Dynamic single-cell sequencing unveils the tumor microenvironment evolution of gastric cancer abdominal wall metastases during radiotherapy.

Although the combination of immunotherapy and radiotherapy (RT) for the treatment of malignant tumors has shown rapid development, the insight of how RT remodels the tumor microenvironment to prime antitumor immunity involves a complex interplay of cell types and signaling pathways, much of which remains to be elucidated. Four tumor samples were collected from the same abdominal wall metastasis site of the patient with gastric cancer at baseline and during fractionated RT for single-cell RNA and T-cell receptor sequencing. The Seurat analysis pipeline and immune receptor analysis were used to characterize the gastric cancer metastasis ecosystem and investigated its dynamic changes of cell proportion, cell functional profiles and cell-to-cell communication during RT. Immunohistochemical and immunofluorescent staining and bulk RNA sequencing were applied to validate the key results. We found tumor cells upregulated immune checkpoint genes in response to RT. The infiltration and clonal expansion of T lymphocytes declined within tumors undergoing irradiation. Moreover, RT led to the accumulation of proinflammatory macrophages and natural killer T cells with enhanced cytotoxic gene expression signature. In addition, subclusters of dendritic cells and endothelial cells showed decrease in the expression of antigen present features in post-RT samples. More ECM component secreted by myofibroblasts during RT. These findings indicate that RT induced the dynamics of the immune response that should be taken into consideration when designing and clinically implementing innovative multimodal cancer treatment regimens of different RT and immunotherapy approaches.

Assessing fluoride treatment and resistance of dental enamel to soft drink erosion in vitro: applications of focus variation 3D scanning microscopy and stylus profilometry.

OBJECTIVES: This study assesses the application of the focus variation 3D microscopy for the evaluation of dental erosion and fluoride treatment for prevention of enamel erosion in vitro. METHODS: Human dental enamel disks were treated with Prevident 5000 (PV, n=15) for 1 week and compared with a reference group (PN, n=15) after orange juice erosion in vitro. A focus variation 3D scanning microscope (IFM) and a stylus type profilometer (SSP) were used to evaluate the erosion depths on enamel. 3D topographic images were taken with vertical resolutions of 0.1 and 0.02 microm. Scratch marks depths from SSP were measured on IFM images. Measurements were compared between the SSP and IFM and between the two study groups. RESULTS: The SSP and IFM measurements of eroded enamel surfaces showed similar trends between the two methods and between the two study groups. The SSP and the IFM measurements were statistically significantly different but correlated with each other. PV group showed consistently lower erosion depth than PN in all profile measures using both SSP and IFM. The stylus tip created scratch marks that were significantly different in depths between the eroded and the reference surfaces in both groups. CONCLUSIONS: The focus variation 3D microscopy is a powerful tool in evaluating surface topography associated with enamel erosion and in assessing the treatment effects of anti-erosive therapies. Topical treatment with Prevident 5000 significantly increased enamel resistance to erosion by orange juice and should be considered as a treatment choice in patients susceptible to acidic dental erosion.