Objective: To establish anin vitromodel of ferroptosis induced by Erastin and RAS-selective lethal 3(RSL3) in hepatoma cells, and to provide theoretical basis for the development of novel therapeutic strategies for HCC. Methods: Hepatoma cells(HCCLM3, HepG2, Hep3B, Huh7 and PLC/PRF/5) in logarithmic growth phase were treated with Erastin(0-40 μmol/L) and RSL3(0-10 μmol/L) at double concentrations respectively. After 24 h, CCK-8 method was used to detect cell viability, draw growth curve, calculate IC50, and HCC cells sensitive to inducers were selected for follow-up experiments. The effect of inducer on the state of hepatoma cells was observed under light microscope, and immunoblotting and flow cytometry were used to verify whether the ferroptotic modelin vitrowas successfully constructed. Results: Huh7, Hep3B and HepG2 cells were sensitive to Erastin and RSL3, but HCCLM3 and PLC/PRF/5 were insensitive to Erastin and RSL3. When the concentration of Erastin and RSL3 reached the maximum, the survival rate was still above 65%. Huh7, Hep3B and HepG2 cells were selected for subsequent experiments. Compared with the control group, the expression of Glutathione peroxidase 4(GPX4), a ferroptotic marker, was down-regulated in a concentration-dependent manner. In Huh7, Hep3B and HepG2 cells, lipid reactive oxygen species(ROS) levels significantly increased after 24 h treatment with 10 μmol/L and 20 μmol/L Erastin, respectively; in Huh7 cells, lipid ROS levels significantly increased after 24 h treatment with 0.5 μmol/L and 1 μmol/L RSL3, respectively; in Hep3B and HepG2 cells, lipid ROS levels significantly increased after 24 h treatment with 1 μmol/L and 2 μmol/L RSL3, respectively, compared with control group. Conclusion Huh7, Hep3B and HepG2 cells are highly sensitive to Erastin and RSL3. Huh7, Hep3B and HepG2 cells treated with 10 μmol/L Erastin for 24 h are good models for simulating ferroptosis induced by Erastinin vitro, Huh7 cells treated with 0.5 μmol/L RSL3 for 24 h and Hep3B and HepG2 cells treated with 1 μmol/L RSL3 for 24 h are good models for simulating ferroptosis induced by RSL3in vitro.

Objective To explore the role of phospholipase A2 receptor 1 (PLA2R1) in the diagnosis,differential diagnosis and evaluation of idiopathic membranous nephropathy (IMN) in adult patients.Methods A total of 242 renal disease patients diagnosed by renal biopsy from March 2015 to January 2016 were enrolled,consisting of 90 IMN,20 secondary membranous nephropathy (SMN),82 IgA nephropathy (IgAN),30 minimal changed disease (MCD),16 focal segmental glomerulosclerosis (FSGS) and 4 membranoproliferative glomerulonephritis (MPGN).Their clinical data including age,sex,serum creatinine (Scr),serum albumin and 24 h urinary protein were collected.Serum PLA2R1 was measured by enzyme linked immunosorbent assay.PLA2R and IgG subclasses in glomeruli were detected by indirect immunofluorescence assay.The positive rate of serum PLA2R1 among those groups and its correlation with clinical-pathological parameters were analyzed.Results Compared with IMN patients,SMN,MCD and FSGS patients were younger (all P ＜ 0.01); IgAN patients were younger and had higher serum albumin and lower 24 h proteinuria (all P ＜ 0.001); MPGN patients had higher Scr (all P ＜ 0.01).The positive rate of serum PLA2R1 was 75.6％ in IMN patients,while it was 0.0％ in non-IMN patients.The distribution between serum PLA2R1 and pathological diagnosis had difference (P ＜ 0.001),their positive coincidence rate was 100％,negative coincidence rate was 87.4％,total coincidence rate was 90.9％ and their consistency was well (Kappa=0.795,P ＜ 0.001).Among IgG subtype comparisons between IMN patients and SMN patients in the glomeruli,only moderate or more positive IgG4 had statistical differences (82.2％ vs 5.0％,P ＜ 0.001); the positive rate of glomerular PLA2R1 was 41.1％ in IMN patients,higher than 10.0％ in SMN patients (P=0.009); positive PLA2R1 with moderate or more positive IgG4 in glomeruli in IMN patients was more than that in SMN patients (40.0％ vs 0.0％,P ＜ 0.001),which could improve the diagnostic specificity of IMN.In IMN patients serum PLA2R1 and glomerular PLA2R1 had statistical differences (P＜0.001).Spearman rank correlation analysis showed that serum PLA2R1 of IMN patients positively correlated with 24 h proteinuria (r=0.315,P=0.002),negatively correlated with serum albumin (r=-0.228,P=0.030) and didn't correlate with Scr (r=0.199,P=0.059).Conclusions Serum PLA2R can be used as the specific indicator for diagnosis,differential diagnosis of IMN and to reflect the severity of IMN in patients.

This paper designed an index for the differences in medical service prices and this index therein named the Hospital Price Difference Index ( HPDI) which is used as a quantitative tool for evaluation. During evalua-tion, the paper measured the levels of prices and the factors of influence in 18 public hospitals listed in the Sichuan Province. The results showed that the effect of regulating the levels of prices was reasonable and effective, but was sig-nificantly affected by the internal and external factors. The internal factors have been found to be the medical and clini-cal technologies, and the grade and scale acted as external ones. This paper suggested that the price department should pay more attention on the levels of prices, and hence made a reasonable reform project for the prices by taking the scale of adjustment of prices into account.