Heteroresistance (HR) to commonly used anti-pseudomonal agents in Pseudomonas aeruginosa has been reported in cross-sectional epidemiological surveys; however, longitudinal studies investigating the long-term epidemiological dynamics and clinical correlates of this adaptive phenotype are still lacking. This retrospective time-series study aims to systematically explore the HR phenotypic profiles and temporal dynamic of clinical P. aeruginosa isolates against piperacillin/tazobactam (TZP) and meropenem (MEM), and further clarify their potential clinical correlates. We comprehensively analyzed 380 non-duplicate clinical strains collected from a tertiary hospital between 2011 and 2024. The disk diffusion and Etest gradient diffusion methods were used for the preliminary determination of HR incidence rates in clinical isolates against six anti-pseudomonal antibiotics. Population analysis profiling (PAP) assays was further performed to assess the temporal dynamics and phenotypic characteristics of HR to TZP MEM. HR was defined as the presence of resistant subpopulations with an MIC at least eightfold higher than that of the dominant population at a frequency of ≥ 1 × 10⁻⁷. Subsequently, univariate and multivariate regression analyses were performed to identify clinical factors associated with HR and non-HR strains. Preliminary screening indicated that 55

Among the intracellular sensors of innate immunity against infection, RIG-I-like receptors (RLRs) serve as cytosolic surveillance sensors that detect viral RNA species. However, the role of kinases in modulating RIG-I in a catalytically-independent manner is not well known. In this study, we report that protein kinase DYRK1B is a novel positive regulator of RIG-I-mediated antiviral innate immunity. Overexpression of DYRK1B markedly amplified RNA virus-induced IFN-I production, whereas CRISPR-mediated knockout of DYRK1B substantially attenuated these antiviral responses. Mechanistic investigation revealed that this regulatory role operates independently of DYRK1B's catalytic kinase function; instead, DYRK1B functions as an adaptor protein that connects TRIM25 with RIG-I, thereby promoting TRIM25-mediated K63-linked polyubiquitination of RIG-I, an essential modification required for RIG-I functional activation. Collectively, our findings reveal that DYRK1B enhances innate immunity against RNA viruses by strengthening the physical association of RIG-I with TRIM25, providing a more profound understanding of the mechanisms involved in antiviral immune regulation.

BACKGROUND:Helicobacter pylori can cause gastritis, peptic ulcers, and even gastric cancer. In addition to conventional T cells, mucosal-associated invariant T (MAIT) cells may have an important immunologic role in mucosal infectious diseases. However, the phenotypic and effector characteristics of gastric MAIT cells and the relationship between mucosal MAIT cells and H. pylori-related diseases have not been established. METHODS:We analyzed the phenotypes and cytokines of gastric and peripheral MAIT cells from H. pylori-infected patients by flow cytometry. CXCL12-mediated chemotaxis of MAIT cells was measured by Transwell assay. Relationship between MAIT cells and inflammation of H. pylori-infected gastric diseases was assessed with immunofluorescence staining and histopathological score. RESULTS:The number of gastric MAIT cells was significantly increased but the frequency of peripheral MAIT cells were decreased in patients with H. pylori infections. The gastric MAIT cells were activated with high expression of CD69 and CD38, and produced several pro-inflammatory cytokines. Furthermore, we observed the expression of CXCR4 and CXCL12 were upregulated in H. pylori-infected stomachs. CXCL12 promoted MAIT cells to migrate, which was blocked by a CXCR4 antagonist. Finally, we demonstrated that the number of gastric MAIT cells was positively correlated with severe inflammation in H. pylori-infected stomachs. CONCLUSIONS:Gastric MAIT cells were accumulated and activated in H. pylori-infected stomachs, which may be involved in the pathologic inflammation of H. pylori-related gastric diseases.

Background: Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive cancer with poor prognosis, characterized by a desmoplastic tumor microenvironment (TME) that limits immune cell infiltration and diminishes response to immunotherapy. Arylacetamide deacetylase (AADAC) is a lipid-processing enzyme, but its role in tumor progression, stromal organization, and immune modulation remains unclear. Methods: We integrated single-cell RNA sequencing (GSE212966) and spatial transcriptomics (GSE235315) to evaluate the association ofAADAC with extracellular matrix (ECM)-rich and immune-restrictive niches in PDAC. We performed spatial co-expression analysis, gene set variation analysis (GSVA) using a core COL6A1-ITGA2-ITGB1 signature, CellChat analysis of ligand-receptor interactions, and functional assays after AADAC knockdown in PANC-1 cells. Results: AADAC expression was significantly upregulated in PDAC epithelium and co-localized with ECM markers (COL6A1, ITGA2, ITGB1), cancer-associated fibroblast (CAF)-associated markers (podoplanin (PDPN), fibroblast activation protein (FAP)), and immunosuppressive genes (TGFB1, ARG1, CD163). Spatial analyses further showed that CD8A distribution was constrained within stromalized, suppressive niches, while AADAC knockdown in PANC-1 cells increased apoptosis and reduced proliferation, migration, and invasion. Conclusions: AADAC expression is associated with ECM-rich and immune-restrictive tumor regions in PDAC. These findings support AADAC as a candidate biomarker of epithelial-stromal-immune interactions and justify further mechanistic studies.

ABSTRACT The contact-induced epithelium gene A (iceA), with allelic variants iceA1 and iceA2, remains a controversial virulence factor in Helicobacter pylori due to inconsistent clinical outcomes. In this study, we analyzed 1,427 H. pylori genomes from the NCBI database and 200 clinical isolates from Shenzhen, China, to elucidate the global distribution of iceA genotypes and their associations with other virulence factors, bacterial gene expression, and the relationship with gastric diseases. We found that iceA genotypes varied significantly across regions, with iceA1 predominating in Asia and Europe, and iceA2 being more prevalent in North America. This suggests adaptive evolution to regional host environments. The association between iceA genotypes and cagA status also exhibited regional patterns. Additionally, iceA1 was linked to higher ureA expression, potentially enhancing acid resistance and colonization potential (P < 0.05). Importantly, patients infected with iceA1 genotype strains were more likely to present with gastric erosion (P < 0.05); however, they showed no association with peptic ulcer disease in any region. These findings highlight the geographical heterogeneity of iceA1 genotypes and their potential role in H. pylori-related gastric diseases.IMPORTANCEOur study found that the iceA1 genotype of Helicobacter pylori, which is relatively common in Asia, is associated with increased urea hydrolysis capacity and shows a specific association with erosive gastritis in Chinese patients. These findings suggest that iceA1 may potentially serve as an early biomarker for the identification of patients at risk of progressive gastric damage. Clinically, detecting iceA1 could improve risk stratification in high-prevalence regions, guiding more targeted monitoring and early intervention strategies. This research supports incorporating bacterial genotyping into clinical practice to optimize personalized treatment and prevent disease progression in susceptible populations.

Introduction Spinocerebellar ataxias (SCAs) types 1, 2, and 3 are the most common autosomal dominant SCAs, characterized by neurodegeneration in the cerebellum, brainstem, and basal ganglia. However, corticospinal tract involvement remains underexplored using combined electrophysiological and neuroimaging approaches. This study aimed to investigate the electrophysiological and imaging characteristics of SCA subtypes. Methods A total of 23 SCA1, 21 SCA2, and 40 SCA3 patients, along with 27 healthy controls, were enrolled. All subjects underwent neurological examination and motor evoked potential (MEP) testing. Additionally, SCA patients underwent diffusion kurtosis imaging (DKI). Correlations between central motor conduction time (CMCT) and corticospinal tract diffusion metrics, as well as clinical parameters, were analyzed. Results All SCAs demonstrated prolonged CMCT in both upper and lower limbs. SCA3 patients exhibited significantly reduced MD values relative to SCA2 (P= 0.0437). Conversely, both SCA1 and SCA2 patients showed significantly lower FA, MK, and KFA values compared to SCA3. In SCA3 patients, lower limb CMCT correlated with DKI metrics (FA: r = -0.4815, P < 0.05; MK: r = -0.4411, P < 0.05) and disease duration (r = 0.5647, P = 0.0003), but not with streamline counts. Conclusion This study identifies distinctive electrophysiological and microstructural corticospinal tract abnormalities across SCA cohorts. The observed correlations of CMCT with DKI parameters and disease duration in SCA3 suggest its potential as a biomarker for monitoring disease progression.

BACKGROUND:Heteroresistance (HR) in Pseudomonas aeruginosa causes misclassification as 'susceptible (S)' on routine antibiotic susceptibility techniques, potentially contributing to subsequent treatment failure. This study aimed to explore clinically relevant risk factors for HR compared with S (S-HR), and resistant (R) compared with HR (HR-R) phenotypes. Additionally, we explored whether integrating medical history and laboratory data can enable rapid and accurate identification of HR and R phenotypes in this pathogen. METHODS:This retrospective study included 420 P. aeruginosa strains collected from 2011 to 2024 in China. The strains were categorized into three groups according to their sensitivity to meropenem: non-heteroresistant susceptible, heteroresistant and non-heteroresistant resistant. Logistic regression, random forest and XGBoost models were constructed using variables identified through LASSO (least absolute shrinkage and selection operator) regression. The models' performance was evaluated via 10-fold cross-validation comparing area under the receiver operating characteristic curve (AUROC), sensitivity and specificity. RESULTS:Multivariate analyses identified central venous catheters as an independent risk factor for S-HR, and malignant solid tumours, pulmonary infections, mechanical ventilation and carbapenem use for HR-R. A discriminating diagnostic model, combining clinical and laboratory data, showed an AUROC of 0.919 for HR-R and 0.856 for S-HR. The calibration plots indicated good alignment between the estimated and observed probabilities. CONCLUSIONS:This study presents a validated two-stage risk assessment model to discriminate the two phases of meropenem heteroresistance in P. aeruginosa. By identifying novel stage-specific risk factors and delivering a practical tool compatible with clinical workflows, this model facilitates the early identification and targeted intervention of HR, offering novel insights into the mechanistic dissection of HR.

Objective This study aimed to evaluate the prognostic value of the Nutritional Risk Index (NRI) and Prognostic Nutritional Index (PNI) in patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) after curative resection. We further sought to elucidate the underlying genomic rationale linking NRI to tumor biology. Methods A retrospective cohort of 146 HBV-related HCC patients who underwent curative hepatectomy was analyzed. Propensity score matching (PSM) was performed to validate the robustness of NRI. Integrated bioinformatics analysis of the TCGA-LIHC cohort was conducted to explore the association between albumin (ALB) expression—the core component of NRI—and HCC driver gene mutations. An NRI-based nomogram was constructed for individualized survival prediction. Results NRI, but not PNI, was identified as an independent prognostic factor for both overall survival (OS) and recurrence-free survival (RFS). After 1:1 PSM, a high NRI (≥ 102.06) remained significantly associated with improved OS (P = 0.0293). Stratified analysis revealed that the prognostic power of NRI was most pronounced in early-stage (TNM Ⅰ+Ⅱ) and non-vascular invasion subgroups. Integrated analysis of TCGA data demonstrated that low ALB expression was robustly associated with TP53 and CTNNB1 mutations (P < 0.05), suggesting a link between NRI and genomic instability. The NRI-based nomogram, incorporating TNM stage, PIVKA-II, and vascular invasion, exhibited favorable discriminative ability (C-index: 0.78) and clinical utility. Conclusion NRI is a reliable and cost-effective prognostic indicator for HBV-related HCC, particularly in early-stage disease. Our findings reveal a "non-nutritional" face of NRI, suggesting that it serves as a serological signature of high-risk molecular subtypes characterized by genomic instability. The NRI-based nomogram provides a practical tool for risk stratification and clinical decision-making.

Only a minority of Helicobacter pylori (H. pylori)-infected individuals progress along Correa's cascade, and classical virulence markers do not fully explain this heterogeneity, motivating genome-wide approaches to quantify strain-level genomic risk associated with advanced lesions. We assembled 528 high-quality H. pylori whole-genome sequences spanning nonatrophic gastritis (NAG), atrophic gastritis (AG), intestinal metaplasia (IM), and gastric cancer (GC) and performed bacterial genome-wide association analyses with explicit adjustment for population structure. We then integrated advanced lesions-associated variants into a random forest model to derive an H. pylori Genomic Risk Score (HpRS), defined as the predicted probability that a strain is associated with advanced lesions (IM/GC) vs. nonadvanced lesions (NAG/AG). Despite phylogenetic analyses revealing that genome-wide clustering was primarily driven by geographic lineage rather than disease stage, HpRS achieved strong discrimination in repeated cross-validation (mean AUC = 0.902, 95% CI: 0.892 to 0.912), remained discriminatory in an internal held-out set (AUC = 0.780), and generalized to two independent cohorts (Bacterial and Viral Bioinformatics Resource Center (BV-BRC): AUC = 0.871; hospital cohort distinguishing IM vs. NAG/AG: AUC = 0.843). Predictive single-nucleotide polymorphisms mapped mainly to core functions (DNA repair, translation, and central and lipid metabolism) and often affected conserved domains, suggesting a polygenic architecture and generating testable functional hypotheses. HpRS provides a proof-of-principle framework for strain-aware risk stratification and may complement future gastric cancer prevention strategies.

ABSTRACT Antibiotic-resistant Helicobacter pylori ( H. pylori ) represents a significant global challenge to the efficacy of eradication therapies, underscoring the urgent need for rapid and precise pretreatment resistance profiling. Traditional genotyping assays frequently neglect synonymous mutations, resulting in false-negative results and reduced diagnostic accuracy. In this study, we developed a single-tube, synonymous codon-optimized multiplex real-time PCR (qPCR) assay for the qualitative detection of H. pylori and the simultaneous identification of mutations associated with resistance to clarithromycin (23S rRNA gene) and levofloxacin ( gyrA gene). The probe design was informed by clinical mutation data and synonymous codon usage patterns from 453 isolates, achieving over 98% coverage of naturally occurring variants. The assay exhibited high sensitivity (limit of detection: 2–20 copies/μL), excellent repeatability (coefficient of variation <5%), and robust specificity. Clinical validation using 316 gastric mucosal specimens demonstrated 97.47% concordance with a commercial infection detection kit and 100% and 98.08% agreement with Sanger sequencing for 23S rRNA and gyrA resistance genotyping, respectively. This assay effectively addresses the critical limitation posed by synonymous mutations, providing a rapid, reliable, and clinically accessible tool to guide personalized H. pylori eradication therapy and promote antimicrobial stewardship. IMPORTANCE Increasing antibiotic resistance in Helicobacter pylori has reduced the effectiveness of eradication therapy and highlights the need for accurate resistance profiling before treatment. However, current molecular assays may miss resistant strains because synonymous mutations in probe-binding regions may interfere with detection and produce false-negative results. In this study, we developed a single-tube multiplex real-time PCR assay that incorporates synonymous codon variation into probe design, enabling simultaneous detection of H. pylori infection and clarithromycin- and levofloxacin-resistance mutations. By using sequence data from 453 clinical isolates, this assay achieved broad coverage of naturally occurring variants and showed high analytical and clinical performance. This approach addresses an important diagnostic blind spot in H. pylori resistance detection and provides a practical tool for pretreatment resistance profiling, individualized therapy selection, and improved antimicrobial stewardship.

Gastric mucosal CD8+ tissue-resident memory T (CD8+TRM) cells are increased in Helicobacter pylori (H. pylori) infected subjects, but the characteristics and chemotactic mechanism of CD8+TRM cells remain unknown. We demonstrated that C-X-C chemokine receptor 5 (CXCR5) was upregulated on gastric mucosal CD8+TRM cells, and gastric CXCL13 was dominantly secreted by dendritic cells and macrophages in H. pylori infected subjects. Gastric mucosal CD8+TRM cells from CagA+ H. pylori infected subjects was increased and could secrete more IFN-γ and TNF-α to engage in local immune response. The number of gastric mucosal CD8+TRM cells and the expression of CXCL13 was elevated in H. pylori infected individuals with the development of gastric diseases. In conclusion, gastric mucosal CD8+TRM cells are increased from CagA+ H. pylori infected subjects and could be recruited through CXCL13-CXCR5 axis, which mainly release IFN-γ and TNF-α in H. pylori related immune response.

The widespread administration of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines has not eliminated breakthrough infections due to immune evasion by viral mutations. Whether individuals with breakthrough infections can elicit effective T cell responses against the circulating Omicron JN.1 variant, and the specific immunodominant characteristics of these responses, remain to be elucidated. To address this, we recruited 93 individuals who had experienced early Omicron subvariant breakthrough infections following a three-dose regimen of inactivated SARS-CoV-2 vaccines. Intracellular cytokine staining was employed to evaluate T cell responses to the JN.1 nucleocapsid (N) protein overlapping peptide pool in peripheral blood mononuclear cells. Immunodominant epitopes were identified using a three-dimensional matrix screening approach. Human leukocyte antigen (HLA) blocking assays and truncated peptides stimulation assays were conducted to define the minimal epitope and HLA restriction. We found that breakthrough infections elicited robust CD4+ and CD8+ T cell responses specific to the JN.1 N protein, characterized by Th1/Tc1-skewed phenotype and preserved polyfunctionality. The immunodominant epitope profile of the N protein has undergone a significant transformation following breakthrough infection. New epitopes targeting mutated sites have emerged, while previously dominant epitopes, such as N-4 and N-66, no longer maintained their immunodominance. Instead, highly conserved epitope N-24 has risen as the most immunodominant target for CD4+ T cells, restricted by HLA-DR, and with the minimal epitope identified as PKDHIGTRNPANNA. Our finding demonstrates that early Omicron strain breakthrough infections induce robust T cell responses targeting the circulating JN.1 subvariant and underscore the immunodominant features of these responses.

Background Spinocerebellar ataxia type 3 (SCA3) is a rare hereditary neurodegeneration disease. The iron distribution of SCA3 is poorly understood, yet quantitative susceptibility mapping (QSM) has rarely been used in SCA3. Methods We prospectively investigated QSM of SCA3 (19 pre-symptomatic and 41 symptomatic) and 37 healthy controls (HCs) recruited from 2018.05 to 2021.01. Group susceptibility was cross-sectionally compared, and the associations between altered brain iron deposition and clinical symptoms, neurofilament light chain (Nfl), and fractional anisotropy of the bilateral corticospinal tracts and cerebellar peduncles were explored. 12 SCA3 participants were followed for at least a year. Results Compared to HCs, bilateral SN were observed with significantly increased susceptibility in pre-symptomatic SCA3. Most of the supratentorial nuclei and the right dental nucleus had increased susceptibility in symptomatic than in pre-symptomatic stage and were partially correlated with symptomatic severity, disease duration, and damaged cerebellar peduncles (p < 0.05) but not Nfl (p > 0.05). The left substantia nigra (SN) demonstrated the highest diagnostic efficacy in identifying pre- (AUC = 0.904) and symptomatic SCA3 (AUC = 0.938). The longitudinal study also confirmed the significant change in the left SN (p < 0.01). Conclusions Our in vivo QSM evidence demonstrates disease-specific patterns for brain iron depositions in SCA3. Brain iron deposition abnormality is an early event of the SCA3's occurrence and development. The left SN might be a critical site for the disease's start and development.

Gut microbiota is essential for the function of peripherally-induced regulatory T (pTreg) cells. However, how commensal bacteria affect thymically derived fat-resident Treg cells that harbor a unique expression of peroxisome proliferator-activated receptor (PPAR)-γ and suppress inflammation in visceral adipose tissue (VAT), is not well defined. Here it is revealed that microbiota depletion causes a drastic decline in Treg cell population in VAT, particularly those expressing ST2 (ST2+ Treg), which are largely restored after gut microbiome reconstruction. Mechanistically, gut microbiota-derived butyrate increases VAT ST2+ Treg cells through binding PPARγ. Butyrate supplementation and high fiber diet increase VAT ST2+ Treg population in obese mice, and ameliorated glucose tolerance and visceral inflammation. Furthermore, human omental adipose Treg cells show positive correlation with fecal butyrate and certain butyrate-producing microbes. This study identifies the critical role of gut microbiota-butyrate-PPARγ axis in maintaining VAT Treg population, pinpointing a potential approach to augment VAT Treg population and ameliorate inflammation.

BACKGROUND:Brucellosis is a zoonotic disease distributed across numerous countries and regions worldwide, presenting with diverse clinical manifestations. The most common complication is spondylitis, which is diagnosed primarily through imaging studies. However, in resource-limited areas, the imaging examinations necessary for diagnosing brucellosis-related spondylitis are often inaccessible. The objective of this study is to establish a simple and readily predictive score within brucellosis patients to screen for spinal involvement. METHODS:We retrospectively collected patient data with brucellosis admitted to the Kashi Affiliated Hospital of Sun Yat-sen University from January 2019 to December 2023, and randomly assigned them into a training cohort and an internal validation cohort. Data of brucellosis patients admitted to The third Affiliated Hospital of Sun Yat-sen University from January 2014 to December 2023 were collected for an external validation cohort. A diagnostic model was constructed by using a nomogram. Calibration plots, receiver operating characteristic curve, and decision curve analyses were employed to evaluate the model's calibration, accuracy, and clinical utility. RESULTS:This study included data from total of 784 patients, of which 210 were diagnosed with Brucella spondylitis. The data was divided into a training cohort (460 patients), an internal validation cohort (198 patients), and an external validation cohort (126 patients). The diagnostic model was formulated using six diagnostic factors: course of disease, age, back pain, joint pain, white blood cell count, and levels of C-reactive proteins. In our study, the AUC values of 0.93 (training), 0.87 (internal validation), and 0.77 (external validation) indicate that the model maintained excellent discriminative ability in the training and internal validation cohorts, and acceptable performance in the external cohort. Decision curve analyses graphically display the significant clinical utility and net benefit of a nomogram. CONCLUSION AND RECOMMENDATION:The diagnostic model for Brucella spondylitis developed in this study has the potential to assist clinicians in resource-limited settings in achieving a rapid and effective diagnosis of the disease. Our model facilitates the identification of brucellosis-related spondylitis patients requiring extended treatment courses, thereby reducing misdiagnosis and missed diagnosis in resource-constrained areas where imaging examinations are difficult to access, and improving the efficiency of healthcare resource utilization.

BACKGROUND:Helicobacter pylori (H. pylori) infection is one of the most important risk factors for chronic gastritis, gastric ulcers, and gastric cancer. Mast cells act as a crucial regulator in bacterial infection. The mechanisms underlying mast cell activation and their role in H. pylori infection remain poorly understood. MATERIALS AND METHODS:In gastric mucosal tissue, the number of mast cells, G-protein-coupled receptor 171 (GPR171) and CCL2 expression were detected by immunohistochemistry (IHC) or immunofluorescence between H. pylori-negative and H. pylori-positive patients. Mast cells were co-cultured with H. pylori, and transcriptome sequencing, RT-qPCR, and Western blotting (WB) were performed to identify receptors involved in mast cell activation. WB, chromatin immunoprecipitation (ChIP), and dual-luciferase reporter assays were conducted to investigate the molecular mechanism by which HIF-1α regulates GPR171 expression. Lentiviral knockdown, ELISA, WB, and IHC were used to evaluate the role of GPR171 during H. pylori infection. An in vivo mouse model of H. pylori infection was employed to assess the effects of GPR171 blockade on CCL2 expression and gastric mucosal inflammation. RESULTS:In the study, we found that mast cell numbers were greatly increased and correlated with the severity of inflammation in H. pylori-infected patients. We found a new receptor, GPR171, was upregulated and involved in mast cell activation upon H. pylori infection. Furthermore, H. pylori infection induced the expression of GPR171 by promoting the activation of hypoxia-inducible factor 1 alpha (HIF-1α), which directly bound to hypoxia response elements in the GPR171 promoter and regulated its transcriptional activity. Blockade or loss of GPR171 in mast cells partially inhibited CCL2 secretion via the ERK1/2 signaling pathway. In the human gastric mucosa, CCL2 derived from mast cells was associated with gastric inflammation during H. pylori infection. In vivo murine studies indicated that H. pylori infection significantly upregulated CCL2 expression, while GPR171 inhibition partially reduced CCL2 levels and alleviated gastric mucosal inflammation. CONCLUSIONS:We provide a novel mechanism that H. pylori activates mast cells to promote gastric inflammation.

Spinocerebellar ataxia type 3 (SCA3) is characterized by extensive supra- and infra-tentorial brain injuries, beginning in the pre-symptomatic stage. However, the alterations and evolution of spontaneous brain activity in SCA3 patients remain poorly understood. This study aimed to investigate the abnormal brain activity of SCA3, classified it into distinct patterns, and explored the association between spontaneous brain activity in affected regions and clinical variables. Ninety-nine SCA3 patients (76 sym- and 23 pre-SCA3 patients, mean age: 39.97 ± 10.83y; 28.74 ± 7.99y) and 49 healthy controls underwent 3.0T MRI and acquired T1 and resting-state functional MRI images. The static and dynamic amplitude of low-frequency fluctuation (s/dALFF) and regional homogeneity (s/dReHo) were used to estimate local brain activity alterations. SCA3 patients exhibited significantly abnormal spontaneous brain activities in multiple regions, classified into four patterns (P < 0.001). Patterns 1 and 2 were found in the pre-symptomatic stage, involving the right opercular part of inferior frontal gyrus, right rolandic operculum, right insula, right angular gyrus, right crus I of cerebellar hemisphere and left superior frontal gyrus. Pattern 3 emerged in symptomatic SCA3, with expanded involvement of supra- to infra-tentorial regions. Pattern 4 showed increased activity in the right cerebellar lobule IX. Negative correlations were found between sReHo value in the right anterior cingulate cortex with the disease duration and severity (P ≤ 0.04). SCA3 patients exhibit different patterns of brain activity damage at different stages. These findings provide new insight into brain dysfunction in SCA3 and suggest potential imaging markers for disease staging and monitoring.

IntroductionNasopharyngeal carcinoma (NPC), one of the most common malignancies of the head and neck, is characterised by a complex pathogenesis and an unfavourable prognosis. Recently, disulfidoptosis, a novel form of cell death, has been proposed. Several studies in recent years have extensively investigated the function of the disulfidoptosis-related SLC7A11 gene in cancer, but the role of its partner protein, SLC3A2, remains unknown unclear in NPC.MethodsGEO database analysis confirmed SLC3A2's prognostic impact on nasopharyngeal carcinoma. ROC, Kaplan-Meier analyses, and stage-specific expression studies showed a strong correlation with poor HNSC prognosis. GO and KEGG analyses pinpointed relevant signaling pathways. In vitro, SLC3A2's influence on cell proliferation, migration, and invasion was evaluated through CCK8, wound healing, colony formation, transwell assays, and cell cycle analysis.ResultsIn this study, we identified the high expression of SLC3A2 in NPC and head and neck squamous cell carcinoma (HNSC) and analyzed its potential mechanism and correlation with patient prognosis. Furthermore, a negative relationship was found between the expression level of SLC3A2 and the extent of immune cell infiltration and immune checkpoint. Differentially expressed genes (DEGs) between the high and low SLC3A2 expression groups were primarily involved in cytokine-cytokine receptor interaction and immune response. Finally, in vitro experiments demonstrated that SLC3A2 stimulates tumor cell proliferation and migration. DiscussionIn conclusion, these results indicated a strong association between SLC3A2 and progression in both NPC and HNSC, suggesting it as a promising biomarker for predicting adverse prognosis in NPC and HNSC patients.

The ongoing evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants highlights the importance of monitoring immune responses to guide vaccination strategies. Although neutralizing antibodies (NAbs) have garnered increasing attention, T-cells are crucial for conferring long-lasting immunity, especially their resilience against viral mutations. However, assessing T-cell responses clinically has been hindered by cost and complexity. In this study, we recruited a cohort of 134 healthy adults, who had been immunized with three doses of the SARS-CoV-2 inactivated vaccine. Cellular immunity elicited by a comprehensive array of overlapping peptides covering the entire sequence of the virus’s structural proteins was assessed by intracellular cytokine staining (ICS). Additionally, a dataset including demographic information, routine blood indices, and immune cell indicators comprising 32 variables was collected. Multivariate analysis revealed age and days post-vaccination as key factors influencing the strength of the T-cell response. Importantly, random forest (RF) and classification and regression tree (CART) algorithms were employed, along with 8 easily accessible indicators to formulate predictive models for the SARS-CoV-2-specific CD4+ and CD8+ T-cell responses. Besides, these models demonstrated substantial accuracy (r > 0.9) in both the training and testing sets. Our findings offer an efficient and economical methodology for evaluating the T-cell reactions in healthy adults following inactivated SARS-CoV-2 vaccination, which is visualizable and easy to use, providing a novel strategy for assessing cellular immunity after vaccination.

OBJECTIVE:Helicobacter pylori infection has been reported to aggravate rheumatoid arthritis (RA), but the relevant mechanism remains unclear. This study aimed to investigate the underlying pathogenic mechanism of H. pylori infection in the progression of RA. METHODS:The Disease Activity Score (DAS-28) and serum anticitrullinated protein antibody (ACPA) levels were compared between H. pylori-negative and H. pylori-positive patients with RA. MH7A cells were stimulated with polyclonal ACPA purified from the peripheral blood of patients with RA. The citrullination levels were assessed by western blot in GES-1 cells and sera. ChIP, luciferase reporter assays, mass spectrometry and ELISA were applied to explore the molecular mechanism of H. pylori infection in RA progression. RESULTS:The DAS-28 and ACPA levels of patients with RA in the H. pylori-positive group were significantly higher than those in the H. pylori-negative group. Polyclonal ACPA derived from H. pylori-positive patients promoted cell proliferation and induced secretion of IL-6 and IL-8. For the first time, we found that H. pylori infection induces cellular protein citrullination by upregulating protein arginine deiminase type 4 (PAD4). Furthermore, we confirmed a direct functional binding of hypoxia-inducible factor 1α on the PADI4 gene promoter. We demonstrated that PAD4 interacts with and citrullinates keratin 1 (K1), and serum and synovial fluid levels of anti-Cit-K1 antibody were markedly increased in H. pylori-infected patients with RA. CONCLUSION:Our findings reveal a novel mechanism by which H. pylori infection contributes to RA progression. Therapeutic interventions targeting H. pylori may be a viable strategy for the management of RA.