Objective:To observe the effects of myricetin on autophagy and mitochondrial fission in the human ovarian cancer SKOV3 cells,and to explore its induction effect on autophagy and promoting effect on mitochondrial fission.Methods: The SKOV3 cells were cultured in vitro.0,20,40,and 60 g·L-1 myricetin were used in control group and low, middle, high doses of myricetin groups for 12 h.The changes of mitochondrial membrane potential were detected by flow cytometry;the morphology of mitochondria was observed by MitoTracker Red;the expression levels of dynamin related protein1 (Drp1) and fission 1 (Fis1) in various groups were detected by Western blotting method;and the expression levels of autophagy related protein LC3 were detected by both Western blotting method and immunofluorescence method.Results:Compared with control group, the ratios of decreased mitochondria in different doses of myricetin groups were significantly increased(t=3.27, t=6.85, t=5.49,P<0.05).Compared with control group, the numbers of mitochondria in different doses of myricetin groups were increased, and the mitochondria looked more like gravel.Compared with control group,the expression levels of Drp1 in different doses of myricetin groups were significantly increased (t=4.35, t=3.28, t=6.17,P<0.05), and the expression levels of Fis1 were increased also(t=8.32, t=6.74, t=9.27).The immunofluorescence results showed that the expression levels of LC3 in different doses of myricetin groups were significantly increased with the increase of myricetin dose compared with control group.The Western blotting results showed that the ratios of LC3-Ⅱ/LC3-Ⅰ in middle and high doses of myricetin groups were significantly increased compared with control group(t=3.28, t=4.21,P<0.05).Conclusion: Myricetin can induce the autophagy of SKOV3 cells, and it can also promote the mitochondrial fission.

Objective To observe protective effects of allicin on heart and lung in aging mice induced by D-galactose, and try to explore the possible mechanism from the aspect of oxidative stress. Methods Fifty male Kunming mice were randomly divided into 5 groups:normal control group ( group C) ,model control group ( group D) and high-,middle-and low-dose of allicin groups ( H,M,L group) . The rats in group D,L,M and H were injected with D-galactose. At the same time,the rats in group L,M and H were injected with allicin. All of the injection lasted for 6 weeks. After that,athletic ability of the mice was evaluated by behavioral experiments. Malondialdehyde ( MDA ) content, superoxide dismutase ( SOD ) activity and total antioxidant capacity ( T-AOC) in the homogenate of heart and lung tissues were measured,and the expression levels of Bax and Bcl-2 were tested in heart and lung. Results Behavioral experiment showed that standing time on Rota Rod System and continuous running time on mice treadmill were significantly shortened in group D as compared with group C ( P<0.05) ,but those were prolonged in the allicin groups. Compared with group C,MDA content in homogenate of myocardium and lung tissues was significantly increased in group D (P<0.05),the activity of SOD was decreased (P<0.05) and T-AOC was decreased (P<0.05) . As compared with group D,MDA content in allicin groups was significantly decreased ( P<0.05) ,SOD activity increased (P<0.05) and A-TOC increased (P<0.05) in a dose-dependent manner. The results of immunohistochemistry showed that the expression of Bax was significantly increased in group D as compared with group C,and the expression of Bcl-2 was decreased. After the intervention of allicin,expression of Bax was decreased in group L,M and H as compared with group D, and expression of Bcl-2 was increased as compared with group D. Expression levels of Bax and Bcl-2 in lung tissues showed the same changing trend. Conclusion Allicin has a potential protect role on heart and lung in D-galactose-induced aging mice by increasing athletic ability,decreasing oxidative stress and decreasing cell apoptosis of myocardium and lung tissues.

OBJECTIVE To observe the effect of the autophagy and endoplasmic reticulum stress (ERS) in vascular smooth muscle cells (VSMCs) with hydrogen peroxide (H2O2). METHODS The VSMCs were incubated with different concentrations of H2O2(50, 100, 200, 400, 600, 800, 1200 and 1600 μmol · L-1)for 12 and 24 h. The cell viability was determined by MTT assay. The cell morphology was observed under an inverted phase contrast microscope. The expression of autophagy related protein ubiquitin binding protein p62 and microtubule-associated protein light chain 3(LC3)was detected by indirect immunofluorescence. Western blotting was used to detect autophagy related protein human coiled-coil myosin-like BCL2-interacting protein (Beclin-1),LC3 and mammalian target of sirolimus Rapamycin(mTOR),as well as the expression of endoplasmic reticulum stress(ERS)related protein glucose regulated proteins 78 ku(GRP78)and C/EBP homologous protein(CHOP). RESULTS MTT results showed that H2O2 inhibited the growth of VSMCs cells. The half inhibitory concentration (IC50) was 597.2±2.3 and(447.4±1.7)μmol·L-1 at 12 and 24 h,respectively. The results of the inverted phase contrast microscope showed that VSMCs in H2O2 group shrinked and turned into smaller round cells. The cell survival rate declined from(97.5 ± 0.1)% in normal control group to(74.4 ± 1.0)% and(56.8 ± 0.9)% in H2O2 400μmol·L-1 at 12 and 24 h, respectively. The results of the laser scanning confocal micro?scope showed that H2O2 400 μmol · L- 1 increased the expression of LC3 and p62 protein in a cytoplasmic time-dependent manner, and increased the colocalization of LC3 and p62,especially at 8 h. The results of Western blotting demonstrated that H2O2 increased the expression of ERS related protein GRP78 and CHOP,autophagy related protein Beclin-1 and LC3Ⅱ/LC3Ⅰ(P<0.01),and decreased mTOR(P<0.01)in VSMCs. CONCLUSION H2O2 induces autophagy through ERS in VSMCs.

Objective To evaluate the effect of five-segemental care model for chronic obstructive pulmonary disease patients with non-invasive mechanical ventilation. Methods A total of 70 community-dwelling COPD patients with non-invasive mechanical ventilation were randomly divided into two groups, with 35 cases in each group. The patients in the control group received routine discharge guidance and follow-up education, while the patients and their family members in the experimental group participated in five-segemental care model within two months after discharge. The frequency of acute exacerbations, lung function and blood gas parameters, as well as the quality of life when discharge and at 6 months after discharge were compared between two groups. Results After 6 month of discharge, the PaCO2 , PaO2 and FEV1/FVC of the experimental group were (61. 3 ± 5. 2)mmHg, (58. 4 ± 4. 7) mmHg, and (66. 8 ± 16. 3)%, which had no significant differences compared with the control groups (t= -0. 159,0. 477, 0. 968;P>0. 05). After 6 month of discharge, the times of acute exacerbation were (2. 5 ± 1. 1) of the experimental group and (3. 4 ± 2. 1) of the control group. There was significant difference between two groups (t= -3. 696,P<0. 01). Conclusions The application of five-segemental care model can improve COPD patients′ self-care knowledge and skills, promote patients′compliance on the application of non-invasive mechanical ventilation, reduce the frequency of acute exacerbations and improve the patients′quality of life.