stemness in pan-cancer.

The immune checkpoint blockade (ICB) response in human cancers is closely linked to the gut microbiota. Here, we report that the abundance of commensal Lactobacillus johnsonii is positively correlated with the responsiveness of ICB. Supplementation with Lactobacillus johnsonii or tryptophan-derived metabolite indole-3-propionic acid (IPA) enhances the efficacy of CD8+ T cell-mediated αPD-1 immunotherapy. Mechanistically, Lactobacillus johnsonii collaborates with Clostridium sporogenes to produce IPA. IPA modulates the stemness program of CD8+ T cells and facilitates the generation of progenitor exhausted CD8+ T cells (Tpex) by increasing H3K27 acetylation at the super-enhancer region of Tcf7. IPA improves ICB responsiveness at the pan-cancer level, including melanoma, breast cancer, and colorectal cancer. Collectively, our findings identify a microbial metabolite-immune regulatory pathway and suggest a potential microbial-based adjuvant approach to improve the responsiveness of immunotherapy.

High-security learning-based optical encryption assisted by disordered metasurface.

Artificial intelligence has gained significant attention for exploiting optical scattering for optical encryption. Conventional scattering media are inevitably influenced by instability or perturbations, and hence unsuitable for long-term scenarios. Additionally, the plaintext can be easily compromised due to the single channel within the medium and one-to-one mapping between input and output. To mitigate these issues, a stable spin-multiplexing disordered metasurface (DM) with numerous polarized transmission channels serves as the scattering medium, and a double-secure procedure with superposition of plaintext and security key achieves two-to-one mapping between input and output. In attack analysis, when the ciphertext, security key, and incident polarization are all correct, the plaintext can be decrypted. This system demonstrates excellent decryption efficiency over extended periods in noisy environments. The DM, functioning as an ultra-stable and active speckle generator, coupled with the double-secure approach, creates a highly secure speckle-based cryptosystem with immense potentials for practical applications.

Targeting pro-inflammatory T cells as a novel therapeutic approach to potentially resolve atherosclerosis in humans.

Atherosclerosis (AS), a leading cause of cardio-cerebrovascular disease worldwide, is driven by the accumulation of lipid contents and chronic inflammation. Traditional strategies primarily focus on lipid reduction to control AS progression, leaving residual inflammatory risks for major adverse cardiovascular events (MACEs). While anti-inflammatory therapies targeting innate immunity have reduced MACEs, many patients continue to face significant risks. Another key component in AS progression is adaptive immunity, but its potential role in preventing AS remains unclear. To investigate this, we conducted a retrospective cohort study on tumor patients with AS plaques. We found that anti-programmed cell death protein 1 (PD-1) monoclonal antibody (mAb) significantly reduces AS plaque size. With multi-omics single-cell analyses, we comprehensively characterized AS plaque-specific PD-1+ T cells, which are activated and pro-inflammatory. We demonstrated that anti-PD-1 mAb, when captured by myeloid-expressed Fc gamma receptors (FcγRs), interacts with PD-1 expressed on T cells. This interaction turns the anti-PD-1 mAb into a substitute PD-1 ligand, suppressing T-cell functions in the PD-1 ligands-deficient context of AS plaques. Further, we conducted a prospective cohort study on tumor patients treated with anti-PD-1 mAb with or without Fc-binding capability. Our analysis shows that anti-PD-1 mAb with Fc-binding capability effectively reduces AS plaque size, while anti-PD-1 mAb without Fc-binding capability does not. Our work suggests that T cell-targeting immunotherapy can be an effective strategy to resolve AS in humans.

Inflammatory dendritic cells restrain CD11b+CD4+ CTLs via CD200R in human NSCLC.

CD4+ cytotoxic T lymphocytes (CD4+ CTLs) are suggested to play a crucial role in inflammatory diseases, including cancer, but their characteristics in human non-small cell lung cancer (NSCLC) remain unknown. Here, using the cell surface marker CD11b, we identify CD11b+CD4+ CTLs as a cytotoxic subset of CD4+ T cells in multiple tissues of NSCLC patients. In addition, tumor-infiltrating CD11b+CD4+ CTLs show a dysfunctional phenotype with elevated expression of CD200 receptor (CD200R), a negatively immunomodulatory receptor. CD4+ regulatory T (Treg) cells restrain the anti-tumor role of CD11b+CD4+ CTLs via CD200. Mechanistically, inflammatory dendritic cells promote the CD200R expression of CD11b+CD4+ CTLs by secreting interleukin-1ß (IL-1ß). Finally, we demonstrate that CD200 blockade can revive the tumor-killing role of CD11b+CD4+ CTLs and prolong the survival of tumor-bearing mice. Taken together, our study identifies CD11b+CD4+ CTLs in NSCLC with decreased cytotoxicity that can be reinvigorated by CD200 blockade, suggesting that targeting CD200 is a promising immunotherapy strategy in NSCLC.

LASSO-derived model for the prediction of lean-non-alcoholic fatty liver disease in examinees attending a routine health check-up.

BACKGROUND: Lean individuals with non-alcohol fatty liver disease (NAFLD) often have normal body size but abnormal visceral fat. Therefore, an alternative to body mass index should be considered for prediction of lean-NAFLD. This study aimed to use representative visceral fat links with other laboratory parameters using the least absolute shrinkage and selection operator (LASSO) method to construct a predictive model for lean-NAFLD. METHODS: This retrospective cross-sectional analysis enrolled 2325 subjects with BMI < 24 kg/m2 from medical records of 51,271 examinees who underwent a routine health check-up. They were randomly divided into training and validation cohorts at a ratio of 1:1. The LASSO-derived prediction model used LASSO regression to select 23 clinical and laboratory factors. The discrimination and calibration abilities were evaluated using the Hosmer-Lemeshow test and calibration curves. The performance of the LASSO model was compared with the fatty liver index (FLI) model. RESULTS: The LASSO-derived model included four variables-visceral fat, triglyceride levels, HDL-C-C levels, and waist hip ratio-and demonstrated superior performance in predicting lean-NAFLD with high discriminatory ability (AUC, 0.8416; 95% CI: 0.811-0.872) that was comparable with the FLI model. Using a cut-off of 0.1484, moderate sensitivity (75.69%) and specificity (79.86%), as well as high negative predictive value (95.9%), were achieved in the LASSO model. In addition, with normal WC subgroup analysis, the LASSO model exhibits a trend of higher accuracy compared to FLI (cut-off 15.45). CONCLUSIONS: We developed a LASSO-derived predictive model with the potential for use as an alternative tool for predicting lean-NAFLD in clinical settings.

Researchers developed a model to predict a type of liver disease called non-alcoholic fatty liver disease (NAFLD) in lean individuals.The model accurately detects NAFLD in lean individuals using factors like visceral fat, triglyceride levels, and waist-to-hip ratio, aiding in identifying the disease in normal-weight people with abnormal fat distribution.

Creating pairs of exceptional points for arbitrary polarization control: asymmetric vectorial wavefront modulation.

Exceptional points (EPs) can achieve intriguing asymmetric control in non-Hermitian systems due to the degeneracy of eigenstates. Here, we present a general method that extends this specific asymmetric response of EP photonic systems to address any arbitrary fully-polarized light. By rotating the meta-structures at EP, Pancharatnam-Berry (PB) phase can be exclusively encoded on one of the circular polarization-conversion channels. To address any arbitrary wavefront, we superpose the optical signals originating from two orthogonally polarized -yet degenerate- EP eigenmodes. The construction of such orthogonal EP eigenstates pairs is achieved by applying mirror-symmetry to the nanostructure geometry flipping thereby the EP eigenmode handedness from left to right circular polarization. Non-Hermitian reflective PB metasurfaces designed using such EP superposition enable arbitrary, yet unidirectional, vectorial wavefront shaping devices. Our results open new avenues for topological wave control and illustrate the capabilities of topological photonics to distinctively operate on arbitrary polarization-state with enhanced performances.

Asymmetric Full-Color Vectorial Meta-holograms Empowered by Pairs of Exceptional Points.

Holography holds tremendous promise in applications such as immersive virtual reality and optical communications. With the emergence of optical metasurfaces, planar optical components that have the remarkable ability to precisely manipulate the amplitude, phase, and polarization of light on the subwavelength scale have expanded the potential applications of holography. However, the realization of metasurface-based full-color vectorial holography remains particularly challenging. Here, we report a general approach utilizing a modified Gerchberg-Saxton algorithm to achieve spatially aligned full-color display and incorporating wavelength information with an image compensation strategy. We combine the Pancharatnam-Berry phase and pairs of exceptional points to address the issue of redundant twin images that generally appear for the two orthogonal circular polarizations and to enable full polarization control of the vectorial field. Our results enable the realization of an asymmetric full-color vectorial meta-hologram, paving the way for the development of full-color display, complex beam generation, and secure data storage applications.

Association between polycyclic aromatic hydrocarbons exposure with red cell width distribution and ischemic heart disease: insights from a population-based study.

This study investigates the association between polycyclic aromatic hydrocarbon (PAH) exposure, red blood cell distribution width (RDW), and ischemic heart disease (IHD) in a sample of 3003 participants from the National Health and Nutrition Examination Survey (NHANES). We hypothesize that RDW may mediate the effect of hydroxylated PAHs (OH-PAH) on IHD. Logistic regression models reveal significant associations between increased urinary PAH metabolite concentrations and IHD, as well as positive associations between PAH metabolites and RDW. Weighted Quantile Sum (WQS) regression and Bayesian Kernel Machine Regression (BKMR) analyses confirm the significant associations of the OH-PAH mixture with IHD and RDW. Mediation analysis demonstrates that RDW partially mediates the relationship between PAH exposure and IHD, accounting for 2-4.6% of the total effects. Our findings highlight the potential underlying mechanisms linking PAH exposure, RDW, and IHD and emphasize the importance of addressing environmental pollutants like PAHs in maintaining cardiovascular health and informing public health policies.

Integrated multi-omics profiling to dissect the spatiotemporal evolution of metastatic hepatocellular carcinoma.

Comprehensive molecular analyses of metastatic hepatocellular carcinoma (HCC) are lacking. Here, we generate multi-omic profiling of 257 primary and 176 metastatic regions from 182 HCC patients. Primary tumors rich in hypoxia signatures facilitated polyclonal dissemination. Genomic divergence between primary and metastatic HCC is high, and early dissemination is prevalent. The remarkable neoantigen intratumor heterogeneity observed in metastases is associated with decreased T cell reactivity, resulting from disruptions to neoantigen presentation. We identify somatic copy number alterations as highly selected events driving metastasis. Subclones without Wnt mutations show a stronger selective advantage for metastasis than those with Wnt mutations and are characterized by a microenvironment rich in activated fibroblasts favoring a pro-metastatic phenotype. Finally, metastases without Wnt mutations exhibit higher enrichment of immunosuppressive B cells that mediate terminal exhaustion of CD8+ T cells via HLA-E:CD94-NKG2A checkpoint axis. Collectively, our results provide a multi-dimensional dissection of the complex evolutionary process of metastasis.

A novel nomogram for predicting the prolonged length of stay in post-anesthesia care unit after elective operation.

BACKGROUND: Prolonged length of stay in post-anesthesia care unit (PLOS in PACU) is a combination of risk factors and complications that can compromise quality of care and operating room efficiency. Our study aimed to develop a nomogram to predict PLOS in PACU of patients undergoing elective surgery. METHODS: Data from 24017 patients were collected. Least absolute shrinkage and selection operator (LASSO) was used to screen variables. A logistic regression model was built on variables determined by a combined method of forward selection and backward elimination. Nomogram was designed with the model. The nomogram performance was evaluated with the area under the receiver operating characteristic curve (AUC) for discrimination, calibration plot for consistency between predictions and actuality, and decision curve analysis (DCA) for clinical application value. RESULTS: A nomogram was established based on the selected ten variables, including age, BMI < 21 kg/m2, American society of Anesthesiologists Physical Status (ASA), surgery type, chill, delirium, pain, naloxone, operation duration and blood transfusion. The C-index value was 0.773 [95% confidence interval (CI) = 0.765 - 0.781] in the development set and 0.757 (95% CI = 0.744-0.770) in the validation set. The AUC was > 0.75 for the prediction of PLOS in PACU. The calibration curves revealed high consistencies between the predicted and actual probability. The DCA showed that if the threshold probability is over 10% , using the models to predict PLOS in PACU and implement intervention adds more benefit. CONCLUSIONS: This study presented a nomogram to facilitate individualized prediction of PLOS in PACU for patients undergoing elective surgery.

Metasurface-empowered snapshot hyperspectral imaging with convex/deep (CODE) small-data learning theory.

Hyperspectral imaging is vital for material identification but traditional systems are bulky, hindering the development of compact systems. While previous metasurfaces address volume issues, the requirements of complicated fabrication processes and significant footprint still limit their applications. This work reports a compact snapshot hyperspectral imager by incorporating the meta-optics with a small-data convex/deep (CODE) deep learning theory. Our snapshot hyperspectral imager comprises only one single multi-wavelength metasurface chip working in the visible window (500-650 nm), significantly reducing the device area. To demonstrate the high performance of our hyperspectral imager, a 4-band multispectral imaging dataset is used as the input. Through the CODE-driven imaging system, it efficiently generates an 18-band hyperspectral data cube with high fidelity using only 18 training data points. We expect the elegant integration of multi-resonant metasurfaces with small-data learning theory will enable low-profile advanced instruments for fundamental science studies and real-world applications.

Varifocal Metalenses: Harnessing Polarization-Dependent Superposition for Continuous Focal Length Control.

Traditional varifocal lenses are bulky and mechanically complex. Emerging active metalenses promise compactness and design flexibility but face issues like mechanical tuning reliability and nonlinear focal length tuning due to additional medium requirements. In this work, we propose a varifocal metalens design based on superimposing light intensity distributions from two orthogonal polarization states. This approach enables continuous and precise focal length control within the visible spectrum, while maintaining relatively high focusing efficiencies (∼41% in simulation and ∼28% in measurement) and quality. In experimental validation, the metalens exhibited flexible tunability, with the focal length continuously adjustable between two spatial positions upon variation of the incident polarization angle. The MTF results showed high contrast reproduction and sharp imaging, with a Strehl ratio of >0.7 for all polarization angles. With compactness, design flexibility, and high focusing quality, the proposed varifocal metalens holds potential for diverse applications, advancing adaptive and versatile optical devices.

Clinical Outcomes and Cost-Effectiveness of Osteoporosis Screening With Dual-Energy X-ray Absorptiometry.

OBJECTIVE: This study aimed to evaluate the clinical outcomes and cost-effectiveness of dual-energy X-ray absorptiometry (DXA) for osteoporosis screening. MATERIALS AND METHODS: Eligible patients who had and had not undergone DXA screening were identified from among those aged 50 years or older at Kaohsiung Veterans General Hospital, Taiwan. Age, sex, screening year (index year), and Charlson comorbidity index of the DXA and non-DXA groups were matched using inverse probability of treatment weighting (IPTW) for propensity score analysis. For cost-effectiveness analysis, a societal perspective, 1-year cycle length, 20-year time horizon, and discount rate of 2% per year for both effectiveness and costs were adopted in the incremental cost-effectiveness (ICER) model. RESULTS: The outcome analysis included 10337 patients (female:male, 63.8%:36.2%) who were screened for osteoporosis in southern Taiwan between January 1, 2012, and December 31, 2021. The DXA group had significantly better outcomes than the non-DXA group in terms of fragility fractures (7.6% vs. 12.5%, P < 0.001) and mortality (0.6% vs. 4.3%, P < 0.001). The DXA screening strategy gained an ICER of US$ -2794 per quality-adjusted life year (QALY) relative to the non-DXA at the willingness-to-pay threshold of US$ 33004 (Taiwan's per capita gross domestic product). The ICER after stratifying by ages of 50-59, 60-69, 70-79, and ≥ 80 years were US$ -17815, US$ -26862, US$ -28981, and US$ -34816 per QALY, respectively. CONCLUSION: Using DXA to screen adults aged 50 years or older for osteoporosis resulted in a reduced incidence of fragility fractures, lower mortality rate, and reduced total costs. Screening for osteoporosis is a cost-saving strategy and its effectiveness increases with age. However, caution is needed when generalizing these cost-effectiveness results to all older populations because the study population consisted mainly of women.

High-Resolution Metalens Imaging with Sequential Artificial Intelligence Models.

An analysis of the optical response of a GaN-based metalens was conducted alongside the utilization of two sequential artificial intelligence (AI) models in addressing the occasional issues of blurriness and color cast in captured images. The optical loss of the metalens in the blue spectral range was found to have resulted in the color cast of images. Autoencoder and CodeFormer sequential models were employed in order to correct the color cast and reconstruct image details, respectively. Said sequential models successfully addressed the color cast and reconstructed details for all of the allocated face image categories. Subsequently, the CIE 1931 chromaticity diagrams and peak signal-to-noise ratio analysis provided numerical evidence of the AI models' effectiveness in image reconstruction. Furthermore, the AI models can still repair the image without blue information. Overall, the integration of metalens and artificial intelligence models marks a breakthrough in enhancing the performance of full-color metalens-based imaging systems.

Extensible benchmarking of methods that identify and quantify polyadenylation sites from RNA-seq data.

The tremendous rate with which data is generated and analysis methods emerge makes it increasingly difficult to keep track of their domain of applicability, assumptions, limitations, and consequently, of the efficacy and precision with which they solve specific tasks. Therefore, there is an increasing need for benchmarks, and for the provision of infrastructure for continuous method evaluation. APAeval is an international community effort, organized by the RNA Society in 2021, to benchmark tools for the identification and quantification of the usage of alternative polyadenylation (APA) sites from short-read, bulk RNA-sequencing (RNA-seq) data. Here, we reviewed 17 tools and benchmarked eight on their ability to perform APA identification and quantification, using a comprehensive set of RNA-seq experiments comprising real, synthetic, and matched 3'-end sequencing data. To support continuous benchmarking, we have incorporated the results into the OpenEBench online platform, which allows for continuous extension of the set of methods, metrics, and challenges. We envisage that our analyses will assist researchers in selecting the appropriate tools for their studies, while the containers and reproducible workflows could easily be deployed and extended to evaluate new methods or data sets.