DNA damage refers to chemical alterations in genomic DNA arising from endogenous or exogenous factors, and delineating the types and levels of these lesions is essential for elucidating the molecular mechanisms and regulatory networks underlying DNA repair. However, current methods are limited to one specific type of damage and often rely on DNA extraction and in vitro processing, which fails to capture the spatial distribution of diverse DNA damages within cellular context. Here we present intracellular strand break end extension-encoded amplification (ISBEA), an in situ imaging strategy that enables simultaneous visualization of multiple DNA lesion types. ISBEA uses a sequential enzymatic workflow in which glycosylases, endonucleases and exonucleases selectively recognize oxidized purines, AP sites and strand breaks, converting them stepwise into uniform 3'-hydroxyl termini. These activated termini undergo programmable extension and serve as primers to initiate DNA-encoded amplification, ultimately generating fluorescence signals present to each damage type. With spatially resolved visualization of diverse DNA damages within cells, ISBEA provides a robust platform for investigating the origins, repair pathways and biological consequences of DNA damage, offering valuable methodological support for advancing the understanding of genome stability maintenance and the development of related diseases.

DNA molecules, with highly variable sequences and inherent programmability, emerge as a promising material for next-generation information storage and data encryption. However, due to the singular encryption method or limited randomness of the secret key, current encryptions remain vulnerable to brute-force attacks and the need for enhanced information security persists. This study introduces a programmable encryption strategy based on long-chain DNA synthesis and sequential encoding. The proposed hairpin-mediated primer exchange reaction (HAMER) system enables the generation of DNA keys and the recording of encoded information. Ultimately, encrypted text and image data can be decoded and retrieved through sequencing with customized access based on user permissions. This approach positions DNA as a high-performance information material and establishes a programmable encryption framework, offering strong potential to meet the confidentiality, integrity, and availability demands of future information security systems.

Cellular structure maintenance and function regulation critically depend on the composition and spatial distribution of numerous membrane proteins. However, current methods face limitations in spatial coverage and data scalability, hindering the comprehensive analysis of protein interactions in complex cellular nanoenvironment. Herein, we introduce proximity-activated DNA scanning encoded sequencing (PADSE-seq), an innovative technique that utilizes flexible DNA probes with adjustable lengths. These dynamic probes are anchored at a single end, enabling free swings within a nanoscale range to perform global scanning, recording, and accumulating of information on diverse proximal proteins in random directions along unrestricted paths. PADSE-seq leverages the autonomous cyclic cleavage of single-stranded DNA to sequentially activate encoded probes distributed throughout the local area. This process triggers strand displacement amplification and bidirectional extension reactions, linking proteins barcodes with molecular barcodes in tandem and further generating millions to billions of amplicons embedded with the combinatorial identifiers for next-generation sequencing analysis. As a proof of concept, we validated PADSE-seq for mapping the distribution of over a dozen kinds of proteins, including HER1, EpCAM, and PDL1, in proximity to HER2 in breast cancer cell lines, demonstrating its ability to decode multiplexed protein proximities at the nanoscale. Notably, we observed that the spatial distribution of proximal proteins around low-abundance target proteins exhibited greater diversity across regions with variable proximity ranges. This method offers a massive access for high-resolution and comprehensive mapping of cellular molecular interactions, paving the way for deeper insights into complex biological processes and advancing the field of precision medicine.

Abstract Objective The present study aims to investigate the protective role of metformin against atrial fibrillation (AF) and stroke in type 2 diabetes (T2DM) patients treated with other antidiabetic medications. Methods We included 10,011 participants with T2DM from the UK biobank database, with a follow-up for new diagnoses of AF and stroke recorded until 2023. We employed logistic regression analysis and competing risk models to assess the association between metformin use and the incidence of AF or stroke. Results The cross-sectional analysis revealed that, compared with the use of other antidiabetics, metformin was associated with a lower prevalence of AF (odds ratio [OR] 0.71, 95% CI: 0.57–0.89) and stroke (OR 0.70, 95% CI: 0.56–0.86). In the competing risk model, compared to other antidiabetics, metformin may reduce the risk of AF (sub-hazard ratio [SHR] 0.86, 95% CI: 0.75–0.98) and stroke (SHR 0.62, 95% CI: 0.44–0.86) (Figure 1). Subgroup analysis indicated a significantly lower incidence of AF or stroke in patients under 65 years of age (SHRs were 0.80, 95% CI:0.67-0.96 for AF and 0.52, 95% CI: 0.34-0.81 for stroke) and in female patients (SHRs were 0.65, 95% CI:0.52-0.81 for AF and 0.53, 95% CI: 0.30-0.91 for stroke). Notably, metformin significantly reduced the incidence of AF in non-insulin users (SHR 0.80, 95% CI: 0.66-0.97) and the incidence of stroke in insulin users (SHR 0.55, 95% CI: 0.35-0.87) (Figure 2). Conclusion Our findings suggest that metformin use, in comparison to other antidiabetic medications, is associated with a reduced risk of AF and stroke, especially in non-insulin users for AF and insulin users for stroke.

With the acceleration of the industrial process and the improvement of people's living standards, a large amount of oily wastewater has been produced, which has threatened the ecological system and human health. Therefore, the treatment of oily wastewater has become a problem and challenge to be solved. Benefit from its excellent three-dimensional porous structure, aerogels are widely used in the sphere of oil-water separation. This paper mainly introduces superwetting aerogel-based oil-water separation materials. Elaborate on its separation mechanism and acquisition pathway. The performance and advantages of polymer-based aerogels, biomass-based aerogels, inorganic-based aerogels and carbon-based aerogels in oil-water separation are analyzed and summarized. Finally, the application prospect of aerogel-based materials in the field of oil-water separation was prospected.

Dual -frequency ultrasound has been widely employed to enhance and regulate acoustic droplet vaporization (ADV) but the role of ultrasonic nonlinear effects on it remains unclear. The main objective of this study is to investigate the influence of nonlinear effects on the control of ADV nucleation under different dual -frequency focused ultrasound conditions. ADV nucleation of PFC nanodroplets activated by nonlinear dual -frequency ultrasound was modeled and parametric studies were conducted to investigate the influence of dual -frequency ultrasound frequency and acoustic power on the degree of nonlinearity (DoN), nucleation rates and dimensions of the nucleation region in a wide parameter range. The results showed that the ultrasonic nonlinearity caused a significant decrease in peak negative pressure due to waveform distortion, which leads to a lower nucleation rate in the nonlinear model compared to that in the linear model. Furthermore, the distributions of nucleation regions were also affected by the interaction between waves of different frequencies and cloud -like spatial distributions were produced, which could be modulated by the dual -frequency ultrasound parameters and have great potentials in the spatial regulation of the ADV and customized treatment protocols in clinical applications. In addition, represented by 1.5 MHz + 3 MHz, such a dual -frequency combination of fundamental and second harmonic could effectively enhance ultrasonic nonlinear effects with relatively lower peak negative pressure and higher DoN. Therefore, nonlinear effect of the dual -frequency ultrasound plays an important role in the ADV regulation, which should be considered in the numerical model and practical applications.

Smart fabrics provide considerable inspiration for the construction of wearable electronic devices due to their excellent skin affinity and breathability. However, the limited tolerance to high humidity environments and the flammable nature of fabrics impedes broader practical applications. In this paper, by an advanced and effective method, phytic acid doped (PA-) polyaniline (PANI) was polymerized in-situ on cotton fabric deposited by carbon black (CB), and then modified by non-fluorine hydrophobic particles, a flexible breathable fabric-based strain sensor (HPCF) with ultrahigh moisture resistance, flame retardancy and environmental resistance was successfully prepared. The obtained sensor shows high sensitivity (16.14 kPa(-1)) in the detection range of 0-2 kPa, fast response/recovery time (82/40 ms) and high cycle stability (>10,000 cycles). Thanks to its excellent hydrophobicity (WCA = 153 degrees) and flame retardancy properties, the material enables the detection of human movement (such as finger/wrist/elbow/knee/foot movement) under a variety of harsh conditions (extremely high temperatures, underwater, acid, alkali, salt and blown sand environments). More importantly, it can be further applied to remote detection of potentially dangerous scenarios. HPCF can successfully transmit the Morse code signals of "SOS" and "HELP" sent by drowning people, and realize water rescue. In case of fire, it serves as a fire warning sensor to realize agile (1 s) fire alarming. The highly sensitive and versatile HPCF fabric sensor enables physical condition monitoring for fire and rescue personnel working in high temperature and humidity environments, which is of great value for all human activities carried out in harsh environments.

Low power density and poor wear resistance seriously hinder the application of a triboelectric nanogenerator (TENG) in daily environments. However, current research has mainly focused on improving the performance of TENGs through inorganic fillers while neglecting the important property of wear resistance. Due to the excellent properties of stearic acid (SA), such as its cheap and easy availability, high electronegativity, wear resistance, hydrophobicity, and lubricity, as well as promoting the formation of electroactive beta phase polyvinylidene fluoride-hexafluoropropylene (PVDF-HFP), SA can be used as an organic multifunctional filler to improve the electrical output performance and wear resistance of TENG while increasing its hydrophobicity. In this work, we doped SA into polytetrafluoroethylene (PTFE)/PVDF-HFP matrix by solution casting method and template method to form SA-doped PTFE/PVDF-HFP(SA-PTFE/PVDF-HFP) composite polymer film, which were assembled with silk to form SA-TENG. Most importantly, the addition of 3 wt % SA not only increases the voltage and current of TENG by 3.7 times and 3.6 times, respectively, but also reduces the coefficient of friction and the amount of wear of TENG by 40.8 and 87.5%, respectively, and increases the hydrophobic angle by 12 to 133(degrees), which ensures the long-term and stable operation of TENG in different environments. In addition, SA-TENG can not only harvest energy from the surrounding environment and charge portable electronic devices but can also be used as a self-powered sensor to monitor human movement. This study not only provides a feasible strategy for organic fillers to solve the bottleneck problem of TENG application but also broadens the application scenarios of TENG.

BACKGROUND:Hepatocellular carcinoma (HCC), the most common type of primary liver cancer, is the fourth leading cause of cancer-related deaths worldwide. Previous evidence shows that the expression of circulating RNA ZFR (circZFR) is upregulated in HCC tissues. However, the molecular mechanism of circZFR in HCC is unclear.METHODS:Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was employed to detect the expression of circZFR, microRNA-624-3p (miR-624-3p) and WEE1 in HCC tissues and cells. RNase R assay and actinomycin D treatment assay were used to analyze the characteristics of circZFR. For functional analysis, the capacities of colony formation, cell proliferation, cell apoptosis, migration and invasion were assessed by colony formation assay, 5-ethynyl-2'-deoxyuridine (EdU) assay, flow cytometry assay and transwell assay. Western blot was used to examine the protein levels of WEE1 and epithelial-mesenchymal transition (EMT)-related proteins. The interactions between miR-624-3p and circZFR or WEE1 were validated by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Xenograft models were established to determine the role of circZFR in vivo.RESULTS:circZFR and WEE1 were upregulated, while miR-624-3p expression was reduced in HCC tissues and cells. circZFR could sponge miR-624-3p, and WEE1 was a downstream gene of miR-624-3p. Knockdown of circZFR significantly reduced the malignant behaviors of HCC and that co-transfection with miR-624-3p inhibitor restored this change. Overexpression of WEE1 abolished the inhibitory effect of miR-624-3p mimic on HCC cells. Mechanistically, circZFR acted as a competitive endogenous RNA (ceRNA) to regulate WEE1 expression by targeting miR-624-3p. Furthermore, in vivo studies have illustrated that circZFR knockdown inhibited tumor growth.CONCLUSIONS:circZFR knockdown reduced HCC cell proliferation, migration and invasion and promoted apoptosis by regulating the miR-624-3p/WEE1 axis, suggesting that the circZFR/miR-624-3p/WEE1 axis might be a potential target for HCC treatment.

As energy scarcity and environmental pollution become increasingly problematic, developing non-polluting and sustainable hydrogen production technologies is crucial, with an emphasis on developing high-efficiency and easily available electrolytic hydrocatalysts. Herein, we present a novel, economical self-supporting electrode, W-MoS2/FeNi2S4/NF, fabricated by a simple hydrothermal method. Benefiting from the insitu growth of uniformly dispersed micro-and nanoparticles with an ultrathin nanosheet-like shape on the surface and heterogeneous engineering and heteroatom doping, the catalytic electrodes possess surface superhydrophilic/underwater superaerophobic properties and superb intrinsic catalytic activity. The superhydrophilic/underwater superaerophobic properties of the catalytic electrodes allow for rapid infiltration of the electrolyte solution into the electrode and accelerate mass transfer while also allowing for the desorption of bubbles from the electrode surface and avoiding the bubble shielding effect, resulting in a significantly increased electrocatalytic rate. With current densities as high as 10 mA cm(-2) for hydrogen evolution reaction (HER) and oxygen evolution reaction (OER), respectively, W-MoS2/FeNi2S4/NF displayed lower overpotentials of 92 and 177 mV. More impressively, only a low cell voltage of merely 1.5 V is sufficient to achieve a current density of 10 mA cm(-2), achieve overall water splitting in an alkaline electrolyte, and exhibit up to 20 h electrochemical durability. Above all, it has been demonstrated that surplus electricity from intermittent energy sources may be used to create eco-friendly hydrogen energy by electrolyzing water, preventing resource waste. This work provides novel insights into the preparation of inexpensive, high-efficiency bifunctional electrocatalysts and new directions for intermittent energy generation for hydrogen production.

Triboelectric nanogenerator (TENG) has been demonstrated as a sustainable energy utilization method for waste mechanical energy and self-powered system. However, the charge dissipation of frictional layer materials in a humid environment severely limits their stable energy supply. In this work, a new method is reported for preparing polymer film as a hydrophobic negative friction material by solution blending poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) and polyvinyl chloride (PVC), doping with titanium dioxide (TiO2) nanoparticles, and further surface patterning modification. The P-TENG composed of the PVDF-HFP/PVC/TiO2 composite film with optimized hydrophobic performance (WCA = 124°) achieved an output voltage of 235 V and a short-circuit current of 35 μA, which is approximately three times that of the bare PVDF-HFP-based TENG. Under charge excitation, the transferred charge of the P-TENG can reach 35 nC. When the external load resistance is 5.5 MΩ, the output peak power density can reach 1.4 W m-2. Meanwhile, the hydrophobic surface layer with a rough surface structure enables the device to overcome the influence of water molecules on charge transfer in a humid environment, quickly recover, and maintain a high output. The P-TENG can effectively monitor finger flexibility and strength and realize real-time evaluation of the exercise state and hand fatigue of the elderly and rehabilitation trainers. It has broad application prospects in self-powered intelligent motion sensing, soft robotics, human-machine interaction, and other fields.

With the rapid development of triboelectric nanogenerators (TENGs), the exploration of self-powered, flexible, and wearable electronic devices has attracted widespread attention. However, the choice of tribomaterials and high humidity environment have a significant impact on the triboelectricity of TENG. Therefore, we prepared a composite fabric (HPC) with superhydrophobic and conductive properties, which was used simultaneously as a tribopositive material and electrode for the construction of promising wearable TENGs. Specifically, the loading of polydopamine, carbon nanotubes, and polypyrrole on the surface of the cotton fabric makes it have not only conductivity but also enhanced tribopositive polarity. Then, cetyltrimethoxysilane was selected to modify it to obtain superhydrophobicity. Compared with the common TENGs with a separate tribolayer and electrode, the integrated HPC-TENG shows the advantages of simpler structure and lighter wear. Moreover, compared with the unmodified fabric-based TENG, the performance of the proposed HPC-TENG is improved by nearly 7.2 times, and the maximum power density can reach 2.6 W m-2. This remarkable output can be attributed to the combination of strong electron-giving groups, high electrical conductivity, and abundant micro- and nanorough structure of the HPC fabric. More importantly, due to the water repellency of the fabric surface, the high output performance can be maintained under high humidity conditions. In addition, HPC-TENG has potential applications as pressure sensors for human motion status monitoring and multichannel sensing for smart game blanket entertainment. The newly designed HPC-TENG offers a new strategy for the development of superhydrophobic fabrics with an electrical conductivity, energy harvesting, and self-powered sensor.

The urgent need to address energy consumption and environmental issues has triggered a strong interest in triboelectric nanogenerators (TENGs), and thus, the exploration of novel materials with environmentally friendly characteristics and low cost for the preparation of high-output TENGs has shown great attraction. Herein, we proposed a method to increase the electron-donating ability of sodium carboxymethyl-cellulose (CMC) by adding polyethyleneimine (PEI) to CMC through simple physical mixing. By systematically adjusting and optimizing the CMC concentration and its content with PEI, it not only greatly improves the tribopositive performance of CMC but also makes the prepared film have excellent flexibility. The obtained CMC/PEI-TENG open-circuit voltage and short-circuit current can reach 385 V and 51 mu A, respectively, with a maximum output power density of 1.056 mW/cm2, which can easily power 320 commercial LEDs. Moreover, the triboelectric output is improved by approximately 1.7 times compared to that of pure phase CMC film-based TENG. In addition, the new TENG shows great potential as a self-powered power source and sensor. The novel CMC-based composite film TENG proposed in this study provides a new idea for the development of environmentally friendly TENGs.

AbstractBackgroundCognitive impairment is one of the serious complications in diabetic patients. Iron accumulation is thought to be related to degenerative diseases and diabetes. Microglia have been shown to have features of iron accumulation in Alzheimer 's disease (AD). Triggering receptor expressed on myeloid cells-1(TREM-1)acts as an activating receptor for myeloid cells and is elevated in several neurodegenerative diseases. It is unclear whether microglial iron overload is involved in diabetic associated cognitive impairment (DACI), and whether there is an association between TREM-1 and iron metabolism.MethodsWe established a model cultured in high glucose (HG) for 72 hours in BV2/HMC3 and another one high fat diet (HFD) combined with STZ in C57 mice. Intracellular mitochondrial changes were observed by Transmission Electron Microscopy, intracellular Fe2 +, GSH/GSSG, MDA and ROS were detected using kits to assess lipid peroxidation in microglia. Prussian blue staining was used to observe iron deposition in the hippocampus. Cognitive performance of mice was assessed by Y-maze and novel object experiments (NOR). Western blot and immunofluorescence were used to observe the expression of iron and antioxidant-related protein (FPN1, FTL, TFR1,GPX4), apoptosis-related protein (BCL-2, BAX and Caspase3), TREM-1, endoplasmic reticulum stress (ERS)-related protein (BIP, P-IRE, IRE, ATF6, P-PERK, PERK, P-eIF2α, eIF2α, ATF4, CHOP).ResultsWe found that hippocampus and microglia undergo iron overload and antioxidant inactivation in HFD/STZ mice. Iron overload, with increase of TREM-1 and antioxidant inactivation, occurred in BV2/HMC3 cultured by HG. Fer-1 could alleviate it by inhibiting HG-induced change of FPN1, GPX4, TFR1, as well as Fe2 +, GSH/GSSG ratio, MDA and ROS. Inhibiting TREM-1 by LP17 could block it, too. Furthermore, PERK/ eIF2α/ATF4/CHOP was obviously down-regulated by LP17 in HG cultured BV2. When LP17 was administered intraperitoneally, cognitive impairment was improved in HFD/STZ mice, iron deposition and antioxidant inactivation in microglia of hippocampus were relieved.ConclusionsIn this study, we found that iron deposition and antioxidant inactivation occurred in microglia of hippocampus in HFD/STZ mice, accompanied by increase of TREM-1, while inhibition of TREM-1 expression alleviated it and improved cognitive dysfunction, which was partially regulated by PERK/ eIF2α/ATF4/CHOP.

Pancreatic ductal adenocarcinoma (PDAC) is one of the lethal malignancies, with limited biomarkers identified to predict its prognosis and treatment response of immune checkpoint blockade (ICB). This study aimed to explore the predictive ability of T cell marker genes score (TMGS) to predict their overall survival (OS) and treatment response to ICB by integrating single-cell RNA sequencing (scRNA- seq ) and bulk RNA- seq data. Multi-omics data of PDAC were applied in this study. The uniform manifold approximation and projection (UMAP) was utilized for dimensionality reduction and cluster identification. The non-negative matrix factorization (NMF) algorithm was applied to molecular subtypes clustering. The Least Absolute Shrinkage and Selection Operator (LASSO)-Cox regression was adopted for TMGS construction. The prognosis, biological characteristics, mutation profile, and immune function status between different groups were compared. Two molecular subtypes were identified via NMF: proliferative PDAC (C1) and immune PDAC (C2). Distinct prognoses and biological characteristics were observed between them. TMGS was developed based on 10 T cell marker genes (TMGs) through LASSO-Cox regression. TMGS is an independent prognostic factor of OS in PDAC. Enrichment analysis indicated that cell cycle and cell proliferation-related pathways are significantly enriched in the high-TMGS group. Besides, high-TMGS is related to more frequent KRAS , TP53 , and CDKN2A germline mutations than the low-TMGS group. Furthermore, high-TMGS is significantly associated with attenuated antitumor immunity and reduced immune cell infiltration compared to the low-TMGS group. However, high TMGS is correlated to higher tumor mutation burden (TMB), a low expression level of inhibitory immune checkpoint molecules, and a low immune dysfunction score, thus having a higher ICB response rate. On the contrary, low TMGS is related to a favorable response rate to chemotherapeutic agents and targeted therapy. By combining scRNA- seq and bulk RNA- seq data, we identified a novel biomarker, TMGS, which has remarkable performance in predicting the prognosis and guiding the treatment pattern for patients with PDAC.

Diabetes-associated cognitive dysfunction (DACD) is considered a significant complication of diabetes and manifests as cognitive impairment. Astrocytes are vital to the brain energy metabolism and cerebral antioxidant status. Ferroptosis has been implicated in cognitive impairment, but it is unclear whether the ferroptosis of astrocytes is involved in the progression of DACD. PPARA/PPARα (peroxisome proliferator-activated receptor alpha) is a transcription factor that regulates glucose and lipid metabolism in the brain. In this study, we demonstrated that high glucose promoted ferroptosis of astrocytes by disrupting iron metabolism and suppressing the xCT/GPX4-regulated pathway in diabetic mice and astrocytes cultured in high glucose. Administration of gemfibrozil, a known PPARα agonist, inhibited ferroptosis and improved memory impairment in db/db mice. Gemfibrozil also prevented the accumulation of lipid peroxidation products and lethal reactive oxygen species induced by iron deposition in astrocytes and substantially reduced neuronal and synaptic loss. Our findings demonstrated that ferroptosis of astrocytes is a novel mechanism in the development of DACD. Additionally, our study revealed the therapeutic effect of gemfibrozil in preventing and treating DACD by inhibiting ferroptosis.

Neuroinflammation caused by microglial activation and consequent neurological impairment are prominent features of diabetes-associated cognitive impairment (DACI). Microglial lipophagy, a significant fraction of autophagy contributing to lipid homeostasis and inflammation, had mostly been ignored in DACI. Microglial lipid droplets (LDs) accumulation is a characteristic of aging, however, little is known about the pathological role of microglial lipophagy and LDs in DACI. Therefore, we hypothesized that microglial lipophagy could be an Achilles's heel exploitable to develop effective strategies for DACI therapy. Here, starting with characterization of microglial accumulation of LDs in leptin receptor-deficient (db/db) mice and in high-fat diet and STZ (HFD/STZ) induced T2DM mice, as well as in high-glucose (HG)-treated mice BV2, human HMC3 and primary mice microglia, we revealed that HG-dampened lipophagy was responsible for LDs accumulation in microglia. Mechanistically, accumulated LDs colocalized with the microglial specific inflammatory amplifier TREM1 (triggering receptor expressed on myeloid cells 1), resulting in the buildup of microglial TREM1, which in turn aggravates HG-induced lipophagy damage and subsequently promoted HG-induced neuroinflammatory cascades via NLRP3 (NLR family pyrin domain containing 3) inflammasome. Moreover, pharmacological blockade of TREM1 with LP17 in db/db mice and HFD/STZ mice inhibited accumulation of LDs and TREM1, reduced hippocampal neuronal inflammatory damage, and consequently improved cognitive functions. Taken together, these findings uncover a previously unappreciated mechanism of impaired lipophagy-induced TREM1 accumulation in microglia and neuroinflammation in DACI, suggesting its translational potential as an attractive therapeutic target for delaying diabetes-associated cognitive decline.Abbreviations: ACTB: beta actin; AIF1/IBA1: allograft inflammatory factor 1; ALB: albumin; ARG1: arginase 1; ATG3: autophagy related 3; Baf: bafilomycin A1; BECN1: beclin 1, autophagy related; BW: body weight; CNS: central nervous system; Co-IP: co-immunoprecipitation; DACI: diabetes-associated cognitive impairment; DAPI: 4',6-diamidino-2-phenylindole; DGs: dentate gyrus; DLG4/PSD95: discs large MAGUK scaffold protein 4; DMEM: Dulbecco's modified Eagle's medium; DSST: digit symbol substitution test; EDTA: ethylenedinitrilotetraacetic acid; ELISA: enzyme linked immunosorbent assay; GFAP: glial fibrillary acidic protein; HFD: high-fat diet; HG: high glucose; IFNG/IFN-γ: interferon gamma; IL1B/IL-1β: interleukin 1 beta; IL4: interleukin 4; IL6: interleukin 6; IL10: interleukin 10; LDs: lipid droplets; LPS: lipopolysaccharide; MAP2: microtubule associated protein 2; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; MWM: morris water maze; NFKB/NF-κB: nuclear factor of kappa light polypeptide gene enhancer in B cells; NLRP3: NLR family pyrin domain containing 3; NOS2/iNOS: nitric oxide synthase 2, inducible; NOR: novel object recognition; OA: oleic acid; PA: palmitic acid; PBS: phosphate-buffered saline; PFA: paraformaldehyde; PLIN2: perilipin 2; PLIN3: perilipin 3; PS: penicillin-streptomycin solution; RAPA: rapamycin; RBFOX3/NeuN: RNA binding protein, fox-1 homolog (C. elegans) 3; RELA/p65: RELA proto-oncogene, NF-kB subunit; ROS: reactive oxygen species; RT: room temperature; RT-qPCR: Reverse transcription quantitative real-time polymerase chain reaction; STZ: streptozotocin; SQSTM1/p62: sequestosome 1; SYK: spleen asociated tyrosine kinase; SYP: synaptophysin; T2DM: type 2 diabetes mellitus; TNF/TNF-α: tumor necrosis factor; TREM1: triggering receptor expressed on myeloid cells 1; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling.

Single-cell RNA sequencing reveals the gene structure and gene expression status of a single cell, which can reflect the heterogeneity between cells. However, batch effects caused by non-biological factors may hinder data integration and downstream analysis. Although the batch effect can be evaluated by visualizing the data, which actually is subjective and inaccurate. In this work, we propose a quantitative method cKBET, which considers the batch and cell type information simultaneously. The cKBET method accesses batch effects by comparing the global and local fraction of cells of different batches in different cell types. We verify the performance of our cKBET method on simulated and real biological data sets. The experimental results show that our cKBET method is superior to existing methods in most cases. In general, our cKBET method can detect batch effect with either balanced or unbalanced cell types, and thus evaluate batch correction methods.

Droplet microfluidics has emerged as a powerful technology to perform high-throughput experiments, while artificial intelligence (AI) serves as a functional tool to analyze a large set of multiplex data. Their convergence creates new opportunities in autonomous system optimization and control, enabling various innovative functions and applications. In this study, we elucidate the basic principles of AI and elaborate on its main functions. The intelligent microfluidic systems applied in droplet generation, material synthesis, and biological analysis are summarized, with their working mechanisms and enabled new functions highlighted. Moreover, we elucidate current challenges in a more widespread combination of AI and droplet microfluidics and offer our perspectives on potential strategies to tackle these challenges. We hope that this review can deepen our understanding of intelligent droplet microfluidics and inspire more functional designs tailored to emerging demands.

Chimeric antigen receptor-modified T cells (CAR T-cells) have shown exhilarative clinical efficacy for hematological malignancies. However, a shared antigen pool between healthy and malignant T-cells remains a concept to be technically and clinically explored for CAR T-cell therapy in T-cell cancers. No guidelines for engineering CAR T-cells targeting self-expressed antigens are currently available. Based on anti-CD70 CAR (CAR-70) T-cells, we constructed CD70 knock-out and wild-type CAR (CAR-70KO and CAR-70WT) T-cells and evaluated their manufacturing and anti-tumor capability. Single-cell RNA sequencing and TCR sequencing were performed to further reveal the underlying differences between the two groups of CAR T-cells. Our data showed that the disruption of target genes in T-cells before CAR transduction advantaged the expansion and cell viability of CAR T-cells during manufacturing periods, as well as the degranulation, anti-tumor efficacy, and proliferation potency in response to tumor cells. Meanwhile, more naïve and central memory phenotype CAR+ T-cells, with higher TCR clonal diversity, remained in the final products in KO samples. Gene expression profiles revealed a higher activation and exhaustion level of CAR-70WT T-cells, while signaling transduction pathway analysis identified a higher level of the phosphorylation-related pathway in CAR-70KO T-cells. This study evidenced that CD70 stimulation during manufacturing process induced early exhaustion of CAR-70 T-cells. Knocking-out CD70 in T-cells prevented the exhaustion and led to a better-quality CAR-70 T-cell product. Our research will contribute to good engineering CAR T-cells targeting self-expressed antigens.