Objective To explore the targets and mechanisms of Tongguanteng Injection in inhibiting the proliferation and migration of cervical cancer.Methods The biological activity of Tongguanteng Injection in inhibiting human cervical cancer SiHa cells was determined by MTT method.Detecting the effect of Tongguanteng Injection on SiHa cell migration through wound healing assay.Using network pharmacology to collect the key targets for treating cervical cancer,and perform molecular docking and enrichment analysis on the targets.Immunohistochemistry and Western blot were used to detect the key proteins to validate the network pharmacology predictions.Result Tongguanteng Injection significantly inhibited the proliferation and migration in a dose-dependent manner in human cervical cancer SiHa cells.Based on the main active ingredients of Marsdenia tenacissima,81 therapeutic targets for cervical cancer were obtained,which may treat cervical cancer by affecting key proteins such as FGF2,MAPK1,and MAPK3.Immunohistochemical results indicated that FGF2,MAPK1 and MAPK3 were expressed in cervical cancer tissues.The western bolt assays showed that Tongguanteng Injection could significantly reduce the FGF2 protein expression.Meanwhile,the MAPK1 and MAPK3 protein expressions were significantly increased.Conclusion Tongguanteng Injection may regulate the FGF2,MAPK1 and MAPK3,effectively impede the proliferation and migration of cervical cancer.

Objective To explore the targets and mechanisms of Tongguanteng Injection in inhibiting the proliferation and migration of cervical cancer.Methods The biological activity of Tongguanteng Injection in inhibiting human cervical cancer SiHa cells was determined by MTT method.Detecting the effect of Tongguanteng Injection on SiHa cell migration through wound healing assay.Using network pharmacology to collect the key targets for treating cervical cancer,and perform molecular docking and enrichment analysis on the targets.Immunohistochemistry and Western blot were used to detect the key proteins to validate the network pharmacology predictions.Result Tongguanteng Injection significantly inhibited the proliferation and migration in a dose-dependent manner in human cervical cancer SiHa cells.Based on the main active ingredients of Marsdenia tenacissima,81 therapeutic targets for cervical cancer were obtained,which may treat cervical cancer by affecting key proteins such as FGF2,MAPK1,and MAPK3.Immunohistochemical results indicated that FGF2,MAPK1 and MAPK3 were expressed in cervical cancer tissues.The western bolt assays showed that Tongguanteng Injection could significantly reduce the FGF2 protein expression.Meanwhile,the MAPK1 and MAPK3 protein expressions were significantly increased.Conclusion Tongguanteng Injection may regulate the FGF2,MAPK1 and MAPK3,effectively impede the proliferation and migration of cervical cancer.

Objective:To investigate the effect and related mechanism of Marsdenia tenacissima combined with XELOX solution on disulfide apoptosis in human colorectal cancer HCT116 cells.Methods:The human colorectal cancer HCT116 cells were cultured in vitro and treated with different concentrations of capecitabine (0, 0.3, 0.6, 1.2, 2.4, 4.8 μg/ml), oxaliplatin (0, 10, 20, 40, 80, 160 μg/ml), Marsdenia tenacissima (0, 15, 30, 60, 120, 240 mg/ml), and the glucose inhibitor BAY-876 (0, 4, 8, 16, 32, 64 μg/ml), respectively. Furthermore, the HCT116 cells were pre-treated with 25 μg/ml of BAY-876, followed by exposure to the specified concentrations of each drug group. HCT116 cells were divided into the following groups: negative control group (no treatment), capecitabine group (4.0 μg/ml), oxaliplatin group (90 μg/ml), Marsdenia tenacissima group (140 mg/ml), XELOX solution group (4.0 μg/ml capecitabine+90 μg/ml oxaliplatin), and Marsdenia tenacissima combined with XELOX solution group (140 mg/ml Marsdenia tenacissima+4.0 μg/ml capecitabine+90 μg/ml oxaliplatin). BAY-876 treatment groups refer to the groups which the glucose inhibitor BAY-876 25 μg/ml was added to each of the above groups. The cell proliferation was assessed using the MTT assay. Apoptosis was determined through Annexin Ⅴ-FITC/PI double staining. The concentration of glucose was quantified using the o-toluidine method. NADPH levels were measured by colorimetry. Cystine uptake fluorescence assay was utilized to quantify the fluorescence intensity of cystine, and cysteine content was determined using cysteine colorimetry. Results:After 0, 0.3, 0.6, 1.2, 2.4, 4.8 μg/ml concentration of capecitabine, 0, 10, 20, 40, 80, 160 μg/ml concentration of oxaliplatin, 0, 4, 8, 16, 32, 64 μg/ml concentration of BAY-876, and after pre-treatment with 25 μg/ml glucose inhibitor BAY-876, HCT116 cells were treated with the above drugs and concentrations, there were statistically significant differences in cell survival rate ( F=644.60, P<0.001; F=417.30, P<0.001; F=1 028.00, P<0.001; F=1 066.00, P<0.001; F=847.70, P<0.001), and with the increase of each drug concentration, the activity of HCT116 cells decreased gradually (all P<0.05). After 0, 15, 30, 60, 120, 240 mg/ml concentration of Marsdenia tenacissima, and after pre-treatment with 25 μg/ml glucose inhibitor BAY-876, HCT116 cells were treated with the above drugs, there were statistically significant differences in cell survival rate ( F=107.50, P<0.001; F=619.70, P<0.001), and with the increase of drug concentration, the activity of HCT116 cells increased first and then decreased (all P<0.05). Annexin Ⅴ-FITC/PI staining showed that the intensity of green, red and yellow fluorescence was weaker in the negative control group. The expression of green fluorescence and red fluorescence was enhanced in each drug group. In capecitabine group, the red fluorescence and yellow fluorescence were larger. The proportion of green fluorescence in oxaliplatin group and Marsdenia tenacissima group was smaller. The proportion of green fluorescence and yellow fluorescence in XELOX solution group was higher than that in capecitabine group and oxaliplatin group. Marsdenia tenacissima combined with XELOX solution group had the highest proportion of green and red fluorescence. After pre-treatment with the glucose inhibitor BAY-876, the green and yellow fluorescence of the cells in each group increased significantly. The green and yellow fluorescence of capecitabine group and oxaliplatin group increased significantly. The fluorescence of XELOX solution group was significantly higher than that of capecitabine group and oxaliplatin group. In Marsdenia tenacissima combined with XELOX solution group, the proportion of fluorescence expressing three colors was the largest. In the negative control group, capecitabine group, oxaliplatin group, Marsdenia tenacissima group, XELOX solution group, Marsdenia tenacissima combined with XELOX solution group, and the groups after BAY-876 pre-treatment, the glucose concentration of HCT116 was (19.91±0.13), (22.82±0.88), (11.87±0.14), (17.93±0.14), (10.53±0.10), (7.56±0.08), (11.44±0.10), (11.73±0.72), (8.98±0.40), (14.25±0.33), (6.77±1.50), and (1.56±0.17) μg/ml, respectively, with statistically significant differences ( F=762.60, P<0.001; F=118.80, P<0.001). Compared with the untreated groups, the intracellular glucose concentration of HCT116 cells treated with BAY-876 was significantly decreased ( t=86.50, P<0.001; t=16.90, P<0.001; t=11.83, P<0.001; t=17.79, P<0.001; t=4.35, P=0.012; t=54.34, P<0.001). NADPH levels in each group were (131.80±2.61), (93.87±1.00), (136.50±3.69), (105.70±0.84), (146.90±2.94), (105.00±2.25), (92.33±0.23), (88.63±0.31), (97.33±2.02), (81.77±1.33), (102.80±1.61), and (85.13±0.45) nmol/gProt, respectively, with statistically significant differences ( F=225.60, P<0.001; F=125.50, P<0.001) ; Compared with the untreated groups, the intracellular NADPH levels of HCT116 cells treated with BAY-876 was significantly decreased ( t=26.11, P<0.001; t=8.62, P<0.001; t=16.13, P<0.001; t=26.38, P<0.001; t=22.78, P<0.001; t=14.97, P<0.001). The fluorescence intensity of cystine in each group was 607.30±8.76, 655.70±6.57, 647.10±19.35, 737.80±6.34, 756.00±8.65, 846.60±11.70, 929.60±6.88, 1 049.00±22.35, 1 021.00±29.49, 1 094.00±16.17, 1 137.00±10.08, and 1 230.00±46.57, respectively, with statistically significant differences ( F=188.00, P<0.001; F=48.32, P<0.001). Compared with the untreated groups, the intracellular fluorescence intensity of cystine of HCT116 cells was significantly increased after treatment with BAY-876 ( t=50.09, P<0.001; t=29.26, P<0.001; t=18.34, P<0.001; t=35.53, P<0.001; t=49.66, P<0.001; t=13.83, P<0.001). The contents of cysteine in each group were (457.00±30.69), (581.20±30.69), (326.40±5.49), (374.20±5.54), (565.30±5.54), (246.80±30.69), (100.30±16.57), (472.90±19.10), (262.70±28.65), (348.70±9.55), (533.40±11.03), (30.23±5.49) μmol/L, respectively, with statistically significant differences ( F=110.00, P<0.001; F=423.50, P<0.001). Compared with the untreated groups, the intracellular contents of cysteine of HCT116 cells treated with BAY-876 was significantly decreased ( t=17.71, P<0.001; t=5.19, P=0.006; t=3.78, P=0.019; t=4.00, P=0.016; t=4.47, P=0.011; t=12.03, P<0.001) . Conclusion:Marsdenia tenacissima combined with XELOX solution can promote HCT116 cell death through disulfide apoptosis.

Objective To investigate the protective effect of lentivirus-mediated silencing information regulation protein 6(SIRT6)overexpression on radioactive lung injury in rats.Methods 72 male 150-200 g Wistar rats were randomly divided into three groups(n=24).The 6 MV-X-ray linear accelerator was used to irradiate the lungs.The rats in each group were injected with normal saline(TNF-α)were measured at 1, 2, 4, and 8 weeks after radiotherapy(Lent-SIRT6), and the levels of tumor necrosis factor-α(TNF-IL-6 and IL-1β were measured by HE staining.The levels of TNF-α, IL-6 and IL-1β in liver tissue were detected by HE staining.The levels of TNF-α, IL-6 and IL-1β in liver tissue were detected by HE staining.Results The alveolar wall and alveolar stroma were normal in the control group.The alveolar wall of the irradiated group was thickened and fibrosis, and a large number of hypertrophic fibrous tissues were found in the alveolar stroma.The alveolar wall of Lent-SIRT6 group was thickened And alveolar stromal fibrosis symptoms were lighter than the irradiation group.Compared with the control group, the levels of serum TNF-α and IL-6, neutrophil count and the levels of TNF-α, IL-6 and IL-1β in the liver tissue were significantly higher than those in the control group (P<0.05).The lentiviruses overexpressing SIRT6 could alleviate the abnormalities caused by radiation (P<0.05).The difference was statistically significant (P<0.05).Conclusion SIRT6 can effectively inhibit the inflammatory reaction, reduce the lung injury symptoms of radiation pneumonia, and have some protective and prevention effect on lung injury.