Amentoflavone promotes ferroptosis by regulating reactive oxygen species (ROS) /5'AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) to inhibit the malignant progression of endometrial carcinoma cells.

It was reported that amentoflavone (AF) had anti-tumor ability. Therefore, this study aimed to investigate the role of AF in endometrial cancer as well as to discuss its underlying mechanism. The viability, proliferation, and apoptosis of endometrial carcinoma cells (KLE) with AF administration were detected by methyl tetrazolium (MTT) assay, clone formation, and terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assays. Thiobarbituric acid reactive substance (TBARS) production and Fe2+ level in AF-treated KLE cells were detected by TBARS assay and Iron assay. The expressions of proliferation- apoptosis-, ferroptosis-, and 5'AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling-related proteins in AF-treated KLE cells were detected by western blot analysis. Reactive oxygen species (ROS) expression in AF-treated KLE cells was determined by ROS assay kit. N-acetyl cysteine (NAC), which is an inhibitor of ROS, was used to confirm whether AF exerted its effects on KLE cells through ROS/AMPK/mTOR signaling. As a result, AF inhibited the viability and proliferation of KLE cells but promoted apoptosis and ferroptosis. The expressions of ROS and AMPK were increased, while mTOR expression was decreased in AF-treated KLE cells. NAC reversed the effects of AF on biological behaviors of KLE cells by inactivating ROS/AMPK/mTOR signaling. In conclusion, AF promoted ferroptosis by activating ROS/AMPK/mTOR to inhibit the viability and proliferation and promoted the apoptosis and ferroptosis of KLE cells.

Forsythoside A Alleviates High Glucose-Induced Oxidative Stress and Inflammation in Podocytes by Inactivating MAPK Signaling via MMP12 Inhibition.

BACKGROUND: Podocyte injury serves an important role during the progression of diabetic nephropathy (DN). The aim of this study was to investigate the effects of forsythoside A (FA) on high glucose (HG)-induced podocyte injury and to identify the possible mechanisms. METHODS: MPC-5 podocytes were cultured under HG conditions. After exposure to different doses of FA, cell viability and apoptosis were respectively evaluated with CCK-8 assay and flow cytometry. Then, the levels of oxidative stress-related markers and inflammatory factors were examined by corresponding kits. Western blot analysis was employed to detect the expression of Nox2, Nox4, COX-2, iNOS and matrix metalloproteinases 12 (MMP12). Subsequently, MMP12 was overexpressed to assess whether the effects of FA on HG-stimulated podocyte injury were mediated by MMP12 and MAPK signaling. RESULTS: Results indicated that FA dose-dependently elevated cell viability, reduced cell apoptosis in HG-induced MPC-5 cells. Additionally, FA significantly inhibited oxidative stress, which could be certified by decreased content of malondialdehyde (MDA), enhanced activities of superoxide dismutase (SOD) and catalase (CAT), and downregulated expression of Nox2 and Nox4. Moreover, notably reduced levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6 were observed in FA-treated MPC-5 cells under HG conditions, accompanied by decreased COX-2 and iNOS expression. Remarkably, FA suppressed MMP12 expression in a dose-dependent manner, and the effects of FA on MPC-5 cells exposed to HG were partially counteracted by MMP12 overexpression. Mechanically, FA inactivated the expression of phospho-ERK (p-ERK), p-p38 and p-JNK, which was restored after MMP12 overexpression. CONCLUSION: These findings demonstrate a protective mechanism of FA by inactivating MAPK signaling via MMP12 inhibition in HG-induced podocyte injury, providing a promising therapeutic candidate for the treatment of DN.

Team-based learning: assessing the impact on anatomy teaching in People's Republic of China.

OBJECTIVES: In this study, the effects of team-based learning (TBL) method on the anatomy course for students in People's Republic of China were assessed. METHODS: The students were randomly divided into the traditional lecture-based teaching group (traditional learning [TL] group, 99 students) and TBL teaching group (98 students). The TBL method required the students to prepare the assigned content in advance and discuss some specific topics in small groups. The test scores and questionnaire were applied to evaluate the effects of the two methods. RESULTS: The students in TBL group had higher examination scores (81.70±8.53 vs 74.41±8.27, F[1,195] =124.6, p<0.01). The ratios of students with excellent (13.27% vs 9.09%, χ2[1] =4.00, p=0.041) and good scores (25.51% vs 16.16%, χ2[1] =4.85, p=0.027) were markedly increased in the TBL than the TL group, and the ratio of students who had just managed to pass was decreased (17.34% vs 32.33% in TL group, χ2[1] =5.91, p=0.015). The students in TBL group significantly achieved some improvement in mutual communication ability (χ2[1] =7.54, p=0.006), expression ability (χ2[1] =4.930, p=0.026), generalization capacity (χ2[1] =4.08, p=0.043), cooperative ability cultivation (χ2[1] =5.04, p=0.024), knowledge extension (χ2[1] =4.50, p=0.034), and enthusiasm mobilization (χ2[1] =4.27, p=0.039). CONCLUSION: TBL could improve not only the test scores of the students, but also their study enthusiasm, initiative learning ability, communication ability, and team awareness.