Journal of Advanced Research

Osteoporosis affects millions of women globally. In this study, we applied bioinformatics methods to screen for novel diagnostic biomarkers of osteoporosis in women using the GSE62402 and GSE56814 datasets. PCSK5, ZNF225, and H1FX were used to construct a diagnostic model. ROC, calibration, and decision curve analyses were performed to assess the diagnostic performance on the training (GSE56814) and external (GSE56815) datasets. The expression level of model genes was validated in GEO datasets. Furthermore, five transcription factors (ETS1, NOTCH1, MAZ, ERG, and FLI1) were identified as common upstream regulators of model genes. PCSK5, ZNF225, and H1FX serve as novel diagnostic biomarkers, providing new insights into the pathogenesis of and treatment strategies for osteoporosis in women.

Staphylococcus aureus plays a central role in the exacerbation of atopic dermatitis (AD), but the population structure and pathogenic determinants of strains colonizing AD patients remain poorly understood. It is unclear whether these strains mirror those circulating in the general community or whether specific clonal lineages are selectively adapted to the AD skin microenvironment. Data addressing this question are scarce, particularly in Portugal. In this study, we investigated the molecular epidemiology and pathogenic traits of S. aureus colonizing skin lesions in adult patients with AD in Portugal. We found that lesion-associated isolates belonged predominantly to the methicillin-susceptible S. aureus MSSA-ST398 clonal type, a lineage that is widely circulating in the Portuguese community, particularly among vulnerable populations, and that has also been implicated in severe human infections. Notably, isolates from this clonal type in AD harboured specific pathogenicity traits associated with skin barrier disruption, including hemolysin and urease production, which may contribute to their success as colonizers in AD. Our findings highlight that S. aureus colonization in AD arises from a dynamic interplay between community-level molecular epidemiology and disease-specific selective pressures. While circulating lineages provide the genetic background diversity, the AD skin microenvironment appears to shape which clones ultimately become dominant. Such an integrated perspective may help to inform future geographically tailored strategies aimed at limiting bacterial burden and preventing disease exacerbation in AD.

Post-acute sequelae of SARS-CoV-2 infection (PASC), commonly referred to as Long COVID, comprise a constellation of persistent, recurrent, or newly emerging symptoms that may endure for months or years following acute infection. Beyond respiratory impairment, PASC is characterized by a wide spectrum of extrapulmonary manifestations, among which neurological and neuropsychiatric symptoms are highly prevalent. Reported features include olfactory dysfunction with loss of smell and taste, fatigue, neuroinflammation, cognitive and memory impairment, depression, and anxiety, with some symptoms persisting up to one year post-infection. Despite increasing recognition of these complications, the molecular mechanisms underlying post-COVID neurological sequelae remain poorly defined. In this study, we employed a label-free quantitative (LFQ) proteomics approach to investigate protein alterations in olfactory neuroepithelium-derived stem cells (ONEs), a unique population of neural progenitors located in the olfactory mucosa at the interface between the respiratory system and both the peripheral and central nervous systems. Due to their anatomical exposure and susceptibility to SARS-CoV-2, ONEs represent a highly relevant translational model for exploring virus-associated neurobiological processes. ONEs derived from healthy donors were incubated with serum from either asymptomatic PCR-positive individuals (AS; n=4) or critically ill hospitalized patients (CR; n=6). Proteomic profiling revealed a distinct differential protein expression pattern in ONEs exposed to CR serum compared with AS serum. Altered pathways were associated with viral infection responses, respiratory and cardiovascular dysfunction, and notably, cerebrovascular and nervous system disorders. These findings highlight the vulnerability of ONEs to systemic factors associated with severe COVID-19 and provide molecular insight into mechanisms potentially contributing to persistent neurological sequelae in PASC. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/710460v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@12cfda5org.highwire.dtl.DTLVardef@c0636borg.highwire.dtl.DTLVardef@bf303eorg.highwire.dtl.DTLVardef@1f861e9_HPS_FORMAT_FIGEXP M_FIG C_FIG

Vitiligo is an autoimmune disorder characterized by melanocyte destruction. We performed a rank-based meta-analysis of six independent transcriptomic studies (115 samples) spanning microarray, bulk and single-cell RNA-seq platforms to identify consensus signatures of lesional skin. Robust Rank Aggregation identified 114 differentially expressed genes (FDR < 0.05) with striking asymmetry: 108 downregulated versus 6 upregulated. Downregulated genes were dominated by melanocyte markers (MLANA, TYRP1, DCT, PMEL, KIT). Upregulated genes included interferon-stimulated genes (OAS1, OAS2, EPSTI1). Pathway-level meta-analysis confirmed uniform suppression of melanogenesis, while immune activation was heterogeneous across datasets. Single-cell data from three included studies confirmed melanocyte depletion. The 108 downregulated genes showed exclusive expression in melanocytes. These include neural genes (PLP1, GPM6B, NRXN3), consistent with melanocytes neural crest origin. We also identified candidate melanocyte markers such as CYB561A3 and QPCT with high melanocyte specificity and consistent downregulation in vitiligo. These findings reveal a robust melanocyte loss signature in vitiligo detectable across all platforms, and study-dependent immune activation possibly influenced by sampling method and disease characteristics.

Nitrogen mustard (NM)-caused severe cutaneous damage lacks effective targeted therapies. Vitamin D3 (VD3) shows promise as a therapy for NM-induced dermal toxicity; however, the underlying mechanisms remain elusive. Herein, we initially confirmed that NM induced gut flora dysbiosis, characterized by a decrease of Akkermansia muciniphila (AKK) abundance, thereby leading to butyrate reduction. Antibiotics (ABX) significantly promoted NM-induced skin injury, whereas fecal microbiota transplantation of the controls feces (HC-FMT) or AKK administration attenuated NM-induced dermal toxicity. HC-FMT or AKK significantly increased butyrate levels in feces and serum of NM-treated mice. Butyrate notably attenuated ABX-caused acceleration of NM-induced skin injury. Meanwhile, NM markedly decreased the expression of -defensins, MMP7, and VDR. NM failed to further decrease AKK abundance and BA contents in intestinal MMP7-deficient mice, which was abolished by human alpha defensin 5 (HD5) overexpression. And intestinal MMP7 deficiency enhanced NM-caused skin injury, which was markedly attenuated by HD5 overexpression, AKK transplantation, or BA supplementation. Moreover, NM also failed to further reduce MMP7 and -defensin expression, AKK abundance, and butyrate levels in intestinal VDR-silenced mice. Finally, VD3 remodeled the gut microbiome particularly enriching AKK, increased butyrate contents and promoted the expression of -defensins, MMP7, and VDR, thereby attenuating NM-induced skin damage. The protective effect of VD3 against NM-caused dermal toxicity was abolished by either ABX or intestinal-specific knockdown of MMP7 or VDR in mice; however, this impairment was reversed by butyrate or AKK. In conclusion, VD3 attenuated NM-caused dermal toxicity by promoting BA production via remodeling the gut microbiota, and this effect was partially mediated by the intestinal VDR--defensin signaling pathway. These highlight that targeting the gut flora or supplementing with BA could be potential therapies for NM-induced dermal toxicity.

Major depressive disorder (MDD) involves dysregulated neuroimmune, metabolic, and oxidative stress (NIMETOX) pathways. Recently, it was shown that NIMETOX pathways should be evaluated in MDD patients stratified for metabolic syndrome (MetS). The current study aims to characterize the metabolic hormone and adipokine profiles of Chinese MDD patients stratified for MetS and to delineate their associations with overall severity of depression (OSOD), suicidal ideation (SI), recurrence of illness (ROI), and physiosomatic symptoms. We enrolled 125 MDD inpatients and 40 healthy controls and measured fasting serum insulin, glucose, glucagon, Glucose-dependent Insulinotropic Polypeptide (GIP), Glucagon-Like Peptide-1 (GLP-1), leptin, secretin, Plasminogen Activator Inhibitor-1 (PAI-1), resistin, ghrelin, and adiponectin, as well as the acute-phase inflammatory (API) response using albumin, transferrin (Tf), and monomeric CRP (mCRP). The results revealed a distinct metabolic hormone and adipokine signature in MDD with significantly lower insulin, glucagon, and PAI-1 levels, alongside an elevated API index (after adjusting for age, MetS, and body mass index). A composite GAP index (ghrelin, adiponectin, PAI-1) correlated negatively with OSOD, SI, ROI, physiosomatic symptoms, and adverse childhood experiences (ACEs). Integrative modeling combining the GAP index, API index, and ACEs achieved an area under the receiver operating characteristic (ROC) curve of 0.864 with an accuracy of 80% for discriminating MDD from controls. In conclusion, the findings delineated that many inpatients with severe MDD suffer from suppressed anabolic hormones and lower adipokine levels coupled with a mild, chronic inflammatory response. The deviations in this "hormonal-immune-metabolic" axis are components of the NIMETOX pathways in MDD and are not associated with MetS.

Syphilis remains a major public health concern. However, current serologic assays are limited in their ability to distinguish active from previously treated disease. We applied tandem mass tag-based quantitative proteomics to plasma from 10 adults with active syphilis and 10 age- and gender-matched non-diseased controls. We identified 54 differentially regulated proteins (36 upregulated, 18 downregulated). Those proteins map to immune and inflammatory responses, acute-phase signaling, coagulation and vascular pathways, and cellular stress processes. Three sets of between 2-5 proteins achieved >99% discrimination between cases and controls. Our exploratory findings support proteomics as a potential tool to develop novel syphilis diagnostics.

BackgroundWhile monosodium urate (MSU) crystal deposition within coronary plaques links hyperuricemia/gout to cardiovascular risk, the existence and functional consequences of MSU deposits within the myocardial interstitium or cardiomyocytes remain unknown. MethodsForensic autopsy myocardial and renal specimens from gout patients (n=4) and controls (n=4) were analyzed for MSU crystals using compensated polarized microscopy, laser capture microdissection coupled with HPLC-MS, and cryo-electron microscopy (cryo-EM) with electron diffraction. Pharmacological (adenine-diet) and genetic (Uox-/-) hyperuricemia murine models were assessed by crystal detection, echocardiography, and correlative histopathology. ResultsMyocardial MSU crystals were identified in all human gout cases (absent in controls), localized perivascularly, interstitially, and intracellularly within cardiomyocytes. Definitive verification was provided by compositional analysis (HPLC-MS m/z 167) and crystal structure confirmation (cryo-EM diffraction matching MSU). Hyperuricemic mice exhibited myocardial MSU deposition, progressive diastolic dysfunction (r =0.7014 vs. crystal burden), and reduced survival associated with higher crystal load. Importantly, myocardial crystal burden correlated more strongly with cardiac impairment than serum uric acid levels. ConclusionThis study provides conclusive morphological, chemical, and crystallographic evidence of intracardiac MSU deposition in gout. It directly links myocardial crystal accumulation to diastolic dysfunction, proposing "uric acid cardiomyopathy" as a novel disease entity. Collectively, these findings achieve a paradigm shift in the perception of gout, redefining it from a peripheral arthropathy to a systemic disorder capable of causing direct myocardial injury.

Patients with HbE/{beta}-thalassemia inheriting the same {beta}-globin mutations display varied clinical manifestations, the mechanism of which is only partially known. The study aimed to decipher the heterogenous basis of HbE/{beta}-thalassemia patients in more details by focusing on both hematological and genetic modifiers influencing the disease severity, which included-(i) HbF and HbE levels using Hb electrophoresis, (ii) {beta}-thalassemia mutations, (iii) anti3.7triplication using Gap-PCR, (iv) individual and cumulative effects of HbF-inducing SNPs in 4 major modifier genes, namely HBG2, BCL11A, HBSB1L_MYB intergenic-region, and HBBP1 which were genotyped using DNA sequencing and Real-time PCR-HRM methods. Accordingly, 130 diagnosed Bangladeshi patients with HbE/{beta}-thalassemia were enrolled and categorized as mild, moderate, and severe as per Mahidol scoring system. c.79G>A+IVS1_5G>C was the most predominant (73.8% of total) mutation pair across all the 3 severity groups, indicating secondary modifiers might influence the severity. Our study found both HbF and HbE protective to HbE/{beta}-thalassemia, as both were inversely related to the severity score (HbF: p<0.0001/r=-0.55; HbE: p<0.0001/r=-0.56). Four SNPs-XmnI-G{gamma}, rs2071348 (HBBP1), rs489544 and rs28384513 (HBS1L_MYB) showed significant association with the elevated HbF levels (p=0.005, 0.0001, 0.0001, 0.004 respectively). The multivariate analysis showed that the risk genotypes with single or combination of 2, 3,and 4 SNPs showed gradually increased risk [Odd Ratio (95%CI)= 2.51, 5.47, 19.5, 39.0, respectively] of less severe phenotype, suggesting that these linked SNP variants had a cumulative effect on both HbF level and clinical severity score. However, low HbE level and copresence of anti3.7triplication were found to nullify the ameliorating effect of multiple SNPs.

Despite important advances in understanding the etiopathology of multiple sclerosis, factors determining disease progression remain partially understood and often difficult to predict. Specific diagnostic and prognostic biomarkers are needed to optimize the risk-benefit ratio of treatment for each patient. The aim of our study was to identify a cerebrospinal fluid proteomic signature associated with diagnosis and short- to mid-term prognosis across the multiple sclerosis continuum. Our multicentric cohort study analyzed CSF samples from 120 patients using a proteomics data-independent acquisition strategy. Differentially expressed proteins were identified across diagnostic groups: 62 patients with multiple sclerosis, 15 patients with clinically isolated syndrome, and 43 healthy controls. We also compared the CSF of patients with no evidence of disease activity with those with disease activity at 2 and 5 years of follow-up. A diagnostic and prognostic classification model was built using iterative cross-validated logistic regression models on shared differentially expressed proteins across these two comparisons. A total of 1,257 proteins were quantified, and 162 differentially expressed proteins were identified across comparisons. We identified a set of ten proteins associated with the diagnosis and prognosis of multiple sclerosis, including previously identified potential biomarkers (CH3L2, IGHG1, IGKC, LAMP2, ADA2), proteins known to be involved in the pathophysiology of multiple sclerosis (A0A8J8YUT9, AT2A2, CO3A1) and two yet unreported proteins (DSC2 and MMRN2). Multivariate models based on these proteins achieved good accuracy for the diagnosis of MS compared with CIS (area under the receiver operating characteristics curve [AUROC] up to 80% using 3 proteins) and prognosis (NEDA vs. EDA; AUROC up to 96% at 2 and 5 years; using 5 proteins). These results, which will require further investigation to validate the new biomarkers, open new perspectives on multiple sclerosis pathophysiology and therapeutic targets.

Schizophrenia (SCZ) and type 2 diabetes mellitus (T2DM) are common comorbid disorders that severely impair patient prognosis and quality of life. This study aimed to explore the association between the methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism and MTHFR promoter methylation in patients with comorbid SCZ and T2DM. A total of 120 participants were enrolled from Liaocheng Fourth Peoples Hospital between January 2025 and June 2025, comprising 30 subjects in each of the four groups: SCZ group, T2DM group, SCZ-T2DM comorbid (SCZ+T2DM) group, and healthy control (CTL) group. Corresponding primers were designed for genetic analysis, and methylation-specific PCR (MSP) was performed to detect the methylation level of the MTHFR promoter. Genotype distribution of the MTHFR C677T polymorphism was consistent with Hardy-Weinberg equilibrium (HWE) (p>0.05). The C677T polymorphism was significantly associated with an elevated risk of SCZ and T2DM comorbidity (p<0.05). Notably, the methylation rate of the MTHFR promoter in the SCZ+T2DM group (95.00%) was not significantly higher than that in the CTL group (90.00%) (p>0.05). In conclusion, the MTHFR gene may serve as a susceptibility gene for SCZ-T2DM comorbidity, whereas MTHFR promoter methylation is not associated with the pathogenesis of this comorbid condition. These results indicate that genetic variation in MTHFR, rather than promoter methylation, contributes critically to the comorbidity of SCZ and T2DM in the Han Chinese population. Our findings may provide novel molecular insights into their shared pathophysiology and inform future clinical strategies for patients with this complex phenotype.

Chronic traumatic encephalopathy (CTE) is a progressive neurodegenerative disease associated with repeated head injuries (RHI) commonly experienced by contact sport athletes, military personnel, and domestic abuse victims. Despite growing recognition of CTE, the molecular mechanisms underlying disease progression remain poorly understood. This study aims to identify proteomic alterations associated with CTE pathology and clinical features to elucidate key biological pathways involved in disease pathogenesis. SomaScan 7k high-throughput proteomics was performed on 204 dorsolateral prefrontal cortex samples from the Boston University CTE Center Brain Bank. We identified differentially expressed proteins associated with CTE, hyperphosphorylated tau (ptau) pathology, duration of contact sports play, dementia status, and Cognitive Difficulty Scale (CDS) scores. Gene set enrichment analysis revealed that proteasome subunit proteins and related pathways were strongly associated with CTE progression and correlated with years of contact sports play. Reduction in ribosomal proteins and pathways was closely associated with ptau burden. Additionally, multiple models demonstrated significant alterations in MAPK-related cell signaling pathways. These findings advance our understanding of CTE progression and identify mechanisms correlated with key pathological features of the disease. Validation of these results could inform the development of diagnostics and treatments for CTE.

ObjectivePreterm birth (PTB) is a leading cause of neonatal morbidity and mortality worldwide, with India alone contributing nearly 27% of the global PTB burden. Although alterations in the vaginal microbiome have been implicated in PTB, its association in the Indian context is underexplored. This study aimed to investigate the association of vaginal microbiome and PTB in Indian women at the time of delivery. Study designThe vaginal swabs were collected at the time of delivery from 72 women (31 term, 41 preterm) admitted to a tertiary care hospital in Western India. Microbial DNA was extracted, and the V3-V4 region of the 16S rRNA gene was sequenced. Community composition, alpha and beta diversity, and differential taxonomic abundance were assessed using bioinformatics pipelines. ResultsAt the time of delivery, there were no significant differences in alpha or beta diversity between term and preterm groups. Principal coordinate and unsupervised clustering analyses showed no group-wise segregation. The relative abundance of individual Lactobacillus species, including L. iners and L. helveticus, did not differ significantly between the two groups. However, a modest difference in the relative abundance of Streptococcus was observed between the two groups after adjustment. ConclusionThis study found no major microbial shifts in the vaginal microbiome associated with preterm birth in this cross sectional cohort of Indian women, suggesting that vaginal dysbiosis at the time of delivery may not be a principal driver of PTB in this population. These findings underscore the need for larger, longitudinal, and ethnically diverse studies using standardized methodologies better to understand the microbiomes role in PTB risk.

AimsTo characterize subtype-associated heterogeneity in type 2 diabetes mellitus (T2DM), particularly normal-weight diabetes, using extracellular vesicle (EV)-associated molecular features in a clinically stratified cohort. MethodsEVs were isolated from plasma using ExoTIC and validated by transmission electron microscopy, nanoparticle tracking analysis, flow cytometry, and Western blotting. EVs from Asian normal-weight (A-NWD), Asian overweight (A-OWD), Non-Hispanic White normal-weight (W-NWD), and Non-Hispanic White overweight (W-OWD) T2DM patients were analyzed by multimodal surface-enhanced Raman spectroscopy (SERS; n=65) and EV-RNA sequencing (n=39). ResultsSERS identified subgroup-associated spectral fingerprints that distinguished the four BMI- and race/ethnicity-defined groups in this cohort. EV-RNA sequencing revealed differential microRNA expression across subgroups, with higher miR-208a and miR-132 in A-OWD and higher miR-484 in A-NWD. Unsupervised analyses also showed partially overlapping EV-associated molecular features between A-NWD and W-OWD, suggesting that BMI-based subgrouping alone may not fully capture shared metabolic states. ConclusionsMultimodal EV profiling identified subgroup-associated spectral and miRNA features in clinically stratified T2DM and provides a framework for studying diabetes heterogeneity, including molecular patterns associated with normal-weight diabetes.

IntroductionEarly-life adversity (ELA) encompasses a range of environmental stressors, including physical, emotional, and social challenges that can affect health during the critical early developmental period. Extensive research has linked ELA to negative long-term health outcomes, yet the underlying biological mechanisms remain poorly understood. The current study investigates how early institutional care changes the epigenetic landscape in young adults. The study also provides insights into role of DNA methylation as a potential mediator for disease susceptibility and altered health trajectories. Materials and MethodsDNA was extracted from blood samples obtained from 111 individuals (71 Controls; 40 ELA) who were part of the EpiPath cohort. DNA methylation was measured using the Infinium Methylation EPIC v2.0 BeadChip. Results3,785 differentially methylated CpG loci were identified in the ELA group in comparison with the control group (FDR <0.05). Pathway enrichment analysis highlighted biological processes involved in metabolic regulation, stress response, and neurodevelopment, with novel pathways such as GTPase-mediated signalling, efferocytosis and glucuronosyltransferase emerging as potential drivers of the ELA phenotype. A subset of 28 CpG loci was used to develop an epigenetic signature, which showed a significant association with the development of chronic diseases in ELA-exposed individuals. ConclusionThis study reinforces Barkers concept of sensitive periods and it underscores the enduring impact of ELA in shaping long-term health outcomes. The persistence of DNA methylation patterns decades after exposure to ELA, and their clear association with the resultant phenotype confirms that stable epigenetic imprints play a potential role in long-term disease risk and resilience.

Neuropsychiatric systemic lupus erythematosus (NPSLE) is a potentially severe complication of systemic lupus erythematosus (SLE), yet its pathogenesis remains largely elusive. By jointly probing the immune dynamics of subjects cerebrospinal fluid (CSF) and peripheral blood, we showed that both innate and adaptive immune responses jointly contribute to the pathogenesis of NPSLE. In particular, we found the remarkable enrichment of BAM-CCL3, a subtype of border-associated macrophages with strong recruitment capacity, implicating its potential role in central nervous system (CNS) inflammation. We also observed pronounced activation of memory B cells and CD4+ regulatory T cells in NPSLE CSF, along with the preferential blood-to-CSF migration and subsequent within-CSF clonal expansion of CD8+ effector memory T cells in NPSLE patients, suggesting a persistent CNS-localized adaptive immune dysregulation. Finally, we developed the single-cell CNS disease CSF-Blood Atlas (scCDCB), a comprehensive collection for CSF and peripheral blood of multiple CNS diseases, which is publicly available at (https://sccdcb.gao-lab.org) to serve as a reference for future research on CNS diseases.

Vitiligo is an acquired pigmentary disorder of the skin and mucus membranes. Previous study has demonstrated that autologous cultured epithelial grafts (ACEG) is an effective treatment for stable vitiligo. However, extraction of full-thickness skin might result in scar formation at donor site, which have hindered the wider application of this technology, especially for patients requiring large-area transplantation. Hair follicle as a source of keratinocyte and melanocyte, could be potential source of cells for preparation of autologous cultured sheet. Through culture system optimization, we have demonstrated maintenance of undifferentiated hair follicle-derived cells in feeder-independent culture system. After expansion, the hair follicle cells were directed to differentiate into a multi-layered, epidermis-like sheet. Cell identity, viability, purity, genomic stability, and antiseptic testing for hair follicle-derived epithelial sheet (HFES) were evaluated to ensure its safety. Immunofluorescence staining showed that basal keratinocytes were the main cell type of the autologous HFES. Optimization of culture conditions leads to increased melanocyte proliferation and functionality. Transcriptomic analysis confirmed upregulation of melanosome maturation genes. The proportions of cells are also similar to composition of cells under physiological conditions. Transplantation of HFES to depigmented areas in patients with stable vitiligo results in skin repigmentation. This technology provides a novel therapeutic option for vitiligo management.

To understand the molecular and cellular mechanisms beyond B-cell depletion with the anti-CD20 monoclonal antibody ocrelizumab, we used comprehensive muti-modal flow cytometry and functional assays in a prospective longitudinal multiple sclerosis (MS) cohort. Ocrelizumab depleted the vast majority of B cells and showed selective effects on different B cells subsets. Analysis of residual/replenished B cells revealed relative enrichment of regulatory B cells like CD27+CD43+ B1 and CD24hiCD38hi transitional B cells, and reduction of CD27+ memory B cell subsets and CD19+IgD+CD27-naive B cells at early time points (1-3 month) and before subsequent infusions at 4-7 months, 11-14 months, and >18 months. CD20+ T cells and peripheral helper T-cells (Tph) were also reduced. RNA sequencing analysis showed B1 cells have significantly higher expression of LGALS1, KCNN4, ITGB1, and IL2RB. Compared to transitional B cells, B1 cells also displayed significantly higher expression of tissue homing molecules ITGAX (CD11c), S100A4, ITGB1, and CXCR3. IL10 signaling pathway is increased in these B cells. Ex vivo B cell functional assays indicated the residual/replenishing B cells were anergic following ocrelizumab, with increased IL10/TNF and IL10/IL6 ratios under BCR stimulation. Ocrelizumab treatment may create a self-reinforcing regulatory circuit: the reduction of Tph cells could alleviate suppression of regulatory B cells, which subsequently expand and further promote regulatory T cell networks via IL2RB, LGALS1, and an increased IL-10 signaling pathway.

An overlap syndrome of myositis and/or myocarditis associated with myasthenia gravis (MG) has emerged as a life-threatening immune-related adverse event (irAE) in cancer patients treated with immune checkpoint inhibitors (ICIs). This syndrome closely resembles a rare form of idiopathic inflammatory myopathy (IIM) seen in a subset of MG patients. In this systematic review, we searched PubMed for reports of concurrent MG and IIM as well as ICI-related overlap syndromes. By integrating clinical, serological, and pathological observations, we delineated a previously unrecognized clinicopathological subtype of myositis that overlaps with MG. This entity is defined by a strong association with striational antibodies (StrAbs) and frequent co- occurrence with thymoma as a paraneoplastic process, and we classify it as StrAb-associated myositis. The idiopathic and ICI-induced forms share similar, though not identical, clinical, serological, and histopathological characteristics. We found that AChR antibody positivity, independent of established clinical risk factors such as respiratory or cardiac involvement, predicted more severe ICI-myotoxicity. Together with supporting evidence, our findings suggest a pathogenic model in which thymoma-driven cytotoxic T-cell responses trigger secondary AChR autoimmunity. These results highlight the potential utility of StrAbs and anti-AChR antibodies as practical biomarkers for diagnosis, risk stratification, and early intervention in patients at risk for severe neuromuscular irAEs.

BackgroundEndometriosis is a heterogeneous disease in which anatomical lesion burden often shows poor correlation with pain severity and quality-of-life impairment. While classification systems such as the revised American Society for Reproductive Medicine (r-ASRM) and Enzian score accurately describe anatomical disease extent, their relationship to symptom burden and reproductive outcomes remains incompletely understood. Objective(s)This study aimed to investigate the relationships between anatomical disease extent, pain severity, quality-of-life impairment, and fertility outcomes across ovarian, deep, and peritoneal endometriosis in a prospective cohort of women undergoing surgical treatment. Study DesignThis prospective observational cohort study included women aged 18-45 years undergoing laparoscopic surgery between 2023 and 2025 at a tertiary endometriosis center. Participants were categorized into ovarian (OE), deep (DE), or peritoneal (PE) endometriosis based on imaging and intraoperative findings. Pain severity was assessed using numeric rating scales across multiple domains, and quality of life was evaluated using the Endometriosis Health Profile (EHP-30+23). Anatomical disease burden was determined using r-ASRM and Enzian classifications. Patients were followed for 12 months after surgery to assess symptom trajectories, pregnancy outcomes, and surgical complications. A subset of lesion samples underwent RNA sequencing to explore molecular signatures associated with pain severity. ResultsA total of 145 women were included (OE n=33, DE n=55, PE n=25, controls n=32). Pain severity showed limited correlation with anatomical staging across subtypes. In contrast, infertility and the need for ureter surgery were strongly associated with higher Enzian scores and structural disease burden. Quality-of-life impairment closely paralleled pain intensity rather than anatomical stage. Transcriptomic analysis identified a molecular signature associated with high pain burden characterized by increased expression of inflammatory mediators (IL6, CCL8, SPP1), endocannabinoid system components (PENK, CNR1) and nociceptive transcription factors (NR4A3, EGR3). Longitudinal follow-up demonstrated substantial postoperative improvement in pain and quality of life independent of pregnancy outcomes. ConclusionsPain severity, quality-of-life impairment, and reproductive dysfunction in endometriosis represent partially independent dimensions of disease activity. While neuroinflammatory mechanisms appear to drive pain and quality-of-life impairment, fertility outcomes and organ-threatening complications are primarily determined by structural disease burden. Integrating anatomical staging with multidimensional symptom assessment and molecular profiling may enable more personalized management strategies for women with endometriosis.