In advanced melanoma, longitudinal disease monitoring remains limited by infrequent biomarker assessment and predominantly categorical imaging readouts. Although circulating tumor DNA (ctDNA) correlates with tumor burden, its behavior under high-frequency sampling and its relationship to quantitative metastatic tumor volume are poorly defined. Clarifying these dynamics is essential to establishing ctDNA as a clinically actionable tool for real-time treatment monitoring.

We retrospectively analyzed 42 patients with unresectable stage III/IV melanoma treated with immune checkpoint inhibitors. Plasma ctDNA was quantified longitudinally across 241 time points using a UMI-based amplicon next-generation sequencing (NGS) assay detecting BRAF, EGFR, KRAS, NRAS, and PIK3CA. We paired ctDNA measurements with radiologic staging and correlated them with response categories (n = 99 time points) and volumetric tumor burden (n = 73 time points) using nonparametric statistics and receiver operating characteristic (ROC) analyses, including stratification by the ctDNA-imaging time interval and descriptive assessment of discordant patterns.

ctDNA levels were significantly associated with radiologic response according to RECIST 1.1, increasing across worsening response categories (Spearman ρ = 0.31, 95% CI 0.20-0.52, p = 0.002). Discriminative performance for identifying progression was moderate (AUC = 0.69). Threshold analyses demonstrated a sensitivity-specificity trade-off, with higher ctDNA levels providing greater specificity (e.g. ≥25 mutant molecules (MM)/mL: specificity ≥ 83%, sensitivity ≤ 30%), while lower thresholds showed limited sensitivity and specificity (e.g. 1-2 MM/mL: ~56% each). ctDNA concentrations were positively associated with total tumor volume (ρ = 0.46, p < 0.0001), with stronger correlations observed for temporally aligned measurements (0-30 days: ρ = 0.56). In contrast, dynamic changes in ctDNA were not significantly correlated with changes in tumor volume (ρ = 0.20, p = 0.25). Organ-associated analyses demonstrated marked heterogeneity in ctDNA shedding, with stronger associations in lymph node and peritoneal metastases and limited detectability in lung, liver, and brain metastases. Concordance between ctDNA dynamics and radiologic response was high in responding disease (CR/PR: 85.0%) but lower in stable or progressive disease (72.9%), with discordant cases associated with tumor heterogeneity, timing differences, and treatment-related factors.

ctDNA reflects radiologic disease status and tumor burden, with increasing levels observed as response worsens. Despite heterogeneity in ctDNA shedding across metastatic sites, its significant correlation with tumor volume, particularly when closely timed to imaging, supports the potential value of ctDNA as a complementary biomarker for longitudinal disease monitoring while highlighting important biological and methodological factors that influence its clinical interpretation.

[18 F]-fluorodeoxyglucose (FDG) positron emission tomography combined with computed tomography (PET-CT) is used for staging and treatment response evaluation in patients with locally advanced cervical cancer. While the maximum standardised uptake value (SUVmax) is a commonly used PET parameter, volume-based parameters (metabolic tumour volume (MTV) and total lesion glycolysis (TLG)) may offer superior predictive value. This study aimed to investigate the predictive value of different treatment response outcomes assessed with PET-CT, using both standard and volume-based PET parameters.

We retrospectively analyzed 133 patients with locally advanced cervical cancer treated with (chemo)radiotherapy who underwent PET-CT pre- and post-treatment. SUVmax, MTV and TLG were semi-automatically extracted from all scans, including tumour, lymph nodes and metastases. Treatment response was classified as complete metabolic response (CMR) (complete resolution of FDG uptake in all lesions), partial metabolic response (PMR) (> 25% reduction of SUVmax/MTV/TLG), progressive metabolic disease (PMD) (> 25% increase of SUVmax/MTV/TLG or appearance of new FDG-avid lesions) or stable disease (SD) (does not qualify for CMR, PMR or PMD). The three PET-parameters were analysed separately. PMR and SD were analysed as one group due to limited number of events. Associations between treatment response and overall survival (OS) as well as recurrence (binary variable) were analysed through Cox and logistic regression models, respectively.

For recurrence (binary variable), the odds ratio (OR) was 1.2 for patients with PMR/SD for SUVmax, MTV and TLG, while in the group of patients with PMD, it was 1.7 for SUVmax and 1.8 for MTV and TLG, using CMR as reference (i.e. for CMR, the OR is 1.0). The hazard ratio (HR) for OS was 4.0 for patients with PMR/SD and 16.0 for those with PMD, based on SUVmax; for MTV and TLG, HRs were 2.9 for patients with PMR/SD and 16.0 for those with PMD, using CMR as reference (HR 1.0). C-index from adjusted Cox models were comparable: 0.84 for SUVmax, and 0.86 for both MTV and TLG.

FDG-PET/CT-based assessment of treatment response is a strong predictor of recurrence and overall survival in locally advanced cervical cancer. SUVmax, MTV, and TLG demonstrated similar predictive performance.

Treatments in the phase of metastatic castration-resistant prostate cancer (mCRPC) aim to prolong survival and reduce the symptom burden. The fast development of life-prolonging therapies makes treatment decision-making (TDM) complex, weighting survival against possible side-effects and quality of life, especially in a real-world context. This study aimed to describe satisfaction with TDM at start of the first, second and third line of life-prolonging treatment and to describe and compare long-term treatment experiences when receiving one or several lines of treatment in patients with mCRPC.

A longitudinal observational study. Participants completed a study-specific questionnaire every three months for up to two years from the start of any first line of life-prolonging treatment after mCRPC. Satisfaction with TDM was measured at the start of each new treatment line while treatment experiences were measured using the first and last completed questionnaire of each participant. Medical data was obtained from medical records. Descriptive statistics were used to present data while inferential statistics were used for analysis of group comparisons and over time changes.

A total of 122 participants (mean age 75.3 years) with an average follow-up time of 18.3 months were included. Satisfaction with TDM was generally high at the start of the first, second and third line of treatment, but lower regarding discussions on how treatment could affect their life situation. Treatment experiences were positive, yet there was a significant decrease from the first to the last follow-up in whether they would choose (p = 0.04) and recommend (p = 0.01) the same treatment to others. For participants who received one line of treatment, a significant decrease was found between the first and the last follow-up (p = 0.02), while no differences were found for those who received two or more lines.

Despite high satisfaction with aspects of TDM and the treatment given it is important to discuss how the treatment can impact the patient's life situation. The slightly decreased satisfaction over time on the treatment received could reflect a possible need to strengthen communication between patients and healthcare professionals, shifting from a disease-centered perspective towards a more person-centered and palliative care approach.

Myeloid-derived suppressor cells (MDSCs) play a crucial role in the tumor microenvironment (TME) of cervical cancer (CC), yet the mechanisms underlying their reprogramming remain poorly understood.

To explore the immune microenvironment change in CC, we applied TCGA-CC immune microenvironment infiltration estimation analysis via Timer 2.0 online datasets. To generate tumor-conditioned MDSCs, the culture medium of MDSCs was supplemented with supernatants from the murine CC cell lines U14 and TC1, respectively. CCK8 assays, transwell migration experiments and qPCR were executed to test the proliferation, migration and iNOS expression. We then conducted proteomics and metabolomics analyses of tumor-conditioned MDSCs.

The tumor immune microenvironment analysis identified MDSCs as key components, predicting poor prognosis in CC. Tumor-conditioned MDSCs presented higher proliferation, migration and iNOS expression. Proteomics and metabolomics analyses showed significant changes in lipid metabolism, especially sphingolipid metabolism. Specifically, the Kng1-sphingosine 1-phosphate axis was identified as a central protein-metabolite regulatory node. Gain- and loss-of-function experiments confirmed that KNG1 modulates multiple cellular processes including proliferation, migration, iNOS expression, and sphingosine 1-phosphate production.

Our findings uncover sphingolipid metabolic reprogramming as a key mechanism in MDSCs-mediated immune suppression, and propose the Kng1-sphingosine 1-phosphate network as a potential therapeutic target for CC treatment.

Hepatocellular carcinoma (HCC) is a major contributor to cancer-related mortality worldwide and is closely linked to oxidative stress and progressive hepatic dysfunction. This study investigated the effects of plumbagin on antioxidant defense systems and hepatic function markers in an N-nitrosodiethylamine-thioacetamide (NDEA-TAA)-induced experimental model of HCC and compared its efficacy with that of sorafenib. Male Wistar rats were assigned to five groups: normal control, plumbagin-only, HCC control, HCC treated with plumbagin, and HCC treated with sorafenib. Hepatocarcinogenesis was induced using NDEA-TAA, after which antioxidant parameters including reduced glutathione (GSH), glutathione-S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA) were evaluated alongside hepatic function markers alanine aminotransferase (ALT) and albumin (ALB). The HCC control group exhibited significant reductions in GSH, GST, GPx, CAT, and SOD activities, accompanied by elevated MDA and ALT levels and decreased ALB concentrations (p ≤ 0.05), indicating severe oxidative stress and hepatic injury. Treatment with plumbagin significantly restored antioxidant enzyme activities, reduced lipid peroxidation, and improved hepatic function relative to the untreated HCC group. These effects were comparable to, and in certain parameters exceeded, those observed following sorafenib administration. The findings demonstrate that plumbagin possesses substantial antioxidative and hepatoprotective properties capable of mitigating oxidative damage and improving liver function in experimental HCC. These findings support further investigation of plumbagin as a redox-modulating candidate in HCC and a basis for future studies incorporating combination treatment, molecular validation and translational models.

The nervous system exhibits remarkable stability, making its repair challenging. Compared to the central nervous system, peripheral nerve injuries demonstrate a greater potential for regeneration. Schwann cells, the unique myelinating glial cells of the peripheral nervous system, possess striking plasticity and support nerve regeneration through phenotypic transformation following injury. While Schwann cells are essential for nerve regeneration after injury, in the context of tumors, their reparative function is exploited to facilitate perineural invasion and the progression of cancer. However, the tumor-promoting effect of Schwann cells is context-dependent, and emerging evidence suggests that under specific conditions, Schwann cells may exert tumor-suppressive functions. This article aims to explore the complex roles of Schwann cells in both physical nerve injury and tumor-induced nerve damage, comparing their similarities and differences and seeking to provide insights for future research and potential therapeutic interventions.

Plasmacytoid dendritic cells (pDCs) have been implicated in both restraining and promoting intrahepatic cholangiocarcinoma (iCCA), leaving their clinical relevance and therapeutic potential unresolved.

Mendelian randomization was used to assess the causal association between circulating pDC levels and iCCA risk. Bulk and single-cell transcriptomic analyses were performed to characterize pDC-related programs and tumor-conditioned states, and multiplex immunofluorescence was used to define spatial distribution and clinical associations in iCCA tissues. To assess reversibility of stress-associated pDC features, IRE1α RNase activity was pharmacologically inhibited with 4µ8C under tumor-conditioned stress in vitro.

Genetically predicted higher circulating pDC levels were associated with lower iCCA risk, consistent with a systemic protective association. In bulk cohorts, higher expression of pDC markers (CLEC4C, NRP1, IL3RA) was associated with an immune-inflamed microenvironment and improved survival in early-stage disease. Single-cell analyses indicated that intratumoral pDCs acquired stress-associated transcriptional programs, including enrichment of endoplasmic reticulum stress and unfolded protein response pathways. In vitro, 4µ8C reduced IRE1α-dependent XBP1 splicing and partially restored type I interferon-linked activation and pDC immunogenic readouts under tumor-conditioned stress. Spatial profiling further showed that higher intratumoral CD303⁺IRF7⁺ pDC activation was associated with advanced stage and poorer overall survival, whereas higher activation in adjacent non-tumor tissues correlated with more favorable outcomes.

Together, these findings support a context-dependent, stress-associated pDC program in iCCA and provide a rationale for further evaluating the IRE1α-XBP1 stress axis as a potential approach to modulate pDC-associated immune states within the tumor microenvironment.

Polycystic ovary syndrome (PCOS) is a complex endocrine-metabolic disorder characterised by hypothalamic-pituitary-gonadal (HPG) axis dysfunction and pathological gonadotropin-releasing hormone (GnRH) hypersecretion. While metformin demonstrates therapeutic efficacy in PCOS, its potential neuroendocrine mechanisms remain incompletely understood. This study aimed to investigate the involvement of hypothalamic signalling in metformin-responsive neuroendocrine regulation in PCOS.

Letrozole-induced PCOS rat models were treated with metformin to assess reproductive and metabolic outcomes. Data-independent acquisition (DIA) proteomics of hypothalamic tissue was performed to identify dysregulated pathways, followed by network analysis to explore candidate regulatory nodes. Mechanistic investigations were conducted in glucosamine-induced insulin-resistant GT1-7 GnRH neurons using pharmacological modulation and siRNA knockdown. In vivo functional relevance was examined using AAV-mediated Epac2 knockdown in the mouse arcuate nucleus.

Metformin improved metabolic and reproductive abnormalities in PCOS rats, including insulin resistance, GnRH/LH hypersecretion, estrous cyclicity disruption, and ovarian morphological alterations. Hypothalamic proteomic and network analyses suggested that metformin-responsive changes were associated with insulin and cAMP-related signalling pathways. In GT1-7 neurons, metformin increased Epac2 expression, restored AKT signalling, and reduced GnRH overproduction under insulin-resistant conditions. Hypothalamic Epac2 knockdown reduced AKT signalling and was associated with systemic insulin resistance, increased GnRH/LH secretion, and preliminary supportive ovarian morphological observations, consistent with a functional contribution of Epac2 to reproductive-metabolic regulation.

These findings support the involvement of hypothalamic Epac2/AKT/GnRH signalling in metformin-responsive neuroendocrine regulation in PCOS. Further studies are required to determine the relative contributions of central and peripheral mechanisms.

Cardiovascular-kidney-metabolic (CKM) syndrome imposes a substantial global burden, yet most studies reduce stage change to a binary outcome that cannot distinguish clinically meaningful trajectories. This study developed and externally validated a multiclass machine learning framework for predicting CKM stage trajectories.

A longitudinal cohort of 2,971 adults with baseline CKM stages 0-3 was used for development, with trajectories classified as Improvement, Stable, Mild Progression and Rapid Progression. Six algorithms were compared by Macro-AUC and decision curve analysis. SHAP values identified consensus predictors for a parsimonious model, externally validated in an independent hospital-based cohort (N = 291).

XGBoost achieved the highest Macro-AUC (0.786, 95% CI 0.757-0.813), with the six algorithms performing comparably. For rapid-progression screening, the primary model reached a sensitivity of 0.767, a specificity of 0.739 and a negative predictive value of 0.949 at a threshold of 0.23, and identified 41.9% of rapid progressors within the highest-risk 10%. Baseline CKM stage was the dominant prognostic determinant, with a stage-only benchmark reaching 0.689. Six consensus predictors were identified: baseline CKM stage, age, fasting glucose, TyG-BMI index, systolic blood pressure and triglycerides. The parsimonious model retained over 98% of full-model discrimination and reached a Macro-AUC of 0.752 externally without retraining.

This framework distinguishes CKM trajectories and is intended for screening rapid progression rather than assigning individuals to a single trajectory. Baseline CKM stage is the principal prognostic determinant, and the six-predictor model preserves discrimination using routinely available measurements, supporting stratified follow-up pending confirmation in larger cohorts.

Very small unruptured intracranial aneurysms (VSUIAs) measuring ≤ 3 mm represent a unique clinical challenge, balancing the low but non-negligible lifetime risk of rupture against the procedural risks of endovascular intervention. This study reports the safety, efficacy, and angiographic outcomes of endovascular coiling for VSUIAs from the ECOSA (Endovascular Coiling of Small Aneurysms) multicenter registry.

We performed a subgroup analysis of the ECOSA multicenter registry of patients with VSUIAs (maximum diameter ≤ 3 mm) who underwent endovascular coiling. Patient demographics, aneurysm characteristics, procedural details, periprocedural complications, clinical outcomes (modified Rankin Scale; mRS), and angiographic follow-up data were analyzed.

A total of 116 patients with 116 VSUIAs underwent endovascular coiling. The mean age was 52.3 years (SD 12.1), and the majority were female (75.0%). The mean aneurysm diameter was 2.66 mm (SD 0.44) and mean neck diameter was 2.1 mm (SD 0.87). Most aneurysms were discovered incidentally (78.4%). Balloon-assisted coiling was used in 14.7% and stent-assisted coiling in 23.3% of cases. The thromboembolic event (TEE) rate was 1.7%, with no symptomatic events. Intraprocedural rupture occurred in 0.9% of cases with no intraprocedural mortality. Among 98 patients with clinical follow-up (median 7.75 months), 93.9% of patients achieved good functional outcomes (mRS 0-2), and all-cause mortality rate was 1.0%. Among 80 patients with angiographic follow-up (median 6 months), adequate occlusion (RROC I-II) was achieved in 86.3%.

Endovascular coiling of VSUIAs ≤ 3 mm is feasible and associated with a low complication rate, high rates of good functional outcomes, and durable angiographic occlusion. These findings support the safety of elective intervention in carefully selected patients with high-risk morphological or clinical features. Larger prospective studies are needed to define optimal patient selection criteria for VSUIAs.