Objective To investigate the effect of macrophage polarization induced by colorectal cancer exosomes on the anti-tumor activity of CD8+T cells.Methods M0 macrophages were co-incubated with PBS,HT-29 cells,and LoVo exosomes(HT-29 and LoVo exo)for 48 h,and termed the PBS,HT-29 exo,and LoVo exo groups.Real-time quantitative polymerase chain reaction was performed to detect M2 macrophage biomarker CD206,Arginase-1,IL-10,and CD163 mRNA expressions as well as M1 macrophage biomarker iNOS and IL-1β mRNA expressions.CD8+T-cells were co-incubated with the macrophages of the aforementioned groups(PBS+M0,HT-29 exo+M0,and LoVo exo+M0 groups)for 48 h.Flow cytometry was performed to detect CD8+T PD-1 expression.Next,the PBS+M0,HT-29 exo+M0,and LoVo exo+M0 groups of CD8+T-cells were incubated with HT-29 or LoVo cells for 24 h,respectively(PBS+M0/CD8+T,HT-29 exo+M0/CD8+T,and LoVo exo+M0/CD8+T groups).Subsequently,the enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of IFN-γ,perforin,and granzyme B in the cell supernatant.The cell lysis rates of HT-29 and LoVo cells were detected through cytotoxicity experiments.Results Compared with the PBS group,CD206,Arginase-1,IL-10,and CD 163 mRNA expressions of macrophages in the HT-29 exo and LoVo exo groups were significantly upregulated(P＜0.05),whereas iNOS and IL-1βmRNA expressions were significantly downregulated(P＜0.05).Compared with the PBS+M0 group,the HT-29 exo+M0 and LoVo exo+M0 groups exhibited significantly increased PD-1 expression(P＜0.05).Compared with the PBS+M0/CD8+T group,the IFN-γ,perforin,and granzyme B levels in the cell culture supernatant of the HT-29 exo+M0/CD8+T and LoVo exo+M0/CD8+T groups were significantly reduced(P＜0.05),and the cell lysis rates of HT-29 and LoVo were significantly reduced(P＜0.001).Conclusion M2 macrophage induced by HT-29 and LoVo exo can inhibit the tumor-killing function of CD8+T cells.

Objective To investigate the effect of macrophage polarization induced by colorectal cancer exosomes on the anti-tumor activity of CD8+T cells.Methods M0 macrophages were co-incubated with PBS,HT-29 cells,and LoVo exosomes(HT-29 and LoVo exo)for 48 h,and termed the PBS,HT-29 exo,and LoVo exo groups.Real-time quantitative polymerase chain reaction was performed to detect M2 macrophage biomarker CD206,Arginase-1,IL-10,and CD163 mRNA expressions as well as M1 macrophage biomarker iNOS and IL-1β mRNA expressions.CD8+T-cells were co-incubated with the macrophages of the aforementioned groups(PBS+M0,HT-29 exo+M0,and LoVo exo+M0 groups)for 48 h.Flow cytometry was performed to detect CD8+T PD-1 expression.Next,the PBS+M0,HT-29 exo+M0,and LoVo exo+M0 groups of CD8+T-cells were incubated with HT-29 or LoVo cells for 24 h,respectively(PBS+M0/CD8+T,HT-29 exo+M0/CD8+T,and LoVo exo+M0/CD8+T groups).Subsequently,the enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of IFN-γ,perforin,and granzyme B in the cell supernatant.The cell lysis rates of HT-29 and LoVo cells were detected through cytotoxicity experiments.Results Compared with the PBS group,CD206,Arginase-1,IL-10,and CD 163 mRNA expressions of macrophages in the HT-29 exo and LoVo exo groups were significantly upregulated(P＜0.05),whereas iNOS and IL-1βmRNA expressions were significantly downregulated(P＜0.05).Compared with the PBS+M0 group,the HT-29 exo+M0 and LoVo exo+M0 groups exhibited significantly increased PD-1 expression(P＜0.05).Compared with the PBS+M0/CD8+T group,the IFN-γ,perforin,and granzyme B levels in the cell culture supernatant of the HT-29 exo+M0/CD8+T and LoVo exo+M0/CD8+T groups were significantly reduced(P＜0.05),and the cell lysis rates of HT-29 and LoVo were significantly reduced(P＜0.001).Conclusion M2 macrophage induced by HT-29 and LoVo exo can inhibit the tumor-killing function of CD8+T cells.

Objective:To investigate the application value of optical coherence tomography (OCT) and intravascular ultrasound (IVUS) in evaluating flow diverter (FD) apposition and endothelialization in aneurysm animal models, and analyze the effect of incomplete stent apposition (ISA) on aneurysm lumen healing and stent endothelialization.Methods:Lateral common carotid artery aneurysm models in swines were established by surgical method and then FD was implanted. Immediately after surgery, OCT and IVUS were used to evaluate the locations and degrees of ISA, and difference between these 2 methods in evaluating FD apposition was compared. DSA was performed at 12 weeks after surgery to evaluate the aneurysm occlusion (Kamran grading) and stent patency. OCT and IVUS were used again to observe the stent endothelial situation; by comparing with histopathologic results, effect of ISA on aneurysm healing and stent endothelialization was analyzed.Results:Lateral common carotid artery aneurysm models in 6 swines were established, and 6 Tubridge FDs were successfully implanted. Compared with IVUS (3 stents, 4 locus), OCT could detect more ISA (6 stents, 14 locus); and the vascular diameter change area (7 locus), aneurysm neck area (4 locus) and the head and tail of FD (3 locus) were the main sites of FD malapposition; average distance between stent wire and vessel wall was (560.14±101.48) μm. At 12 weeks after surgery, DSA showed that 1 patient had a little residual contrast agent at the aneurysm neck (Kamran grading 3), and the remaining 5 had complete aneurysm occlusion (Kamran grading 4). One FD had moderate lumen stenosis, and the other 5 FDs had lumen patency. OCT indicated mostly disappeared acute ISA; ISA proportion decreased to 21.4 % (3/14), including 2 in the aneurysm neck and 1 in the partial stent. Histopathological results showed bare stent woven silk, without obvious endothelial coverage; in one FD with luminal stenosis, intimal hyperplasia was mainly composed of vascular smooth muscle cells.Conclusion:In carotid artery aneurysm model with FD implantation, OCT can detect more ISA than IVUS; most acute ISA have good outcome at 12 th week of follow-up, while severe ISA can cause delayed FD endothelialization and delayed aneurysm occlusion.

Objective:To explore the value of radiomics model based on intratumoral and peritumoral early dynamic contrast-enhanced (DCE) MRI for identifying benign and malignant in breast imaging reporting and data system (BI-RADS) 4 tumors.Methods:A total of 191 patients diagnosed with BI-RADS 4 breast tumors by breast MRI examination with clear pathological diagnosis from January 2016 to December 2020 in the First Affiliated Hospital of Bengbu Medical College were analyzed retrospectively, including 77 benign and 114 malignant cases, aged 23-68 (46±10) years. The one-slice image with the largest area of the lesion of the second stage DCE-MRI images was selected to outline the region of interest, and automatically conformal extrapolated by 5 mm to extract the intra-tumoral and peritumoral radiomics features. The included cases were randomly divided into training and testing cohorts in the ratio of 8∶2. The statistical and machine learning methods were used for feature dimensionality reduction and selection of optimal radiomics features, and logistic regression was used as the classifier to establish the intratumoral, peritumoral, and intratumoral combined with peritumoral radiomics models. The independent risk factors that could predict the benignity and malignancy of breast tumors were retained as clinical-radiological characteristics by univariate and multivariate logistic regression to establish a clinical-radiological model. Finally, the intratumoral and peritumoral radiomics features were combined with clinical-radiological features to develop a combined model of the three. The receiver operating curve was used to analyze the predictive performance of each model and calculate the area under the curve (AUC),the AUC was compared by DeLong test. The stability of the three-component combined diagnostic model was tested by 10-fold cross-validation, and the model was visualized by plotting nomogram and calibration curves.Results:In the training cohort, the AUC of the three-component combined model for identifying benign and malignant BI-RADS 4 breast tumors was significantly higher than that of the intratumoral radiomics model ( Z=3.38, P<0.001), the peritumoral radiomics model ( Z=4.01, P<0.001), the intratumoral combined with peritumoral radiomics model ( Z=3.11, P=0.002), and the clinical-radiological model ( Z=3.24, P=0.001). And the AUC, sensitivity, specificity, accuracy, and F1-score of the three-component combined model were 0.932, 91.2%, 86.9%, 87.0% and 0.89, respectively. In the testing cohort, the three-component combined model also had the highest AUC value (0.875), and diagnostic sensitivity, specificity, accuracy and malignancy F1-score were 95.7%, 62.5%, 76.9%, and 0.89, respectively. The AUC calculated by 10-fold cross-validation was 0.90 (0.85-0.92), and the predicted curve of the three-component combined model in the calibration curve was in good agreement with the ideal curve. Conclusion:The three-component combined diagnostic model based on the intratumoral and peritumoral radiomics features and clinical-radiological features of early DCE-MRI has good performance and stability for identifying the benign and malignant in BI-RADS 4 breast tumors, and it can provide guidance for clinical decision non-invasively.