INTRODUCTION:The comorbidity rate of depression and anxiety disorders is as high as 50%, with patients suffering from both conditions facing an elevated risk of suicide. Given the challenges in clinical diagnosis, there is an urgent need for objective diagnostic biomarkers to accurately identify patients with comorbid depression and anxiety disorders. METHODS:The case-control study enrolled 77 participants, including healthy controls (n = 38) and patients with comorbid depression and anxiety disorders (n = 39). Serum levels of short peptides were assessed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Binary logistic regression and receiver operating characteristic analysis (ROC) were employed to identify potential biomarkers, while decision curve analysis (DCA) was performed to evaluate the clinical utility of the candidate cyclic pentapeptide. RESULTS:A total of 25 differentially expressed short peptides were identified, among which 19 were significantly downregulated, and 6 were upregulated. Logistic regression analysis revealed that reduced levels of cyclo (D-trp-D-asp-pro-D-val-leu) were associated with an increased risk of comorbid depression and anxiety disorders. The area under the ROC curve (AUC) for this cyclic pen-tapeptide in diagnosing comorbid depression and anxiety disorders was 0.941 (95% CI, 0.884-0.997; P < 0.0001). DCA demonstrated that the cyclic pentapeptide provides meaningful net benefits when the threshold probability exceeded 0.11. Notably, this cyclic pentapeptide is an endothelin A receptor antagonist, which is implicated in vasoconstriction, hypertension, and anxiety, and elevated blood pressure was indeed observed in the patient cohort. DISCUSSION:The identified cyclic pentapeptide serves as a potential diagnostic tool, linking the pathophysiology of comorbid depression and anxiety disorders to the endothelin system. This association offers a novel integrative perspective on the co-occurring psychiatric and cardiovascular manifestations observed in these patients. CONCLUSION:In conclusion, this study identifies a specific cyclic pentapeptide as a promising serum biomarker that may facilitate the objective diagnosis of patients with comorbid depression and anxiety disorders. Its biological role as an endothelin A receptor antagonist not only provides a potential tool for clinical differentiation but also suggests a novel pathophysiological link to cardiovascular comorbidities, thereby offering new directions for elucidating the underlying mechanisms of these conditions.

Background: Sepsis-associated liver injury (SALI) is a severe and life-threatening complication, yet the mechanisms underlying gut–liver crosstalk during sepsis remain poorly defined. Clinical observations suggest that gut dysbiosis may contribute to organ dysfunction, but causal pathways linking specific microbial taxa to liver injury are unclear. Methods: We assessed the relationship between gut microbial composition and liver dysfunction in patients with sepsis. In mice, we performed mono-colonization with Prevotella copri and used cecal ligation and puncture to induce sepsis. Hepatic macrophage origin was tracked using Kaede photoconversion lineage tracing. CCR9-dependent immune trafficking was evaluated through genetic and pharmacologic blockade. Findings: In patients with sepsis, increased Prevotella abundance correlated with liver dysfunction and disease severity. In mice, P. copri mono-colonization exacerbated septic liver injury and increased hepatic macrophage accumulation. Lineage tracing demonstrated that these macrophages were predominantly recruited from the gut. CCR9-dependent trafficking was essential for this process, as CCR9 blockade reduced macrophage recruitment, inflammation, and liver damage. Gut-to-liver macrophage migration occurred only when sepsis-induced intestinal barrier injury was present. Interpretation: These findings identify a previously unrecognized gut–liver immune axis in which P. copri promotes CCR9-dependent recruitment of gut-derived macrophages to aggravate SALI. Targeting CCR9-mediated immune trafficking may represent a potential therapeutic strategy for preventing or mitigating liver injury in sepsis..

Endometriosis (EM) is an intractable gynecological condition with uncertain etiology. Gut microbiota have been increasingly implicated in EM, yet the underlying mechanisms remain unclear. Here, we identify a gut microbe-metabolite-uterus axis that drives EM pathogenesis. In our EM cohort, we found that the gut bacterium Olsenella profusa (O. profusa) and metabolite lysophosphatidylcholine (LPC) (16:0) were significantly enriched in patients with EM and showed positive correlation. In a mouse model of EM, oral gavage with O. profusa accelerated ectopic lesion growth through producing LPC (16:0) via its phospholipase, an effect recapitulated by LPC (16:0) alone. Mechanistically, LPC (16:0) promotes EM by inducing endometrial stromal cell proliferation and migration through the secreted phosphoprotein 1 (SPP1)-PI3K-AKT pathway. Moreover, O. profusa and LPC (16:0) show potential as early diagnostic biomarkers. This study suggests a causal role for the gut O. profusa-LPC (16:0) metabolic axis in EM progression and identifies promising targets for diagnosis and therapy.

Early antibiotic exposure increases late-onset sepsis (LOS) risk in preterm infants, potentially via gut dysbiosis. Analyzing 4,938 longitudinal fecal samples from preterm infants in China, the US, and the UK, we identified a differential pace of gut microbiota development among preterm infants. Delayed maturation correlated with over one-third of LOS risk associated with early antibiotic exposure. Deficiency of a bacterial DL-endopeptidase represented a hallmark of delayed microbiota development and correlated with elevated LOS risk. Supplementation with DL-endopeptidase-producing Enterococcus faecium or Limosilactobacillus reuteri activated the NOD2 receptor via muramyl dipeptide (MDP), regulated macrophage differentiation and polarization, restrained hyperinflammation via cylindromatosis (CYLD) induction, and protected neonatal mice from LOS. A pilot randomized controlled trial showed that L. reuteri supplementation enhanced fecal NOD2 activation in preterm infants. These findings link microbiota immaturity and reduced DL-endopeptidase activity to antibiotic exposure and LOS risk and highlight a candidate biomarker that warrants further validation for clinical translation.

Stroke-induced gut dysbiosis exacerbates brain injury via the toll-like receptor 4 (TLR4) signaling pathway. However, the role of intestinal-specific TLR4 in ischemic stroke remains unclear. This study aimed to investigate the role of intestinal-specific TLR4 in the acute phase of post-stroke recovery and to explore the underlying gut-brain mechanisms. A murine ischemic stroke model was established, and intestinal-specific interventions were performed using TLR4 inhibitors, agonists, and conditional knockout mice. Post-stroke brain injury, systemic inflammation, and gut barrier function were systematically evaluated. Transcriptomic profiling and 16 S rRNA sequencing were integrated to elucidate the regulatory mechanisms of intestinal TLR4 signaling in stroke outcomes. Pharmacological inhibition of intestinal TLR4 ameliorated, whereas its activation exacerbated, both cerebral and intestinal injury following stroke. Intestinal epithelial-specific TLR4 knockout (TLR4flox/flox; VilCre) significantly reduces cerebral infarction, improves neurological function, alleviates neuronal damage, decreases microglial activation, and preserves intestinal barrier integrity after stroke. Compared to brain-specific TLR4 knockout mice (TLR4flox/flox; Emx1Cre), intestinal epithelial-specific TLR4 knockout mice exhibit greater efficacy in alleviating brain injury, reducing systemic and neuroinflammation and protecting the gut barrier following stroke. RNA sequencing reveals upregulation of gamma-aminobutyric acid type A (GABAA) receptor signaling in the colon of TLR4flox/flox; VilCre mice. The protective effects of intestinal epithelial-specific TLR4 knockout are abolished by GABAA receptor inhibition, highlighting the role of the intestinal TLR4-GABAA axis in stroke recovery. Intestinal epithelial-specific TLR4 knockout reshapes the gut microbiota composition after stroke, and the altered microbial taxa, such as Bacteroides and Prevotella, are closely associated with improved neurological outcomes and upregulation of colonic GABAA receptors. This study identifies intestinal TLR4 as a key determinant of stroke outcomes during the acute phase and establishes the intestinal TLR4-GABAA signaling axis as a central mechanistic pathway mediating gut-brain crosstalk after stroke.

Oxylipins derived from n-6 and n-3 polyunsaturated fatty acids (PUFAs) are crucial signaling molecules involved in various physiological and pathological processes. However, the quantification of these metabolites remains challenging due to their low abundance, high structural similarity, and poor electrospray ionization efficiency. Furthermore, existing methods often suffer from complex sample preparation and the lack of cost-effective, reliable internal standards. Therefore, establishing a highly sensitive, automated, and standardized analytical platform for comprehensive oxylipin profiling is the prerequisite for elucidating their complex roles in disease pathogenesis. To address these challenges, we developed a highly sensitive and precise liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, incorporating automated sample preparation through magnetic solid-phase extraction and chemical isotope derivatization using 4- (aminomethyl)-N,N-dimethylaniline-d0/d6 (4-AND-d0/d6) reagents. This approach enables the quantification of 69 oxylipins derived from n-6 and n-3 PUFAs, achieving a remarkable 55- to 1000- fold increase in MS detection sensitivity compared to non-derivatized analysis. Additionally, a retention index (RI)-based predictive model was established to facilitate the screening and identification of unknown regio-isomers. As a proof of concept, this method was applied to quantify oxylipins in serum from a neonatal hypoxic-ischemic encephalopathy (HIE) rat model and to evaluate its applicability in complex biological matrices. Our method offers a sensitive, automated, and reliable tool for oxylipin profiling, with potential applications in biomarker discovery and lipid mediator research. This work presents the first integrated LC-MS/MS strategy combining automated magnetic-bead-assisted extraction and chemical isotopic derivatization for oxylipin analysis, enabling the highly sensitive quantification of 69 target analytes. This study is significant for establishing a robust retention index-based predictive model that facilitates the screening and identification of unknown regio-isomers, offering a reliable and high-throughput analytical solution for clinical biomarker discovery and lipid mediator profiling.

PURPOSE:We evaluated the quality of blood culture (BC) in critically ill patients in 19 tertiary comprehensive teaching hospitals in Guangdong Province, and analyzed the factors affecting the quality. METHODS:BC specimen data of 37,552 critically ill patients from January 1, 2021 to December 31, 2021 were screened, providing a total of 137,560 BC bottles. The quality indicators of BCs were analyzed. RESULTS:The positive and submission rates of BCs in critically ill patients varied among different hospitals. Of the most common and clinically significant isolates in positive BCs, there were 10 bacterial strains (n = 5572) and 1 fungal strain (n = 617), with median turnaround times for antimicrobial susceptibility test (AST) of 2.86 (2.19-3.62) days and 3.72 (3.00-4.58) days, respectively. BC specimen collection mainly occurred between 06:00 and 24:00, with reception and loading peaking from 08:00 and 24:00. Gram-staining and AST reports were concentrated between 10:00 and 10:59, and organism identification reports were between 09:00 and 09:59. CONCLUSION:There were differences in the quality of BC in critically ill patients among different hospitals and BCs had significant delays at night during the pre-analytical stage and specimen reporting times. These indicate the need for further optimization of related workflows.

Gestational diabetes mellitus (GDM) is a prevalent pregnancy complication. Both environmental changes and gut dysbiosis have been associated with increased risk of GDM. While environmental exposures may influence gut microbiome, the exact mechanisms linking these factors to GDM risk remain poorly understood. The study aimed to investigate the relationships between environmental exposures, gut microbiome and GDM risk. A population-based cohort study was conducted in Guangzhou, China from January 2017 to September 2017, enrolling 260 women with GDM and 508 normal pregnancies. Ambient pollution and meteorological exposures during different pregnancy windows were estimated for each participant. We identified the changes in gut microbiome characteristics related to environmental exposures. Temperature exposure during the preconception period was most strongly associated with alterations in the gut microbiome observed during the second trimester, particularly in women who developed GDM. Multivariate logistic regression and interaction analyses showed potential relationships between temperature exposure, gut microbiome and GDM risk. These findings contribute to the understanding of the relationship between environmental exposures, gut microbiome and GDM, providing insights into public health strategies to shield pregnant women from the detrimental effects of environmental exposures.

Ischemic stroke is a highly prevalent disease with limited therapeutic options, and emerging evidence suggests that the gut microbiota influences stroke pathophysiology. However, whether the gut microbiota affects individual susceptibility to ischemic stroke remains unclear. Here, middle cerebral artery occlusion was performed to stratify mice into ischemic stroke-sensitive (SEN: reaching humane endpoints within day 1) and ischemic stroke-resistant (RES: surviving to day 7) groups based on post-stroke survival. SEN mice exhibited more severe brain injury than RES mice, accompanied by increased systemic inflammation and elevated intestinal permeability. Fecal microbiota from SEN or RES donors was transplanted into antibiotic-treated recipients. Mice receiving SEN microbiota developed significantly worse outcomes compared with those receiving RES microbiota. Multi-omic analyses of cecal contents, including 16S rRNA gene sequencing and liquid chromatography-mass spectrometry/mass spectrometry-based untargeted metabolomics, identified reduced butyrate-producing bacteria and altered glutathione metabolism as potential contributors to stroke susceptibility. Collectively, these findings demonstrate that gut microbiota and microbial metabolites modulate susceptibility to ischemic stroke. IMPORTANCE:The role of the gut microbiota in determining susceptibility to ischemic stroke has remained poorly defined. This study demonstrates that microbiota dysbiosis and metabolite alterations functionally increase vulnerability to stroke injury, highlighting the gut microbiome as a potential target for risk stratification and preventive interventions. Modulating the gut microbiota may therefore represent a novel strategy for reducing stroke susceptibility.

Chronic alcohol consumption disrupts gut-liver homeostasis not only by inducing direct hepatotoxic injury, but also by perturbing host-microbial defense mechanisms that normally protect the liver from metabolic and inflammatory stress. We show that hepatocyte-specific deletion of Nod2 exacerbates ethanol-induced steatosis, oxidative stress, and mitochondrial dysfunction, establishing NOD2 as a critical protective factor in alcohol-associated liver disease (ALD). Importantly, beyond its direct hepatotoxic effects, ethanol exposure simultaneously diminishes this protective NOD2 pathway by limiting microbiota-derived ligand availability. Guided by this functional deficit, clinical metagenomic analysis (n = 1516) revealed that alcohol consumption is associated with a selective depletion of gut microbial DL-endopeptidase, a rate-limiting enzyme for NOD2 ligand generation, which inversely correlated with liver injury severity. Mice receiving fecal microbiota from donors with low DL-endopeptidase activity showed increased susceptibility to ALD. Importantly, supplementation with a NOD2 ligand or its clinical analogue, mifamurtide, restored mitochondrial homeostasis and alleviated liver injury. Together, these findings identify the gut microbial DL-endopeptidase-NOD2 axis as a key protective mechanism against ethanol-induced liver injury and a promising therapeutic target in alcohol-associated liver disease.

BACKGROUND:Stroke induces complex pathophysiological responses that extend beyond the brain, yet the mechanisms through which peripheral signals influence stroke recovery remain largely unclear. OBJECTIVE:Here, we identify a novel gut-brain neural circuit that promotes stroke recovery via kynurenic acid (KYNA) signalling. DESIGN:In a training cohort (30 patients with acute ischaemic stroke (AIS) and 30 controls), untargeted metabolomics profiled intestinal metabolites and the key metabolite KYNA was validated in an independent cohort (100 patients with AIS and 100 controls) using targeted metabolomics and assessed for its 3-month prognostic value. In stroke mouse models, KYNA was administered to evaluate therapeutic effects. Mechanistic studies combined neuronal calcium imaging, enteric neuron receptor manipulation, vagotomy, neuronal tracing, electrophysiology and immunofluorescence to delineate the KYNA-mediated gut-brain neural circuit regulating stroke recovery. RESULTS:Our study demonstrates a significant reduction of intestinal KYNA in patients with AIS and validates its prognostic value for neurological recovery at 3 months poststroke in both the training and validation cohorts. Oral KYNA supplementation markedly improves poststroke cerebral injury by activating G protein-coupled receptor 35 (GPR35) on enteric neurons, initiating vagal nerve signalling. Mechanistically, KYNA-GPR35 interaction activates vagal afferents, transmitting signals through the nucleus tractus solitarius to hippocampal and hypothalamic regions. This GPR35-vagus nerve signalling pathway, further validated with the selective GPR35 agonist Zaprinast, confers neuroprotection by shifting microglial polarisation towards the anti-inflammatory M2 phenotype and enhancing neuronal α7 nicotinic acetylcholine receptor activity. CONCLUSION:KYNA acts through an intestinal GPR35-vagus neural pathway to influence stroke recovery, highlighting this gut-brain signalling axis as a promising therapeutic avenue.

Biological aging has been associated with altered risk of aging-related diseases, but the contribution of the gut microbiota to this process remains poorly understood. Here, we constructed an interpretable gut microbiota age clock using metagenomic data from 8115 fecal samples across five continents. We discovered a key microbial perturbation occurring at 56-60 years of chronological age, which was validated in an independent cohort of 2263 metagenomes. This perturbation was associated with a decline in ecological stability and substantial changes in the abundance of core species. Notably, the association between gut microbiota age and diseases was identified to be significantly altered before and after this inflection time. Moreover, within-species analyses uncovered phylogenetic divergence for seven age-related species, such as Escherichia coli, alongside functional alterations in older individuals, including enhanced cell motility, carbohydrate metabolism and horizontal gene transfer. Overall, our global gut microbiome atlas uncovers a critical age transition phase, highlighting opportunities for microbiota-based therapies and offering novel insights into evolutionary dynamics during aging.

Abstract Background Gut dysbiosis has been increasingly implicated in post-stroke cognitive impairment (PSCI), yet the causal contribution and therapeutic potential of gut microbiota-derived metabolites remain unclear. This study aimed to identify key microbiota-derived metabolites involved in PSCI and to elucidate their underlying mechanisms. Results We found that both PSCI patients and middle cerebral artery occlusion (MCAO) mice exhibited distinct gut microbial alterations, characterized by a marked reduction in tryptophan-metabolizing bacteria and indole-3-propionic acid (IPA), a gut microbiota-derived tryptophan metabolite. Exogenous IPA administration alleviated PSCI-like phenotypes in MCAO mice. Mechanistically, IPA preserved tyrosine hydroxylase-positive (Th⁺) fibers and catecholamine levels in the dorsal hippocampus. Further analyses showed that IPA binds to the adaptor protein Ywhab, promotes ERK activation, and enhances neuronal survival, thereby counteracting neuronal apoptosis-associated inflammation and subsequent Th⁺ fiber degeneration. Conclusion These findings identify IPA as a gut microbiota-derived neuromodulator that mitigates PSCI by preserving dorsal hippocampal catecholaminergic transmission. IPA may therefore serve as a promising predictive biomarker and therapeutic candidate for PSCI.

BACKGROUND:Gut microbiota has been widely recognised as playing a critical role in maintaining immune imbalance and the development of rheumatoid arthritis (RA). As key roles mediating interkingdom crosstalk among plants, microbiomes and mammals, plant-derived exosome-like nanoparticles (ELNs) could use lipids and microRNA components to precisely modulate gene expression of gut microbiota, showing potential as a dietary intervention for RA treatment. OBJECTIVE:We aimed to investigate gut microbiota-immune interactions inducing immune dysregulation in RA and explore potential applications of edible plant-derived ELNs for RA treatment through gut microbiota manipulation. DESIGN:Combinations of microbial analysis of clinical cohort, metabolomics, in vivo and in vitro examination were performed to establish potential gut-immune mechanisms for interventions. Several representative edible plants ELNs were chosen to compare the modulation effects based on the above mechanism. Small RNA sequencing and lipidomic analysis were performed to identify key components and reveal the related mechanisms mediating therapeutic effects. RESULTS:Ruminococcus gnavus was significantly enriched in RA and aggravated arthritis through secreting phenylethylamine (PEA) to induce excessive neutrophil extracellular traps (NETs) formation. Among several plants ELNs, Pueraria lobata-derived ELNs (Pu-ELNs) were preferentially taken up by R. gnavus and decreased PEA production. Mechanistically, the lipid components of Pu-ELNs induced intestinal accumulation of ELN-derived gma-miR4412, which reduces phenylalanine decarboxylase (PDC) expression, relieving the arthritis aggravation caused by R. gnavus through acting on the PEA-Bruton's tyrosine kinase (BTK)-NETs axis. CONCLUSIONS:Our findings suggest the crucial role of R. gnavus in aggravating RA and underscore the application of plant-derived ELNs for microbiota manipulation.

Sepsis remains a leading cause of morbidity and mortality in neonatal and pediatric intensive care units (NICU/PICU). Given the rapid clinical deterioration of these vulnerable populations, shifting from reactive diagnosis to proactive prediction of sepsis onset is crucial. Multidimensional hematological parameters may provide new insights for risk stratification. While routine complete blood count (CBC) is widely used, the predictive potential of high-dimensional hematological cell morphology features that extend beyond traditional CBC remains under-investigated in large-scale pediatric cohorts. This multicenter study utilized a large-scale dataset from the CALM2302 project, encompassing 17,066 critically ill neonates and children across 30 Chinese hospitals between October 2020 and October 2023. We developed population-specific CatBoost models for NICU and PICU to predict sepsis onset. To mitigate class imbalance, random undersampling was applied to the training sets. From a candidate pool of 1,924 parameters (37 routine CBC and 1,887 research features), optimized signatures of approximately 200 features were selected for each population. Model performance was evaluated by the area under the ROC curve (AUC) across internal and dual external validation cohorts. Furthermore, SHAP (SHapley Additive exPlanations) analysis was performed to ensure model interpretability and identify key biological predictors. Among the total cohort, 10,347 infants were categorized into the NICU cohort (sepsis incidence: 7.48

ABSTRACT Emerging evidence highlights the potential impact of intratumoral microbiota on cancer. However, the microbial composition and function in glioma remains elusive. Consequently, our study aimed to investigate the microbial community composition in glioma tissues and elucidate its role in glioma development. We parallelly performed microbial profiling, transcriptome sequencing, and metabolomics detection on tumor and adjacent normal brain tissues obtained from 50 glioma patients. We employed immunohistochemistry, multicolor immunofluorescence, and fluorescence in situ hybridization (FISH) staining to observe the presence and location of bacteria. Furthermore, an animal model was employed to validate the impact of key bacteria on glioma development. Six genera were found to be significantly enriched in glioma tissues compared to adjacent normal brain tissues, including Fusobacterium, Longibaculum, Intestinimonas, Pasteurella, Limosilactobacillus, and Arthrobacter. Both bacterial RNA and lipopolysaccharides (LPS) were observed in glioma tissues. Integrated microbiomics, transcriptomics, and metabolomics revealed that genes associated with intratumoral microbes were enriched in multiple synapse-associated pathways and that metabolites associated with intratumoral microbes were (R)-N-methylsalsolinol, N-acetylaspartylglutamic acid, and N-acetyl-l-aspartic acid. Further mediation analysis suggested that the intratumoral microbiome may affect the expression of neuron-related genes through bacteria-associated metabolites. In addition, both in vivo and in vitro models of glioma show that Fusobacterium nucleatum promotes glioma proliferation and upregulates CCL2, CXCL1, and CXCL2 levels. Our findings shed light on the intricate interplay between intratumoral bacteria and glioma.IMPORTANCEOur study adopted a multi-omics approach to unravel the impact of intratumoral microbes on neuron-related gene expression through bacteria-associated metabolites. Importantly, we found bacterial RNA and LPS signals within glioma tissues, which were traditionally considered sterile. We identified key microbiota within glioma tissues, including Fusobacterium nucleatum (Fn). Through in vivo and in vitro experiments, we identified the crucial role of Fn in promoting glioma progression, suggesting that Fn could be a potential diagnostic and therapeutic target for glioma patients. These findings offer valuable insights into the intricate interplay between intratumoral bacteria and glioma, offering novel inspiration to the realm of glioma biology.

BACKGROUND:Chronic kidney disease (CKD) is a major global public health issue, with a steadily increasing incidence. Urinary protein detection serves as a crucial indicator for the diagnosis, monitoring and management of CKD. However, current methods for urinary protein measurement, such as urine dipstick tests, colorimetric assays, and 24-h total urine protein analysis, have certain limitations that restrict their routine application in CKD screening and follow-up. Therefore, there is an urgent need for a simple, convenient, and rapid diagnostic approach for kidney function assessment. RESULTS:In this study, we have developed and validated a novel method for urine protein quantification based on dried droplet morphology analysis. Our approach demonstrates robust performance across a wide range of protein concentrations and is resilient to common interfering substances and variations in sample processing. It's worth noting that while our method shows excellent agreement with the colorimetric assay, it offers several potential advantages. These include reduced sample volume requirements, simplified sample preparation, and rapid analysis time. These factors could make our method particularly suitable for point-of-care testing or resource-limited settings where traditional laboratory infrastructure may be unavailable. SIGNIFICANCE:The novel method for protein quantification in urine based on the morphology of a dried droplet uses only one drop of urine specimen. Combined with machine learning models, by identifying protein content in urine droplet drying patterns without the need for staining or antibody binding, may provide a more convenient alternative to current techniques for the assessment of proteinuria. Simple, low-cost, and fast, the system can be used as a powerful tool for CKD surveillance at the point of care.

Colorectal cancer (CRC) remains a formidable global health challenge and is associated with dismal survival outcomes and high mortality among patients diagnosed at advanced stages. Despite advancements in early screening and therapeutic interventions, the outcomes of patients with advanced-stage CRC remain suboptimal, as these patients continue to exhibit a persistently low 5-year survival rate. Palliative radiotherapy (RT) is crucial for advanced CRC patients, but radioresistance remains a significant clinical challenge. This resistance is attributed to multiple mechanisms, such as genetic heterogeneity, dysregulated DNA damage repair and tumor microenvironment metabolic disorders. Recent studies have shown that noncoding RNAs (ncRNAs), mainly microRNAs, long ncRNAs (lncRNAs) and circular RNAs, play pivotal roles in regulating CRC radiosensitivity through diverse mechanisms, such as epithelial-mesenchymal transition, epigenetic reprogramming, posttranscriptional regulation and oncogenic signaling pathway activation. For example, microRNAs such as miR-141-3p and miR-630 enhance CRC radiosensitivity by targeting oncogenic pathways. In addition, lncRNAs, including the lncRNAs HOTAIR and LINC00630, influence the radiosensitivity of CRC through interactions with the DNA damage repair machinery and epigenetic modulators, respectively. In addition, circ_0124554 acts as a competitive endogenous RNA to regulate oncogenic signaling. ncRNAs also serve as potential biomarkers for predicting radiosensitivity and prognosis. This review synthesizes the current evidence on the ncRNA-mediated regulatory networks that influence CRC radiosensitivity, emphasizing their potential as therapeutic targets to overcome RT resistance and improve outcomes in advanced CRC. By bridging mechanistic insights with clinical applications, this work aims to guide future research and the implementation of precision RT strategies.

The increasing use of tissue clearing techniques underscores the urgent need for cost-effective and simplified deep imaging methods. While traditional inverted confocal microscopes excel in high-resolution imaging of tissue sections and cultured cells, they face limitations in deep imaging of cleared tissues due to refractive index mismatches between the immersion media of objectives and sample container. To overcome these challenges, the RIM-Deep was developed to significantly improve deep imaging capabilities without compromising the normal function of the confocal microscope. This system facilitates deep immunofluorescence imaging of the prefrontal cortex in cleared macaque tissue, extending imaging depth from 2 mm to 5 mm. Applied to an intact and cleared Thy1-EGFP mouse brain, the system allowed for clear axonal visualization at high imaging depth. Moreover, this advancement enables large-scale, deep 3D imaging of intact tissues. In principle, this concept can be extended to any imaging modality, including existing inverted wide-field, confocal, and two-photon microscopy. This would significantly upgrade traditional laboratory configurations and facilitate the study of connectomes in the brain and other tissues.