Innovative technologies for the fabrication of 3D/4D smart hydrogels and its biomedical applications - A comprehensive review.

Repairing and regenerating damaged tissues or organs, and restoring their functioning has been the ultimate aim of medical innovations. 'Reviving healthcare' blends tissue engineering with alternative techniques such as hydrogels, which have emerged as vital tools in modern medicine. Additive manufacturing (AM) is a practical manufacturing revolution that uses building strategies like molding as a viable solution for precise hydrogel manufacturing. Recent advances in this technology have led to the successful manufacturing of hydrogels with enhanced reproducibility, accuracy, precision, and ease of fabrication. Hydrogels continue to metamorphose as the vital compatible bio-ink matrix for AM. AM hydrogels have paved the way for complex 3D/4D hydrogels that can be loaded with drugs or cells. Bio-mimicking 3D cell cultures designed via hydrogel-based AM is a groundbreaking in-vivo assessment tool in biomedical trials. This brief review focuses on preparations and applications of additively manufactured hydrogels in the biomedical spectrum, such as targeted drug delivery, 3D-cell culture, numerous regenerative strategies, biosensing, bioprinting, and cancer therapies. Prevalent AM techniques like extrusion, inkjet, digital light processing, and stereo-lithography have been explored with their setup and methodology to yield functional hydrogels. The perspectives, limitations, and the possible prospects of AM hydrogels have been critically examined in this study.

Neuronal population activity in the olivocerebellum encodes the frequency of essential tremor in mice and patients.

Essential tremor (ET) is the most prevalent movement disorder, characterized primarily by action tremor, an involuntary rhythmic movement with a specific frequency. However, the neuronal mechanism underlying the coding of tremor frequency remains unexplored. Here, we used in vivo electrophysiology, optogenetics, and simultaneous motion tracking in the Grid2dupE3 mouse model to investigate whether and how neuronal activity in the olivocerebellum determines the frequency of essential tremor. We report that tremor frequency was encoded by the temporal coherence of population neuronal firing within the olivocerebellums of these mice, leading to frequency-dependent cerebellar oscillations and tremors. This mechanism was precise and generalizable, enabling us to use optogenetic stimulation of the deep cerebellar nuclei to induce frequency-specific tremors in wild-type mice or alter tremor frequencies in tremor mice. In patients with ET, we showed that deep brain stimulation of the thalamus suppressed tremor symptoms but did not eliminate cerebellar oscillations measured by electroencephalgraphy, indicating that tremor-related oscillations in the cerebellum do not require the reciprocal interactions with the thalamus. Frequency-disrupting transcranial alternating current stimulation of the cerebellum could suppress tremor amplitudes, confirming the frequency modulatory role of the cerebellum in patients with ET. These findings offer a neurodynamic basis for the frequency-dependent stimulation of the cerebellum to treat essential tremor.

Prognostic and Predictive Significance of Ki67 in Primary Non-metastatic or Recurrent Acral Melanoma: Evidence from a Multicenter Retrospective Study.

BACKGROUND: The purpose of this work was to investigate the prognostic significance of Ki67 in acral melanoma (AM). PATIENTS AND METHODS: Ki67 values in primary lesions (pKi67) of 481 patients with primary non-metastatic AM (primary cohort) from three tertiary hospitals and in recurrent lesions (rKi67) of 97 patients (recurrent cohort) were recorded. The associations of p/rKi67 with clinicopathological features and prognosis were analyzed. RESULTS: In the primary cohort, high pKi67 group tended to have more ulceration, pT4, lymph node metastasis (LNM), nodal macrometastases, and recurrence (all P < 0.05). Logistic regression analysis revealed that pKi67 was significantly associated with pT4 and LNM (P = 0.004 and 0.027, respectively). Furthermore, both 5-year overall survival (OS) and recurrence-free survival (RFS) rates in high pKi67 group were significantly worse than those in moderate and low pKi67 groups (OS 47.8% versus 55.7 versus 76.8%, P = 0.002; RFS: 27.1 versus 42.8 versus 61.8%, P < 0.001). Similarly, in the recurrent cohort, the 5-year survival after recurrence (SAR) rates in high rKi67 group was significantly worse than those in moderate and low rKi67 groups (31.7 versus 47.4 versus 75%; P = 0.026). Stratified analysis also indicated a significant survival difference among pKi67 groups within various subgroups. Most importantly, multivariate Cox analysis demonstrated that pKi67 could be independently associated with OS and RFS, as well as rKi67 for SAR (all P < 0.05). CONCLUSIONS: A high Ki67 value was significantly associated with adverse pathological and prognostic features in both primary and recurrent AM cohorts. Ki67 should be routinely evaluated to guide risk stratification and prognostic prediction.

Exciton polariton condensation from bound states in the continuum at room temperature.

Exciton-polaritons (polaritons) resulting from the strong exciton-photon interaction stimulates the development of novel low-threshold coherent light sources to circumvent the ever-increasing energy demands of optical communications1-3. Polaritons from bound states in the continuum (BICs) are promising for Bose-Einstein condensation owing to their theoretically infinite quality factors, which provide prolonged lifetimes and benefit the polariton accumulations4-7. However, BIC polariton condensation remains limited to cryogenic temperatures ascribed to the small exciton binding energies of conventional material platforms. Herein, we demonstrated room-temperature BIC polariton condensation in perovskite photonic crystal lattices. BIC polariton condensation was demonstrated at the vicinity of the saddle point of polariton dispersion that generates directional vortex beam emission with long-range coherence. We also explore the peculiar switching effect among the miniaturized BIC polariton modes through effective polariton-polariton scattering. Our work paves the way for the practical implementation of BIC polariton condensates for integrated photonic and topological circuits.

Phase and Composition Engineering of Self-Intercalated 2D Metallic Tantalum Sulfide for Second-Harmonic Generation.

Self-intercalation in two-dimensional (2D) materials is significant, as it offers a versatile approach to modify material properties, enabling the creation of interesting functional materials, which is essential in advancing applications across various fields. Here, we define ic-2D materials as covalently bonded compounds that result from the self-intercalation of a metal into layered 2D compounds. However, precisely growing ic-2D materials with controllable phases and self-intercalation concentrations to fully exploit the applications in the ic-2D family remains a great challenge. Herein, we demonstrated the controlled synthesis of self-intercalated H-phase and T-phase Ta1+xS2 via a temperature-driven chemical vapor deposition (CVD) approach with a viable intercalation concentration spanning from 10% to 58%. Atomic-resolution scanning transmission electron microscopy-annular dark field imaging demonstrated that the self-intercalated Ta atoms occupy the octahedral vacancies located at the van der Waals gap. The nonperiodic Ta atoms break the centrosymmetry structure and Fermi surface properties of intrinsic TaS2. Therefore, ic-2D T-phase Ta1+xS2 consistently exhibit a spontaneous nonlinear optical (NLO) effect regardless of the sample thickness and self-intercalation concentrations. Our results propose an approach to activate the NLO response of centrosymmetric 2D materials, achieving the modulation of a wide range of optoelectronic properties via nonperiodic self-intercalation in the ic-2D family.

Boosting exciton mobility approaching Mott-Ioffe-Regel limit in Ruddlesden-Popper perovskites by anchoring the organic cation.

Exciton transport in two-dimensional Ruddlesden-Popper perovskite plays a pivotal role for their optoelectronic performance. However, a clear photophysical picture of exciton transport is still lacking due to strong confinement effects and intricate exciton-phonon interactions in an organic-inorganic hybrid lattice. Herein, we present a systematical study on exciton transport in (BA)2(MA)n-1PbnI3n+1 Ruddlesden-Popper perovskites using time-resolved photoluminescence microscopy. We reveal that the free exciton mobilities in exfoliated thin flakes can be improved from around 8 cm2 V-1 s-1 to 280 cm2V-1s-1 by anchoring the soft butyl ammonium cation with a polymethyl methacrylate network at the surface. The mobility of the latter is close to the theoretical limit of Mott-Ioffe-Regel criterion. Combining optical measurements and theoretical studies, it is unveiled that the polymethyl methacrylate network significantly improve the lattice rigidity resulting in the decrease of deformation potential scattering and lattice fluctuation at the surface few layers. Our work elucidates the origin of high exciton mobility in Ruddlesden-Popper perovskites and opens up avenues to regulate exciton transport in two-dimensional materials.

Two birds with one stone: One-pot concurrent Ta-doping and -coating on Ni-rich LiNi0.92Co0.04Mn0.04O2 cathode materials with fiber-type microstructure and Li+-conducting layer formation.

A novel scalable Taylor-Couette reactor (TCR) synthesis method was employed to prepare Ta-modified LiNi0.92Co0.04Mn0.04O2 (T-NCM92) with different Ta contents. Through experiments and density functional theory (DFT) calculations, the phase and microstructure of Ta-modified NCM92 were analyzed, showing that Ta provides a bifunctional (doping and coating at one time) effect on LiNi0.92Co0.04Mn0.04O2 cathode material through a one-step synthesis process via a controlling suitable amount of Ta and Li-salt. Ta doping allows the tailoring of the microstructure, orientation, and morphology of the primary NCM92 particles, resulting in a needle-like shape with fine structures that considerably enhance Li+ ion diffusion and electrochemical charge/discharge stability. The Ta-based surface-coating layer effectively prevented microcrack formation and inhibited electrolyte decomposition and surface-side reactions during cycling, thereby significantly improving the electrochemical performance and long-term cycling stability of NCM92 cathodes. Our as-prepared NCM92 modified with 0.2 mol% Ta (i.e., T2-NCM92) exhibits outstanding cyclability, retaining 84.5 % capacity at 4.3 V, 78.3 % at 4.5 V, and 67.6 % at 45 â„ƒ after 200 cycles at 1C. Even under high-rate conditions (10C), T2-NCM92 demonstrated a remarkable capacity retention of 66.9 % after 100 cycles, with an initial discharge capacity of 157.6 mAh g-1. Thus, the Ta modification of Ni-rich NCM92 materials is a promising option for optimizing NCM cathode materials and enabling their use in real-world electric vehicle (EV) applications.

Comparison of Micro-flow Imaging and Contrast-Enhanced Ultrasound in Ultrasound-Guided Microwave Ablation of Benign Thyroid Nodules.

OBJECTIVE: The study described here was aimed at ascertaining the utility of micro-flow imaging (MFI) during ultrasound (US)-guided microwave ablation (MWA) of thyroid nodules by contrasting its effectiveness with that of contrast-enhanced ultrasound (CEUS). METHODS: Seventy-three patients with eighty-eight thyroid nodules who underwent US-guided MWA were included in our study from January 2020 to June 2023. Thirty-five patients underwent CEUS during the MWA process, and thirty-eight patients underwent MFI during the MWA process. We compared the two groups' baseline characteristics, tumor volume (V), volume reduction rate (VRR), complications and clinical characteristics. RESULTS: Both groups exhibited similar outcomes with respect to V and VRR at 1, 3, 6, 12 and 18 mo after MWA (p > 0.05). Consistency was observed with respect to post-operative complications, supplementary ablation times and surgical duration (p > 0.05). It is worth noting that the MFI group had lower treatment costs compared with the CEUS group (11,337.64 ± 80.93 yuan for the MFI group versus 12,971.23 ± 254.89 yuan for the CEUS group, p < 0.05). CONCLUSION: In the MWA procedure for thyroid nodules, MFI is similar to CEUS with respect to safety and efficacy. Simultaneously, it offers the advantage of reducing surgical expenses, which lessens the economic burden for patients.

The mechanisms of ferroptosis and its role in atherosclerosis.

Ferroptosis is a newly identified form of non-apoptotic programmed cell death, characterized by the iron-dependent accumulation of lethal lipid reactive oxygen species (ROS) and peroxidation of membrane polyunsaturated fatty acid phospholipids (PUFA-PLs). Ferroptosis is unique among other cell death modalities in many aspects. It is initiated by excessive oxidative damage due to iron overload and lipid peroxidation and compromised antioxidant defense systems, including the system Xc-/ glutathione (GSH)/glutathione peroxidase 4 (GPX4) pathway and the GPX4-independent pathways. In the past ten years, ferroptosis was reported to play a critical role in the pathogenesis of various cardiovascular diseases, e.g., atherosclerosis (AS), arrhythmia, heart failure, diabetic cardiomyopathy, and myocardial ischemia-reperfusion injury. Studies have identified dysfunctional iron metabolism and abnormal expression profiles of ferroptosis-related factors, including iron, GSH, GPX4, ferroportin (FPN), and SLC7A11 (xCT), as critical indicators for atherogenesis. Moreover, ferroptosis in plaque cells, i.e., vascular endothelial cell (VEC), macrophage, and vascular smooth muscle cell (VSMC), positively correlate with atherosclerotic plaque development. Many macromolecules, drugs, Chinese herbs, and food extracts can inhibit the atherogenic process by suppressing the ferroptosis of plaque cells. In contrast, some ferroptosis inducers have significant pro-atherogenic effects. However, the mechanisms through which ferroptosis affects the progression of AS still need to be well-known. This review summarizes the molecular mechanisms of ferroptosis and their emerging role in AS, aimed at providing novel, promising druggable targets for anti-AS therapy.

Dissolving microneedle system containing Ag nanoparticle-decorated silk fibroin microspheres and antibiotics for synergistic therapy of bacterial biofilm infection.

Most cases of delayed wound healing are associated with bacterial biofilm infections due to high antibiotic resistance. To improve patient compliance and recovery rates, it is critical to develop minimally invasive and efficient methods to eliminate bacterial biofilms as an alternative to clinical debridement techniques. Herein, we develop a dissolving microneedle system containing Ag nanoparticles (AgNPs)-decorated silk fibroin microspheres (SFM-AgNPs) and antibiotics for synergistic treatment of bacterial biofilm infection. Silk fibroin microspheres (SFM) are controllably prepared in an incompatible system formed by a mixture of protein and carbohydrate solutions by using a mild all-aqueous phase method and serve as biological templates for the synthesis of AgNPs. The SFM-AgNPs exert dose- and time-dependent broad-spectrum antibacterial effects by inducing bacterial adhesion. The combination of SFM-AgNPs with antibiotics breaks the limitation of the antibacterial spectrum and achieves better efficacy with reduced antibiotic dosage. Using hyaluronic acid (HA) as the soluble matrix, the microneedle system containing SFM-AgNPs and anti-Gram-positive coccus drug (Mupirocin) inserts into the bacterial biofilms with sufficient strength, thereby effectively delivering the antibacterial agents and realizing good antibiofilm effect on Staphylococcus aureus-infected wounds. This work demonstrates the great potential for the development of novel therapeutic systems for eradicating bacterial biofilm infections.

Doping-Induced Surface and Grain Boundary Effects in Ni-Rich Layered Cathode Materials.

In this work, the effects of dopant size and oxidation state on the structure and electrochemical performance of LiNi0.8 Co0.1 Mn0.1 O2 (NCM811) are investigated. It is shown that doping with boron (B) which has a small ionic radius and an oxidation state of 3+, leads to the formation of a boron oxide-containing surface coating (probably Li3 BO3 ), mainly on the outer surface of the secondary particles. Due to this effect, boron only slightly affects the size of the primary particle and the initial capacity, but significantly improves the capacity retention. On the other hand, the dopant ruthenium (Ru) with a larger ionic radius and a higher oxidation state of 5+ can be stabilized within the secondary particles and does not experience a segregation to the outer agglomerate surface. However, the Ru dopant preferentially occupies incoherent grain boundary sites, resulting in smaller primary particle size and initial capacity than for the B-doped and pristine NCM811. This work demonstrates that a small percentage of dopant (2 mol%) cannot significantly affect bulk properties, but it can strongly influence the surface and/or grain boundary properties of microstructure and thus the overall performance of cathode materials.

The impact of serum estradiol and progesterone levels during implantation on obstetrical complications and perinatal outcomes in frozen embryo transfer.

BACKGROUND: This study sought to evaluate obstetric complications and perinatal outcomes in frozen embryo transfer (FET) using either a natural cycle (NC-FET) or a hormone therapy cycle (HT-FET). Furthermore, we investigated how serum levels of estradiol (E2) and progesterone (P4) on the day of and 3 days after embryo transfer (ET) correlated with clinical outcomes in the two groups. METHODS: We conducted a retrospective, single-center study from January 1, 2015, to December 31, 2019. The study included couples who underwent NC-FET or HT-FET resulting in a singleton live birth. Serum levels of E2 and P4 were measured on the day of and 3 days after ET. The primary outcomes assessed were preterm birth rate, low birth weight, macrosomia, hypertensive disorders in pregnancy, gestational diabetes mellitus, postpartum hemorrhage, and placenta-related complications. RESULTS: A total of 229 singletons were included, with 49 in the NC-FET group and 180 in the HT-FET group. There were no significant differences in obstetric complications and perinatal outcomes between the two groups. The NC-FET group had significantly higher serum levels of P4 (17.2 ng/mL vs 8.85 ng/mL; p < 0.0001) but not E2 (144 pg/mL vs 147 pg/mL; p = 0.69) on the day of ET. Additionally, 3 days after ET, the NC-FET group had significantly higher levels of both E2 (171 pg/mL vs 140.5 pg/mL; p = 0.0037) and P4 (27.3 ng/mL vs 11.7 ng/mL; p < 0.0001) compared with the HT-FET group. CONCLUSION: Our study revealed that although there were significant differences in E2 and P4 levels around implantation between the two groups, there were no significant differences in obstetric complications and perinatal outcomes. Therefore, the hormonal environment around implantation did not appear to be the primary cause of differences in obstetric and perinatal outcomes between the two EM preparation methods used in FET.

Functional characterization of four antenna-biased chemosensory proteins in Dioryctria abietella reveals a broadly tuned olfactory DabiCSP1 and its key residues in ligand-binding.

The orientation of the oligophagous cone-feeding moth Dioryctria abietella (Lepidoptera: Pyralidae) to host plants primarily relies on olfactory-related proteins, particularly those candidates highly expressed in antennae. Here, through a combination of expression profile, ligand-binding assay, molecular docking and site-directed mutagenesis strategies, we characterized the chemosensory protein (CSP) gene family in D. abietella. Quantitative real-time PCR (qPCR) analyses revealed the detectable expression of all 22 DabiCSPs in the antennae, of which seven genes were significantly enriched in this tissue. In addition, the majority of the genes (19/22 relatives) had the expression in at least one reproductive tissue. In the interactions of four antenna-dominant DabiCSPs and different chemical classes, DabiCSP1 was broadly tuned to 27 plant-derived odors, three man-made insecticides and one herbicide with high affinities (Ki < 6.60 µM). By contrast, three other DabiCSPs (DabiCSP4, CSP6 and CSP17) exhibited a narrow odor binding spectrum, in response to six compounds for each protein. Our mutation analyses combined with molecular docking simulations and binding assays further identified four key residues (Tyr25, Thr26, Ile65 and Val69) in the interactions of DabiCSP1 and ligands, of which binding abilities of this protein to 12, 15, 16 and three compounds were significantly decreased compared to the wildtype protein, respectively. Our study reveals different odor binding spectra of four DabiCSPs enriched in antennae and identifies key residues responsible for the binding of DabiCSP1 and potentially active compounds for the control of this pest.

Increased risk of chronic fatigue syndrome following infection: a 17-year population-based cohort study.

BACKGROUND: Previous serological studies have indicated an association between viruses and atypical pathogens and Chronic Fatigue Syndrome (CFS). This study aims to investigate the correlation between infections from common pathogens, including typical bacteria, and the subsequent risk of developing CFS. The analysis is based on data from Taiwan's National Health Insurance Research Database. METHODS: From 2000 to 2017, we included a total of 395,811 cases aged 20 years or older newly diagnosed with infection. The cases were matched 1:1 with controls using a propensity score and were followed up until diagnoses of CFS were made. RESULTS: The Cox proportional hazards regression analysis was used to estimate the relationship between infection and the subsequent risk of CFS. The incidence density rates among non-infection and infection population were 3.67 and 5.40 per 1000 person-years, respectively (adjusted hazard ratio [HR] = 1.5, with a 95% confidence interval [CI] 1.47-1.54). Patients infected with Varicella-zoster virus, Mycobacterium tuberculosis, Escherichia coli, Candida, Salmonella, Staphylococcus aureus and influenza virus had a significantly higher risk of CFS than those without these pathogens (p < 0.05). Patients taking doxycycline, azithromycin, moxifloxacin, levofloxacin, or ciprofloxacin had a significantly lower risk of CFS than patients in the corresponding control group (p < 0.05). CONCLUSION: Our population-based retrospective cohort study found that infection with common pathogens, including bacteria, viruses, is associated with an increased risk of developing CFS.

Research trends and hotspots of exercise therapy in Panvascular disease: A bibliometric analysis.

Panvascular diseases are a group of vascular system diseases, mainly including the heart, brain, neck, and other parts of the vascular lesions. As a non-pharmacological intervention, exercise therapy could prevent and treat Panvascular diseases. However, few bibliometric analyses of exercise therapy in Panvascular disease exist. This study aimed to analyze the trends and hotspots over the past decade and provide insights into the latest state of the art in global research, thereby contributing to further research in the field. We systematically searched the Web of Science Core Collection (WOSCC) for articles on exercise therapy and Panvascular disease. The acquired information from the reports was analyzed using CiteSpace and VOSviewer software to assess and forecast this field hottest areas and trends. The final analysis included 294 articles by our specified inclusion criteria. The number of publications has gradually increased over the past decade. Stroke was one of the most studied Panvascular diseases. China and the University of Sao Paulo were the country es and institutions that contributed the most to the field. Mary M. McDermott was the most prolific researcher, and the Journal of Vascular Surgery published the most articles. The 6-minute walk test, skeletal muscle, oxidative stress, and supervised exercise therapy were hot topics from 2019 to 2023. In conclusion, exploring exercise therapy programs and exercise mechanisms for Panvascular diseases has been ongoing. This study revealed the current status and trends of research in the field and identified hot topics. It was helpful for scholars to understand exercise therapy critical role in treating and preventing Panvascular diseases and provided a reference for clinical decision-making and further research.

Development of lung tissue models and their applications.

The lungs are important organs that play a critical role in the development of specific diseases, as well as responding to the effects of drugs, chemicals, and environmental pollutants. Due to the ethical concerns around animal testing, alternative methods have been sought which are more time-effective, do not pose ethical issues for animals, do not involve species differences, and provide easy investigation of the pathobiology of lung diseases. Several national and international organizations are working to accelerate the development and implementation of structurally and functionally complex tissue models as alternatives to animal testing, particularly for the lung. Unfortunately, to date, there is no lung tissue model that has been accepted by regulatory agencies for use in inhalation toxicology. This review discusses the challenges involved in developing a relevant lung tissue model derived from human cells such as cell lines, primary cells, and pluripotent stem cells. It also introduces examples of two-dimensional (2D) air-liquid interface and monocultured and co-cultured three-dimensional (3D) culture techniques, particularly organoid culture and 3D bioprinting. Furthermore, it reviews development of the lung-on-a-chip model to mimic the microenvironment and physiological performance. The applications of lung tissue models in various studies, especially disease modeling, viral respiratory infection, and environmental toxicology will be also introduced. The development of a relevant lung tissue model is extremely important for standardizing and validation the in vitro models for inhalation toxicity and other studies in the future.

Trajectory in biological metal-organic frameworks: Biosensing and sustainable strategies-perspectives and challenges.

The ligand attribute of biomolecules to form coordination bonds with metal ions led to the discovery of a novel class of materials called biomolecule-associated metal-organic frameworks (Bio-MOFs). These biomolecules coordinate in multiple ways and provide versatile applications. Far-spread bio-ligands include nucleobases, amino acids, peptides, cyclodextrins, saccharides, porphyrins/metalloporphyrin, proteins, etc. Low-toxicity, self-assembly, stability, designable and selectable porous size, the existence of rigid and flexible forms, bio-compatibility, and synergistic interactions between metal ions have led Bio-MOFs to be commercialized in industries such as sensors, food, pharma, and eco-sensing. The rapid growth and commercialization are stunted by absolute bio-compatibility issues, bulk morphology that makes it rigid to alter shape/porosity, longer reaction times, and inadequate research. This review elucidates the structural vitality, biocompatibility issues, and vital sensing applications, including challenges for incorporating bio-ligands into MOF. Critical innovations in Bio-MOFs' applicative spectrum, including sustainable food packaging, biosensing, insulin and phosphoprotein detection, gas sensing, CO2 capture, pesticide carriers, toxicant adsorptions, etc., have been elucidated. Emphasis is placed on biosensing and biomedical applications with biomimetic catalysis and sensitive sensor designing.

Comprehensive analysis of circulating cell-free RNAs in blood for diagnosing non-small cell lung cancer.

Early screening and detection of non-small cell lung cancer (NSCLC) is crucial due to the significantly low survival rate in advanced stages. Blood-based liquid biopsy is non-invasive test to assistant disease diagnosis, while cell-free RNA is one of the promising biomarkers in blood. However, the disease related signatures have not been explored completely for most cell-free RNA transcriptome sequencing (cfRNA-Seq) datasets. To address this gap, we developed a comprehensive cfRNA-Seq pipeline for data analysis and constructed a machine learning model to facilitate noninvasive early diagnosis of NSCLC. The results of our study have demonstrated the identification of differential mRNA, lncRNAs and miRNAs from cfRNA-Seq, which have exhibited significant association with development and progression of lung cancer. The classifier based on gene expression signatures achieved an impressive area under the curve (AUC) of up to 0.9, indicating high specificity and sensitivity in both cross-validation and independent test. Furthermore, the analysis of T cell and B cell immune repertoire extracted from cfRNA-Seq have provided insights into the immune status of cancer patients, while the microbiome analysis has revealed distinct bacterial and viral profiles between NSCLC and normal samples. In our future work, we aim to validate the existence of cancer associated T cell receptors (TCR)/B cell receptors (BCR) and microorganisms, and subsequently integrate all identified signatures into diagnostic model to improve the prediction accuracy. This study not only provided a comprehensive analysis pipeline for cfRNA-Seq dataset but also highlights the potential of cfRNAs as promising biomarkers and models for early NSCLC diagnosis, emphasizing their importance in clinical settings.

Advanced Detection Method for Dengue NS1 Protein Using Ultrasensitive ELISA with Thio-NAD Cycling.

Dengue fever, a mosquito-borne disease in tropical and subtropical climates caused by the dengue virus (DENV), has become a major social and economic burden in recent years. However, current primary detection methods are inadequate for early diagnosis of DENV because they are either time-consuming, expensive, or require training. Non-structural protein 1 (NS1) is secreted during DENV infection and is thus considered a suitable biomarker for the development of an early detection method. In the present study, we developed a detection method for the NS1 protein based on a previously reported thio-NAD cycling ELISA (i.e., ultrasensitive ELISA) and successfully achieved a LOD of 1.152 pg/mL. The clinical diagnosis potential of the detection system was also evaluated by using 85 patient specimens, inclusive of 60 DENV-positive and 25 DENV-negative specimens confirmed by the NAAT method. The results revealed 98.3% (59/60) sensitivity and 100% (25/25) specificity, which was in almost perfect agreement with the NAAT data with a kappa coefficient of 0.972. The present study demonstrates the diagnostic potential of using an ultrasensitive ELISA as a low-cost, easy-to-use method for the detection of DENV compared with NAAT and could be of great benefit in low-income countries.