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 study the occurrence site characteristics of middle cerebral artery aneurysm (MCAA ) in volume CT digital subtraction angiograhy(VCTDSA) .Methods The image characteristics in 72 cases of MCAA from May 2009 to January 2011 were retrospectively analyzed ,according to the running of middle cerebral artery ,the occurrence sites of aneurysm were divided into four categories :M1 segment ,M2 bifurcation ,M2 distal and M3-M5 segment ,the aneurysm number was conducted the statistics and the image characteristics were analyzed ;two neuroradiologists adopted the double-blind method to measure the MCAA angle in bifurcation of M2 segment and compared it with the bifurcation angle in the normal middle cerebral artery ,the difference between them were statistically analyzed .Results (1)M1 segment aneurysms were 7 cases(9 .70% ) ,M2 bifurcation aneurysms were 58 ca-ses(80 .56% ) ,M2 distal aneurysms were 5 cases(6 .94% ) and M3-M5 segment aneurysms were 2 cases(2 .78% ) .(2)The angle in M2 bifurcation of the normal middle cerebral artery was(99 .30 ± 22 .96)° ,M2 bifurcation aneurysm angle was(139 .26 ± 27 .61)° , the difference between them showing statistical significance (P<0 .01) .(3)The difference between left and right of M 2 bifurcation angle in the normal middle cerebral artery had no statistical significance (P>0 .05) .(4)The M2 bifurcation angle in ruptured aneu-rysm was(133 .98 ± 30 .24)° ,which in unruptured aneurysms was(144 .53 ± 21 .81)° ,the difference between them had no statistical significance(P>0 .05) .Conclusion MCAA mainly occurred in M2 bifurcation .There is significant difference in M2 bifurcation an-gle between the aneurysm group and non-aneurysm group ,M2 bifurcation angle is increased ,the probability of aneurysm occurrence is increased .