Functional/dissociative seizure (FDS) disorder presents diagnostic challenges due to its complex semiology and the lack of positive diagnostic biomarkers. Current evidence suggests that specific patterns of brain activity and distinct dynamics in the brain network may underlie the neurobiological mechanisms of FDS. We retrospectively analyzed ictal EEG recordings from 46 patients with FDS, in three different conditions: during resting wakefulness (RW), during resting while having eyes closed (EC), and during functional/dissociative seizures of variable semiology (FDS). The occipital EEG in the alpha band (8-13 Hz) was assessed in terms of power and functional connectivity, quantified by debiased weighted phase lag index (dwPLI). Heart rate was also analyzed across conditions. Statistical comparisons were performed via paired permutation testing with Benjamini-Hochberg correction for multiple comparisons, complemented by Bayes factors for the key FDS-EC comparisons. EEG during FDS exhibited significantly higher occipital alpha power than RW (p = 0.0003) and levels not significantly different from EC, with moderate Bayesian evidence for the absence of a difference (BF01 = 6.1). Alpha power was elevated across all FDS subtypes and was, paradoxically, numerically highest in hypermotor seizures, although subtype differences did not survive correction for multiple comparisons. Control analyses showed that the alpha power did not significantly differ across video-rated states of eyes during FDS (open, intermittently closed, closed), including when conditions were matched for eye state. Functional whole-brain connectivity in the alpha band was intermediate between the two resting conditions, being higher during FDS than during RW but lower than during EC. Heart rate was highest during FDS, with a non-significant tendency toward higher rates in hypermotor than atonic FDS. No significant correlation was found between occipital alpha power and heart rate during FDS. Our findings suggest that FDS are characterized by a paradoxical hypersynchronization in the EEG alpha band, similar to EC, despite concurrent motor activity. Increased functional alpha connectivity, simultaneous with elevated heart rate during FDS, suggests that FDSs involve altered brain network dynamics rather than simple alpha power suppression by voluntary motor activity. These findings may aid in developing positive biomarkers for FDS diagnosis.
Previous studies have shown that the level of oxidative stress in individuals with recurrent pregnancy loss (RPL) is significantly higher than in fertile individuals. Alpha-lipoic acid (ALA) has been considered an antioxidant that can improve male fertility. This study aimed to investigate the antioxidant effect of ALA on sperm parameters and DNA fragmentation index (DFI) in couples with a history of RPL. In this randomized, double-blind clinical trial, 64 couples with a history of RPL, who were referred to the Milad Infertility Center, Mashhad, Iran, between February 2021 and March 2022, initially enrolled. Of these, 40 couples completed the three-month follow-up and were included in the final analysis. The intervention group received ALA and folic acid for three months, while the control group received a placebo and folic acid for the same duration. Baseline sperm parameters, including concentration, motility, and morphology, were assessed alongside the sperm DFI before and three months after the intervention. There were no significant differences in baseline characteristics between the two groups (P>0.05). After three months, however, sperm concentration, motility and morphology significantly improved in the intervention group. In contrast, while sperm concentration improved in the control group, no significant changes were observed in sperm motility or morphology. There were no significant differences between the two groups regarding sperm parameters before and after the intervention (P>0.05). DFI showed a significant reduction in the intervention group compared to the control group (P<0.05). ALA supplementation appears to positively influence sperm parameters and reduce DFI within individuals in couples with a history of RPL. While within-group improvements were noted, between-group differences in sperm concentration, motility, and morphology were not statistically significant. These findings suggest a potential role for antioxidant therapy in the management of male infertility, though further studies are needed to confirm efficacy (registration number: IRCT20181030041503N4).
Antibody-drug conjugates (ADCs) are an effective treatment for recurrent/refractory testicular germ cell tumors (rrTGCTs). In this exploratory study, we report disease stabilization achieved with the ADC Mirvetuximab soravtansine (MIRV) in a heavily pretreated patient with rrTGCT and compare this ADC with a novel antibody-polymer-drug conjugate (APDC) targeting folate receptor α (FRα), MIRV-P-MMAE. APDC MIRV-P-MMAE with a drug-to-antibody ratio ∼23, was synthesized using orthogonal attachment of a highly hydrophilic copolymer to the hinge region of anti-FRα antibody. Its efficacy was compared with MIRV in vitro in TGCT cells and in vivo in a metastatic model of human testicular choriocarcinoma (CHC). Furthermore, we evaluated FRα expression in TGCT and assessed efficacy of MIRV in an rrTGCT patient. Our investigations revealed the highest antiproliferative effect of MIRV-P-MMAE in FRα-positive CHC cells, JAR and JEG3, with IC50 values of 19.8 pM and 22.5 pM, respectively. These IC50 values are 210- and 70-fold lower than those of MIRV. Cytotoxicity was also confirmed in SuSa, PA1, and NCR-G1 cell lines. High efficacy was demonstrated in animals intravenously injected with JAR-luc cells, in which APDC treatment significantly prolonged survival (p = 0.0002) and completely cured 3 of 8 animals. Our data showed FRα membrane staining in 5,1% of TGCT samples, but confirmed cytoplasmic positivity in all 5 viable rrTGCT post-chemotherapy patient samples, suggesting that APDC with high DAR could be explored in rrTGCT patients with low expression of the target antigen. Our data warrant further exploration of FRα-targeting APDC as a novel approach for treating FRα-expressing rrTGCT.
This methodological study was conducted to evaluate the Turkish validity and reliability of the "Instrument of Self-care for Prevention of Diabetic Foot" developed by Herrera-Guerra and Bautista-Arellano. This research is of methodological type. The study was conducted with 202 diabetic individuals followed up at a university hospital in Türkiye. Translation-back translation was used to assess linguistic validity, and the opinions of 11 experts were consulted for content validity. Confirmatory Factor Analysis (CFA) was applied to assess construct validity, and the Diagonally Weighted Least Squares (DWLS) estimator was used in the analyses. For reliability analysis, internal consistency coefficients (Cronbach's alpha and McDonald's omega) were calculated, and a test-retest reliability test was performed on 26 patients at four-week intervals. The content validity index was found to be 0.98. CFA results showed that the three-factor structure of the scale (Self-Care Monitoring, Self-Care Maintenance, and Self-Care Management) is suitable for the Turkish sample and that the model fit indices are at an excellent level (GFI = 0.989, CFI = 0.995, RMSEA = 0.055, SRMR = 0.061). Cronbach's alpha coefficients for the sub-dimensions of the scale were found to be in the range of 0.859-0.885, and McDonald's omega coefficients were in the range of 0.869-0.887. In the test-retest analysis, it was determined that there was no systematic deviation between the measurements in the Bland-Altman plots and that there was a very high level of positive correlation between test and retest scores (r = 0.992-0.994). As a result of the research, it was determined that the Instrument of Self-care for Prevention of Diabetic Foot is a valid and reliable scale in Turkish. This tool can be used as a guide in nursing care in terms of preventing diabetic foot ulcers and monitoring self-care of patients with diabetes in the Turkish population.
Poor sleep is linked to adverse health outcomes. Animal studies suggest the gut microbiome may influence sleep, but human findings remain inconsistent. We examined associations of self-reported insomnia symptoms, daytime sleepiness, and actigraphy-measured sleep with gut microbiome diversity and composition in older adults. We studied 869 Baltimore Longitudinal Study of Aging participants with self-reported sleep and shotgun metagenomic sequencing; 332 also had actigraphy. We tested associations of sleep with alpha diversity, beta diversity, and species composition using regression, PERMANOVA, and ANCOM-BC2, adjusting for age, sex, BMI, physical activity, education, and depressive symptoms. Participants had mean age 70.7 years; 54.8% were female and 66.9% White. Trouble falling asleep ≥5 times/week was associated with higher Shannon diversity (β=0.41 SD; 95% CI: 0.09, 0.73) and Pielou's evenness, but not richness metrics. No actigraphy-measured sleep variables were associated with alpha or beta diversity. Beta diversity analyses suggested excessive sleepiness (1-2 o 3-4 times/week) was associated with different microbial composition, though variance explained was small. In species-level analyses, frequent insomnia symptoms or excessive sleepiness were associated with depleted or undetected Eubacterium sp. CAG:251. In exploratory actigraphy models, each doubling of sleep efficiency was associated with higher Eubacterium sp. CAG:251 prevalence (PR=2.15; 95% CI: 1.47, 3.14), while each 30-minute increase in wake after sleep onset was associated with lower prevalence (PR=0.49; 95% CI: 0.29, 0.81). Global diversity findings were limited and inconsistent, whereas subjective and objective sleep disturbances converged on Eubacterium sp. CAG:251. Findings are exploratory and require longitudinal replication.
Carbapenem-resistant Enterobacterales (CRE) pose a critical threat in Intensive Care Units (ICUs) within low- and middle-income countries (LMICs), with prevalence exceeding 20% and mortality reaching 50%. Despite high clinical demand, routine screening remains unstandardized in Vietnam due to resource constraints and a lack of validated tools. This study aimed to develop and validate a psychometric instrument to evaluate healthcare workers' (HCWs) perspectives across three domains: barriers, feasibility and readiness. A cross-sectional validation study was conducted using total population census sampling (n=97, total front-line staff) across three hospitals in Vietnam (Viet Tiep, Kien Thuy, and Thuy Nguyen). Following expert panel review (S-CVI/Avg = 0.89) and a 10.3% pilot test (n=10), construct validity and reliability were evaluated via Exploratory and Confirmatory Factor Analysis (EFA/CFA). Group differences were analyzed using Fisher's Exact and Chi-square tests. The barriers subscale was excluded due to low internal consistency (alpha < 0.60), yielding a refined 16-item, two-factor instrument (6.1:1 observation-to-item ratio). The final tool demonstrated excellent reliability (alpha: 0.84-0.89). CFA confirmed a robust model fit for the Feasibility and Readiness structure [GFI/CFI = 0.95; RMSEA = 0.062]. HCWs reported high Readiness but moderate Feasibility. Significant occupational divergences emerged: 88.9% of nurses viewed sampling as simple (Item R4) compared to only 45.8% of laboratory technicians (p = 0.003). Furthermore, an experience-perception paradox was identified (p < 0.05); mid-career staff (5-10 years) reported the lowest feasibility scores (50.0% high score) compared to junior and senior cohorts. The psychometric validation resulted in a methodologically sound 16-item tool tailored for LMICs. The divergence between clinical readiness and laboratory feasibility highlights critical implementation bottlenecks. Sustainable transition to routine CRE surveillance requires resource-optimized targeted workflows to reduce frontline workloads, alongside peer-led governance that leverages senior expertise to mitigate mid-career burnout.
Falls are common in individuals with dementia and pose a significant problem in acute hospital care. Aging and dementia affect functional brain networks including the default mode network (DMN), dorsal attention network (DAN) and salience network (SAN). The DAN and SAN are essential for the rapid detection of responses to unexpected stimuli. We hypothesize that global efficiency within these networks is reduced in patients with dementia who experience falls compared with those who do not. We analyzed 347 inpatients with dementia from a neurological clinic in Vienna, Austria. Of 153 available EEGs, only those with normal findings were included. FC was calculated using the phase-locking-value across delts, theta, alpha, and beta bands. The characteristic path length of the DAN, SAN and DMN was computed. Fallers showed reduced global efficiency in the DAN. FC between the right rostral middle frontal gyrus and left middle frontal gyrus was decreased in delta, theta, and alpha bands - regions involved in visual stimulus processing and executive control. Reduced DAN connectivity may impair the ability to respond to unexpected stimuli, increasing fall risk in individuals with dementia. This supports the role of attention network dysfunction in motoric cognitive risk syndrome. Falls are a significant problem in dementia, not well understood so far. Identifying network level dysfunction in dementia related falls offers a novel biomarker for risk stratification and possible targets for future interventions.
The Trier Social Stress Test (TSST) is a well-established laboratory stress-inducing paradigm. However, its implementation requires a research environment with dedicated personnel (usually at a research institution). When participants cannot travel to a research institution (due to age, mobility, or geographical restrictions), a home version of the TSST is needed. We have developed the TSST-home, which shares the essential characteristics with the laboratory version but is implemented at the participant's home. In this pilot study, we aimed to investigate the feasibility of the TSST-home and its ability to induce stress responses in comparison to a non-stressful control condition. Seventeen healthy men (mean age and range: 24 years; 18-30 years) were exposed, in randomized order, to the TSST-home and the placebo-TSST-home (control condition) in their respective homes. During each session we assessed perceived stress, salivary cortisol, salivary alpha-amylase, heart rate to account for the multifaceted nature of acute stress responses. Conservative and liberal cortisol responder rates (≥2.5 and ≥ 1.5 nmol/l increase) in the TSST-home condition were 70% and 82%, respectively. The TSST-home and placebo-TSST-home differed significantly in their trajectories of stress markers with stronger stress-induced responses across all stress measures in the TSST-home condition. Findings suggest that the TSST-home elicits reliable and valid acute stress responses across different stress markers comparable to those found in the original laboratory-based TSST. This tool can be used to investigate stress responses of participants who cannot otherwise participate in laboratory-based studies at research institutions.
Although the association between diabetes and cutaneous squamous cell carcinoma (cSCC) is well recognized, the specific role of insulin resistance (IR) as an independent driver of cSCC pathogenesis remains underexplored. This review synthesized emerging evidence on the ultraviolet (UV)-independent molecular mechanisms by which IR promotes cSCC initiation and progression. Hyperinsulinemia activates the insulin-like growth factor-1 receptor (IGF-1R), which triggers both the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) and phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathways, stimulating keratinocyte proliferation and suppressing apoptosis. In parallel, hyperglycemia-driven formation of advanced glycation end products (AGEs) and oxidative stress cause deoxyribonucleic acid (DNA) damage and impair tumor suppressor functions, notably that of tumor protein p53 (TP53). The resulting reactive oxygen species (ROS) activate nuclear factor-kappa B (NF-κB), establishing a chronic inflammatory milieu that remodels the tumor microenvironment through cytokines, including interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α), and by upregulating matrix metalloproteinases (MMPs). These processes collectively facilitate the malignant transformation of actinic keratosis (AK) to invasive cSCC. The analysis in the present study identified novel therapeutic targets and reaffirmed the importance of further studies on microbiome interactions and lifestyle interventions for IR-associated cSCC.
Methamphetamine (METH) is a widely misused psychostimulant whose pulmonary toxicity is poorly characterised. Omega-3 fatty acids (Omega-3 FAs) are nutritional lipids with established antioxidant and anti-inflammatory properties, yet their capacity to protect against METH-induced lung injury has not been studied. To investigate the effects of chronic METH exposure on pulmonary oxidative stress, antioxidant defences, inflammation, apoptosis, and energy metabolism, and to evaluate whether omega-3 FA supplementation attenuates selected markers of METH-induced pulmonary injury. Twenty male Wistar rats were allocated to four groups (n = 5): Control, METH (10 mg/kg/day), Omega-3 (300 mg/kg/day EPA/DHA), and METH + Omega-3. All treatments were administered once daily by oral gavage for 60 consecutive days. Lung homogenates were analysed for pro-oxidant, antioxidant, inflammatory, apoptotic, and metabolic markers. Pulmonary histopathology was evaluated using H&E-stained sections. METH significantly increased MDA, H2O2, XO, uric acid, DFI, MPO, LDH, IL-1β, TNF-α, caspase-3, pyruvate, and lactate while reducing GPx, GSH, GST, total thiol, NO, and Ca2+-ATPase (all p < 0.05). Histopathology revealed pneumonitis with perivascular inflammatory infiltration in METH-only animals. Omega-3 co-supplementation significantly attenuated several injury markers, with statistically confirmed rescue of MPO, DFI, LDH, Ca2+-ATPase, lactate, and GPx. Attenuation of MDA, H2O2, IL-1beta, TNF-alpha, caspase-3, and uric acid was observed numerically but did not reach statistical significance versus METH alone. The pyruvate and lactate accumulation in METH animals, combined with Ca2+-ATPase suppression, indicates a shift toward anaerobic glycolysis consistent with mitochondrial respiratory dysfunction; omega-3 co-supplementation partially corrected this bioenergetic profile. The pyruvate and lactate accumulation in METH animals, combined with Ca2+-ATPase suppression, indicates a shift toward anaerobic glycolysis consistent with mitochondrial respiratory dysfunction; omega-3 co-supplementation partially corrected this bioenergetic profile. Omega-3 fatty acids attenuated METH-induced pulmonary oxidative stress, inflammation, apoptosis, and metabolic disruption in this preclinical model. These findings justify further dose-response and mechanistic studies before any clinical translation can be considered. not applicable.
Emotion recognition is an important component for enabling machines to perceive and respond to human emotions. Existing electroencephalogram (EEG)-based emotion recognition methods often rely on single-band signals or purely spatial representations, which may fail to capture complementary spectral information and left-right hemispheric asymmetry. To address these limitations, we propose a multi-band spatial asymmetry convolutional neural network (MBSACNN) for EEG-based emotion recognition. After baseline signal removal, EEG signals are decomposed into four frequency bands, namely theta, alpha, beta, and gamma. Based on the international 10-20 electrode system, two types of 3D inputs are constructed: the original EEG matrix (OEM) and the spatial asymmetric EEG matrix (SAEM). OEM preserves the original spatial distribution of multi-band EEG activities, whereas SAEM explicitly encodes the signed differences between symmetric electrode pairs to represent hemispheric asymmetry. A 2D CNN is then used to extract spatial-temporal features from the dual-input representations. Experiments on the DEAP dataset show that MBSACNN achieves average accuracies/F1-scores of 97.07%/97.19% for arousal and 96.61%/96.89% for valence, with accuracy standard deviations of 1.46% and 1.53%, respectively. The proposed model outperforms representative conventional and deep-learning baselines, including DT, MLP, CNN-RNN, DGCNN, 4D-CRNN, BiDCNN, EmT, and miMamba. Ablation analyses further demonstrate that multi-band decomposition, spatial asymmetry modeling, the signed asymmetric operation, and the 4 × 4 convolution kernel jointly contribute to the accuracy and stability of MBSACNN.
Background and objective Sounds carrying inaudible high-frequency components (HFCs) above 20 kHz can alter brain activity, autonomic tone, and subjective state, a cluster of responses termed the hypersonic effect. However, only single physiological domains have been examined till now; therefore, whether central, autonomic, and subjective indices move together under one stimulus in the same participants remains unresolved. This study measured electroencephalographic (EEG), heart rate variability (HRV), and subjective responses simultaneously to characterize the layered response to HFC-containing sound. Methods Twenty healthy adult men completed a participant-blind crossover protocol in which the order of the two conditions was randomized and counterbalanced (10 received the wide-band sound first, 10 the audible-only sound first): an audible-band sound limited to ≤20 kHz (low frequency (LF)) and a wide-band sound containing the HFC (low frequency plus high frequency (LFHF)). Each condition comprised five minutes of pre-exposure rest, 10 minutes of sound presentation (analyzed as Audio1 and Audio2), and five minutes of post-exposure rest (Post). EEG centro-parieto-occipital (CPO) Alpha-2 relative power, normalized HRV indices, and 20 semantic-differential (SD) ratings were analyzed as changes from the pre-exposure baseline. Aligned rank transform analysis of variance (ART-ANOVA; condition × time) was used, with epoch-specific Wilcoxon signed-rank condition contrasts (Bonferroni correction, m = 3); effect sizes are the rank-biserial correlation (r). Results EEG Alpha-2 showed a trend-level main effect of condition (ART-ANOVA F = 3.59, p = 0.074), with no effect of time or condition-by-time interaction. The condition-by-time interaction was not significant; in pre-specified epoch comparisons, the condition contrast was significant during the second half of presentation (Audio2: Bonferroni-corrected p = 0.025, r = 0.657, indicating a large effect) and showed a trend after presentation (Post: corrected p = 0.089, r = 0.552). For HRV, relative low-frequency power (LF%) was significantly lower under LFHF at Post (corrected p = 0.025, r = -0.657, indicating a large effect; median difference -10.1 percentage points, 95% confidence interval -17.0 to -2.9), indicating a reduction in the relative low-frequency component of normalized HRV. None of the 20 SD items differed between conditions (all p > 0.10). Conclusions Under these conditions, HFC-containing sound was associated with a trend-level increase in posterior Alpha-2 activity, most evident in pre-specified comparisons during the second half of presentation, and a lower relative LF component of normalized HRV after exposure, without significant differences in subjective ratings. These preliminary findings suggest a possible central-to-autonomic pattern that can be quantified using noninvasive physiological recordings and support further study of HFC-containing sound as an environmental, non-pharmacological approach.
Posttraumatic stress disorder (PTSD) is increasingly conceptualized as involving stress-related alterations in large-scale neural systems supporting memory-related processes, alongside broader cognitive and emotional regulation. The present study examined resting-state, frequency-specific oscillatory activity and functional connectivity in PTSD using magnetoencephalography (MEG). Guided by the tri-network model, we tested whether PTSD would be associated with altered cortical oscillatory dynamics and reduced synchrony within and between the default mode, salience, and central executive networks, systems suggested to be implicated in memory and cognitive control. Eyes-open resting-state MEG data were acquired from trauma-exposed Canadian Armed Forces (CAF) members and Veterans with PTSD (n = 57) and trauma-exposed control (TEC) members and Veterans (n = 54). PTSD showed increased delta-band activity (1-3 Hz) in occipital, parietal, and temporal regions, alongside reduced gamma-band activity above 30 Hz in frontal and limbic regions. Reduced alpha-band synchrony (8-14 Hz) was also seen within and between regions of the default mode, salience, and central executive networks. These findings indicate disrupted temporal coordination across cortical systems that support memory function and cognitive control processes in PTSD. Such alterations may reflect impaired integration and regulation of internally generated information, consistent with core clinical features of PTSD. Resting-state MEG measures, therefore, provide frequency-specific markers of large-scale neural coordination differences associated with trauma-related psychopathology.
Mitochondrial dysfunction is a cardinal, causative, and convergent hallmark in both Alzheimer's disease (AD) and Parkinson's disease (PD). However, therapeutics that target the process of mitophagy, the selective removal of damaged mitochondria, are relatively undeveloped. Prior work has largely centered around post-translational modifications of the PINK1-Parkin signaling pathway while ignoring the key need for sustained protein synthesis of Parkin. In this review, we explore an innovative transcriptional circuit involving the gut microbiome, AMP-activated protein kinase (AMPK), sirtuin 1 (SIRT1), and mitophagy: gut-derived metabolites, such as Urolithin A (UA), activate AMPK and SIRT1, both of which converge to deacetylate and phosphorylate peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α). The transcription of the mitophagy protein, Parkin, is then driven by activation of PGC-1α. This UA/AMPK/SIRT1/PGC-1α/Parkin/mitophagy pathway is disrupted in multiple layers in AD and PD; this includes impaired gut function, lowering the level of UA produced in the body, proteinopathy leading to reduced PGC-1α activity, and decreased transcription of Parkin. Therapeutic targets of these various nodes include UA, PGC-1α activator ZLN005, and SIRT1 activators, such as resveratrol or nicotinamide riboside. By shifting the paradigm from post-translational activation to transcriptional restoration of Parkin, this gut-brain metabolic axis offers a unifying, testable, and therapeutically tractable framework for mitigating mitophagy failure in AD and PD.
The prognostic value of on-treatment hepatitis B virus (HBV) RNA, hepatitis B core-related antigen (HBcrAg), and quantitative hepatitis B surface antigen (HBsAg) for hepatocellular carcinoma (HCC) risk in patients with chronic hepatitis B (CHB) remains uncertain. We evaluate the predictive roles of these viral markers in patients with CHB receiving long-term entecavir therapy. This prospective, multicenter study enrolled patients with CHB-related cirrhosis undergoing long-term entecavir treatment and HCC surveillance in Taiwan. Serum HBV RNA, HBcrAg, and HBsAg levels were measured at enrollment. Multivariable regression identified predictors of HBV RNA positivity and HCC development. Overall, 451 patients were included. After a median of 6.2 years of antiviral therapy, the median levels of HBV RNA, HBcrAg, and HBsAg were 0.9 log10 copies/mL, 3.2 log10 U/mL, and 2.4 log10 IU/mL, respectively. During a median 3.3-year follow-up, 55 developed HCC. 57.2% of patients had detectable HBV RNA. HBV RNA detectability was associated with older age, lower alanine aminotransferase (ALT), HBeAg positivity, and higher HBsAg and HBcrAg levels. In multivariable analysis, higher fibrosis-4 (FIB-4) index (adjusted subdistribution hazard ratio [aSHR], 1.10; 95% confidence interval [CI] 1.00-1.21) and elevated alpha-fetoprotein (AFP) (aSHR, 1.01; 95% CI 1.00-1.01) independently predicted HCC development, whereas the three viral markers did not. After a 6-year entecavir therapy, 57.2% of patients with CHB-related cirrhosis had detectable HBV RNA. Year-6 HBV RNA, HBcrAg, and HBsAg levels were not predictive of HCC, while FIB-4 index and AFP remained independent predictors.
Selective therapeutic gene activation in tumors remains challenging because tumor-enriched promoters often exhibit basal activity in non-target cells and may not fully capture intratumoral heterogeneity. In addition, direct promoter-driven expression of cytotoxic or tumor-suppressive payloads may be vulnerable to promoter leakiness, which can compromise specificity and safety. Here, we developed SLICER, a two-vector adeno-associated virus (AAV)-delivered Cre-loxP gene circuit that integrates an alpha-fetoprotein (AFP) promoter-driven sensor with post-transcriptional gating through microRNA (miRNA) recognition elements (MREs) for miR-21 and miR-122. This dual-layer design separates tumor-context sensing from downstream payload activation, converting context-dependent Cre accumulation into recombination-gated payload expression from the actuator vector. In AFP-active liver tumor cell models, including HepG2, Huh7, and PLC/PRF/5, SLICER induced robust Cre expression and loxP-dependent luciferase activation, whereas non-target/comparator cells showed minimal background activity and limited functional toxicity. Mutation of miR-21 and/or miR-122 recognition elements increased Cre accumulation and reporter output, supporting an MRE-dependent gating mechanism that contributes to circuit specificity. A pro-apoptotic BAX payload triggered caspase-dependent apoptosis in target cells, which was partially attenuated by Z-VAD-FMK, while non-target cells remained largely unaffected. Perturbation of miRNA inputs further tuned circuit output, consistent with miRNA-guided regulation. SLICER also accommodated modular tumor-suppressor payloads, including TP53 and PTEN, supporting actuator-layer interchangeability. In vivo, systemic AAV delivery of SLICER-TP53 suppressed tumor growth and reduced tumor burden in both HepG2 and Huh7 xenograft models, accompanied by increased intratumoral P53 expression. Together, these findings establish SLICER as a modular proof-of-concept platform for combinatorial transcriptional and post-transcriptional control of therapeutic gene activation in AFP-active liver tumor contexts, while supporting further development of logic-gated gene circuits for more precise tumor-selective payload delivery.
Weeds significantly threaten rice production. While herbicides are the most efficient weed-control method, prolonged use accelerates resistant weed emergence. 4-Hydroxyphenylpyruvate dioxygenase (HPPD)-inhibiting herbicides offer potential for managing resistant weeds in rice, but the availability of resistant rice mutants is limited. This study aimed to optimize the method of DNA shuffling to conduct directed evolution of the rice HPPD gene, discover novel resistance mutations, and generate herbicide-resistant rice germplasm. We developed an efficient DNA shuffling method utilizing flanking sequences and segmented amplification to construct random mutation libraries of rice HPPD and maize (Zea mays L.) HPPD genes. Mutant libraries were screened via chromogenic reaction in Escherichia coli. CRISPR-mediated adenine base editing was adopted to generate rice mutants, and hydroponic assays were conducted to evaluate herbicide resistance. The crystal structure of rice HPPD was determined by X-ray crystallography, while mutant structures and ligand docking were simulated with AlphaFold3. Flanking sequences enhanced single-stranded DNA amplification efficiency, and segmented amplification reduced nonsense mutations. A resistant mutant, OsHPPD-mHelix, was obtained. Enzyme kinetics analysis demonstrated that its Ki for mesotrione was increased approximately 2.5-fold compared to that of wild-type OsHPPD, while its enzyme activity was not significantly affected. Using the mutation sites contained in OsHPPD-mHelix as hotspots, genome-edited rice was further generated. It was found that the mesotrione resistance of rice carrying the E423G mutation was approximately 2.2-fold higher than that of wild-type rice. Structural and molecular dynamics analyses suggested the E423G mutation promotes a closed terminal α-helix conformation, with free energy landscapes indicating reduced mesotrione binding stability, potentially explaining the resistance mechanism. This study provides an efficient DNA shuffling technology and novel herbicide-resistant rice germplasm, enriching the toolbox for rice gene-directed evolution and resistant variety cultivation.
Menopause marks the natural end of reproductive phase of women caused by decline in levels of estrogen, progesterone and other reproductive hormones. Asparagus racemosus (Shatavari) is a medicinal herb particularly known to be effective in alleviating women's reproductive health. Shatavarin IV, a steroidal saponin and a primary bioactive component in this herb, acts as a phytoestrogen by modulating the ER-alpha/ER-beta signalling, the TrkB-BDNF axis, and the Hypothalamic-Pituitary-Gonadal (HPG) and Hypothalamic-Pituitary-Adrenal (HPA) axes. In this study, the efficacy of SheVari4® on alleviating the menopause-specific endocrine dysfunction and subsequent impaired quality of life (QOL) was assessed through a randomized double-blind placebo-controlled parallel arm trial conducted on 60 pre-, peri-, and post-menopausal women. Sixty women (age 50.0 ± 7.4 years) randomized in two equal groups were administered with 100 mg SheVari4® or placebo capsules once daily for 8 weeks. Assessment of Menopause-Specific Quality of Life (MENQOL) along with domain scores across vasomotor, psychosocial, physical, and sexual function constituted primary endpoint analysis; changes in serum estradiol E2, Follicle Stimulating Hormone (FSH), progesterone, cortisol, Anti Mullerian Hormone (AMH), Sex Hormone-Binding Globulin (SHBG) and free testosterone were assessed as secondary endpoint analysis. Safety of the formulation was evaluated in terms of body mass index, blood biochemistry and liver function test. SheVari4 induced a 42.8% reduction in MENQOL versus placebo. Significant improvements in functioning (vs. placebo) were observed, respectively, across vasomotor (53.1% vs. 14.5%), psychosocial (53.6% vs. 9.9%), physical (39.2% vs. 6.5%) and sexual (29.4% vs. 10.7%) domains. The formulation also increased E2 (+36.44% vs. -2.08%) and progesterone (+0.30% vs. -2.88%) coupled with marked reduction in levels of FSH (-38.18% vs. +12.97%) and cortisol (-26.87% vs. +17.16%). Concomitantly, AMH, SHBG, and free testosterone were also favorably modulated. Blood biochemistry, liver function tests, and vital signs confirmed the broad-spectrum safety of SheVari4®. SheVari4® significantly improved menopause-specific QOL and endocrine parameters in peri- and post-menopausal women, with a well-established safety profile.
The Glasgow Sleep Effort Scale (GSES) assesses the extent to which individuals engage in conscious, deliberate attempts to control their sleep. The present study aimed to conduct a reliability generalization meta-analysis to estimate the overall internal consistency of the GSES, to examine whether these estimates vary as a function of study and sample characteristics, and to assess the prevalence of reliability induction practices in the literature. This systematic review and meta-analysis was preregistered on the Open Science Framework. A search was conducted in PubMed, Scopus, Web of Science, and PsycINFO from inception to November 12, 2025, using the term "Glasgow Sleep Effort Scale". Two reviewers independently screened records using predefined eligibility criteria. Studies reporting Cronbach's alpha for the GSES total score were included. We conducted a random-effects meta-analysis using restricted maximum likelihood estimation. Thirty-four articles contributed 40 independent estimates (total N = 16,022). The pooled estimate was α = 0.81 (95% CI [0.79, 0.82]), indicating good overall internal consistency, although significant heterogeneity was observed (I2 ≈ 85%). Publication year significantly moderated internal consistency estimates, whereas language, mean age, sex, sample type, study design, and methodological quality did not. Reliability induction was observed in nearly 58% of the initially retrieved studies, most commonly due to the omission of reliability coefficients. The GSES shows good internal consistency. Its frequent use underscores the need to report sample-specific reliability estimates.
Di-(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer with well-documented hepatotoxic effects mediated through oxidative stress, inflammation, apoptosis, and ferroptosis-related molecular alterations. This study investigated the hepatoprotective potential of free fisetin (FSN) and fisetin-loaded liposomal nanoparticles (FSN-LNPs) against DEHP-induced liver injury in rats, with special emphasis on the comparative efficacy of the nanoformulation. Sixty adult male Sprague Dawley rats were randomly allocated into six groups: control, FSN, FSN-LNPs, DEHP, DEHP + FSN, and DEHP + FSN-LNPs. Following 28 days of oral administration, liver function indices, lipid profile, oxidative stress markers, inflammatory mediators, apoptotic and ferroptosis-related biomarkers, along with histopathological and ultrastructural alterations, were evaluated. DEHP exposure significantly impaired liver function, as evidenced by elevated serum ALT, AST, and ALP levels, along with disrupted protein synthesis and dyslipidemia. These biochemical changes were accompanied by increased oxidative stress, NF-κB activation, elevated pro-inflammatory cytokines (TNF-α and IL-6), and activation of apoptotic (caspase-3, Bax) signaling, together with ferroptosis-associated molecular alterations characterized by increased ACSL4 expression, iron accumulation, and lipid ROS levels, accompanied by reduced GPX4 expression, alongside suppression of antioxidant defenses (SOD, CAT, GSH, Nrf2/HO-1). Histopathological and ultrastructural analyses further highlighted severe hepatic injury in DEHP-treated rats. Co-treatment with FSN alleviated these alterations, while FSN-LNPs demonstrated significantly superior protective effects across most evaluated parameters. Notably, FSN-LNPs restored several biomarkers to near-normal levels and markedly improved hepatic architecture and ultrastructure. In conclusion, FSN-LNPs provided enhanced hepatoprotection against DEHP-induced toxicity compared with free FSN, which was associated with attenuation of oxidative stress and inflammation and modulation of apoptosis- and ferroptosis-related pathways, likely due to improved bioavailability and cellular delivery. To the best of our knowledge, this is the first study to evaluate fisetin in a liposomal nanoparticle formulation for protection against DEHP-induced liver injury, highlighting its promising therapeutic potential while providing a foundation for future mechanistic and translational investigations.