Evodiamine activates autophagy and inhibits the proliferation in colon cancer cell / 中草药

Objective To investigate the effect of evodiamine (Evo) on the autophagy and proliferation of colon cancer HCT-116 cells and the underlying mechanism. Methods The effect of Evo on proliferation of HCT-116 cells was detected by CCK-8 method. After being processed with Evo (3 and 6 μmol/L) for 48 h, the number of autophagic vesicles were detected by MDC method. The amount of ROS in HCT-116 cells was measured by DHE assay, and the protein related with autophagy and AMPK/mTOR pathway was detected by Western blotting. After the treatment of Evo (6 μmol/L) combined with autophagy inhibitor 3-MA (3-methyladenine) or apoptosis inhibitor Z-DEVD-FMK respectively for 48 h, Western blotting was used to detect the expression of autophagy and apoptosis-related protein in HCT-116 cells. Results Compared with the control group, Evo inhibited the proliferation of HCT-116 cells in a dose-dependent manner; After treated with Evo (3 and 6 μmol/L) for 48 h, the amount of intracellular ROS and autophagic vesicles were increased, the protein expression levels of LC3, p-AMPK, and mTOR were increased while the expression of p62 was increased. After being treated with Evo and autophagy inhibitor, the protein expression of LC3 was decreased while activated Caspase-3 was increased; Combination of Evo and apoptosis inhibitor increased the expression of LC3 and inhibited the expression of activated Caspase-3. Conclusion Evo can activate autophagy of HCT-116 cells through AMPK/mTOR pathway and inhibit the proliferation, and the effect of autophagy and apoptosis on cells are complementary.

Morphological and molecular biological peculiarities of the experimental autoimmune prostatitis rat model / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the morphological and molecular biological peculiarities of the experimental autoimmune prostatitis (EAP) rat model made by SC purified prostate protein twice with immune adjuvant.</p><p><b>METHODS</b>Male rats were intradermally immunized with a saline extract of male rat prostate glands (RPG) in Freund's complete adjuvant (FCA) and Pertussis-Diphtheria-Tetanus vaccine 0.5 ml i.p. at the 0 and 30th day, and the concentrations of the extract were respectively 5 mg/ml, 10 mg/ml and 15 mg/ml. At the 45th day, the rats were sacrificed and the morphological and molecular biological changes of the prostate specimens were observed to determine the effective concentration of RPG for a successful model.</p><p><b>RESULTS</b>The expression of inflammation genes such as TNF-alpha, IL-1beta, IL-2 and iNOS obviously increased in the high-dosage model group; LM, EM and in situ hybridization revealed appearant chronic inflammation response, but this was not the case in the other two dosage groups.</p><p><b>CONCLUSION</b>15 mg/ml RPG mixed with FCA (1:1) 1.0 ml SC with Pertussis-Diphtheria-Tetanus vaccine 0.5 ml i.p. was an effective dosage for the successful model in our experiment.</p>

Vascular smooth muscle cell proliferation and apoptosis of the aortic wall during the development of abdominal aortic aneurysm / 中国普通外科杂志

Objective To explore vascular smooth muscle cell(SMC) proliferation and cell apoptosis during the development of abdominal aortic aneurysm(AAA). Methods The animal model of AAA was established in Wistar rats and the specimens were harvested at the 3rd day,and 1、2、3 and 4 week after the model initiation. In situ end-labeling of DNA fragments (TUNEL) was used to detect SMC apoptosis and immunohistochemical staining was applied to investigate the expression of SMC apoptosis markers(bcl-2,bax),proliferating cell nuclear antigen (PCNA) and ?-actin. Results TUNEL-positive and PCNA-positive SMC reached the maximum at 2～3 week and 1 week respectively;The count of TUNEL-positive SMC was less than PCNA-positive SMC during the period of day 3 to 1 week and that was vice versa from 2nd to 4th week with SMC amount significantly decreased;Bcl-2 and bax protein was strongly expressed at 1 week and 3 week after operation(all P