Optical microscopy is essential for exploring biological and material structures, with resolution determining the level of observable detail. The advent of super-resolution fluorescence microscopy has broken the diffraction limit, achieving nanoscale resolution. However, traditional assessment methods, such as the Rayleigh criterion and point spread function (PSF) width measurement, rely on empirical judgments and diffraction-limited models, rendering them inadequate for modern super-resolution imaging. This review systematically traces the evolution of resolution assessment methodologies, from classical criteria to advanced strategies tailored for various super-resolution modalities. We first discuss Fourier-based quantitative methods. Fourier ring correlation (FRC) and its 3D counterpart, Fourier shell correlation (FSC), objectively determine resolution by evaluating the statistical correlation of two independent image reconstructions in frequency space. These methods offer robustness against noise and provide a global resolution metric, but they require data independence and are computationally intensive. They have become the prevailing standards in electron and super-resolution microscopy. Subsequently, we examine adaptations for specific super-resolution techniques. For single-molecule localization microscopy (SMLM) techniques such as PALM and STORM, the Fourier image resolution (FIRE) method extends FRC by incorporating a physical model that accounts for localization precision and labeling density. For stimulated emission depletion (STED) microscopy and other nonlinear techniques, assessment strategies differ. While PSF shrinkage measurements using fluorescent beads are useful for system calibration, evaluating the effective resolution directly on biological samples is more practical. This is typically performed via linewidth analysis of known structures (e.g., microtubules) or edge-spread function measurements, capturing the effects of photobleaching and sample-induced aberrations. A major paradigm shift is parameter-free resolution estimation based on decorrelation analysis. This method analyzes the autocorrelation decay of a single image’s Fourier spectrum to identify the cutoff spatial frequency without requiring dual datasets or user-defined thresholds. Its high efficiency and broad applicability have been validated across widefield, confocal, STED, SIM, and SMLM modalities. Optimized rendering strategies for SMLM data further enhance its accuracy, and it is emerging as a tool for real-time optimization of experimental parameters. The review also addresses the “gold standard” of resolution validation using well-defined nanostructures, such as DNA origami and nuclear pore complexes, which provide ground truth for verifying resolution claims and detecting artifacts. In the era of artificial intelligence, deep learning plays a dual role: it powerfully enhances image resolution but also introduces challenges, as models may generate “hallucinations” or false details. This underscores the need for new validation metrics to verify the physical fidelity of AI-generated content. Finally, we outline future directions: developing unified cross-modality standards, enabling real-time dynamic resolution monitoring for live-cell imaging, creating techniques for generating local resolution maps to capture sample heterogeneity, and integrating intelligent error correction to ensure data veracity. By providing a comprehensive overview of resolution assessment progress and challenges, this review aims to equip researchers with the knowledge to select appropriate tools, thereby fostering rigorous quantitative imaging in the life and material sciences.

With the rising incidence of diabetes， diabetic kidney disease （DKD） has become a significant global health burden. Although current prevention and treatment strategies can partially delay the progression of DKD， the risk of patients advancing to end-stage renal disease remains high. Since the concept of the "gut-kidney axis" was first introduced at the International Congress on Dialysis in 2011， research on the role of gut microbiota in the pathogenesis of DKD has received increasing attention. This review summarizes the current research on gut microbiota， explores the mechanisms through which it contributes to DKD development， and outlines clinical approaches for DKD prevention and treatment based on the "gut-kidney axis" theory. Evidence indicates that dietary interventions， intake of probiotics or prebiotics， use of metformin and novel antidiabetic drugs， and application of traditional Chinese medicine （TCM） compound formulas can effectively improve gut microbiota composition， influence metabolite production， and restore the intestinal mucosal barrier. These interventions can further regulate intestinal innate immunity and inflammatory responses， thereby modulating the progression of DKD. Despite challenges posed by the traditional oral administration of water-decocted TCM compound formulas and the complexity of their ingredients， increasing evidence suggests that TCM may indirectly affect the occurrence and development of DKD by modulating gut microbiota. This finding provides a new perspective on the potential mechanisms of TCM in DKD treatment and may offer novel strategies for DKD prevention and therapy.

ObjectiveCerebrospinal fluid (CSF) plays a crucial role in maintaining the homeostasis of the central nervous system (CNS). CSF rapidly exchanges with interstitial fluid (ISF) via the glymphatic system within the brain parenchyma. CSF-ISF circulation and its associated mechanisms are often referred to as the brain lymphatic system. This system is connected directly to meningeal lymphatic vessels (mLVs), jointly performing the function of clearing metabolic waste from the CNS. Emerging evidence indicates that this system is closely associated with the onset and progression of neurodegenerative diseases (NDs) such as Alzheimer’s disease (AD). Importantly, abnormal CSF circulation is not only a downstream consequence of AD pathology, but also a risk factor. In AD, the dynamics of CSF flow within the CNS are diminished, immune dysregulation occurs, and this may increase the risk of AD by exacerbating the burden of amyloid β-protein (Aβ). In the mouse model of AD, impaired CSF flow compromises this clearance function, leading to cognitive deficits. Clinically, acupuncture at cognition-related acupoints is commonly used for the prevention and treatment of AD. However, whether its therapeutic effects are mediated through the modulation of CSF dynamics remains unclear. This study aimed to evaluate the impact of acupuncture on CSF flow and investigate its acupoint specificity. MethodsMice were randomly assigned to experimental groups for the different electroacupuncture groups with the following acupoints: Baihui point (GV 20), Ear point, Neiguan point (PC 6), and Tianshu point (ST 25). Wild-type mice on a C57BL/6J background were used as controls. Fluorescent tracer was injected into the cisterna magna to label CSF flow. Fluorescence imaging was employed to assess the distribution of CSF within the brain before and after acupuncture stimulation. ResultsFollowing tracer injection into the cisterna magna, fluorescence signals rapidly reached the cerebellum and medulla—the regions closest to the injection site. Fluorescence intensity was higher in ventral brain regions compared to dorsal regions, likely due to greater vascular density in ventral areas facilitating CSF-ISF exchange. Electroacupuncture at the GV 20 produced the most pronounced enhancement of CSF across the whole brain, while stimulation at the ST 25 primarily augmented flow within subcortical regions. In contrast, electroacupuncture at the Ear point or the PC 6 had no observable effect on CSF in mice. ConclusionElectroacupuncture promotes CSF flow into the brain parenchyma in an acupoint-specific manner, with GV 20 exhibiting the most pronounced enhancement of CSF dynamics. These findings suggest that acupuncture-mediated facilitation of CSF flow may represent a potential therapeutic strategy for preventing or delaying age-related cognitive decline.

ObjectiveCerebrospinal fluid (CSF) plays a crucial role in maintaining the homeostasis of the central nervous system (CNS). CSF rapidly exchanges with interstitial fluid (ISF) via the glymphatic system within the brain parenchyma. CSF-ISF circulation and its associated mechanisms are often referred to as the brain lymphatic system. This system is connected directly to meningeal lymphatic vessels (mLVs), jointly performing the function of clearing metabolic waste from the CNS. Emerging evidence indicates that this system is closely associated with the onset and progression of neurodegenerative diseases (NDs) such as Alzheimer’s disease (AD). Importantly, abnormal CSF circulation is not only a downstream consequence of AD pathology, but also a risk factor. In AD, the dynamics of CSF flow within the CNS are diminished, immune dysregulation occurs, and this may increase the risk of AD by exacerbating the burden of amyloid β-protein (Aβ). In the mouse model of AD, impaired CSF flow compromises this clearance function, leading to cognitive deficits. Clinically, acupuncture at cognition-related acupoints is commonly used for the prevention and treatment of AD. However, whether its therapeutic effects are mediated through the modulation of CSF dynamics remains unclear. This study aimed to evaluate the impact of acupuncture on CSF flow and investigate its acupoint specificity. MethodsMice were randomly assigned to experimental groups for the different electroacupuncture groups with the following acupoints: Baihui point (GV 20), Ear point, Neiguan point (PC 6), and Tianshu point (ST 25). Wild-type mice on a C57BL/6J background were used as controls. Fluorescent tracer was injected into the cisterna magna to label CSF flow. Fluorescence imaging was employed to assess the distribution of CSF within the brain before and after acupuncture stimulation. ResultsFollowing tracer injection into the cisterna magna, fluorescence signals rapidly reached the cerebellum and medulla—the regions closest to the injection site. Fluorescence intensity was higher in ventral brain regions compared to dorsal regions, likely due to greater vascular density in ventral areas facilitating CSF-ISF exchange. Electroacupuncture at the GV 20 produced the most pronounced enhancement of CSF across the whole brain, while stimulation at the ST 25 primarily augmented flow within subcortical regions. In contrast, electroacupuncture at the Ear point or the PC 6 had no observable effect on CSF in mice. ConclusionElectroacupuncture promotes CSF flow into the brain parenchyma in an acupoint-specific manner, with GV 20 exhibiting the most pronounced enhancement of CSF dynamics. These findings suggest that acupuncture-mediated facilitation of CSF flow may represent a potential therapeutic strategy for preventing or delaying age-related cognitive decline.

Objectives: . Idiopathic sudden sensorineural hearing loss (ISSNHL) is a significant cause of hearing loss. Intratympanic steroid injection (ITSI) is commonly used as an initial or salvage treatment; however, the lack of a standardized treatment protocol has resulted in variability in clinical practice. In addition, no efficient prediction model currently exists to support personalized management. Therefore, this study aimed to develop tailored management strategies for ISSNHL using a machine-learning model. Methods: . This retrospective multicenter cohort study was conducted between January 2015 and December 2020, with data analysis performed between January 2021 and March 2024. Patients were selected based on the International Classification of Diseases, 10th Revision criteria for ISSNHL, along with relevant medication and procedure codes. Patients with pure-tone audiogram results not meeting ISSNHL criteria, better initial hearing in the affected ear, an identifiable etiology, no post-treatment audiogram, or delayed treatment (>6 weeks) were excluded. We included 770 patients diagnosed with ISSNHL who received ITSI. The primary outcome was the area under the receiver operating characteristic curve for prediction performance. Recovery status was determined using the last pure-tone audiogram. Modeling was conducted on the Quanta for Medical Care AI platform using five machine-learning algorithms and a nested cross-validation framework, in which feature selection and hyperparameter tuning were performed in the inner folds and model performance was evaluated in the outer folds. Results: . A random forest classifier outperformed the other models in predicting hearing outcomes, achieving an area under the receiver operating characteristic curve of 0.788. Time to ITSI was the most influential treatment-related factor, with ITSI administered within 10 days of hearing loss being associated with better outcomes. This model can be used to provide personalized prognostic estimates under different treatment protocols. Conclusion . The machine-learning-based prediction model facilitates personalized treatment strategies and timely treatment adjustments for ISSNHL, thereby optimizing the likelihood of complete recovery.

Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

ObjectiveTo analyze the composition of respiratory virus pathogen spectrum and epidemiological characteristics among adult influenza-like illness (ILI) cases in Putuo District of Shanghai from 2015 to 2024, to summarize the epidemic patterns of respiratory viruses across different time periods and populations, and to provide scientific evidence for the prevention and control of adult respiratory virus infections in Putuo District of Shanghai. MethodsSurveillance data of 2 273 adult ILI cases from sentinel hospitals under the comprehensive surveillance system for acute respiratory infections in Putuo District of Shanghai from 2015 to 2024 were retrospectively collected. Descriptive epidemiological methods were adopted to analyze the temporal and population distribution characteristics of respiratory viruses, including influenza virus (IFV), respiratory syncytial virus (RSV), adenovirus (ADV), and human metapneumovirus (HMPV). The mixed infection status among different age groups was compared, and the distribution of influenza virus subtypes was described. SPSS 22.0 and WPS Office software were used for statistical analyses. ResultsThe overall detection rate of respiratory viruses in 2 273 adult ILI cases was 43.51%. The detection rate in females (45.48%) was significantly higher than that in males (41.25%), with statistical significance (χ²=4.113, P=0.043). Individuals aged 60 years old and above had the highest detection rate (47.41%) (χ²=8.430, P=0.014). The detection rates in winter (50.41%) and summer (48.42%) were higher than those in other seasons (χ²=47.028, P<0.001). The overall viral detection rate decreased after 2020 (χ²=208.411, P<0.001). Detection rates of various viruses varied by year, with IFV and ADV showing high rates from 2015 to 2019 (all P<0.001), while human coronavirus (HCoV) and rhinovirus/enterovirus (HRV/EV) exhibited higher rates from 2020 to 2022 (all P<0.001). Co-infection with IFV and HCoV was the most common, followed by co-infection with IFV and ADV, together accounting for 50.00% of all mixed infections. During the surveillance period, the detection rate of IFV was significantly higher than that of other respiratory viruses. Influenza B/Victoria lineage predominated in 2021, followed by successive predominance of influenza A(H1N1), A(H3N2), and the co-circulation of the two subtypes from 2022 to 2023. Influenza A(H1N1) re-emerged as the predominant subtype in the first half of 2024. ConclusionFrom 2015 to 2024, respiratory virus infections in adults prevailed in winter and summer, and people aged 60 years old and above were the key high-risk population. The detection rates of respiratory viruses declined after 2020. IFV was the predominant pathogen detected, and different influenza virus subtypes circulated alternately each year.

ObjectiveTo systematically evaluate the measurement content, psychometric properties and quality of evidence of patient-reported outcome measures (PROMs) for adolescents with spinal dysfunction based on International Classification of Functioning, Disability and Health-Children and Youth (ICF-CY) framework, Consensus-based Standards for the Selection of Health Measurement Instruments (COSMIN) and modified GRADE approach. MethodsDatabases including CNKI, Wanfang Data, PubMed, Embase and Web of Science were systematically searched for studies assessing psychometric properties of PROMs targeting adolescents with spinal dysfunction. The methodological quality of included studies was evaluated using COSMIN Risk of Bias checklist, the psychometric properties were rated according to COSMIN criteria, and evidence quality was graded using modified GRADE approach. Item content of each PROM was coded following the ICF-CY linking rules. ResultsA total of 23 studies involving eight PROMs were included. Brace Questionnaire (BrQ), Italian Spine Youth Quality of Life Questionnaire (ISYQOL), Quality of Life Profile for Spinal Deformities (QLPSD), Spinal Appearance Questionnaire (SAQ), Scoliosis Japanese Questionnaire-27 (SJ-27) and Scoliosis Research Society-22 Revised Questionnaire (SRS-22r) were rated as grade B, while EuroQol Five-Dimension Youth Five-Level Questionnaire (EQ-5D-Y-5L) and Young Spine Questionnaire (YSQ) were rated as grade C. The eight PROMs were linked to six to 24 ICF-CY categories. BrQ covered the largest number of categories (n = 24), followed by SJ-27 (n = 19) and SRS-22r (n = 16). Except for SAQ which mainly focused on dimension of body functions and structures, all other PROMs covered both dimension of body functions and structures, and activities and participation, while BrQ, ISYQOL, SRS-22r and YSQ also covered dimension of environmental factors. Major methodological limitations included insufficient evidence for content validity, inappropriate factor-analysis approaches, incomplete reporting for test-retest reliability, and overall scarcity of evidence concerning measurement error, minimal important change (MIC) and responsiveness. ConclusionThe quality of PROMs in adolescents with spinal dysfunction varies. BrQ, ISYQOL, QLPSD, SAQ, SJ-27 and SRS-22r show application potential, yet further high-quality studies are required to supplement their psychometric evidence. EQ-5D-Y-5L and YSQ exist high-quality insufficient evidence. Selection of PROMs should comprehensively consider target population, measurement purpose, content coverage and quality of evidence. Further researches are needed to improve reporting quality.

Acute burns and chronic wounds frequently fail to heal owing to various reasons. Most drugs currently used for wound therapy in clinical practice have notable drawbacks, making their application a substantial concern. For instance, anti-inflammatory drugs can exert multisystem toxicity, and cellular therapies are costly and difficult to retain. In recent years, natural functional proteins derived from animals and plants have gained increasing attention owing to their unique biological activities, low cost, and broad application prospects in wound therapy. Herein, we isolated a new protein (JH015Y) from amphibians and demonstrated its excellent wound repair and regeneration properties compared with those of epidermal growth factor, both in vitro and in vivo. JH015 protein increased the proliferative ability of human keratinocytes and skin fibroblasts by 47.73 and 41.40%, respectively. In vivo, the medium-dose (0.5 mg/dose) groups of JH015Y protein demonstrated accelerated wound healing from day 4, with wound healing rates 1.26, 1.27, and 1.14 times that of the blank group in acute wounds, burn wounds, and diabetic ulcer, respectively. Histological analysis of Masson-stained sections indicated that the JH015Y protein contributed to collagen deposition on the wound surface, markedly reduced inflammatory cell infiltration, and exhibited low biological toxicity. Accordingly, the JH015Y protein is a promising biotherapeutic agent for accelerated wound repair and regeneration.