Clear cell renal cell carcinoma (ccRCC) progression is closely associated with pro-tumorigenic neutrophil extracellular traps (NETs) within the tumor microenvironment; however, the upstream molecular mechanisms remain poorly defined. Through integrated multi-omics analysis of ccRCC cohorts, we identified ITPKB as a key downregulated gene associated with NET formation. Subsequent functional validation, including dual-luciferase reporter assays, confirmed that ITPKB is a direct target of the miR-301b-3p. Mechanistically, the loss of ITPKB prevents the ubiquitination-mediated degradation of PARP1, leading to its stabilization and accumulation. Stabilized PARP1 acts as a transcriptional co-activator for NF-κB, triggering a robust secretion of IL-8, which recruits neutrophils and induces their transformation into NETs. These NETs facilitate tumor cell migration and adhesion while conferring resistance to tyrosine kinase inhibitors (sunitinib and sorafenib) and anti-PD-1 therapy. In vivo, the treatment with DNase1 to disrupt NETs effectively reversed this therapeutic resistance. Our findings unveil the miR-301b-3p/ITPKB/PARP1/IL8/NETs axis as a novel mechanistic link between tumor-intrinsic signaling and neutrophil-driven immunosuppression, providing a solid experimental foundation for developing combination therapeutic strategies for advanced ccRCC.

Research question What is the critical age for the onset of deterioration in pregnancy outcomes associated with advancing maternal age in frozen embryo transfer (FET) cycles following preimplantation genetic testing (PGT)? Design This retrospective analysis included 6354 women (aged 20-45 years) who underwent PGT with single blastocyst transfer within 1 year of oocyte retrieval at a single centre, utilizing data collected from February 2017 to December 2024. Only the first FET cycle after oocyte retrieval was included, and fertilization was performed via intracytoplasmic sperm injection. Maternal characteristics were examined by transfer age; outcomes included clinical pregnancy rate (CPR), live birth rate (LBR), implantation rate, implantation failure rate (IFR) and abortion rate. Locally estimated scatterplot smoothing (LOESS) curves were used to visualize continuous trends. Restricted cubic spline (RCS) curves were used to assess age-related associations and non-linear effects. Threshold effect analysis identified the pivotal age at which endometrial aging has an adverse effect on outcomes. Results Advancing maternal age at embryo transfer was associated with declining CPR and LBR, and increasing IFR and abortion rate. Non-linear associations between maternal age at embryo transfer and clinical pregnancy (P non-linear = 0.009), live birth (P non-linear = 0.026), and implantation failure (P non-linear=0.009) were identified based on RCS analysis. Each year beyond 33 years of age was associated with a relative 5.8% decrease in the likelihood of clinical pregnancy [adjusted hazard ratio (aHR) = 0.942, 95% CI 0.908-0.977; P = 0.001], a relative 7.2% decrease in the probability of live birth (aHR = 0.928, 95% CI 0.895-0.963; P = 0.006), and a relative 6.1% increase in the risk of implantation failure (aHR = 1.061, 95% CI 1.023-1.101; P = 0.001). Conclusions Endometrial aging may be associated with significantly worsened pregnancy outcomes in women aged >33 years.

Prostate cancer (PCa) ranks among the most common and deadly malignancies worldwide. The clinical treatment of advanced prostate cancer is particularly challenging due to acquired drug resistance. Autophagy and lysosome-related pathways are key drivers of this resistance. Targeting the lysosome represents a potential therapeutic strategy for PCa. In this study, we identified Heat Shock Protein Family A Member 8 (HSPA8) as a critical functional node of Aloperine (ALO). ALO suppresses autophagic flux, disrupts lysosomal homeostasis, and induces lysosomal vacuolation in cancer cells by inhibiting the function of HSPA8, impairing chaperone-mediated autophagy (CMA)-mediated ATP6V1A degradation. The resulting pathological accumulation and enhanced V1-V0 association of the V-ATPase complex drive pronounced lysosomal hyperacidification and severe osmotic swelling. This biochemical and physical stress is associated with lysosomal membrane permeabilization (LMP) and downstream loss of lysosomal integrity. Furthermore, we reveal that ALO-induced vacuolation triggers a compensatory upregulation of cholesterol biosynthesis to buffer membrane expansion; preemptively disrupting this adaptive response with the DHCR7 inhibitor AY9944 yields significant synergistic lethality. Collectively, our findings reveal the specific cytotoxic mechanism of ALO and demonstrate that pharmacological targeting of the HSPA8-CMA-ATP6V1A axis is a valuable strategy for inducing lethal lysosomal vacuolation in advanced PCa.

Purpose:This study aimed to develop international expert consensus statements on chronic endometritis (CE) through a comprehensive literature review of existing evidence and a modified Delphi approach. Methods:Ten panelists of the Japan Society of Reproductive Medicine Female Reproductive Tract Special Interest Group on CE performed a comprehensive review of the literature and derived statements with related comments on six specific domains of interest on CE. A two-round modified e-Delphi questionnaire was conducted among 31 international experts to evaluate the statements. Results:Twenty-three out of 24 statements, including 43 detailed items, on epidemiology (associated diseases and risk factors), symptomatology (asymptomatic or oligosymptomatic nature with subtle and nondescript gynecologic manifestations), etiology and pathogenesis (inflammation, infection, and clinical course), microbiology (associated pathogens), diagnosis (histopathology, immunohistochemistry, hysteroscopy, and microbiome analysis), and treatment (antibiotics, surgery, and multidrug resistance) finally reached a consensus. Notably, for the histopathologic diagnosis of CE, a threshold of ≥ 5 endometrial stromal plasma cells/10 high-power fields received an agreement rate of 81%. Conclusions:This comprehensive literature review and Delphi study provide a real-world clinical perspective on CE from diverse international experts. These consensus statements have the potential to lay a foundation for diagnostic criteria and/or clinical guidelines on CE.

Abstract Introduction Hysteroscopy remains the gold standard for diagnosis and treatment of intrauterine pathologies, though its invasive nature, cost, and potential complications warrant careful consideration.Discussions about hysteroscopy before embryo transfer are abundant, while there is limited research available on the effect of hysteroscopy before Intrauterine insemination (IUI), especially Intrauterine insemination with donor sperm (IUI-D), and the existing evidence about its necessityin IUI cycles remains controversial. Materials and methods Propensity score matching (PSM) was used to adjust the baseline features at a ratio of 1:3. After PSM, there were 369 patients in the hysteroscopy group and 956 patients in the control group. We statistically analyzed all intrauterine abnormalities encountered during diagnostic hysteroscopy prior to IUI-D and compared the pregnancy outcomes between patients who underwent diagnostic hysteroscopy and those who did not. Simultaneously, pregnancy outcomes were compared between individuals with normal and abnormal hysteroscopic findings within the hysteroscopy group through logistic regression analysis. The primary outcome was live birth following the first IUI-D cycle. Results The live birth rate was 17.89% (66 of 369) in the hysteroscopy group compared with 20.92% (200 of 956) in the control group ( P = 0.216). Within the hysteroscopy group, pregnancy outcomes were similar between women with normal and abnormal findings in subgroup analysis. Conclusions The decision to perform diagnostic hysteroscopy prior to IUI-D should be individualized, balancing potential benefits against the risks of complications and treatment delay.

Abnormal tumor vasculature creates a permissive microenvironment that fuels the malignant progression of bladder cancer (BCa). While Leucine-rich alpha-2 glycoprotein 1 (LRG1) is known to regulate angiogenesis, its specific role in remodeling the BCa microenvironment remains poorly defined.We integrated single-cell RNA sequencing (scRNA-seq) with bulk transcriptomic datasets to identify key cellular subclusters. Functional validation was performed using subcutaneous and orthotopic BCa mouse models, neutrophil depletion, DNase I treatment, and clinical specimens. The molecular interactome was mapped via pull-down assays, mass spectrometry, and confocal imaging.LRG1 is significantly upregulated in BCa and correlates with hematogenous metastasis and poor prognosis. Mechanistically, tumor-derived LRG1 binds to Annexin A2 (ANXA2) on neutrophils through its LRR domain, impeding the mitochondrial translocation of Akt and triggering mtROS-dependent release of Neutrophil Extracellular Traps (NETs). These NETs directly cause vascular destabilization by stripping mural cell coverage. Blockade of the LRG1-NETosis axis induces vascular normalization, effectively overcoming microenvironmental barriers to increase drug delivery and T-cell infiltration, thereby profoundly sensitizing BCa to cisplatin and anti-PD-1 therapy.The LRG1-neutrophil-NETosis axis is a critical driver of vascular dysfunction and therapeutic resistance in BCa. Targeting this axis represents a promising translational strategy to improve clinical outcomes.

BACKGROUND:Intrauterine adhesions (IUAs), or Asherman's syndrome, are common complications of intrauterine surgery, such as hysteroscopic myomectomy, leading to menstrual disturbances and infertility. Postoperative inflammation is thought to drive adhesion formation, but the predictive value of specific inflammatory biomarkers remains unclear. METHODS:We conducted a prospective observational study of 150 women aged 19-45 years undergoing hysteroscopic resection of submucosal fibroids. Serum C-reactive protein (CRP), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels were measured preoperatively, 48 h postoperatively, and at 3 months. IUAs were evaluated at 12 weeks by second-look hysteroscopy and graded by the American Fertility Society criteria. Logistic regression and receiver operating characteristic analysis were performed. RESULTS:Among 135 patients who completed follow-up, 40 (30.8%) developed IUAs (20 mild, 12 moderate, and eight severe). Postoperative (48-h) serum IL-6 and CRP levels increased significantly in patients who developed adhesions compared with those who did not (P < 0.01), while postoperative TNF-α differences were less pronounced. IL-6 showed the strongest correlation with adhesion severity (ρ = 0.53) and was an independent predictor of moderate-to-severe IUA risk (adjusted odds ratio [OR] = 1.25 per pg/mL, 95% confidence interval [CI] 1.10-1.42). CRP was also predictive (OR = 1.30 per mg/L, 95% CI 1.05-1.60). IL-6 had the best discriminative ability (area under the curve [AUC] = 0.80), outperforming CRP (AUC = 0.72) and TNF-α (AUC = 0.65). CONCLUSION:Perioperative elevations in IL-6 and CRP are strongly associated with adhesion formation after hysteroscopic myomectomy. These markers might support risk-stratified postoperative surveillance in future multicenter studies.

BackgroundAssisted reproductive treatment for women with endometriosis and diminished ovarian reserve (DOR) is challenging, and the optimal controlled ovarian stimulation (COS) protocol for this population remains uncertain. This study compared four commonly used COS regimens for IVF/ICSI in women with EM and DOR.MethodsIn this retrospective cohort (2014-2023), 859 IVF/ICSI cycles in women aged 40 years or younger with EM and DOR were allocated to four COS protocols (ultra-long, long, short, antagonist). The primary outcomes were cumulative live birth rate (CLBR) and live birth rate (LBR) per started cycle; clinical pregnancy rate (CPR) was secondary. Between-protocol differences were assessed using multivariable logistic regression, adjusting for age, infertility duration, history of endometrioma cystectomy, basal FSH, LH, E2, AMH, antral follicle count, BMI, and COS protocol, with additional stabilized inverse-probability-weighting and 1:1:1:1 propensity score matching used to address confounding by indication.ResultsThe analysis included a total of 859 cycles, which were allocated into four treatment groups: the short protocol (n=295), the long protocol (n=145), the ultra-long protocol (n=202), and the antagonist protocol (n=217). Among these, the ultra-long protocol yielded the highest CLBR of 48.5%, LBR of 36.6%, and CPR of 41.1%. After PSM, the advantage difference of the ultra-long protocol is more significant. A subsequent multivariable logistic regression analysis confirmed that the ultra-long protocol was significantly more effective for achieving a live birth than the other protocols in the study: long protocol (OR: 0.577, P = 0.042), short protocol (OR: 0.608, P = 0.028), and antagonist protocol (OR: 0.486, P = 0.004). Similarly, the short (OR: 0.559, P = 0.010) and antagonist (OR: 0.484, P = 0.004) protocols were less effective for CPR.ConclusionFor infertile women aged 40 years or younger with endometriosis and DOR undergoing fresh embryo transfers, the ultra-long protocol appears to be more beneficial than the long, short, and antagonist protocols in terms of cumulative live birth, live birth, and clinical pregnancy rates.

Endometriosis affects approximately 10

Endometriosis (EM) is driven by immune dysregulation and macrophage dysfunction, yet the underlying mechanisms remain unclear. Here, metabolomic profiling revealed excessive itaconate accumulation in EM lesions, primarily due to elevated cis-aconitate decarboxylase 1 (ACOD1) expression in ectopic stromal cells (ESCs). ESC-derived itaconate was internalized by peritoneal macrophages, where it suppressed pro-inflammatory activity and phagocytosis, thereby facilitating ESC survival and dissemination. Mechanistically, itaconate exerted dual regulatory effects on macrophages: it activated NRF2 signaling to repress the transcription of pro-inflammatory genes, and it enhanced lysosomal acidification, thereby reducing lysosomal calcium release, which in turn inhibited p38-MAPK activation and further attenuated pro-inflammatory gene expression. In vivo, ACOD1 inhibition restored macrophage function and reduced lesion burden, while exogenous 4-octyl itaconate aggravated disease progression. These findings define a novel "ESC-ACOD1-itaconate-macrophage" axis that mediates immunosuppression in EM and identify ACOD1 as potential therapeutic targets.

The limited clinical efficacy of immune checkpoint inhibitors (ICIs) remains a major challenge in the treatment of bladder cancer (BCa). Here, we report that a ketogenic diet (KD) and its principal circulating metabolite, β-hydroxybutyrate (β-HB), suppress bladder tumor growth and enhance responses to PD-L1 inhibitor. In a syngeneic MB49 model, KD reduced tumor growth compared with normal diet (ND). KD increased plasma β-HB and was associated with higher intratumoral PD-L1 expression. Exogenous β-HB supplementation under ND recapitulated the antitumor effect of KD, further enhancing anti-PD-L1 efficacy. Mechanistically, β-HB increased intracellular reactive oxygen species (ROS) and disrupted mitochondrial membrane potential in bladder cancer cells, leading to ATP depletion and enhanced apoptosis. Besides, β-HB also upregulated PD-L1 in vitro and in vivo through GPR109A-JAK2-STAT3 signaling axis. Immune profiling of treated tumors showed increased infiltration and effector function of CD8+ T cells and NK cells and decreased immunosuppressive populations, changes that were further amplified when combined with anti-PD-L1. Together, our results indicate that KD and β-HB exert both tumor intrinsic and immune modulatory effects that sensitize bladder tumors to PD-L1 inhibitor, supporting further evaluation of ketogenic interventions as adjuvants to immunotherapy in BCa.

The periconception period is critical for embryo development, pregnancy outcomes, and offspring health. During this stage, oviductal and uterine fluids facilitate embryo-maternal interactions and support early embryonic development. Using PANDORA-seq, we identify a diverse repertoire of small non-coding RNAs in female mouse oviduct fluid and uterine fluid during preimplantation, with tRNA-derived small RNAs and rRNA-derived small RNAs being predominant. Maternal high-fat diet during preimplantation period significantly alters tsRNA and rsRNA expression in oviduct fluid and uterine fluid compared to normal diet, disrupting blastocyst metabolic gene expression. While implantation remained unaffected, these alterations impair mid-gestation embryonic and placental growth, resulting in reduced birth weight and length, as well as metabolic disorders in offspring. Furthermore, transfecting embryos with uterine fluid-derived sncRNAs altered by maternal high-fat diet mimics the in vivo effects. These findings suggest that tsRNAs and rsRNAs in reproductive fluids may reflect maternal metabolic status and transmit dietary information to the early embryo, which might influence pregnancy outcomes and offspring health.

Surgically-induced ampullary hydrosalpinx is a potential complication following salpingostomy for ectopic pregnancy, yet its impact on subsequent IVF/ICSI outcomes remains unclear. This study aimed to investigate the efficacy of surgical correction of this specific hydrosalpinx type. This retrospective cohort study analyzed 374 patients diagnosed with ampullary hydrosalpinx via hysterosalpingography or hysterosalpingo-contrast sonography after previous salpingostomy for tubal ectopic pregnancy. Participants were divided into a surgically treated group (n = 86) and an untreated group (n = 74) for frozen embryo transfer (FET) cycles, with additional fresh transfer cycle data included. Primary outcomes included implantation rate, clinical pregnancy rate, live birth rate, and miscarriage rate. In frozen embryo transfer cycles, the surgically treated group was linked to a markedly lower incidence of early miscarriage compared to the untreated group (5.6

Re-transurethral resection of bladder tumors (reTURBTs) improves staging and local control in high-risk bladder cancer, posing the clinical challenge of determining the most effective adjuvant intravesical regimen. In this multicenter retrospective study, 174 patients undergoing reTURBT between 2010 and 2024 received either Bacillus Calmette-Guérin (BCG) or pirarubicin/epirubicin (THP/EPI) intravesical therapy, delivered as post-reTURBT instillations alone (PIT) or combined with interval instillations before and after reTURBT (IIT + PIT). We compared recurrence-free survival (RFS) and adverse events across regimens and performed subgroup analyses by tumor characteristics. IIT + PIT significantly prolonged RFS: BCG IIT + PIT conferred a 2.93-fold lower recurrence risk versus BCG PIT (p = 0.03), while THP/EPI IIT + PIT achieved a 2.61-fold reduction versus THP/EPI PIT (p < 0.001). The greatest benefit appeared in high-grade, cT1 lesions. Our findings endorse IIT + PIT—particularly with BCG—as an effective strategy to reduce recurrence in high-risk bladder cancer.

Currently, bladder reconstruction using gastrointestinal segments is considered as the gold standard for bladder repair or augmentation. However, postoperative complications including bowel dysfunction, metabolic abnormalities, chronic urinary tract infection and stone formation are common. This study aims to evaluate the utility of the compound graft for bladder patch repair by seeding mesothelial cells onto autologous granulation tissue. 22 Fr silastic tubes were implanted subcutaneously in 18 male rabbits. The animals were equally divided into two groups. In the experimental group, nine animals underwent omentum biopsies and mesothelial cells were cultured and expanded in vitro. Two weeks after the implantation, the autologous granulation tissue encapsulating the silastic tubes were harvested. The tissue was cut longitudinally and trimmed to 2 × 2 cm2. In the experimental group, mesothelial cells were seeded onto the outer surface of the tissue and cocultured for 7 days to construct a compound graft. In each animal, a full-thickness defect of 2 × 2 cm2 in the anterior wall of the bladder was created. The defect was repaired with the constructed compound graft in the experimental group and with the granulation tissue directly in the control group. The bladder was harvested and analyzed grossly and histologically at 1, 2 and 6 months postoperatively. In addition, urodynamics were performed 6 months postoperatively to evaluate the function of the bladder. In the experimental group, no severe fibrosis and shrinkage were observed postoperatively and the mucosa at the grafts site appeared sleek and normal in color. Histologically, multilayers of urothelium surrounded by increasingly organized smooth muscles were observed. In contrast, severe contracture and fibrosis of the grafts occured in the control group. Gross examination of bladder mucosa revealed ulcer and stone formation at the site of the grafts. Histologically, accumulation of inflammatory cells and fibroblasts, and extensive scarring occurred. Six months after surgery, the urodynamic results showed that the experimental group had a larger bladder capacity (83.33 ± 2.08 ml vs. 76.33 ± 3.22 ml, p = 0.034), a lower maximum intravesical pressure (18.77 ± 1.79 mmHg vs. 24.02 ± 1.22 mmHg, p = 0.014) and a better bladder compliance (5.64 ± 0.8 ml/mmHg vs. 3.87 ± 0.13 ml/mmHg, p = 0.019), compared to the control group. Mesothelial cell-seeded granulation tissue can be successfully used for bladder patch repair in male rabbits. With further investigations, this technique may be considered for clinical use in human. Keywords · ··

BACKGROUND:Intrauterine adhesion (IUA) has become the subject of an expanding corpus of academic research. This study endeavors to perform a bibliometric analysis of the IUA literature to elucidate key research domains and trends over the past two decades. METHODS:A thorough search was conducted within the Web of Science Core Collection, concentrating on publications pertaining to intrauterine adhesion from January 2005 to December 2024. Bibliometric analysis and visualization were carried out utilizing Microsoft Excel 2019, VOSviewer, R-bibliometrix, and CiteSpace. RESULTS:Our analysis identified 1,488 relevant publications in this field, revealing a notable upward trajectory in annual publication rates. The People's Republic of China and the United States were identified as principal contributors to IUA research. The Journal of Minimally Invasive Gynecology was recognized as the most prolific journal in this area. Dabao Xu emerged as the most prolific author in terms of publication output and H-index. The analysis of keyword co-occurrence underscored an increasing emphasis on the mechanisms and therapeutic strategies for IUA. Keywords such as "endometrial regeneration," "fibrosis," "exosomes," "platelet-rich plasma," "mesenchymal stem cells," and "extracellular vesicles" indicate emerging trends and potential focal points for ongoing scientific research into IUA. CONCLUSION:This bibliometric analysis delineates the evolution of IUA research from surgical management to biological regeneration, highlighting the pivotal roles of stem cell biology and bioengineering. Future progress depends on translating these preclinical advances into clinical practice through international collaboration and standardized trials.

Inflammatory responses within the ovarian microenvironment are increasingly recognized as significant disruptors of ovarian function, yet their specific effects on early follicular development, particularly the activation of primordial follicles, remain poorly understood. In this study, we employ a mouse model of transient lipopolysaccharide (LPS)-induced inflammation to mimic the inflammatory conditions associated with chronic pelvic inflammatory disease (PID) and systemic infections. We demonstrate that LPS stimulation triggers the premature activation of primordial follicles, leading to a depletion of the ovarian reserve. This finding underscores the detrimental impact of inflammation on ovarian health. However, we also identify a protective mechanism mediated by the cGAS-STING pathway, a central regulator of innate immunity and cellular stress responses. Activation of the cGAS-STING pathway effectively inhibits LPS-induced primordial follicle activation, thereby preserving ovarian function. Further mechanistic investigations reveal that the Integrated Stress Response (ISR), a downstream effector of STING signaling, plays a critical role in this protective process. The ISR, orchestrated by kinases such as PERK, modulates cellular homeostasis under stress conditions by phosphorylating eukaryotic initiation factor 2α (eIF2α). Our data show that STING activation induces ISR signaling, which in turn suppresses the overactivation of primordial follicles. To explore the therapeutic potential of this pathway, we utilized STING and ISR agonists, which successfully mitigated LPS-induced primordial follicle activation and preserved ovarian reserve in our experimental model. These findings highlight the dual role of inflammation in ovarian biology: while acute inflammatory stimuli can disrupt follicular quiescence, the cGAS-STING-ISR axis serves as a critical regulatory network to counteract these adverse effects. Our study not only elucidates the molecular mechanisms underlying inflammation-induced ovarian dysfunction but also identifies STING and ISR as promising therapeutic targets for preserving ovarian reserve in women exposed to chronic inflammatory conditions. These insights have significant clinical implications, offering potential strategies to protect ovarian function in patients with inflammatory diseases or those undergoing treatments that compromise ovarian health.