Phenotypic comparison and the potential antitumor function of immortalized bone marrow-derived macrophages (iBMDMs).

Introduction: Macrophages are an important component of innate immunity and involved in the immune regulation of multiple diseases. The functional diversity and plasticity make macrophages to exhibit different polarization phenotypes after different stimuli. During tumor progression, the M2-like polarized tumor-associated macrophages (TAMs) promote tumor progression by assisting immune escape, facilitating tumor cell metastasis, and switching tumor angiogenesis. Our previous studies demonstrated that functional remodeling of TAMs through engineered-modifying or gene-editing provides the potential immunotherapy for tumor. However, lack of proliferation capacity and maintained immune memory of infused macrophages restricts the application of macrophage-based therapeutic strategies in the repressive tumor immune microenvironment (TIME). Although J2 retrovirus infection enabled immortalization of bone marrow-derived macrophages (iBMDMs) and facilitated the mechanisms exploration and application, little is known about the phenotypic and functional differences among multi kinds of macrophages. Methods: HE staining was used to detect the biosafety of iBMDMs, and real-time quantitative PCR, immunofluorescence staining, and ELISA were used to detect the polarization response and expression of chemokines in iBMDMs. Flow cytometry, scratch assay, real-time quantitative PCR, and crystal violet staining were used to analyze its phagocytic function, as well as its impact on tumor cell migration, proliferation, and apoptosis. Not only that, the inhibitory effect of iBMDMs on tumor growth was detected through subcutaneous tumor loading, while the tumor tissue was paraffin sectioned and flow cytometry was used to detect its impact on the tumor microenvironment. Results: In this study, we demonstrated iBMDMs exhibited the features of rapid proliferation and long-term survival. We also compared iBMDMs with RAW264.7 cell line and mouse primary BMDMs with in vitro and in vivo experiments, indicating that the iBMDMs could undergo the same polarization response as normal macrophages with no obvious cellular morphology changes after polarization. What's more, iBMDMs owned stronger phagocytosis and pro-apoptosis functions on tumor cells. In addition, M1-polarized iBMDMs could maintain the anti-tumor phenotypes and domesticated the recruited macrophages of receptor mice, which further improved the TIME and repressed tumor growth. Discussion: iBMDMs can serve as a good object for the function and mechanism study of macrophages and the optional source of macrophage immunotherapy.

Diagnostic Value of Artificial Intelligence-Assistant Diagnostic System Combined With Contrast-Enhanced Ultrasound in Thyroid TI-RADS 4 Nodules.

OBJECTIVES: This study evaluated the diagnostic value of artificial intelligence-assistant diagnostic system combined with contrast-enhanced ultrasound in The American College of Radiology Thyroid Imaging, Reporting and Data System (ACR TI-RADS) 4 category thyroid nodules. METHODS: Thyroid nodules that were evaluated as ACR TI-RADS 4 by conventional ultrasound were selected, all of which had pathological or fine needle aspiration (FNA) results. All nodules were examined by contrast-enhanced ultrasound (CEUS) and artificial intelligence (AI) analysis. The sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of AI, CEUS and their combined diagnosis were compared; Analyzed and compared the diagnostic efficiency of AI, CEUS and their combined diagnosis. RESULTS: A total of 148 thyroid nodules were included in 140 patients, including 58 malignant nodules and 89 benign nodules. The sensitivity of combined diagnosis was significantly higher than that of AI or CEUS alone (P < .05). The NPV of AI, CEUS and combined diagnosis were statistically significant (P < .05). There was no significant difference in the diagnostic efficacy between AI and CEUS (P > .05), but there was a significant difference in NPV between AI and combined diagnosis (P < .05). The AUC of the combined diagnosis was 0.859, which was higher than that of AI, CEUS alone. CONCLUSIONS: AI has a high diagnostic efficiency, which was helpful for radiologists to make rapid assessment. AI combined CEUS can significantly improve the diagnostic sensitivity and NPV, which was beneficial for the early detection of malignant nodules.