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Objective:To investigate the effects of anemoside B4 on apoptosis of retinal cells in diabetic rats.Methods:Sixty Sprague-Dawley rats were randomized into three groups: the normal control(control), diabetic rats(DM)and diabetic rats treated with Anemoside B4(B4)groups(n=20, each group). Rats in the DM and B4 groups were rendered diabetic with an intraperitoneal injection of streptozotocin(STZ, 60 mg/kg). After 3 days of successful modeling, rats in the B4 group were intraperitoneally injected with anemoside B4(5 mg/kg), twice/day, for 8 weeks, while rats in the control and DM groups were injected with an equivalent volume of normal saline.After 8 weeks of anemoside B4 and normal saline injection, rats were sacrificed and retinas were harvested for examination.Paraffin sections of retina were stained with the hematoxylin-eosin(H-E)method for morphological evaluation.Protein levels of Bax and Bcl-2 were detected by using Western blot.The expression of caspase-3 mRNA was detected with quantitative PCR.Results:H-E staining results showed the control group had intact retinal structure and clear morphological features, whereas disordered retinal structure, thinner layers, and sparse and disorganized cells were seen in the DM group.However, retinal structure and morphology were improved after treatment with anemoside B4.Compared with the control group, the protein expression of Bcl-2 was lower( t=57.81, P<0.01), the protein expression of Bax was higher( t=10.47, P<0.01), and the Bcl-2/Bax ratio was lower( t=23.98, P<0.01)in the DM group.Compared with the DM group, the protein expression of Bcl-2 was higher( t=41.07, P<0.01), the protein expression of Bax was lower( t=6.811, P<0.01), and the Bcl-2/Bax ratio was higher( t=14.70, P<0.01)in the B4 group.Caspase-3 mRNA expression was higher in the DM group than in the control group( t=7.916, P<0.01), but was lower in the B4 group compared with the DM group( t=6.221, P<0.01). Conclusions:Anemoside B4 can inhibit the apoptosis of retinal cells by up-regulating Bcl-2 expression and down-regulating Bax and caspase-3 expression in diabetic rats.
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Objective@#To explore the relationship between psychological resilience and cognitive bias towards school violence in grade 3-5 primary school students in Luzhou city, so as to provide scientific basis for prevention and control of school violence in primary school students.@*Methods@#Students from grade 3-5 in primary schools in Luzhou were selected through stratified cluster random sampling method and were investigated with questionnaire survey.@*Results@#A total of 5 976 valid questionnaires were included, with an average score of psychological resilience (40.08±8.05) and an average score of school violence cognition (62.55±6.38). Multivariate results showed that psychological resilience was an independently associated with school violence perception (OR=1.04, P<0.01). The awareness of campus violence increased with resilience score. In addition, public school (OR=0.45) was associated with low awareness of school violence; senior grades (OR=1.77), girls (OR=1.20), and a greater number of friends(OR=1.37), student cadre(OR=1.37), middle/upper score in class(OR=2.13), no game playing(OR=1.33), no off-campus wandering(OR=1.78), timely parenting (OR=1.45) was associated with high awareness of school violence(P<0.05).@*Conclusion@#Psychological resilience positively correlates with cognition bias towards school violence. The higher the psychological resilience, the more positive perception of campus violence. Family, school and community-based interventions to enhance the resilience of students, increasing awareness towards school violence and ultimately reducing potential adverse impacts of school violence.
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Aim To investigate the effect of small in-terfering RNA(siRNA) targeting DNMT1 gene on cell proliferation, apoptosis and histone modulation in acute lymphoid leukemia cell line, Molt-4. Methods The small interfering RNA targeting DNMT1 gene was transfected into Molt-4 cells by LipofectamineTM 2000. The DNMT1 mRNA and protein level were detected by RT-PCR and Western blot. Cell proliferation was de-termined by MTT. Cell apoptosis was measured by Flow Cytometry. The expression of Bcl-2, procaspase-3, P15, histone methylation and histone acetylation was detected by Western blot. Results DNMT1 was suppressed by siRNA targeting DNMT1 in a concentra-tion-dependent manner. DNMT1 siRNA suppressed cells proliferation and induced apoptosis in Molt-4 cells. Apoptotic rate was (4. 27 ± 1. 42)% , (15. 25 ± 1. 54)% , (35. 63 ± 2. 54)% , (66. 27 ± 3. 02)%after transfecting with DNMT1 siRNA at 0, 30, 60, 120 nmol·L - 1 for 24 hours, P < 0. 05. The expres-sion of Bcl-2, procaspase-3 was suppressed and P15 was promoted after transfecting of DNMT1 siRNA. DN-MT1 siRNA downregulated histone methylated H3K9 and upregulated histone methylated H3K4. The altera-tion of histone acetylation of H3 was not seen. Conclu-sion DNMT1 siRNA suppresses DNMT1 efficiently in Molt-4 cells. The depletion of DNMT1 downregulates histone methylation of H3K9, and upregulates histone methylation of H3K4. It inhibits cell growth and in-duces cell apoptosis in Molt-4 cell line.
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Aim To study the protective effects of Ganoderma lucidum polysaccharides peptide(GLPP)on ECV304 from oxidative injury.Methods Cultured ECV304 were injured by oxygen free radicals derived from tBOOH.Various concentrations of GLPP(12.5,25,50,100 mg?L-1)were added into culture medium.The survival rate of cells was measured by MTT assay.The morphological change of cells and injury of mitochondria were examined under the light and electron microscopes.The percentage of apoptosis of ECV304,labeled with AnnexinV/PI,was measured by flow cytometry.Results GLPP(12.5,25,50,100 mg?L-1)could reduce oxidative injury induced by tBOOH in ECV304 cells.The survival rate of cells treated with GLPP increased.The light microscopic examination showed that the injured cells decreased in GLPP-treated groups.Under the electron microscope it was found that GLPP(50 mg?L-1,incubated for 24 h)could protect the organelle such as mitochondria from oxidative injury and cells from apoptosis by tBOOH.The result of flow cytometry showed that the total percentage of apoptosis in control,GLPP and injury treated group was 2.24%?0.43%,24?6.4%(P
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Aim To study the effects of Ganoderma polysaccharide peptide (GLPP) on the production of nitric oxide (NO) in mice peritoneal macrophages induced by LPS and its mechanism. Methods The effects of GLPP on the NO releasing from mice peritoneal macrophages induced by LPS were detected by using Griess reagent and the expression of iNOS by GLPP in mice peritoneal macrophages was detected by immunohistochemical method.Result The production of NO was increased by GLPP.(25～200 mg?kg -1) ig for five days or by GLPP(3.125～200 mg?L -1) in vitro but the effects of LPS on the production of NO was not influenced significantly. The expression of iNOS was increased by GLPP(25～200 mg?kg -1) ig for five days or GLPP(3.125～200 mg?L -1) in vitro.Conclusion GPP in vivo or in vitro could increase the production of NO in mice peritoneal macrophages. It might take effects by enhancing the synthesis of iNOS.
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AIM: To investigate the free radical scavenging activity of Ganoderma lucidum polysaccharides peptide (GLPP) on peritoneal macrophages in mice. METHODS: Alloxan and tert butylhydroperoxide (tBOOH) were used as an oxidant to injury peritoneal macrophages in vivo in mice or in vitro, respectively. 2', 7' dichlorodihydrofluorescein diacetate (DCHF DA) was used as fluorescent probe. The fluorescence from cells was observed under the laser confocal microscope. Time series scan of conforcal microscope was used to observe the changes of fluorescence by GLPP in mice peritoneal macrophages over time. RESULTS: The results of confocal microscopy showed that GLPP (100 mg?kg -1 , ig for 5 d ) lowed fluorescence in the mice macrophages injured by alloxan (75 mg?kg -1 iv). GLPP (10 mg?L -1 ) also lowed fluorescence in the mice macrophages injured by tBOOH ( 7.76 ?10 -5 mol?L -1 ) in vitro as well. Time series scan showed that GLPP (10 mg?L -1 ) lowed fluorescence in the mice macrophages at rest state or during the respiratory burst induced by PMA (50 nmol?L -1 ). CONCLUSION: GLPP shows antioxidant effects and might have free radical scavenging effects on peritoneal macrophages in mice.
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Aim To study the protective effects of Ganoderma lucidum polysaccharides peptide(GLPP)on the human umbilical vein endothelial cells(HUVECs)injured by reactive oxygen species(ROS),derived from tert-butylhydroperoxide(t-BOOH).Methods Vascular endothelial cells were obtained from infant umbilical cord for primary cultivation,identified by endothelial cell adhesion molecule-1(CD31)by fluorescence microscope.HUVECs were injured by ROS,derived from t-BOOH.The survival rate of cells was measured by Cell Counting Kit-8 assay.Hoechst 33258 nucleus staining was used to observe the early apoptosis injury by ROS.And the activation of Caspase-3 was detected by spectrophotometry.Electron microscopy was used to show the morphological changes in HUVECs.Results GLPP(6.125,12.5,25,50,100 mg?L-1)could inhibit the apoptosis of HUVECs by ROS.The survival rate of HUVECs was increased by GLPP and the percentage of cell apoptosis was decreased by it too.GLPP decreased the activation of Caspase-3 of HUVECs up-regulated by ROS.Electron microscope showed that the organelle such as mitochondria injury by ROS could be relieved by GLPP.Conclusion GLPP can prevent human umbilical vein endothelial cells from oxidative injury.