Objective:To explore the effects of combining APR-246 with irradiation for enhancing anti-tumor immune response against 4T1 breast cancer cells, and to develop multiple tumor treatment strategies.Methods:The control group, APR-246 group, irradiation group and irradiation combined APR-246 group were used both in the cell experiment and tumor-bearing mice experiment. The inhibitory effect of APR-246 on the proliferation of 4T1 cells was assessed by using Cell Counting Kit-8. The effect of APR-246 with irradiation on the survival rate of 4T1 cells using clone formation assay was measured. The levels of reactive oxygen species (ROS) and lipid peroxidation (LPO) in tumor cells using a 2’, 7’-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe and a lipid peroxidation sensor, the tumor inhibition rates of different groups of tumor bearing mice were compared, and the proportions of CD4+ and CD8+ T cells and the ratio of M1/M2 macrophages were determined in the tumor microenvironment by flow cytometry.Results:Compared with irradiation group, 2, 4, 6 Gy irradiation combined APR-246 group significantly reduced the survival rates of 4T1 cells ( t = 2.89, 4.15, 2.62, P < 0.05), the 6 Gy irradiation combined APR-246 group significantly increased the levels of ROS ( t = 16.95, P < 0.05) and LPO ( t = 6.09, P < 0.05) in 4T1 cells, and significantly increased the apoptosis rate of 4T1 cells ( t = 10.99, P < 0.05). Meanwhile, from the 16 th day of tumor inoculation, the 10 Gy irradiation combined APR-246 group showed significantly inhibited tumor growth ( t = 2.38-2.91, P < 0.05) and significantly increased proportions of CD4+ and CD8+ T cells ( t = 9.96, 6.28, P < 0.05) and M1/M2 ratio ( t = 15.30, P < 0.05) in tumor tissues. Conclusions:APR-246 combined with irradiation can effectively increase ROS and LPO levels in 4T1 cells, promote tumor cell apoptosis, and induce anti-tumor immune response, thus potentially inhibiting the growth of 4T1 cells.

Objective:To explore the effects of combining APR-246 with irradiation for enhancing anti-tumor immune response against 4T1 breast cancer cells, and to develop multiple tumor treatment strategies.Methods:The control group, APR-246 group, irradiation group and irradiation combined APR-246 group were used both in the cell experiment and tumor-bearing mice experiment. The inhibitory effect of APR-246 on the proliferation of 4T1 cells was assessed by using Cell Counting Kit-8. The effect of APR-246 with irradiation on the survival rate of 4T1 cells using clone formation assay was measured. The levels of reactive oxygen species (ROS) and lipid peroxidation (LPO) in tumor cells using a 2’, 7’-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe and a lipid peroxidation sensor, the tumor inhibition rates of different groups of tumor bearing mice were compared, and the proportions of CD4+ and CD8+ T cells and the ratio of M1/M2 macrophages were determined in the tumor microenvironment by flow cytometry.Results:Compared with irradiation group, 2, 4, 6 Gy irradiation combined APR-246 group significantly reduced the survival rates of 4T1 cells ( t = 2.89, 4.15, 2.62, P < 0.05), the 6 Gy irradiation combined APR-246 group significantly increased the levels of ROS ( t = 16.95, P < 0.05) and LPO ( t = 6.09, P < 0.05) in 4T1 cells, and significantly increased the apoptosis rate of 4T1 cells ( t = 10.99, P < 0.05). Meanwhile, from the 16 th day of tumor inoculation, the 10 Gy irradiation combined APR-246 group showed significantly inhibited tumor growth ( t = 2.38-2.91, P < 0.05) and significantly increased proportions of CD4+ and CD8+ T cells ( t = 9.96, 6.28, P < 0.05) and M1/M2 ratio ( t = 15.30, P < 0.05) in tumor tissues. Conclusions:APR-246 combined with irradiation can effectively increase ROS and LPO levels in 4T1 cells, promote tumor cell apoptosis, and induce anti-tumor immune response, thus potentially inhibiting the growth of 4T1 cells.

Objective:To investigate the clinical value of liquid based cytology in the diagnosis of early non-small cell lung cancer (NSCLC).Methods:From October 2018 to October 2019, 120 patients with early NSCLC who were confirmed diagnossis by histopathology were selected from Xixi Hospital of Hangzhou and Tongde Hospital of Zhejiang Province.All patients underwent fiberbronchoscopy.All the specimens were performed liquid-based cell wax block and HE staining.Two or more experienced histopathological doctors read the histological samples and then gave pathological reports.The cytological samples were first screened by the primary cytological diagnosis doctors, and then read by the deputy director or chief physician.The cytological pathological reports were given in combination with the clinical practice.Results:HE staining in liquid-based cytology showed that squamous cell carcinoma had many different sizes and shapes, single scattered, polygonal and round, even distribution of chromatin, and no obvious nucleolus.HE staining of adenocarcinoma in liquid-based cytology often gathered solid mass, rich cytoplasm, regular cell size, round nucleus, fine chromatin, and obvious nucleolus.HE staining of small cell cancer in liquid-based cytology mostly scattered, the cytoplasm was rare as naked nucleus, the nucleus was deeply stained, some were angular or short shuttle shaped.The sensitivity and specificity of liquid based cytology in the diagnosis of NSCLC were higher than those of traditional smear (χ 2=4.874, 4.512, all P<0.05). The positive rates of squamous cell carcinoma (72.88%) and adenocarcinoma (85.29%) in liquid based cytology were significantly higher than those in traditional cytology (54.24% and 61.76%) (χ 2=4.427, 6.031, all P<0.05), but there was no statistically significant difference in positive rate of small cell cancer between the two groups (χ 2=1.333, P>0.05). The positive rate of stage Ⅰa in liquid based cytology (86.48%) was higher than that in traditional cytology (64.86%), there was statistically significant difference (χ 2=4.698, P<0.05), but there were no statistically significant differences in the positive rates of stage Ⅰb and stage Ⅱa between the two groups (all P>0.05). Conclusion:Liquid based cytology is of great value in the diagnosis of early NSCLC, which can increase the sensitivity and specificity of diagnosis.