Metabolic-associated fatty liver disease (MAFLD), which is previously known as non-alcoholic fatty liver disease (NAFLD), represents a major health concern worldwide with limited therapy. Here, we provide evidence that ferroptosis, a novel form of regulated cell death characterized by iron-driven lipid peroxidation, was comprehensively activated in liver tissues from MAFLD patients. The canonical-GPX4 (cGPX4), which is the most important negative controller of ferroptosis, is downregulated at protein but not mRNA level. Interestingly, a non-canonical GPX4 transcript-variant is induced (inducible-GPX4, iGPX4) in MAFLD condition. The high fat-fructose/sucrose diet (HFFD) and methionine/choline-deficient diet (MCD)-induced MAFLD pathologies, including hepatocellular ballooning, steatohepatitis and fibrosis, were attenuated and aggravated, respectively, in cGPX4-and iGPX4-knockin mice. cGPX4 and iGPX4 isoforms also displayed opposing effects on oxidative stress and ferroptosis in hepatocytes. Knockdown of iGPX4 by siRNA alleviated lipid stress, ferroptosis and cell injury. Mechanistically, the triggered iGPX4 interacts with cGPX4 to facilitate the transformation of cGPX4 from enzymatic-active monomer to enzymatic-inactive oligomers upon lipid stress, and thus promotes ferroptosis. Co-immunoprecipitation and nano LC-MS/MS analyses confirmed the interaction between iGPX4 and cGPX4. Our results reveal a detrimental role of non-canonical GPX4 isoform in ferroptosis, and indicate selectively targeting iGPX4 may be a promising therapeutic strategy for MAFLD.

OBJECTIVE： To investigate the substance basis and mechanism of Xiaochaihu decoction in treatment of sepsis， and to provide reference for clinical application and R&D of the decoction. METHODS： Based on TCM integrative pharmacology platform （TCMIP）， chemical component analysis of Xiaochaihu decoction， disease target prediction， gene function and pathway enrichment analysis were all performed. The multi-dimensional network relationship of “TCM-chemical components-core targets-key pathways” was established， and the mechanism of Xiaochaihu decoction in treatment of sepsis was investigated. RESULTS： A total of 224 predicted chemical ingredients of Xiaochaihu decoction （including saikoside， ginsenoside， glycyrrhizin， etc.） interacted with 118 key targets about sepsis， including PF4， MYD88， TLR4， CD14， NOS3， etc. Its anti-sepsis mechanism involved nervous system， endocrine system， immune response and energy metabolism， etc. CONCLUSIONS： Based on “neuronal- endocrine-immune-metabolism”， Xiaochaihu decoction achieved its role in regulating sepsis by multi-level， multi-channel and multi-channel. This research may reveal the potential mechanism of Xiaochaihu decoction for sepsis， and the prescription provide theoretical basis for further experimental research of pharmacodynamic substance basis and mechanism of action．

Objective To investigate the display status of ultrasonography imaging check in central nervous system (CNS) in infants of early pregnancy and the diagnostic value of CNS malformation in infants of early pregnancy.Methods Gestational weeks of 2 751 enrolled subjects were divided according to the ultrasonic measurement of the crown rump length (CRL):11-11 +6 weeks group,12-12+6 weeks group,and 13-13 + 6 weeks group,prenatal ultrasound were performed to examine fetal CNS anatomy in infants of early pregnancy,record the display status in each groups of infants and analyze the relationship between the display situation and gestational age.Results Fourteen cases of fetal CNS malformation (20 malformations) in total were found by prenatal ultrasound,and the incidence of CNS malformation was about 5.09％ (14/2 571).Wherein,12 cases of early pregnancy were diagnosed,and 2 cases of middle pregnancy were diagnosed.The sensitivity of ultrasound of early pregnancy in the diagnosis of fetal CNS malformation was 85.71％.In the group of research,the ultrasound display ratios of 11-11+6 weeks group,12-12+6 weeks group and 13-13+6 weeks group were 96.73％,97.94％,98.06％,respectively.There was no significant difference in early pregnancy fetal CNS display ratio among groups (x2 =1.56,v =2,x2＜ x0.05.2 =5.99,P ＞ 0.05).Conclusions The display rate of CNS structure in infants of early pregnancy (11-13+6 weeks)is higher,and is not affected by gestational weeks.Prenatal ultrasound can effectively diagnose CNS severe malformation in infants of early pregnancy.

Objective To investigate the effects of hyperbaric oxygen (HBO) on the osteogenic differentiation of adipose-derived stem cells.Methods Adipose-derived stem cells were isolated,cultured and passed to next generations.The fourth adipose-derived stem cells were collected and seeded to the 96-well plates (2 × 105 cells/well) for culture in osteogenic induction medium,and were randomly divided into the control group and the hyperbaric oxygen (HBO) group.In the entire experiment,the control group was only kept in the 5％ CO2 incubator at a temperature of 37 ℃,while the HBO group was treated with HBO for 24 hours a day at the same culture conditions as the control group.(The HBO treatment cells were placed in the HBO chamber,which was washed with pure oxygen at a ventilation rate of 10 L/min for 15min.When oxygen flow rate was down-regulated to 2-3 L/min,the chamber was compressed for 20-25 min to a pressure of 2.5 ATA,which was maintained for 60 minutes.Then,uniform decompression was implemented for 20-25 minutes.At days 7,14 and 21,alkaline phosphatase activity of the cells was detected by alkaline phosphatase kit.At days14 and 21,osteocalcin concentration was detected by using the osteocalcin quantitative test kit,and at day 21,the calcified nodules were stained with Alizarin red.The data obtained were analyzed by 2 independent samples t test,paired sample t test and repeated test ANOVA.Results The cultured cells could express such mesenchymal stem cell markers as CD90 and CD105,but could not express hematopoietic markers such as CD34.After 7,14 and 21 days of culture,the activity of alkaline phosphatase in the HBO group was significantly increased,as compared with that of the control group A (P ＜ 0.01).Following culture for 14 and 21 days,the expression level of osteocalcin in the HBO group was significantly elevated,as compared with that of the control group (P ＜ 0.05).At day 21,the detection results by alizarin red staining showed that the number of calcified nodules for the HBO group was increased markedly as compared with that of the control group,and the calcified size became larger.Conclusions Treatment of HBO could increase osteogenic differentiation of adipose-derived stem cells.

Objective To investigate the effects of hyperbaric oxygen (HBO) on the osteogenic differentiation of adipose-derived stem cells.Methods Adipose-derived stem cells were isolated,cultured and passed to next generations.The fourth adipose-derived stem cells were collected and seeded to the 96-well plates (2 × 105 cells/well) for culture in osteogenic induction medium,and were randomly divided into the control group and the hyperbaric oxygen (HBO) group.In the entire experiment,the control group was only kept in the 5％ CO2 incubator at a temperature of 37 ℃,while the HBO group was treated with HBO for 24 hours a day at the same culture conditions as the control group.(The HBO treatment cells were placed in the HBO chamber,which was washed with pure oxygen at a ventilation rate of 10 L/min for 15min.When oxygen flow rate was down-regulated to 2-3 L/min,the chamber was compressed for 20-25 min to a pressure of 2.5 ATA,which was maintained for 60 minutes.Then,uniform decompression was implemented for 20-25 minutes.At days 7,14 and 21,alkaline phosphatase activity of the cells was detected by alkaline phosphatase kit.At days14 and 21,osteocalcin concentration was detected by using the osteocalcin quantitative test kit,and at day 21,the calcified nodules were stained with Alizarin red.The data obtained were analyzed by 2 independent samples t test,paired sample t test and repeated test ANOVA.Results The cultured cells could express such mesenchymal stem cell markers as CD90 and CD105,but could not express hematopoietic markers such as CD34.After 7,14 and 21 days of culture,the activity of alkaline phosphatase in the HBO group was significantly increased,as compared with that of the control group A (P ＜ 0.01).Following culture for 14 and 21 days,the expression level of osteocalcin in the HBO group was significantly elevated,as compared with that of the control group (P ＜ 0.05).At day 21,the detection results by alizarin red staining showed that the number of calcified nodules for the HBO group was increased markedly as compared with that of the control group,and the calcified size became larger.Conclusions Treatment of HBO could increase osteogenic differentiation of adipose-derived stem cells.