Background: Endometriosis (EMS) is an estrogen-dependent disease in which endometrial glands and stroma arise outside the uterus. Current studies have suggested that the number and function of immune cells are abnormal in the abdominal fluid and ectopic lesion tissues of patients with EMS. The developed CIBERSORT method allows immune cell profiling by the deconvolution of gene expression microarray data.Methods: By applying CIBERSORT, we assessed the relative proportions of immune cells in 68 normal endometrial tissues (NO), 112 eutopic endometrial tissues (EU) and 24 ectopic endometrial tissues (EC). The obtained immune cell profiles provided enumeration and activation status of 22 immune cell subtypes. We obtained associations between the immune cell environment and EMS r-AFS stages. Macrophages were evaluated by immunohistochemistry (IHC) in 60 patients with ovarian endometriomas.Results: Total natural killer (NK) cells were significantly decreased in EC, while plasma cells and resting CD4 memory T cells were increased in EC. Total macrophages in EC were significantly increased compared to those of EU and NO, and M2 macrophages were the primary macrophages in EC. Compared to those of EC from patients with r-AFS stage I ~ II, M2 macrophages in EC from patients with stage III ~ IV were significantly increased. IHC experiments showed that total macrophages were increased in EC, with M2 macrophages being the primary subtype.Conclusions: Our data demonstrate that deconvolution of gene expression data by CIBERSORT provides valuable information about immune cell composition in EMS.

Although ovarian cancer, a gynecological malignancy, has the highest fatality rate, it still lacks highly specific biomarkers, and the differential diagnosis of ovarian masses remains difficult to determine for gynecologists. Our study aimed to obtain ovarian cancer-specific protein candidates from the circulating small extracellular vesicles (sEVs) and develop a protein panel for ovarian cancer screening and differential diagnosis of ovarian masses. In our study, sEVs derived from the serum of healthy controls and patients with cystadenoma and ovarian cancer were investigated to obtain a cancer-specific proteomic profile. In a discovery cohort, 1119 proteins were identified, and significant differences in the protein profiles of EVs were observed among groups. Then, 23 differentially expressed proteins were assessed using the parallel reaction monitoring in a validation cohort. Through univariate and multivariate logistic regression analyses, a novel model comprising three proteins (fibrinogen gamma gene (FGG), mucin 16 (MUC16), and apolipoprotein (APOA4)) was established to screen patients with ovarian cancer. This model exhibited an area under the receiver operating characteristic curve (AUC) of 0.936 (95% CI, 0.888–0.984) with 92.0% sensitivity and 82.9% specificity. Another panel comprising serum CA125, sEV-APOA4, and sEV-CD5L showed excellent performance (AUC 0.945 (95% CI, 0.890–1.000), sensitivity of 88.0%, specificity of 93.3%, and accuracy of 89.2%) to distinguish malignancy from benign ovarian masses. Altogether, our study provided a proteomic signature of circulating sEVs in ovarian cancer. The diagnostic proteomic panel may complement current clinical diagnostic measures for screening ovarian cancer in the general population and the differential diagnosis of ovarian masses.

Endometriosis, a chronic immune-mediated inflammatory disease, remains elusive in its pathogenesis. Given vitamin D (VD)’s pivotal role in modulating innate and adaptive immune responses, we sought to elucidate how VD modulates inflammatory responses in endometriosis. We isolated primary human ectopic endometrial stromal cells (EESCs) from ectopic endometrium of ovarian endometrioma, alongside Ishikawa cells, and subjected them to treatment with lipopolysaccharide (LPS), a potent inducer of inflammation, alongside varying concentrations of 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃), the biologically active form of VD, and its analog TEI-9647 (25-dehydro-1α-hydroxyvitamin D₃-26,23-lactone). Our results revealed that 1,25(OH)₂D₃ significantly reversed LPS-induced cell proliferation, migration, and inflammatory factor production in EESCs and Ishikawa cells, and induced apoptosis. Additionally, 1,25(OH)₂D₃ inhibited the expression and nuclear translocation of phosphorylated p65 in LPS-activated EESCs and Ishikawa cells. Furthermore, 1,25(OH)₂D₃ counteracted LPS-induced suppression of VD receptor (VDR)/IκBα and enhancement of Toll-like receptor 4 (TLR4)/pyrin domain (PYD)-containing protein 3 (NLRP3) activation, while the addition of TEI-9647 reversed VD’s regulatory effects on the NF-κB pathway. In vivo experimental results showed that 1,25(OH)₂D₃ significantly inhibited lesion growth, suppressed NF-κB pathway activation, and corresponding inflammatory phenotypes in a rat model of endometriosis. Collectively, these results underscore the potential of 1,25(OH)₂D₃ as a therapeutic target for endometriosis via VDR-dependent endometrial homeostasis regulation, suppressing LPS-mediated inflammatory responses and NF-κB signaling pathway through VDR activation and IκBα stabilization.

A nanodrug for targeted delivery of artesunate was synthesized to inhibit angiogenesis, serving as a non-hormonal therapy option for endometriosis.

Multifunctional small‐molecule theranostic agents hold significant clinical potential for non‐invasive endometriosis (EMS) management. Current EMS treatment faces challenges due to imprecise lesion localization and therapy‐associated side effects. Herein, an integrated theranostic probe, cRGDyK‐ICG‐Lys‐DTPA@Gd (cRGD‐ILD), designed for concurrent precise lesion localization and targeted phototherapy in EMS, is developed. This molecular probe integrates three key functional components: the DTPA@Gd complex provides contrast for magnetic resonance imaging, the ICG fragment enables fluorescence imaging and photoacoustic imaging while serving as the phototherapeutic agent, and the cRGD peptide targets integrin α _v β ₃ receptors to facilitate precise accumulation in ectopic endometrial tissue. Specifically, cRGD‐ILD leverages its small size and receptor‐mediated transcytosis to penetrate neovascularized tissue and target ectopic endometrial cells. Under 808 nm laser irradiation, the ICG moiety generates reactive oxygen species and heat, exerting combined photodynamic and photothermal therapeutic effects that effectively suppress lesion progression in both autografted EMS rats and xenografted nude mice. Notably, composed of clinically available and FDA‐approved constituents, the probe demonstrates favorable biocompatibility and biosafety for in vivo applications. Overall, this well‐designed molecular probe enables precise multimodal imaging guidance and localized phototherapy for EMS.

The increasing global prevalence of cesarean scar endometriosis necessitates a thorough understanding of the risk factors for postoperative recurrence, as this is crucial for developing preventive strategies and informed decision-making. To obtain insight into the clinical risk factors for postoperative recurrence of cesarean scar endometriosis following open lesion resection. The cohort for this study comprised 272 women, including 26 patients with postoperative recurrence and 246 without recurrence. Various parameters, including baseline characteristics, preoperative, intraoperative, and postoperative conditions, and follow-up information, were analyzed. A comparison of these parameters was made between patients with and without postoperative recurrence. Time-to-recurrence analyses were conducted using Cox's univariate and multivariate proportional hazard analyses, the Kaplan-Meier method, and the log-rank test. The results revealed significant differences between patients with and without postoperative recurrence in terms of visual analog scale for abdominal pain (P=.008), method of surgery (P<.001), and incision length (P=.002). The Cox proportional hazard model identified the visual analog scale for abdominal pain ≥4 as a significant risk factor for postoperative recurrence (hazard ratio, 3.72 [95% confidence interval, 1.65–8.43]; P=.002). In addition, patients who received removal of scar, excision of mass, and exploration underneath the scar (named as integrated excision) had a lower risk of recurrence than those who received local excision of mass (hazard ratio, 0.14 [95% confidence interval, 0.04–0.48]; P=.002). Furthermore, older patients (aged ≥35 years) were found to have a lower risk of postoperative recurrence than those <35 years (hazard ratio, 0.35 [95% confidence interval, 0.12–1.04]; P=.058). In addition, the depth of involvement was identified as a meaningful factor in postoperative recurrence for patients with local excision of mass, as determined by the log-rank test (P=.018). The study highlights that the visual analog scale for abdominal pain ≥4 is a risk factor for the recurrence of cesarean scar endometriosis after open lesion resection. Furthermore, the surgical method of integrated excision was identified as a protective factor.