Label-free detection of ConA-induced T-lymphocyte activation at single-cell level by microfluidics.

Lymphocyte activation is critical in regulating immune responses. The resulting T-cell proliferation has been implicated in the pathogenesis of a variety of autoimmune diseases, such as SLE and rheumatoid arthritis. ConA (concanavalin A)-induced activation has been widely used in the T lymphocytes model of immune-mediated liver injury, autoimmune hepatitis, and so on. In those works, it usually requires fluorescent labeling or cell staining to confirm whether the cells are transformed successfully after medicine treatment to figure out efficacy/pharmacology. The detection preparation steps are time-consuming and have limitations for further proteomic/genomic identifications. Here, a label-free microfluidic method is established to detect lymphocyte activation degree. The lymphocyte and ConA-activated lymphocyte were investigated by a microfluidic device. According to where single cells in the sample were captured in the designed channel, lymphocyte and ConA-activated samples are differentiated and characterized by population electric field factors, 2.08 × 104 and 2.21 × 104 V/m, respectively. Furthermore, salidroside, a herbal medicine that was documented to promote the transformation, was used to treat lymphocyte cells, and the treated cell population is detected to be 2.67 × 104 V/m. The characterization indicates an increasing trend with the activation degree. The result maintains a high consistency with traditional staining methods with transformed cells of 15.8%, 28.8%, and 48.3% in each cell population. Dielectrophoresis is promising to work as a tool for detecting lymphocyte transformation and medical efficacy detection.

An insoluble cellulose nanofiber with robust expansion capacity protects against obesity.

An imbalance between energy intake and energy expenditure predisposes obesity and its related metabolic diseases. Soluble dietary fiber has been shown to improve metabolic homeostasis mainly via microbiota reshaping. However, the application and metabolic effects of insoluble fiber are less understood. Herein, we employed nanotechnology to design citric acid-crosslinked carboxymethyl cellulose nanofibers (CL-CNF) with a robust capacity of expansion upon swelling. Supplementation with CL-CNF reduced food intake and delayed digestion rate in mice by occupying stomach. Besides, CL-CNF treatment mitigated diet-induced obesity and insulin resistance in mice with enhanced energy expenditure, as well as ameliorated inflammation in adipose tissue, intestine and liver and reduced hepatic steatosis, without any discernible signs of toxicity. Additionally, CL-CNF supplementation resulted in enrichment of probiotics such as Bifidobacterium and decreased in the relative abundances of deleterious microbiota expressing bile salt hydrolase, which led to increased levels of conjugated bile acids and inhibited intestinal FXR signaling to stimulate the release of GLP-1. Taken together, our findings demonstrate that CL-CNF administration protects mice from diet-induced obesity and metabolic dysfunction by reducing food intake, enhancing energy expenditure and remodeling gut microbiota, making it a potential therapeutic strategy against metabolic diseases.

FXR activation remodels hepatic and intestinal transcriptional landscapes in metabolic dysfunction-associated steatohepatitis.

The escalating obesity epidemic and aging population have propelled metabolic dysfunction-associated steatohepatitis (MASH) to the forefront of public health concerns. The activation of FXR shows promise to combat MASH and its detrimental consequences. However, the specific alterations within the MASH-related transcriptional network remain elusive, hindering the development of more precise and effective therapeutic strategies. Through a comprehensive analysis of liver RNA-seq data from human and mouse MASH samples, we identified central perturbations within the MASH-associated transcriptional network, including disrupted cellular metabolism and mitochondrial function, decreased tissue repair capability, and increased inflammation and fibrosis. By employing integrated transcriptome profiling of diverse FXR agonists-treated mice, FXR liver-specific knockout mice, and open-source human datasets, we determined that hepatic FXR activation effectively ameliorated MASH by reversing the dysregulated metabolic and inflammatory networks implicated in MASH pathogenesis. This mitigation encompassed resolving fibrosis and reducing immune infiltration. By understanding the core regulatory network of FXR, which is directly correlated with disease severity and treatment response, we identified approximately one-third of the patients who could potentially benefit from FXR agonist therapy. A similar analysis involving intestinal RNA-seq data from FXR agonists-treated mice and FXR intestine-specific knockout mice revealed that intestinal FXR activation attenuates intestinal inflammation, and has promise in attenuating hepatic inflammation and fibrosis. Collectively, our study uncovers the intricate pathophysiological features of MASH at a transcriptional level and highlights the complex interplay between FXR activation and both MASH progression and regression. These findings contribute to precise drug development, utilization, and efficacy evaluation, ultimately aiming to improve patient outcomes.

Effects of thyroid hormone on monochromatic light combinations mediate skeletal muscle fiber pattern in broilers.

It has been shown that monochromatic green light and blue light promote skeletal muscle development in early (P0-P26) and later growth stages (P27-P42), respectively. This study further investigated the effects of monochromatic light combinations on myogenesis and myofiber types transformation in broilers. Here, a total of 252 chicks were exposed to monochromatic light [red (R), green (G), blue (B), or white light (W)], and monochromatic light combination [green and blue light combination (GB), blue and green light combination (BG), red and blue combination (RB)] until P42. Compared with other groups, GB significantly increased body weight, and muscle organ index, both proportions of larger-size myofibers and oxidative myofibers in the pectoralis major (PM) and gastrocnemius muscle (GAS). Meanwhile, GB up-regulated the abundance of oxidative genes MYH7B and MYH1B, transcription factors PAX7 and Myf5, antioxidant proteins Nrf2, HO-1, and GPX4, and the activities of antioxidant enzymes CAT, GPx, and T-AOC, but down-regulated the abundance of glycolytic related genes MYH 1A, MyoD, MyoG, Mstn, Keap1, TNFa, and MDA levels. Consistent with the change of myofiber pattern, GB significantly reduced serum thyroid hormone (TH) levels, up-regulated skeletal muscle deiodinase DIO3 expression and down-regulated deiodinase DIO2 expression, which may directly lead to the reduction of intramuscular TH levels to affect myofiber types transformation. In contrast, the proportion of fast glycolytic muscle fibers increased in the RR with increasing TH levels. After thyroidectomy, the above parameters were inversed and resulted in no significant difference of each color light treatment group. These data suggested that GB significantly increased the proportion of oxidative muscle fibers and antioxidant capacity in skeletal muscle of broilers, which was regulated by TH-DIO2/DIO3 signaling pathway.

NGS panel enhance precise diagnosis of myeloid neoplasms under WHO-HAEM5 and International Consensus Classification: An observational study.

This study aimed to assess hematological diseases next-generation sequencing (NGS) panel enhances the diagnosis and classification of myeloid neoplasms (MN) using the 5th edition of the WHO Classification of Hematolymphoid Tumors (WHO-HAEM5) and the International Consensus Classification (ICC) of Myeloid Tumors. A cohort of 112 patients diagnosed with MN according to the revised fourth edition of the WHO classification (WHO-HAEM4R) underwent testing with a 141-gene NGS panel for hematological diseases. Ancillary studies were also conducted, including bone marrow cytomorphology and routine cytogenetics. The cases were then reclassified according to WHO-HAEM5 and ICC to assess the practical impact of these 2 classifications. The mutation detection rates were 93% for acute myeloid leukemia (AML), 89% for myelodysplastic syndrome (MDS), 94% for myeloproliferative neoplasm (MPN), and 100% for myelodysplasia/myeloproliferative neoplasm (MDS/MPN) (WHO-HAEM4R). NGS provided subclassified information for 26 and 29 patients with WHO-HAEM5 and ICC, respectively. In MPN, NGS confirmed diagnoses in 16 cases by detecting JAK2, MPL, or CALR mutations, whereas 13 "triple-negative" MPN cases revealed at least 1 mutation. NGS panel testing for hematological diseases improves the diagnosis and classification of MN. When diagnosed with ICC, NGS produces more classification subtype information than WHO-HAEM5.

NAD+ supplementation improves mitochondrial functions and normalizes glaucomatous trabecular meshwork features.

Glaucoma is characterized by pathological elevation of intraocular pressure (IOP) due to dysfunctional trabecular meshwork (TM), which is the primary cause of irreversible vision loss. There are currently no effective treatment strategies for glaucoma. Mitochondrial function plays a crucial role in regulating IOP within the TM. In this study, primary TM cells treated with dexamethasone were used to simulate glaucomatous changes, showing abnormal cellular cytoskeleton, increased expression of extracellular matrix, and disrupted mitochondrial fusion and fission dynamics. Furthermore, glaucomatous TM cell line GTM3 exhibited impaired mitochondrial membrane potential and phagocytic function, accompanied by decreased oxidative respiratory levels as compared to normal TM cells iHTM. Mechanistically, lower NAD + levels in GTM3, possibly associated with increased expression of key enzymes CD38 and PARP1 related to NAD + consumption, were observed. Supplementation of NAD + restored mitochondrial function and cellular viability in GTM3 cells. Therefore, we propose that the aberrant mitochondrial function in glaucomatous TM cells may be attributed to increased NAD + consumption dependent on CD38 and PARP1, and NAD + supplementation could effectively ameliorate mitochondrial function and improve TM function, providing a novel alternative approach for glaucoma treatment.

Microfluidics identify moso bamboo and henon bamboo by leaf protoplast subpopulations with single-cell analysis.

Current techniques identifying herbal medicine species require marker labeling or lack systematical accuracy (expert authentication). There is an emerging interest in developing an accurate and label-free tool for herbal medicine authentication. Here, a high-resolution microfluidic-based method is developed for identifying herbal species by protoplast subpopulations. Moso bamboo and henon bamboo are used as a model to be differentiated based on protoplast. Their biophysical properties factors are characterized to be 7.09 (± 0.39) × 108 V/m2 and 6.54 (± 0.26) × 108 V/m2, respectively. Their biophysical distributions could be distinguished by the Cramér-von Mises criterion with a 94.60% confidence level. The subpopulations of each were compared with conventional flow cytometry indicating the existence of subpopulations and the differences between the two species. The subsets divided by a biophysical factor of 8.05(± 0.51) × 108 V/m2 suggest good consistency with flow cytometry. The work demonstrated the possibility of microfluidics manipulation on protoplast for medication safety use taking advantage of dielectrophoresis. The device is promising in developing a reliable and accurate way of identifying herbal species with difficulties in authentication.

VEGF-C and 5-Fluorouracil Improve Bleb Survival in a Rabbit Glaucoma Surgery Trabeculectomy Model.

Purpose: To evaluate VEGF-C-induced lymphoproliferation in conjunction with 5-fluorouracil (5-FU) antimetabolite treatment in a rabbit glaucoma filtration surgery (GFS) model. Methods: Thirty-two rabbits underwent GFS and were assigned to four groups (n = 8 each) defined by subconjunctival drug treatment: (a) VEGF-C combined with 5-FU, (b) 5-FU, (c) VEGF-C, (d) and control. Bleb survival, bleb measurements, and IOP were evaluated over 30 days. At the end, histology and anterior segment OCT were performed on some eyes. mRNA was isolated from the remaining eyes for RT-PCR evaluation of vessel-specific markers (lymphatics, podoplanin and LYVE-1; and blood vessels, CD31). Results: Qualitatively and quantitatively, VEGF-C combined with 5-FU resulted in blebs which were posteriorly longer and wider than the other conditions: vs. 5-FU (P = 0.043 for longer, P = 0.046 for wider), vs. VEGF-C (P < 0.001, P < 0.001) and vs. control (P < 0.001, P < 0.001). After 30 days, the VEGF-C combined with 5-FU condition resulted in longer bleb survival compared with 5-FU (P = 0.025), VEGF-C (P < 0.001), and control (P < 0.001). Only the VEGF-C combined with 5-FU condition showed a negative correlation between IOP and time that was statistically significant (r = -0.533; P = 0.034). Anterior segment OCT and histology demonstrated larger blebs for the VEGF-C combined with 5-FU condition. Only conditions including VEGF-C led to increased expression of lymphatic markers (LYVE-1, P < 0.001-0.008 and podoplanin, P = 0.002-0.011). Expression of CD31 was not different between the groups (P = 0.978). Conclusions: Adding VEGF-C lymphoproliferation to standard antimetabolite treatment improved rabbit GFS success and may suggest a future strategy to improve human GFSs.

Polysaccharide extracted from Sarcandra glabra residue attenuate cognitive impairment by regulating gut microbiota in diabetic mice.

Diabetic encephalopathy (DE), characterized by cognitive impairment, currently lacks targeted treatment. Previous studies have shown that Sarcandra glabra extracted residue polysaccharide (SERP) exhibited hypoglycemic effects either in vitro or in streptozotocin-induced diabetes mice. However, the therapeutic effect of SERP on DE was not elucidated. This study investigated the therapeutic effect of SERP on DE and its underlying mechanism. Our results revealed that SERP regulates glucose and lipid metabolism, improves cognitive function, and exhibits diminished activity post-antibiotic intervention. Importantly, we discovered a novel mechanism by which SERP modulates the gut microbiota, specifically enriching Bacteroidales S24-7, resulting in elevated levels of butyric acid in the intestine. This regulation modulates the intestinal endocrine cell lipid metabolism level, restores damaged intestinal barriers and neural epithelial circuits, thus exhibiting cure effects. Our findings suggest that SERP could become a candidate for treating DE, potentially involving the regulation mechanism of the "microbiota-gut-brain axis". This study underscores the unique therapeutic efficacy of SERP in managing DE, offering fresh drug candidates and innovative treatment strategies for this challenging condition.

Tree-ring widths of Pinus tabulaeformis Carr reveal variability of winter half-year precipitation on the north-south transition zone in central China over the past 220 years.

Long-term, high-resolution regional drought records contribute to understanding the impacts of drought on environmental and social systems in central China. Here, we develop a regional tree-ring width chronology of Pinus tabulaeformis Carr from the northern slope of Funiu Mountains on the north-south transition zone in central China. Monthly correlation analyses showed that temperature and humidity in current May and June are main limiting factors on tree growth. Despite that, the highest correlation with tree growth was found to be precipitation from previous December to current June (PreDJ, 0.718, p < 0.001), which was chosen for reconstruction. The reconstructed PreDJ revealed six drought periods and five wet periods over the past 220 years, and the recent dry spell would likely to continue. Spectral analyses indicated that the reconstructed PreDJ was closely related to the El Nino-Southern Oscillation (ENSO, 2-7a) and 35a climatic oscillation of Bruckner, and was also affected by the Quasi-Biennial Oscillation (QBO). Wavelet analyses showed that the quasi-cycle of 2-7a persisted over the past 220 years and strengthened after the 1980s, and the QBO signals appeared from the 1860s to 1970s and wear off thereafter, and 35a cycle only appeared during 1820-1920. Spatial analysis found that the reconstructed PreDJ had good spatial representation of precipitation in the central-eastern China. Therefore, the results of this study provide reliable information for understanding long-term drought impacts on environmental conditions and socioeconomic development in central China.

Discovery of novel and selective farnesoid X receptor antagonists through structure-based virtual screening, preliminary structure-activity relationship study, and biological evaluation.

Farnesoid X receptor (FXR) is a bile acids receptor and plays a crucial role in regulating bile acids, lipids, and glucose metabolism. Previous research suggests that inhibiting FXR activation can be beneficial in reducing cholesterol and low-density lipoprotein cholesterol (LDL-C) levels, offering potential treatment options for metabolic syndrome with lipid disorders. Herein, we report p-acetylaminobenzene sulfonate derivatives as a novel scaffold of FXR antagonists by multistage screening. Among these derivatives, compound F44-A13 exhibited a half-maximal inhibitory concentration of 1.1 µM. Furthermore, compound F44-A13 demonstrated effective inhibition of FXR activation in cellular assays and exhibited high selectivity over eleven other nuclear receptors. Besides, compound F44-A13 significantly suppressed the regulation of FXR target genes Shp, Besp, and Cyp7a1, while reducing cholesterol levels in human hepatoma HepG2 cells. Pharmacological studies conducted on C57BL/6 mice further confirmed that compound F44-A13 had beneficial effects in reducing cholesterol, triglycerides, and LDL-C levels. These findings highlight that F44-A13 is a highly selective FXR antagonist that might serve as a useful molecule for further FXR studies as well as the development of FXR antagonists for the potential treatment of metabolic diseases with lipid disorders.

Dioxygen compatible electron donor-acceptor catalytic system and its enabled aerobic oxygenation.

The photochemical properties of Electron Donor-Acceptor (EDA) complexes present exciting opportunities for synthetic chemistry. However, these strategies often require an inert atmosphere to maintain high efficiency. Herein, we develop an EDA complex photocatalytic system through rational design, which overcomes the oxygen-sensitive limitation of traditional EDA photocatalytic systems and enables aerobic oxygenation reactions through dioxygen activation. The mild oxidation system transfers electrons from the donor to the effective catalytic acceptor upon visible light irradiation, which are subsequently captured by molecular oxygen to form the superoxide radical ion, as demonstrated by the specific fluorescent probe, dihydroethidine (DHE). Furthermore, this visible-light mediated oxidative EDA protocol is successfully applied in the aerobic oxygenation of boronic acids. We believe that this photochemical dioxygen activation strategy enabled by EDA complex not only provides a practical approach to aerobic oxygenation but also promotes the design and application of EDA photocatalysis under ambient conditions.

Discovery of 3-oxo-1,2,3,4-tetrahydropyrido[1,2-a]pyrazin derivatives as SARS-CoV-2 main protease inhibitors through virtual screening and biological evaluation.

The COVID-19 caused by SARS-CoV-2 has led to a global pandemic that continues to impact societies and economies worldwide. The main protease (Mpro) plays a crucial role in SARS-CoV-2 replication and is an attractive target for anti-SARS-CoV-2 drug discovery. Herein, we report a series of 3-oxo-1,2,3,4-tetrahydropyrido[1,2-a]pyrazin derivatives as non-peptidomimetic inhibitors targeting SARS-CoV-2 Mpro through structure-based virtual screening and biological evaluation. Further similarity search and structure-activity relationship study led to the identification of compound M56-S2 with the enzymatic IC50 value of 4.0 µM. Moreover, the molecular simulation and predicted ADMET properties, indicated that non-peptidomimetic inhibitor M56-S2 might serve as a useful starting point for the further discovery of highly potent inhibitors targeting SARS-CoV-2 Mpro.

Obesity induced disruption on diurnal rhythm of insulin sensitivity via gut microbiome-bile acid metabolism.

The disruption of the diurnal rhythm has been recognized as a significant contributing factor to metabolic dysregulation. The important role of gut microbiota and bile acid metabolism has attracted extensive attention. However, the function of the gut microbiota-bile acid axis in regulating the diurnal rhythms of metabolic homeostasis remains largely unknown. Herein, we aimed to investigate the interplay between rhythmicity of host metabolism and gut microbiota-bile acid axis, as well as to assess the impact of obesity on them. We found that high fat diet feeding and Leptin gene deficiency (ob/ob) significantly disturbed the rhythmic patterns of insulin sensitivity and serum total cholesterol levels. The bile acid profiling unveiled a conspicuous diurnal rhythm oscillation of ursodeoxycholic acid (UDCA) in lean mice, concomitant with fluctuations in insulin sensitivity, whereas it was absent in obese mice. The aforementioned diurnal rhythm oscillations were largely desynchronized by gut microbiota depletion, suggesting the indispensable role of gut microbiota in diurnal regulation of insulin sensitivity and bile acid metabolism. Consistently, 16S rRNA sequencing revealed that UDCA-associated bacteria exhibited diurnal rhythm oscillations that paralleled the fluctuation in insulin sensitivity. Collectively, the current study provides compelling evidence regarding the association between diurnal rhythm of insulin sensitivity and gut microbiota-bile acid axis. Moreover, we have elucidated the deleterious effects of obesity on gut microbiome-bile acid metabolism in both the genetic obesity model and the diet-induced obesity model.

Preparation of La2Zr2O7 composite oxides with fluorite/pyrochlore phases by excluding element influences for catalyzing oxidative coupling of methane.

To elucidate the effect of the A2B2O7 phase on the oxidative coupling of methane (OCM) while excluding elemental influences, La2Zr2O7 compounds with a disordered defect fluorite (La2Zr2O7-F) structure and an ordered pyrochlore phase (La2Zr2O7-P) have been synthesized. Irrespective of their element composition, the catalytic performance of La2Zr2O7-F exceeds that of La2Zr2O7-P. Furthermore, the La2Zr2O7-F surface has more oxygen vacancies/defects than the La2Zr2O7 surface because La2Zr2O7-F exhibits a higher lattice disorder degree and lower B-O bond strength, which leads to the formation of more reactive oxygen anions (O2- and O22-) and basic sites for OCM. Isotopic exchange results have testified that surface-active oxygen sites are generated due to the gaseous O2 adsorption/activation occurring on the surface vacancies via both simple and multiple hetero-exchange mechanisms. In conclusion, crystal structure is the primary factor that governs the catalytic performance of A2B2O7 compounds, with the disordered defect fluorite phase being the most optimal structure for OCM.

Functional screening and rational design of compounds targeting GPR132 to treat diabetes.

Chronic inflammation due to islet-residing macrophages plays key roles in the development of type 2 diabetes mellitus. By systematically profiling intra-islet lipid-transmembrane receptor signalling in islet-resident macrophages, we identified endogenous 9(S)-hydroxy-10,12-octadecadienoic acid-G-protein-coupled receptor 132 (GPR132)-Gi signalling as a significant contributor to islet macrophage reprogramming and found that GPR132 deficiency in macrophages reversed metabolic disorders in mice fed a high-fat diet. The cryo-electron microscopy structures of GPR132 bound with two endogenous agonists, N-palmitoylglycine and 9(S)-hydroxy-10,12-octadecadienoic acid, enabled us to rationally design both GPR132 agonists and antagonists with high potency and selectivity through stepwise translational approaches. We ultimately identified a selective GPR132 antagonist, NOX-6-18, that modulates macrophage reprogramming within pancreatic islets, decreases weight gain and enhances glucose metabolism in mice fed a high-fat diet. Our study not only illustrates that intra-islet lipid signalling contributes to islet macrophage reprogramming but also provides a broadly applicable strategy for the identification of important G-protein-coupled receptor targets in pathophysiological processes, followed by the rational design of therapeutic leads for refractory diseases such as diabetes.

Single-cell detection of DMSO promoted HL-60 differentiation toward granulocyte based on DC-iDEP for medicine screening.

The most common form of leukemia in adults is acute leukemia. Drug differentiation control is an extremely critical treatment for acute leukemia. Unfortunately, current techniques detecting differentiation control experience long time and complex steps of verification hindering the pace of medicine discovery: flow cytometry and RT-PCR are highly accurate and efficient at a cost of inconvenient fluorescent labeling or a high risk of contamination; conventional staining leads to cell death unavailable for further pharmacological tests. There is a great interest in developing simple, fast, and non-invasive techniques to screen medicine. DC-iDEP is an emerging label-free identification technique taking advantage of the whole cell native biophysical property for sorting cell populations. Here, HL-60 cell line has been used as a model to study the differentiation process toward granulocytes and medicine efficacy. The results showed that DEP succeeded in detecting the DMSO promoted HL-60 differentiation degree by the weighted average characterization factor. This factor is related to the single cell biophysical property, which accumulates to generate differences in each population with distinct constitutions. Furthermore, cichoric acid was investigated to be capable of promoting DMSO-induced differentiation efficiently. Using the change induced by cichoric acid, the HL-60 medicine screening application has been first attempted based on DEP. A rapid, label-free medicine screening method has been established to monitor HL-60 differentiation toward granulocyte and has great potential for medicine screening.

Role of m6A modification in immune microenvironment of digestive system tumors.

Digestive system tumors are huge health problem worldwide, largely attributable to poor dietary choices. The role of RNA modifications in cancer development is an emerging field of research. RNA modifications are associated with the growth and development of various immune cells, which, in turn, regulate the immune response. The majority of RNA modifications are methylation modifications, and the most common type is the N6-methyladenosine (m6A) modification. Here, we reviewed the molecular mechanism of m6A in the immune cells and the role of m6A in the digestive system tumors. However, further studies are required to better understand the role of RNA methylation in human cancers for designing diagnostic and treatment strategies and predicting the prognosis of patients.

Enzymatic Catalysis in Size and Volume Dual-Confined Space of Integrated Nanochannel-Electrodes Chip for Enhanced Impedance Detection of Salmonella.

Nanochannel-based confinement effect is a fascinating signal transduction strategy for high-performance sensing, but only size confinement is focused on while other confinement effects are unexplored. Here, a highly integrated nanochannel-electrodes chip (INEC) is created and a size/volume-dual-confinement enzyme catalysis model for rapid and sensitive bacteria detection is developed. The INEC, by directly sandwiching a nanochannel chip (60 µm in thickness) in nanoporous gold layers, creates a micro-droplet-based confinement electrochemical cell (CEC). The size confinement of nanochannel promotes the urease catalysis efficiency to generate more ions, while the volume confinement of CEC significantly enriches ions by restricting diffusion. As a result, the INEC-based dual-confinement effects benefit a synergetic enhancement of the catalytic signal. A 11-times ion-strength-based impedance response is obtained within just 1 min when compared to the relevant open system. Combining this novel nanoconfinement effects with nanofiltration of INEC, a separation/signal amplification-integrated sensing strategy is further developed for Salmonella typhimurium detection. The biosensor realizes facile, rapid (<20 min), and specific signal readout with a detection limit of 9 CFU mL-1 in culturing solution, superior to most reports. This work may create a new paradigm for studying nanoconfined processes and contribute a new signal transduction technique for trace analysis application.