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.

ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

ObjectiveTo evaluate the predictive value of epicardial adipose tissue （EAT） parameters for microvascular obstruction （MVO） formation in patients with ST segment elevation myocardial infarction （STEMI） using cardiac magnetic resonance quantification. MethodsA total of 139 STEMI patients were included in this study， and various parameters such as EAT thickness， volume， and mass index were measured utilizing cardiac magnetic resonance. All included patients were divided into MVO group and non-MVO group according to whether MVO occurred. Differences in EAT related parameters between two groups were compared and correlation analysis was applied to evaluate the correlation between quantitative indicators of EAT and indicators such as infarct size and ejection fraction. Logistic regression analysis was used to identify the relevant risk factors for MVO formation. Receiver Operating Characteristic （ROC） curve analysis was performed to assess the predictive value of the epicardial adipose tissue （EAT） quality index and other indicators for the occurrence of MVO. ResultsCompared with non MVO group， patients in MVO group presented with higher peak troponin T levels， increase of neutrophil lymphocyte ratio （NLR） and C-reactive protein （CRP）， larger infarct size and compromised left ventricular ejection fraction （LVEF） （P<0.05）. Total EAT volume， EAT mass index， left atrioventricular EAT volume， left atrioventricular EAT mass index and thickness of EAT in the left atrioventricular groove were significantly higher in patients with MVO. Multivariate Logistic regression analysis demonstrated that NLR， peak troponin T levels and left atrioventricular EAT mass index were independent predictors of MVO. The ROC curve suggested that the left atrioventricular EAT mass index had the highest predictive power for MVO formation in STEMI patients. ConclusionThe parameters of EAT quantified by cardiac magnetic resonance serve as imaging biomarkers for predicting MVO formation in STEMI patients. These metrics enable risk stratification post-myocardial infarction and facilitate early identification of high-risk individuals， thereby supporting personalized therapeutic decision-making.

Ultra performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry/mass spectrometry(UPLC-Q-TOF-MS/MS), network pharmacology, and animal experiments were integrated o explore the blood glucose-lowering effects and mechanisms of Poria aqueous extract. Firstly, the active components of Poria aqueous extract were identified by UPLC-Q-TOF-MS/MS. Subsequently, network pharmacology was employed to predict the blood glucose-lowering components and mechanisms of Poria aqueous extract. Finally, a rat model of diabetes mellitus, 16S rDNA sequencing, and Western blot were employed to investigate the blood glucose-lowering effect and mechanism of Poria aqueous extract. A total of 39 triterpenoids were identified in the Poria aqueous extract, among them, 25-hydroxypachymic acid, 25α-hydroxytumulosic acid, 16α-hydroxytrametenolic acid, polyporenic acid C, and tumulosic acid may be the main active ingredients for treating diabetes. The Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis revealed that Poria might exert its therapeutic effects through multiple pathways such as NOD-like receptor signaling pathway, nuclear factor-kappa B(NF-κB) signaling pathway, and tumor necrosis factor(TNF) signaling pathway. The results of animal experiments demonstrated that Poria aqueous extract significantly reduced the levels of blood glucose and lipids and regulated the intestinal flora in diabetic rats. The main affected taxa included g＿Escherichia-Shigella, g＿Corynebacterium, g＿Prevotella＿9, g＿Prevotellaceae＿UCG-001, and g＿Bacteroidota＿unclassified. In addition, Poria aqueous extract lowered the levels of D-lactic acid and lipopolysaccharide, alleviated colonic mucosal damage, significantly down-regulated the protein levels of NOD-like receptor pyrin domain-containing protein 3(NLRP3), NF-κB, and TNF-α, and significantly up-regulated the protein levels of zonula occludens 1 and occludin in diabetic rates. Poria aqueous extract may play a role in treating diabetes mellitus by repairing the intestinal flora disturbance, protecting the intestinal barrier function, and inhibiting the NF-κB/NLRP3 signaling pathway. The results provide a scientific basis for clinical application and expansion of indications of Poria.
Animals ; Rats ; Network Pharmacology ; Tandem Mass Spectrometry ; Male ; Drugs, Chinese Herbal/pharmacology* ; Chromatography, High Pressure Liquid ; Blood Glucose/drug effects* ; Rats, Sprague-Dawley ; Hypoglycemic Agents/administration & dosage* ; Poria/chemistry* ; Diabetes Mellitus, Experimental/metabolism* ; NF-kappa B/genetics* ; Gastrointestinal Microbiome/drug effects* ; Humans

This study employed bioinformatics to screen the feature genes related to efferocytosis in diabetic kidney disease(DKD) and explores traditional Chinese medicine(TCM) regulating these feature genes. The GSE96804 and GSE30528 datasets were integrated as the training set, and the intersection of differentially expressed genes and efferocytosis-related genes(ERGs) was identified as DKD-ERGs. Subsequently, correlation analysis, protein-protein interaction(PPI) network construction, enrichment analysis, and immune infiltration analysis were performed. Consensus clustering was conducted on DKD patients based on the expression levels of DKD-ERGs, and the expression levels, immune infiltration characteristics, and gene set variations between different subtypes were explored. Eight machine learning models were constructed and their prediction performance was evaluated. The best-performing model was evaluated by nomograms, calibration curves, and external datasets, followed by the identification of efferocytosis-related feature genes associated with DKD. Finally, potential TCMs that can regulate these feature genes were predicted. The results showed that the training set contained 640 differentially expressed genes, and after intersecting with ERGs, 12 DKD-ERGs were obtained, which demonstrated mutual regulation and immune modulation effects. Consensus clustering divided DKD into two subtypes, C1 and C2. The support vector machine(SVM) model had the best performance, predicting that growth arrest-specific protein 6(GAS6), S100 calcium-binding protein A9(S100A9), C-X3-C motif chemokine ligand 1(CX3CL1), 5'-nucleotidase(NT5E), and interleukin 33(IL33) were the feature genes of DKD. Potential TCMs with therapeutic effects included Astragali Radix, Trionycis Carapax, Sargassum, Rhei Radix et Rhizoma, Curcumae Radix, and Alismatis Rhizoma, which mainly function to clear heat, replenish deficiency, activate blood, resolve stasis, and promote urination and drain dampness. Molecular docking revealed that the key components of these TCMs, including β-sitosterol, quercetin, and sitosterol, exhibited good binding activity with the five target genes. These results indicated that efferocytosis played a crucial role in the development and progression of DKD. The feature genes closely related to both DKD and efferocytosis, such as GAS6, S100A9, CX3CL1, NT5E, and IL33, were identified. TCMs such as Astragali Radix, Trionycis Carapa, Sargassum, Rhei Radix et Rhizoma, Curcumae Radix, and Alismatis Rhizoma may provide a new therapeutic strategy for DKD by regulating efferocytosis.
Humans ; Computational Biology ; Diabetic Nephropathies/physiopathology* ; Protein Interaction Maps ; Medicine, Chinese Traditional ; Drugs, Chinese Herbal ; Phagocytosis/genetics* ; Efferocytosis

As a member of the Citrus genus of the Rutaceae family, Citrus changshan-huyou(CSHY) is mainly produced in Quzhou city, Zhejiang province. Modern research shows that different medicinal parts of CSHY(immature fruit, mature fruit peel, flower buds, leaves, seeds, etc.) are abundant in flavonoids, terpenes, coumarins, phenolic acids, and volatile oils. Their pharmacological activities include respiratory system protection, liver protection, anti-inflammation, anti-hyperlipidemia, anti-hyperglycemia, and antioxidation. Based on the summarization of 374 chemical components in different medicinal parts of CSHY identified in the past 20 years, this study reviewed their pharmacological actions and mechanisms and further analyzed the current status of quality control of different medicinal parts of CSHY, aiming to provide reference for the resource development and exploitation and the quality control research of different medicinal parts of CSHY.
Citrus/chemistry* ; Drugs, Chinese Herbal/pharmacology* ; Humans ; Plants, Medicinal/chemistry* ; Quality Control ; Animals

OBJECTIVES: To evaluate the causal association between circulating levels of zinc, magnesium, and other minerals and autism spectrum disorder (ASD). METHODS: A two-sample Mendelian randomization (MR) analysis was performed using summary statistics from large-scale genome-wide association studies of European populations, including 18 382 ASD cases and 27 969 controls. Genetic data for iron, calcium, and magnesium were obtained from the UK Biobank, and data for zinc and selenium were sourced from an Australian-British cohort. A total of 351 genetic instrumental variables were selected. Causal inference was performed using inverse-variance weighting as the primary analysis method. Sensitivity analyses were performed by Cochran's Q test and MR-PRESSO global test to assess the robustness of the findings. RESULTS: No statistically significant causal effect was observed for circulating zinc, magnesium, calcium, selenium, or iron levels on ASD risk (all P>0.05). The odds ratios and 95% confidence intervals from the inverse-variance weighting analysis were 0.934 (0.869-1.003) for zinc, 1.315 (0.971-1.850) for magnesium, 1.055 (0.960-1.159) for calcium, 1.015 (0.953-1.080) for selenium, and 0.946 (0.687-1.303) for iron. Sensitivity analysis revealed significant heterogeneity in the causal association between circulating calcium and ASD (P=0.006), while the effect estimate remained stable after MR-PRESSO correction (P=0.487). The causal effect estimates for the remaining minerals demonstrated good robustness. CONCLUSIONS This study did not find significant evidence supporting a causal association between circulating zinc, magnesium, calcium, selenium, or iron levels and ASD risk, providing important clues for the etiology of ASD and precision nutritional interventions.
Humans ; Mendelian Randomization Analysis ; Autism Spectrum Disorder/genetics* ; Magnesium/blood* ; Zinc/blood* ; Minerals/blood* ; Genome-Wide Association Study ; Selenium/blood*

OBJECTIVE: To retrospectively analyze the clinical characteristics, prognosis and prognostic factors of patients with light-chain (AL) amyloidosis, so as to provide reference for the diagnosis and treatment of AL amyloidosis. METHODS: Clinical data of 52 patients diagnosed with AL amyloidosis at two hospitals from January 2017 to November 2022 were collected. The clinical characteristics, differences in clinical indexes between the deceased group and the survival group were analyzed. Kaplan-Meier curves were used for overall survival (OS) analysis, and Cox regression models were used to analyze the factors affecting the prognosis. RESULTS: The median age of the 52 patients at diagnosis was 61(41-81) years old, and 63.5% of the patients were male. Heart (69.2%) and kidney (67.3%) were the most involved organs, and 67.3% of the patients had two or more organs involved. Most patients (71.2%) received chemotherapy regimens containing bortezomib, including 5 patients (9.6%) who received treatment with daratumumab in combination with bortezomib. The proportion of male patients (81.0%), the proportion of patients with cardiac involvement (95.2%), and the proportion of patients with Mayo 2012 stage ≥III (95.2%), as well as the levels of hs-cTnI and NT-proBNP in the deceased group were significantly higher than those in the survival group ( P < 0.05). The median OS time of the enrolled patients was 33.4(2.6-60.2) months, with 1-year, 2-year, 3-year and 5-year OS rates of 83.7%, 79.3%, 58.9% and 32.7%, respectively. The Kaplan-Meier survival curve analysis revealed that patients with male gender (P =0.040), NT-proBNP ≥3 600 ng/L ( P < 0.001), Mayo 2012 stage ≥III ( P < 0.001), and cardiac involvement (P =0.008) had poor prognosis and shorter overall survival (OS) time. The multivariate regression analysis showed that Mayo 2012 stage ≥III was an independent risk factor for prognosis. CONCLUSION In recent years, the survival rate of patients with AL amyloidosis has improved significantly, but the 5-year survival rate is still relatively low. Cardiac biomarkers (NT-proBNP and hs-cTnI) and Mayo 2012 stage at diagnosis continue to provide important prognostic information. Bortezomib-based regimens were used as the primary treatment in most patients, and the addition of daratumumab is becoming increasingly common.
Humans ; Retrospective Studies ; Prognosis ; Middle Aged ; Male ; Female ; Aged ; Immunoglobulin Light-chain Amyloidosis/diagnosis* ; Adult ; Aged, 80 and over ; Kaplan-Meier Estimate