Tanshinone IIA protects intestinal epithelial cells from ferroptosis through the upregulation of GPX4 and SLC7A11

Background:Inflammatory bowel disease(IBD)is a chronic inflammatory disease of the gastrointestinal tract.The destruction of the intestinal epithelial barrier is one of the major pathological processes in IBD pathology.Growing evidence indicated that epithelial cell ferroptosis is linked to IBD and is considered a target process.Methods:RAS-selective lethal 3(RSL3)was used to induce ferroptosis in intestinal epithelial cell line No.6(IEC-6)cells,and cell ferroptosis and the effects of tanshinone IIA(Tan IIA)were determined by cell counting kit-8(CCK-8),reactive oxygen species(ROS)staining,Giemsa staining and transmission electron microscope(TEM).The cell viability of natural product library compounds was determined by CCK-8.The expression of ferroptosis-related genes were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and western blot.Results:Treatment of IEC-6 cells results in the accumulation of ROS and typical morphological characteristics of ferroptosis.RSL3 treatment caused rapid cellular cytotoxicity which could be reversed by ferrostatin-1(Fer-1)in IEC-6 cells.Natural product library screening revealed that Tan IIA is a potent inhibitor of IEC-6 cell ferroptosis.Tan IIA could significantly protect the RSL3-induced ferroptosis of IEC-6 cells.Furthermore,the ferroptosis suppressors,glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),and miR-17-92 were found to be early response genes in RSL3-treated cells.Treatment of IEC-6 cells with Tan IIA resulted in upregulation of GPX4,SLC7A11,and miR-17-92.Conclusion:Our study demonstrated that Tan IIA protects IEC-6 cells from ferroptosis through the upregulation of GPX4,SLC7A11,and miR-17-92.The findings might provide a theoretical grounding for the future application of Tan IIA to treat or prevent IBD.

Expression and clinical significance of PTEN protein in osteosarcoma

Objective： To investigate the expression and significance of cancer inhibitory gene PTEN protein in osteosar-coma. To analyze the level of its expression in different histological classification of osteosarcoma. To determine the possibility of taking PTEN protein as a marker gene for diagnosing osteosarcoma. To observe the clinical value of PTEN expression levels as a reference index for osteosarcoma classification. Methods： 43 specimens collected from osteosarcoma excision were studied. 30 specimens collected during the same period from benign lesion of bone （osteochondroma） operation were taken as the control group. Immunohistochemistry staining （ElivisonTM two steps method） was used to detect the expression of PTEN protein in 43 cases of osteosarcoma. SPSS 10.0 was used in statistical analysis. Results： Immunohistochemistry staining showed that the positive reaction of PTEN protein was all oriented to cytoplasm, which were brown or yellowish- brown granules. By way of X^2 test, the significant difference of the positive expressions of PTEN protein between bone benign lesion and osteosarcoma （X^2 = 7.976, P 〈 0.01） was observed. Osteosarcoma with different degrees of histodifferentiation showed different level expression of PTEN protein. There was significant difference between well-differentiated osteosarcoma （grades Ⅰ-Ⅱ） and poorly-differentiated osteosarcoma （grade Ⅲ） statistically （P 〈 0.01）. The level of expression of PTEN was negatively correlated to the histological grade of osteosarcoma. There was great significance statistically （rs=-0.4922, P 〈 0.01）. Conclusion： PTEN protein may be used as candidate gene of cancer inhibitory gene： PTEN protein is a cancer suppressor gene protein which has expression in bone tumors. It might not only be used in the study of pulmonary carcinoma and neurogliocytoma, but also in the study of bone tumor; the expression of PTEN is related to benignancy or malignancy of bone tumor and their degree of differentiation. The expression of PTEN is positively correlated with degree of differentiation.

Meta analysis of the effect of distal or local point selection on acupuncture efficacy

To evaluate the effect of distal point selection, local point selection, and distal-local point selection on acupuncture efficacy. Methods： According to the requirements of evidence-based medicine（EBM）, the literature on acupuncture with distal and local point selection in recent 20 years included in domestic and overseas medical databases was retrieved, and meta-analysis of the included articles was carried out. Results： Thirty-two articles were included finally, totaling 2829 patients. The results of meta-analysis indicated that the curative effect of distal-local point combination was superior to that of simple distal point selection or local point selection during acupuncture–moxibustion treatment for diseases, and there was no obvious difference in the curative effect between simple distal point selection and local point selection. According to the comprehensive analysis of 18 included articles, the difference in total effective rate between distal point selection and local point selection was not statistically significant, i.e. the curative effect of distal point selection and local point selection was equivalent [OR=0.83,95%CI（0.83,1.18）, Z=1.04, P0.01]. According to the comprehensive analysis of 20 included articles, the difference in total effective rate between local point selection and distal-local point selection was statistically significant, and the results indicated that the curative effect of distal-local point selection was superior to that of local point selection [OR=0.32, 95%CI（0.23,0.44）, Z=6.90, P0.01]. According to the comprehensive analysis of 8 included articles, the difference in total effective rate between distal point selection and distal-local point selection was statistically significant, and the results indicated that the curative effect of distal-local point selection was superior to that of distal point selection [OR=0.20,95%CI（0.10,0.40）, Z=4.50, P0.01]. In addition, the analysis of publication bias of this study indicated that publication bias might exist. Since the quality of included articles was generally lower, the above conclusion still needed to be supported by more evidence-based medicine proofs with high quality. Conclusion： The curative effect of distal-local point combination was superior to that of simple distal point selection or local point selection during acupuncture treatment for diseases, and there was no obvious difference in the curative effect between simple distal point selection and local point selection.

Specific bFGF targeting of KIM-1 in ischemic kidneys protects against renal ischemia-reperfusion injury in rats

Renal ischemia-reperfusion(I/R)injury is one of the major causes of acute kidney injury.However,there is still no effective treatment for this disease.Basic fibroblast growth factor(bFGF)has been reported to be beneficial for recovery from ischemic diseases.It is vital to increase the local concentration and reduce the diffusion of bFGF in vivo for renal I/R injury therapy.A targeted growth factor delivery system that responds to specific biological signals in the regenerative environment to guide release has been highlighted in tissue repair.In the present study,a specific peptide was fused with bFGF and called bFGF-kidney injury targeting(KIT-bFGF),and this compound specifically targeted kidney injury molecule-1 both in hypoxic renal HK-2 cells in vitro and ischemic kidneys in vivo after intravenous injection.When administered to rat models of renal I/R injury,KIT-bFGF attenuated renal tubule damage and fibrosis,and promoted functional recovery compared to the effects of native bFGF and the control.We also investigated the mechanism by which KIT-bFGF activated the ERK1/2 and Akt signaling pathways to significantly reduce apoptosis and protect against ischemic injury in the kidney.These results demonstrated that targeted delivery of KIT-bFGF could be an effective strategy for the treatment of renal I/R injury.