Objective To explore whether nano-vesicles derived from M1 macrophages (M1-NVs) can reprogram M2 macrophages into M1 phenotype and further affect the development of endometriosis (EMS). Methods Extracellular vesicles (EVs) were isolated from macrophage culture supernatant by differential centrifugation. Immunofluorescence cytochemistry was used to detect the expression of vimentin, CD31 and F4/80 to identify endometrial stromal cells (EMS-ESCs), HUVECs and polarized peritoneal macrophages of EMS patients. M1-NVs were prepared by filtering cell suspension through (5, 1, 0.4, 0.22)μm polycarbonate membrane filters after syringe aspiration at 0-4 DegreesCelsius. Flow cytometry was used to analyze the polarization of RAW264.7 mouse peritoneal macrophages in vitro, and reverse transcription PCR (RT-qPCR) was employed to detect mRNA expression of VEGF, CD86, interleukin-6 (IL-6), IL-1β, tumor necrosis factor α (TNF-α), arginase 1 (Arg1), CD163, CD206, and IL-10. PKH67-labeled M1-NVs were co-cultured with EMS-ESCs, HUVECs and macrophages. And tubule formation experiments were conducted to assess the impact of M1-NVs on the tubule formation of HUVECs. Transwell^TM invasion and migration assays were employed to evaluate changes in the migration and invasion abilities of EMS-ESCs. Results By monitoring the contents of NVs, it was found that NVs contained much more protein and other bioactive particles than the same amount of EVs; immunofluorescence staining results showed that PKH67 labeled M1-NVs were internalized by EMS-ESCs, HUVECs and macrophages when co-cultured. The results of flow cytometry and RT-qPCR multi-target analysis showed that after treatment with different concentrations of M1-NVs or M0-NVs, 20 μg/mL of M1-NVs could effectively reprogram M2 macrophages into M1 macrophages compared with M0-NVs. Transewell^TM results showed that compared with the blank group and M0-NVs group, the number of EMS-ESCs migrating from the upper chamber to the lower chamber after M1-NV treatment was significantly reduced, while the number of EMS-ESCs treated with M2NVs increased significantly. The invasion situation was similar to the migration situation, indicating that M1-NVs directly or indirectly inhibited invasion, migration and tubule formation of EMS-ESCs in vitro. Conclusion M1-NVs reprogrammes M2 macrophages into M1 macrophages by internalization of primary cells and macrophages, thereby inhibiting invasion, migration and angiogenesis of EMS-ESCs, and further hindering the occurrence and development of EMS.
Female ; Humans ; Animals ; Mice ; Endometriosis ; Macrophages ; Macrophages, Peritoneal ; Coculture Techniques

【Objective】 To compare the diagnostic performance of gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid (Gd-EOB) enhanced magnetic resonance imaging (MRI) and multi-detector computed tomography (MDCT) in detecting liver metastases from metastatic colorectal cancer (mCRC). 【Methods】 We made a retrospective collection of 128 patients diagnosed with mCRC from May 2019 to June 2022 at Haikou Hospital, Xiangya School of Medicine, Central South University and Xijing Hospital, Air Force Military Medical University. All patients underwent Gd-EOB MRI and MDCT imaging. Three radiologists judged the accuracy, sensitivity, specificity, positive predictive value and negative predictive value of the two modalities for colorectal liver metastases, respectively. 【Results】 Of the 128 patients diagnosed with mCRC, a total of 462 lesions were obtained, with 424 positive and 38 negative metastases confirmed by pathology. In the interpretation of physician A, Gd-EOB MRI judged 404 positive and 38 negative liver metastases, with accuracy of 95.67%, sensitivity of 95.28%, specificity of 100.00%, a positive predictive value of 100%, and a negative predictive value of 65.52%. MDCT judged 337 positive and 37 negative liver metastases, with accuracy of 80.95%, sensitivity of 79.48% and specificity of 97.37%, a positive predictive value of 99.70%, and a negative predictive value of 29.84%. In the interpretation of physician B, Gd-EOB MRI judged 403 positive and 36 negative liver metastases, with accuracy of 95.02%, sensitivity of 95.05%, specificity of 94.74%, a positive predictive value of 99.51%, and a negative predictive value of 64.91%. MDCT judged 335 positive and 35 negative liver metastases, with accuracy of 80.09%, sensitivity of 79.01%, specificity of 92.11%, a positive predictive value of 99.11%, and a negative predictive value of 28.23%. In the interpretation of physician C, Gd-EOB MRI judged 406 positive and 38 negative liver metastases, with accuracy of 96.10%, sensitivity of 95.75%, specificity of 100.00%, a positive predictive value of 100.00%, and a negative predictive value of 67.86%. MDCT judged 352 positive and 34 negative liver metastases, with accuracy of 83.55%, sensitivity of 83.02%, specificity of 89.47%, a positive predictive value of 98.88%, and a negative predictive value of 32.08%. Gd-EOB MRI judged the nature of liver metastases with higher accuracy, sensitivity and negative predictive value than MDCT, and had better agreement with pathological examination results in the judgment of physician A and physician C (Kappa=0.770, 0.788). In physician B’s judgment, the agreement with pathological findings was fair (Kappa=0.731), while the agreement between the results of MDCT examination and pathological findings was poor (Kappa=0.379, 0.378 and 0.400). 【Conclusion】 Gd-EOB MRI has higher accuracy, sensitivity and positive predictive rate than MDCT in diagnosing colorectal liver metastasis, and has higher diagnostic performance. Therefore, it can provide more valuable reference information for clinical differential diagnosis. Subcapsular lesions, peribiliary metastases and hepatic steatosis can reduce the diagnostic performance of MDCT, while Gd-EOB MRI detection can provide more accurate results than MDCT.