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OBJECTIVE@#To investigate whether hydrogen-rich water exerts a protective effect against cellular injury by affecting the level of autophagy after oxygen glucose deprivation/reoxygenation (OGD/R) in a mouse hippocampal neuronal cell line (HT22 cells).@*METHODS@#HT22 cells in logarithmic growth phase were cultured in vitro. Cell viability was detected by cell counting kit-8 (CCK-8) assay to find the optimal concentration of Na2S2O4. HT22 cells were divided into control group (NC group), OGD/R group (sugar-free medium+10 mmol/L Na2S2O4 treated for 90 minutes and then changed to normal medium for 4 hours) and hydrogen-rich water treatment group (HW group, sugar-free medium+10 mmol/L Na2S2O4 treated for 90 minutes and then changed to medium containing hydrogen-rich water for 4 hours). The morphology of HT22 cells was observed by inverted microscopy; cell activity was detected by CCK-8 method; cell ultrastructure was observed by transmission electron microscopy; the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin-1 was detected by immunofluorescence; the protein expression of LC3II/I and Beclin-1, markers of cellular autophagy, was detected by Western blotting.@*RESULTS@#Inverted microscopy showed that compared with the NC group, the OGD/R group had poor cell status, swollen cytosol, visible cell lysis fragments and significantly lower cell activity [(49.1±2.7)% vs. (100.0±9.7)%, P < 0.01]; compared with the OGD/R group, the HW group had improved cell status and remarkably higher cell activity [(63.3±1.8)% vs. (49.1±2.7)%, P < 0.01]. Transmission electron microscopy showed that the neuronal nuclear membrane of cells in the OGD/R group was lysed and a higher number of autophagic lysosomes were visible compared with the NC group; compared with the OGD/R group, the neuronal damage of cells in the HW group was reduced and the number of autophagic lysosomes was notably decreased. The results of immunofluorescence assay showed that the expressions of LC3 and Beclin-1 were outstandingly enhanced in the OGD/R group compared with the NC group, and the expressions of LC3 and Beclin-1 were markedly weakened in the HW group compared with the OGD/R group. Western blotting assay showed that the expressions were prominently higher in both LC3II/I and Beclin-1 in the OGD/R group compared with the NC group (LC3II/I: 1.44±0.05 vs. 0.37±0.03, Beclin-1/β-actin: 1.00±0.02 vs. 0.64±0.01, both P < 0.01); compared with the OGD/R group, the protein expression of both LC3II/I and Beclin-1 in the HW group cells were notably lower (LC3II/I: 0.54±0.02 vs. 1.44±0.05, Beclin-1/β-actin: 0.83±0.07 vs. 1.00±0.02, both P < 0.01).@*CONCLUSIONS@#Hydrogen-rich water has a significant protective effect on OGD/R-causing HT22 cell injury, and the mechanism may be related to the inhibition of autophagy.
Subject(s)
Mice , Animals , Oxygen/metabolism , Beclin-1/pharmacology , Glucose/metabolism , Actins , Sincalide , Autophagy/physiology , Hydrogen/pharmacology , Reperfusion Injury , ApoptosisABSTRACT
Objective:To analyze and screen the key genes of sepsis secondary to pulmonary infection by bioinformatics, and to provide theoretical basis for the basic research of the disease and find an ideal animal model program.Methods:Experiment 1 (bioinformatics analysis): gene expression data sets of pulmonary infection secondary sepsis patients and multiple sepsis animal models were screened by Gene Expression Omnibus (GEO) Database, and gene differences were analyzed by R software. Differential genes were analyzed by gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Correlation analysis was conducted between differential genes and clinical symptoms in the data set of pulmonary infection secondary sepsis, and the correlation heat map between differential genes and clinical symptoms was drawn. Key genes were screened by weighted gene co-expression network analysis (WGCNA) and protein-protein interaction network analysis (PPIN) clustering. Experiment 2 (sepsis animal model preparation): male mice weighing 21-25 g were randomly divided into the key genes group and the control (Sham) group. And cecal ligation and puncture (CLP) was used to establish mouse sepsis model, while the mice in sham group were performed by exposure of cecum. And all the mice were scarified 24 hours after surgery to extract the total RNA from lung tissue, real time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to detect mRNA expression of key genes.Results:Experiment 1 (bioinformatics analysis): 319 differential genes were showed by GSE 134364 and GSE 65682 data set analysis of pulmonary infection secondary sepsis. And there was no genetic difference between community acquired pneumonia (CAP) and hospital acquired pneumonia (HAP) in patients with pulmonary infection secondary to sepsis. Obvious differences existed between differential genes in animal models, and there was no common differential gene. Differential genes in patients and animal models were similarly enriched in GO function, mainly in cell differentiation, regulation of cell process, and regulation of cellular response to stimuli, there were significant differences in pathway enrichment, among which, CLP animal models showed higher consistency with patients. The key genes obtained by WGCNA and PPIN analysis were MAPK14, NLRC4 and LCN2. Experiment 2 (sepsis animal model preparation): animal experiment results showed that the mRNA expressions of MAPK14, NLRC4 and LCN2 in lung tissue of CLP model mice were significantly up-regulated compared with the sham group.Conclusions:MAPK14, NLRC4 and LCN2 are key genes involved in the regulation of biological processes of pulmonary sepsis secondary to infection, and are potential research directions of this disease. What's more, CLP animal model can better reflect the biological characteristics of patients with pulmonary infection secondary sepsis, and is one of the ideal animal model schemes for pulmonary infection secondary sepsis.
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As China enters the era of organ donation after the citizen's death, the organ donation cases with potential malignant tumor history increase gradually. Guide to the Quality and Safety of Organs for Transplantation (6th Edition) by European Committee was translated into Chinese in 2019. This article interprets Chapter 9 of the guideline "Risk of Malignant Tumor Transmission" to assist organ donation coordinator and transplant team in screening donors at risk of malignant tumor transmission in our country and to reduce the risk of donor-derived tumor transmission.
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Objective@#To analyze the influence of sexuality education on primary school students’ attitude towards homosexuality, and provide practical teaching evidence of intervening campus bullying based on sexual orientation.@*Methods@#A primary school offering school-based sexuality education curriculums in Beijing was selected as an experimental school, another school with similar location, nature and scale, quality of teachers and teaching environment was selected as a control school. The graduates in experimental school received 6 years of sexuality education in grade 1 to grade 6 of primary school. A survey was conducted in Jun. 2018 among all the graduates in grade 6 of the two schools through an attitude scale on homosexuality adopting cluster sampling.@*Results@#The scores of attitude towards homosexuality in experimental school and control school were (42.85±15.46) and (60.17±12.54) respectively, with significantly statistical difference(t=-9.47, P<0.01). There were interaction effects on attitudes of primary school students towards homosexuality between gender and experience of sexuality education (P<0.05). There was no significant difference in attitude towards homosexuality between male and female students in control school(t=0.58, P>0.05), but in experimental school, the attitude scores of male and female students respectively were (47.64±14.31) and (38.13±15.19), with significantly statistical difference(t=3.47, P<0.01). There was no significant difference in attitude towards different sexual orientation between experimental school students and control school students(t=0.10，1.53，P>0.05).@*Conclusion@#Sexuality education curriculum can help improve attitude towards homosexuality of primary school students, which has better intervention effects on female students than male students, attitudinal difference in gay and lesbian has not formed in primary school students.
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Objective To investigate the functional polarization of T cells in children with obstructive sleep apnea hypopnea syndrome (OSAHS).Methods Forty five cases of OSAHS children were chosen to undergo tonsillectomy and adenoidectomy.Preoperative analyses of peripheral blood Th1 /Th2 ratio and Tc1 /Tc2 ratio were compared with those of normal children and those of postoperative 6 months for observation of the change of T cells.Th1/Th2 ratio and Tc1/Tc2 ratio were detected by flow cytometry.Results The preoperative ratio of Th1 cells in children with OSAHS was significantly lower than normal children [(9.66 ± 3.73) ％ vs (13.34 ± 4.36) ％,P ＜ 0.05],while the preoperative ratio of Th2 cells was significantly higher than normal children [(3.42 ± 1.64) ％ vs (2.53 ± 1.97) ％,P ＜ 0.05],which made the Thl/Th2 ratio lower than normal children (3.51 ± 1.53 vs 5.41 ±2.17,P ＜0.05).At the 6th month after tonsillectomy and adenoidectomy,the postoperative ratio of Th1 cells was significantly higher than preoperative ratio [(12.99 ± 3.68) ％ vs (9.66 ± 3.73) ％,P ＜ 0.05],while the postoperative ratio of Th2 cells was lower than preoperative ratio [(2.62 ± 1.88) ％ vs (3.42 ± 1.64) ％,P ＜ 0.05],which made the postoperative Th1 /Th2 ratio higher than preoperative ratio (5.15 ± 1.86 vs 3.51 ± 1.53,P ＜ 0.05).There was no significant difference in Tc1 /Tc2 ratio between the OSAHS children and normal children.Conclusions Functional polarization of T cells was abnormal in OSAHS children accompanied by the predomination of Th2 cells and the normal Tc cell polarization.Functional polarization of T cells can return to normal at the 6th month after tonsillectomy and adenoidectomy.