Abstract Androgen deprivation therapy (ADT) remains the standard treatment for advanced prostate cancer (PCa); however, most patients ultimately progress to lethal castration-resistant PCa (CRPC). Emerging evidence implicates RNA N⁶-methyladenosine (m⁶A) modification as a key regulator of cancer biology, yet its role in CRPC remains poorly understood. As a critical adaptor in the m⁶A methyltransferase complex, RNA-binding motif protein 15 (RBM15) directs m⁶A deposition to specific mRNA targets. Here, we identified RBM15 as the key methyltransferase member significantly upregulated in CRPC tissues and strongly correlated with poor patient survival. Functionally, RBM15 overexpression reduces PCa cell sensitivity to enzalutamide, whereas its knockdown suppresses tumor growth and invasion. Mechanistically, RBM15 is an androgen-responsive protein whose expression increases upon chronic androgen deprivation. It catalyzes m⁶A methylation at position A1384 of damaged DNA binding protein 1 (DDB1) mRNA, leading to YTHDF2-dependent transcript decay and reduced DDB1 protein levels. Lower DDB1 impairs K48-linked polyubiquitination of the androgen receptor (AR), thereby stabilizing AR and amplifying AR signaling. Importantly, AR transcriptionally activates RBM15, forming a feed-forward loop that drives CRPC progression. Collectively, our findings establish RBM15 as a central epitranscriptomic driver of CRPC and identify the RBM15–DDB1–AR axis as a promising therapeutic target. Dual inhibition of RBM15 and AR may offer a novel strategy to overcome treatment resistance in advanced PCa.

Background: Copper overload in plaque macrophages has long been associated with atherosclerosis, however, the molecular mechanisms underlying its pathological accumulation and its causal role in driving inflammatory responses and lipid dysregulation remain incompletely defined. Identifying the key regulator that couples copper homeostasis to immune and metabolic crosstalk is critical for understanding plaque vulnerability and developing targeted therapies. Methods: We performed integrated analyses combining human clinical cohorts (n = 399), bulk and single-cell RNA sequencing, and multi-omics profiling of carotid atherosclerotic lesions. Mechanistic studies were conducted in human PBMC-derived macrophages and THP-1 cells using oxLDL-induced foam-cell models, with CRISPR-based gene editing, pharmacological inhibitors, acyl-biotin exchange assays, and chromatin immunoprecipitation. In vivo validation was carried out in ApoE⁻/⁻ mice fed a high-fat diet with copper supplementation or the chelator tetrathiomolybdate (ATTM). Findings: We identify SLC31A1 as a critical upstream regulator that is both transcriptionally upregulated via oxLDL-triggered TGF-β–SMAD2 signaling and post-translationally modified by ZDHHC18-mediated S-palmitoylation at Cys189. This palmitoylation anchors SLC31A1 to the plasma membrane, preventing its copper-induced degradation and sustaining pathological copper influx. The resultant copper overload triggers cuproptosis and mitochondrial dysfunction, leading to mitochondrial DNA release into the cytosol, which activates the cGAS–STING–IKKα NFκB inflammatory cascade. NFκB subsequently transactivates the scavenger receptor CD36, amplifying lipid uptake and foam-cell formation, thereby establishing a feed-forward loop that couples inflammation to lipid dysregulation. In vivo, copper supplementation accelerates atherosclerosis in ApoE⁻/⁻ mice, while ATTM treatment effectively disrupts this axis and attenuates plaque progression without systemic toxicity. Clinically, serum copper levels serve as a robust biomarker for vulnerable plaques (AUC = 0.762). Interpretation: Our findings establish SLC31A1 as a critical regulator that bridges copper homeostasis mitochondrial integrity, innate immune activation, and lipid metabolism in atherosclerosis, and position the SLC31A1 palmitoylation–copper cGAS–STING–CD36 pathway as a promising therapeutic node for high-risk vascular disease.

Objective Beta-ketothiolase deficiency (BKTD) is a rare genetic metabolic disorder caused by variants in the ACAT1 gene. It can induce severe metabolic acidosis, which may be life-threatening. This study reports two critically ill children with newly diagnosed BKTD and includes a literature review to comprehensively depict the clinical, biochemical, and genetic profiles of BKTD patients in China. Methods We retrospectively analyzed clinical, biochemical, and Sanger sequencing data of two newly diagnosed BKTD children from our hospital, and systematically reviewed 74 additional Chinese cases from the literature, totaling 76 patients. Results Both newly diagnosed cases presented with severe metabolic crises. Patient 1 was found to have a globally novel compound heterozygous variant of ACAT1 (c.1222G>A and c.414A>C). Patient 2, who presented with pre-existing developmental delay and growth failure, achieved normal growth and development following metabolic intervention. The retrospective cohort study indicated that 38.0% (19/50) of the patients had neurological involvement, but only a few had permanent sequelae. Regarding screening markers, the detection rate of 2-methyl-3-hydroxybutyric acid (2M3HB) was 100% (66/66), while that of 3-hydroxybutyrylcarnitine (C4OH acylcarnitine) was 96.3% (26/27). A total of 77 variants were identified in 70 sequenced patients, confirming that c.622C>T and c.1124A>G were the most frequent variant hotspots in the Chinese population, differing from those reported in other countries. Conclusion Acute episodes of BKTD are often accompanied by neurological involvement, but this neurological involvement is mostly transient and reversible. Blood C4OH acylcarnitine can serve as an effective supplement to traditional screening methods. This study discovered new variations that expand the variant spectrum of ACAT1 and identified the ACAT1 gene variant hotspots in the Chinese population. Overall, this study comprehensively and systematically depicts the disease profile of BKTD patients in China, providing valuable baseline data for clinical diagnosis and genetic counseling in the country.

Background:Central obesity represents a well-established risk factor for stress urinary incontinence (SUI), with sagittal abdominal diameter-to-height ratio (SADHtR) recognized as a superior anthropometric indicator of visceral adiposity. However, the evidence between the SADHtR and SUI remains scarce. This study aimed to elucidate the association between SADHtR and SUI of adult women. Methods:This cross-sectional study incorporated 4,846 women ≥20 years from the 2011-2016 cycles of the National Health and Nutrition Examination Survey (NHANES). Multivariable-adjusted logistic and linear regression models were used to investigate the relationship between SADHtR and the prevalence and severity of SUI. Smooth curve fitting methods and a two-piecewise linear regression model were employed to examine potential nonlinear relationships. Subgroup analyses and interaction tests were performed to identify potential effect modifiers. Propensity score matching (PSM) was employed to balance confounders, strengthening the robustness of the findings. Results:A positive association was observed between SADHtR and both the prevalence and severity of SUI after adjusted for potential confounders [odds ratio (OR) =3.59; 95% confidence interval (CI): 2.75-4.69; P<0.001] (β=0.71; 95% CI: 0.23-1.19; P=0.004). The relationship between SADHtR and the prevalence of SUI exhibited non-linear dose-response, with an inflection point at SADHtR =0.14. Subgroup analyses demonstrated consistent associations across all examined cohorts and showed that the positive relationship was more pronounced among young women. Conclusions:This study demonstrated a nonlinear positive association between SADHtR and SUI risk. Our findings establish SADHtR as a valuable anthropometric measure for SUI risk evaluation.

Olfactory dysfunction (OD) is a common sequela of SARS-CoV-2 infection, but its prevalence and associated factors beyond 2 years remain unclear. In this multicenter observational study, we assessed 155 recovered COVID-19 patients approximately three years after infection and included 170 age-matched healthy controls as a reference group. Demographic, clinical, psychological, and sleep-related data were collected through structured interviews. Olfactory function at the 3-year assessment was objectively evaluated using Toyota-Takagi (T&T) olfactometry. Paired baseline-to-follow-up T&T data were available only for a small exploratory subgroup of nine patients and were analyzed descriptively. At follow-up, 7 of 155 recovered patients (4.5%) met our T&T-based definition of persistent quantitative olfactory dysfunction, all of whom were older than 50 years. Emotional and sleep disturbances were also common, with descriptive trends toward higher frequencies among women and older individuals. Exploratory analyses suggested that insomnia (AIS > 6; OR 5.35, 95% CI 1.07-26.60; p = 0.033) and anxiety (HAMA ≥ 7; OR 10.54, 95% CI 1.21-91.82; p = 0.04) were associated with persistent T&T-defined quantitative OD, although the small number of outcome events limited statistical precision. These findings indicate that a small proportion of COVID-19 survivors have persistent objective OD 3 years after infection and that persistent OD is associated with anxiety and insomnia.

QuestionDoes letrozole improve semen-based outcomes in men with spermatogenic failure?FindingsIn this randomized clinical trial of 296 men with spermatogenic failure, 14.3% of participants treated with letrozole (2.5 mg daily) plus vitamins C and E achieved an upgrade in World Health Organization sperm concentration category at 3 months compared with 5.4% in those receiving vitamins C and E alone, a statistically significant difference.MeaningIn this study, letrozole improved sperm concentration categories among men with spermatogenic failure, potentially enabling less invasive reproductive management. This randomized clinical trial evaluates whether treatment with letrozole improves sperm concentration categories among men with spermatogenic failure in China. ImportanceSpermatogenic failure (SPGF) is a severe form of male infertility with limited evidence-based medical treatment options. Aromatase inhibitors represent a promising therapeutic strategy for SPGF, but high-quality evidence is lacking.ObjectiveTo evaluate the efficacy and safety of letrozole for improving sperm concentration categories in men with SPGF.Design, Setting, and ParticipantsThis multicenter, open-label, assessor-blinded randomized clinical trial was conducted at 10 male infertility centers in China from July 2023 to March 2024. Men with SPGF (nonobstructive azoospermia [NOA], cryptozoospermia, or severe oligozoospermia) were enrolled. Follow-up for the primary outcome was completed in June 2024. Data were analyzed from July 2024 to March 2025.InterventionsParticipants were randomized to receive letrozole (2.5 mg daily) plus vitamins C and E or vitamins C and E alone (control group) for 3 months.Main Outcomes and MeasuresThe primary outcome was World Health Organization Sperm Concentration Categories (WHO-SCC) upgrade rate at 3 months after randomization. The secondary outcomes were WHO-SCC grades, Dutch Society of Obstetrics and Gynecology Total Motile Sperm Count Categories (NVOG-TMSCC) upgrade rate, semen parameters, and reproductive hormone levels.ResultsAmong 296 participants (mean [SD] age, 30.2 [3.9] years; 218 [73.6%] with NOA; 147 randomized to the letrozole group and 149 to the control group), 247 (83.4%) completed the trial, and all 296 were included in the primary analysis. WHO-SCC upgrade rates were 14.3% (21 of 147 participants) with letrozole vs 5.4% (8 of 149 participants) with control (risk difference, 9.2% [95% CI, 2.5%-15.8%]; P = .01). Participants in the letrozole group had higher odds of achieving a better WHO-SCC grade than those in the control group (common odds ratio, 2.65 [95% CI, 1.28-5.47]; P = .008). The NVOG-TMSCC upgrade rate was also higher with letrozole. Letrozole significantly increased serum gonadotropins and testosterone while decreasing estradiol. There were no significant between-group differences in semen parameters. Decreased libido (18 [12.2%] vs 8 [5.4%]; P = .04) was more frequent with letrozole than with control.Conclusions and RelevanceIn this randomized clinical trial of men with spermatogenic failure, letrozole significantly improved sperm concentration categories and was well tolerated, supporting its use as an option to downstage infertility severity and potentially enable less invasive reproductive management.Trial Registrationchictr.org.cn Identifier: ChiCTR2300073861

Emerging evidence suggests that microglia exhibit dual regulatory roles in the pathogenesis of Parkinson’s disease (PD); however, their precise function in α-synuclein clearance remains incompletely understood. Here, we provide compelling evidence that α-synuclein preformed fibrils (α-syn PFF) impair lysosomal acidification in microglia, leading to defective autophagic flux and disrupted α-syn degradation. This dysfunction further promotes the secretion of microglial extracellular vesicles (EVs), exacerbating disease pathology. Mechanistic investigations uncover that α-syn PFF directly interacts with ATP6V0C, a pivotal V0 subunit of V-ATPase. This interaction sterically hinders V0-V1 domain assembly, disrupting proton pump complex formation and reducing ATP6V0C expression. Functionally, ATP6V0C overexpression rescues lysosomal acidification deficits and facilitates α-syn degradation in vitro, while in vivo, ATP6V0C overexpression alleviates neurotoxicity and reduces phosphorylated α-syn aggregation in α-syn PFF mouse models. Further investigation identifies the PI3K-AKT-mTOR-TFEB pathway as a key regulatory axis of ATP6V0C-mediated lysosomal acidification in microglia. Notably, both TFEB activation and mTOR inhibition restore lysosomal acidity and upregulate ATP6V0C expression, thereby enhancing α-syn clearance. These findings establish the TFEB-ATP6V0C axis as a key determinant of microglial proteostasis, proposing targeted activation of this pathway as a promising strategy to mitigate PD progression.

Background Diabetes mellitus-induced erectile dysfunction (DMED) is a common and debilitating vascular-neurogenic complication of diabetes. The poor responsiveness of DMED patients to phosphodiesterase type-5 inhibitors underscores the need for therapies capable of restoring both endothelial integrity and cavernous nerve function. The Slit2/Roundabout (Robo) pathway, a conserved regulator of axon guidance and vascular remodeling, is well positioned to coordinate neurovascular repair; however, its involvement in DMED remains largely undefined.Methods Single-cell RNA sequencing from embryonic genital tubercle to postnatal stages was used to map fibroblast-endothelial-neuronal signaling and identify Slit2 as a key secreted ligand. Diabetic and control mouse corpus cavernosum tissues were analyzed for endothelial and neuronal Slit2-Robo1/4 activity. In vitro, recombinant Slit2-N was applied to cells under high glucose to assess proliferation, migration, tube/sprout formation, and neurite outgrowth; Robo1/4 knockdown tested receptor dependency. In vivo, AAV-Slit2 was delivered intracavernously to diabetic mice, with erectile function assessed by sodium-nitroprusside perfusion and electrostimulation, and structural remodeling evaluated by endothelial, neural, and fibrosis markers.Results Single-cell profiling identified Slit2 as a fibroblast-derived developmental cue for corpus cavernosum morphogenesis. Diabetic mice exhibited markedly reduced endothelial and neuronal Slit2-Robo signaling. Slit2-N restored angiogenic and neuroregenerative functions in vitro under high glucose, enhancing endothelial activity and neurite growth; all effects were lost after Robo1/4 silencing. In vivo, AAV-Slit2 re-established local Slit2 signaling, improved penile perfusion and erectile responses, enhanced endothelial proliferation and nerve regeneration, and reduced fibrosis, indicating substantial neurovascular recovery.Conclusions Slit2/Robo signaling is a key regulatory axis disrupted in DMED. Local restoration of Slit2 via AAV delivery drives synchronized neurovascular regeneration and effectively rescues erectile function in diabetic mice. These findings establish Slit2 as a promising therapeutic target for reversing the underlying pathophysiology of DMED rather than merely providing symptomatic relief.

Idiopathic pulmonary fibrosis (IPF) is a life-threatening interstitial lung disease characterized by the abnormal activation of pulmonary fibroblasts. In our study, we demonstrated that FoxM1 is highly expressed in activated pulmonary fibroblasts, and its nuclear translocation plays a crucial role in conferring resistance to FasL-induced apoptosis in pulmonary fibroblasts. Disruption of FoxM1 function was shown to restore the ability to resolve fibrosis in mice treated with bleomycin. Mechanistic investigations revealed that a decrease in SIRT3 expression leads to increased acetylation of FoxM1, which is essential for the activation of pulmonary fibroblasts in vitro. Further, downregulation of SIRT3 expression enhances the stability of FoxM1, thereby accelerating bleomycin-induced pulmonary fibrosis through the activation of pulmonary fibroblasts. Importantly, treatment with nicotinamide riboside was found to suppress the activation of pulmonary fibroblasts and protect mice from bleomycin-induced pulmonary fibrosis by activating SIRT3. In summary, our findings highlight a critical role of the SIRT3/FoxM1 axis in regulating the activation of pulmonary fibroblasts. These insights suggest potential therapeutic strategies against pulmonary fibrosis, focusing on modulating this pathway for effective treatment. This work opens new avenues for the development of targeted therapies aimed at mitigating the progression of IPF.

Existing diagnostic technologies for bladder cancer (BC) suffer from low sensitivity, low specificity, or a lack of validation. Therefore, validated, non-invasive diagnostic biomarkers with high sensitivity and specificity for early detection of BC are needed to complement and improve upon the limitations of existing diagnostic methods. We used low-pass whole genome sequencing (LP-WGS) technology to detect copy number variations (CNVs) in small extracellular vesicle (sEV) DNA isolated from urine samples of patients. Based on these results, we constructed and validated a diagnostic model to differentiate between benign and malignant bladder lesions. We conducted a receiver operating characteristic analysis and calculated the area under the curve (AUC) to evaluate the performance of the diagnostic model. The urine sEV-DNA LP-WGS data revealed CNV differences between benign and malignant samples. The diagnostic model achieved an AUC of 0.953, a sensitivity of 86.7%, and a specificity of 100% in the training cohort and an AUC of 0.985, a sensitivity of 90%, and a specificity of 100% in the validation cohort. Even at the lowest coverage depth of 0.01X, the performance of the diagnostic model remained relatively robust. Notably, the performance of this diagnostic model surpassed that of the biomarker neuron-specific enolase (sensitivity: 85.7% vs. 64.3%; specificity: 100% vs. 87.5%) and urinary cytology (sensitivity: 100% vs. 66.7%; specificity: 100% vs. 94.1%). Our study demonstrates that urine sEV-DNA exhibits high discriminatory power in distinguishing between benign and malignant bladder lesions, making it a promising tool for auxiliary diagnosis of BC.

OBJECTIVES:The external physical vibration lithecbole (EPVL) is a new device. It is clinically employed to improve the stone-free rate (SFR). However, it is not widely accepted in clinical practice due to the lack of high-level evidentiary support and a standard protocol. This study aimed to evaluate the effectiveness and safety of external physical vibration lithecbole (EPVL) as an adjunct to extracorporeal shock wave lithotripsy (ESWL) or retrograde intrarenal surgery (RIRS) for upper urinary tract stones. METHODS:We systematically searched PubMed, Web of Science, Embase, the China National Knowledge Infrastructure (CNKI), and the Cochrane Library from inception to 13 January 2026, for randomized controlled trials (RCTs) comparing EPVL plus standard care with standard care alone, and we pooled relative risks (RR) with 95% confidence intervals (CI). RESULTS:Nine RCTs involving 1418 patients were analyzed. Compared with standard care alone, EPVL significantly increased early stone-free rates at about 1 week (RR 1.44, 95% CI 1.18-1.77, p < 0.001) and 2 weeks (RR 1.40, 95% CI 1.20-1.63, p < 0.001) after ESWL or RIRS, with greater benefit for lower-pole (RR 1.56, 95% CI 1.25-1.96, p < 0.001) and renal pelvic stones (RR 1.54, 95% CI 1.10-2.14, p = 0.01). EPVL was associated with a lower rate of overall complications (RR 0.58, 95% CI 0.46-0.73, p < 0.001); specifically, it was linked to lower risks of hematuria (RR 0.64, 95% CI 0.48-0.86, p = 0.002) and urinary tract infection-related findings (RR 0.28, 95% CI 0.14-0.57, p < 0.001). CONCLUSIONS:Adjunctive EPVL improves short-term stone clearance after ESWL or RIRS without adding measurable risk and may also be associated with a reduced incidence of complications. Where available, EPVL can be considered as a non-invasive option to enhance fragment clearance, particularly for lower-pole renal stones. Further multicenter trials are needed to confirm long-term outcomes and generalizability. TRIAL REGISTRATION:PROSPERO CRD42024600537.

KMT2C (lysine methyltransferase 2C), also known as mixed-lineage leukemia 3 (MLL3), is a member of the KMT2 family of histone methyltransferases that catalyzes histone H3 lysine 4 monomethylation (H3K4me1), a hallmark of active enhancer elements. Operating within COMPASS-like complexes (Complex of Proteins Associated with Set1) and in association with the ASCOM coactivator complex (ASC-2–containing complex), KMT2C plays a central role in maintaining enhancer and super-enhancer integrity, thereby sustaining lineage-specific transcriptional programs. Across gastrointestinal malignancies, KMT2C is recurrently altered, predominantly through truncating loss-of-function variants, splice-disrupting events, and structural alterations that impair protein function. Importantly, the biological impact of KMT2C alteration is highly context dependent, shaped by mutation class, co-occurring genomic lesions, and tissue-specific transcriptional circuitry. Rather than inducing linear dysregulation of individual signaling pathways, KMT2C deficiency preferentially destabilizes enhancer and super-enhancer networks, leading to large-scale transcriptional rewiring. Disruption of enhancer modules that enforce cellular identity and homeostasis is frequently accompanied by activation of stress-adaptive and metabolic programs. Concurrently, defects in homologous recombination and replication-stress responses promote genomic instability, while attenuation of cell-cycle checkpoints and senescence barriers facilitates epithelial–mesenchymal transition, stem-like plasticity, and invasive or metastatic behavior. Beyond tumor-intrinsic effects, KMT2C dysfunction can reshape the tumor immune microenvironment through altered antigenic burden, inflammatory signaling, senescence-associated secretory programs, and dynamic stromal interactions, ultimately giving rise to heterogeneous therapeutic vulnerabilities. Clinically, KMT2C alteration has been linked to tumor mutational burden (TMB), microsatellite instability (MSI), immune infiltration patterns, and outcomes following immune checkpoint blockade (ICB). In parallel, KMT2C-associated DNA repair deficiencies provide a mechanistic basis for synthetic-lethal strategies involving poly(ADP-ribose) polymerase (PARP) inhibitors and inhibitors of ataxia telangiectasia and Rad3-related protein (ATR) or checkpoint kinase 1 (CHK1), including rational combinations with epigenetic therapies. In this review, we integrate evidence from hepatocellular carcinoma, pancreatic ductal adenocarcinoma, cholangiocarcinoma, colorectal cancer, gastric cancer, esophageal cancer, and gallbladder cancer within a unified framework that links KMT2C domain architecture to enhancer-network destabilization, phenotypic state transitions, and clinical manifestations. We further propose a functional evaluation paradigm that reframes discrete KMT2C variants as graded states of epigenetic deficiency, coupled with a closed-loop validation strategy integrating tissue-based profiling, liquid biopsy monitoring, and spatial multi-omics analyses.

Objective To evaluate the protective effects of Luteolin (LUT) against hyperoxaluria-induced renal injury and calcium oxalate (CaOx) crystal deposition, and to explore the underlying molecular mechanisms. Method The targets related to LUT and kidney stones were screened in a variety of databases, and the potential targets and pathways were identified by network pharmacology. Subsequently, the interaction between LUT and the core targets was verified by molecular docking and molecular dynamics simulation. Finally, a glyoxylate-induced kidney stone mouse model and high oxalate-induced HK2 cells were used to verify the effect and potential mechanism of LUT on kidney stone formation. Results Network pharmacology identified 223 intersecting targets between kidney stones and LUT, with KEGG enrichment highlighting the PI3K/Akt signaling pathway. Molecular docking revealed a strong binding affinity between LUT and p85α (-6.947 kcal/mol), and molecular dynamics simulations confirmed complex stability after 25 ns. In vivo, LUT significantly reduced renal calcium oxalate (CaOx) crystal deposition and alleviated tissue injury in the mouse model. In vitro, LUT effectively inhibited oxalate-induced PI3K/Akt activation and inflammatory cytokine production in HK-2 cells. Furthermore, CETSA analysis suggested a potential target engagement between LUT and p85α. Conclusion This study suggests a protective effect of LUT against kidney stone formation at multiple levels. Our results indicate that LUT attenuates renal calcium crystal deposition, potentially through the inhibition of the PI3K/Akt signaling pathway. These findings provide new insights into the use of natural products for the prevention of nephrolithiasis.

Background/Objectives: Diabetes mellitus-induced erectile dysfunction (DMED) is characterized by severe endothelial and smooth-muscle dysfunction and reduced responsiveness to conventional therapy. Guanxinning tablet (GXN), a traditional Chinese medicine compound containing Salvia miltiorrhiza (Danshen) and Ligusticum stratum (Chuanxiong), has vascular-protective and anti-oxidative properties, but its effect on DMED remains unclear. This study evaluated whether GXN improves erectile function in a streptozotocin (STZ)-induced type 1 diabetic rat model and explored the possible involvement of PI3K/Akt/FOXO1 signaling. Methods: STZ-induced diabetic rats with ED were treated with GXN. Artery pressure (AP) and intracavernous pressure (ICP) were measured to evaluate erectile dysfunction. Western blots, immunohistochemistry, immunofluorescence, biochemical assays, TUNEL staining, RNA-seq, and in vitro HCMEC assays were performed to evaluate erectile effector cell function, oxidative stress, fibrosis, apoptosis, and PI3K/Akt/FOXO1-associated changes. Results: GXN improved erectile function in DMED rats and was associated with improved endothelial and smooth-muscle-related markers, reduced oxidative stress, attenuated fibrosis, and decreased apoptosis. RNA-seq and protein validation suggested that GXN treatment was associated with activation of PI3K/Akt/FOXO1-related signaling. In HCMECs, LY294002 (a PI3K inhibitor) partially reversed the GXN-associated improvement in cell viability and apoptosis-related markers. Conclusions: GXN ameliorated erectile dysfunction and tissue injury in STZ-induced type 1 diabetic rats, which were associated with PI3K/Akt/FOXO1 signaling. These findings support the potential of GXN as a traditional Chinese medicine compound for treating DMED.

The pathogenicity of variants of uncertain significance in the LRRK2 gene remains underexplored. Investigating the LRRK2 variant spectrum in a large Chinese population cohort can provide deeper insights into its pathogenic mechanisms. This study examined the LRRK2 gene variants in 20,519 Chinese individuals, including 7,562 Parkinson’s disease (PD) patients, 3,077 Essential tremor (ET) patients, and 9880 healthy controls. We conducted a genetic analysis of low-frequency and common non-synonymous variants in the LRRK2 gene across the cohorts. A total of 287 low-frequency non-synonymous LRRK2 variants were identified in the PD and control cohorts. Among these, six reported pathogenic variants (p.R1325Q, p.R1441C, p.R1441H, p.V1447M, p.G2019S, p.I2020T) and three reported likely pathogenic variants (p.R1067Q, p.N1437D, p.R1728H) were enriched in PD cases, with a frequency of 0.71%. In contrast, only one pathogenic variant (p.R1325Q) and one likely pathogenic variant (p.R1067Q) were observed in healthy controls (0.11%), and the ET cohort exhibited similar variant distribution to controls (0.19%). Burden analysis and association analysis revealed novel likely pathogenic variants, including p.A312V, p.M968K, and p.R1320S as candidates. These novel variants were significantly more frequent in PD patients (0.79%) compared to healthy controls (0.20%) or ET patients (0.42%). Additionally, seven common missense variants of LRRK2 were identified, and significant associations with PD for p.A419V, p.R1628P, and p.G2385R were confirmed, but no common variants were linked to ET. This study provides the first comprehensive characterization of the LRRK2 variant spectrum in a large Chinese population, underscoring the pivotal role of LRRK2 in PD pathogenesis but not in ET. These findings advance the understanding of LRRK2 in neurodegenerative disorders and lay a foundation for personalized therapeutic strategies based on genetic profiling.

BACKGROUND:Hyperlipidemia-related erectile dysfunction (HLED) responds poorly to first-line treatments such as phosphodiesterase type 5 inhibitors (PDE5Is), emphasizing the need for alternative therapeutic strategies. Rutaecarpine (RUT), a bioactive alkaloid, exhibits significant anti-inflammatory and anti-fibrotic effects. However, its therapeutic potential and mechanisms in HLED remain unexplored. OBJECTIVES:To evaluate the therapeutic effects and underlying mechanisms of RUT on HLED in vivo and in vitro. METHODS:In vivo, 48 male Sprague-Dawley rats were allocated into four groups: control, HLED, HLED+RUT-LD (low dose), and HLED+RUT-HD (high dose). Hyperlipidemia was induced through a high-fat diet, and RUT was administered orally. Erectile function was assessed using cavernous manometry, followed by analysis of penile tissues. In vitro, NR8383s were treated with lipopolysaccharide to induce M1 polarization and co-cultured with corpus cavernosum fibroblasts (CCFBs) to evaluate the impact of RUT on inflammation and fibrosis. RESULTS:RUT treatment partially improved erectile function in HLED rats. RUT reduced macrophage M1 polarization and fibrosis in penile tissues. In vitro, RUT inhibited LPS-induced M1 macrophage polarization and reduced fibrosis-related phenotype in co-cultured CCFBs. CONCLUSION:RUT could enhance erectile function in HLED rats by inhibiting macrophage M1 polarization and associated corpus cavernosum fibrosis. This study suggested that RUT could be a promising drug for treating HLED.

Periodic limb movements (PLM) refer to periodic episodes of repetitive and stereotypical limb movements, predominantly affecting the lower extremities. The prevalence of PLM is estimated at 17.6% to 86.7% in patients with Parkinson's disease (PD), which is much higher than 4% to 11% in the general population. PD is a chronic neurodegenerative movement disorder characterized by both motor and non-motor symptoms (NMS), and PLM represents one of the common non-motor manifestations. PLM in PD has garnered increasing attention due to its high prevalence, association with reduced sleep quality, elevated risk of comorbidities, unclear pathophysiology, and limited treatment options. This review outlines the epidemiology and risk factors for PLM in PD patients. We explore several underlying mechanisms including iron deficiency, dopaminergic dysregulation, and sympathetic nervous activity. Treatment strategies for PLM comorbid with PD are broadly categorized into non-pharmacological and pharmacological therapies where dopamine agonists can alleviate symptoms of both PD and PLM. Clarifying the relationship between PLM and PD is essential, as it may lead to expanded novel treatment approaches and improved quality of life for affected patients.

BACKGROUND:Erectile dysfunction (ED) affects more than half of male patients with diabetes. Diabetes mellitus-related ED (DMED) remains challenging to manage due to hyperglycemia-induced apoptosis and fibrosis of corpus cavernosum smooth muscle cells (SMCs). Exosomes secreted by normal SMCs (NSMC-EXOs) have emerged as promising therapeutic vectors. This study investigated the therapeutic role and molecular mechanism of NSMC-EXO-carried microRNA-30a-5p (miR-30a-5p) in DMED. METHODS:Erectile function in DMED rats treated with NSMC-EXOs was assessed via electrophysiological testing. Exosomal miRNA profiles were characterized by sequencing, and potential target genes were identified through bioinformatic prediction. The interaction between miR-30a-5p and Calm1 was experimentally validated. Expression levels of miR-30a-5p, Calm1, and markers of apoptosis and fibrosis were examined in vivo and in vitro. MiR-30a-5p mimics and inhibitors were employed to confirm the regulatory mechanism. RESULTS:Administration of NSMC-EXOs significantly restored erectile function in DMED rats. Sequencing revealed a marked enrichment of miR-30a-5p in NSMC-EXOs compared with exosomes from high-glucose-treated SMCs. Calm1 was identified as a direct target of miR-30a-5p, with involvement in apoptotic and fibrotic signaling pathways. Both in vivo and in vitro studies demonstrated that NSMC-EXO-derived miR-30a-5p was efficiently internalized by SMCs, suppressed Calm1 expression, and inhibited activation of the Calm1-AMPK-JNK and Calm1-CaMK2-TGFβ1 cascades. Consequently, apoptosis and fibrosis of corpus cavernosum tissue under hyperglycemic conditions were alleviated. Importantly, these protective effects were abolished following miR-30a-5p inhibition, underscoring its pivotal role. CONCLUSION:NSMC-EXO-derived miR-30a-5p protects against DMED by directly targeting Calm1 and attenuating pro-apoptotic and pro-fibrotic signaling. These findings provide mechanistic insight supporting the development of exosome-based therapeutic approaches for DMED.