OBJECTIVETo investigate the effect of maslinic acid preconditioning against injuries of rat cortical neurons induced by oxygen-glucose deprivation (OGD).

METHODSThe cortical neurons were isolated from Sprague-Dawley rat embryos at 15-17 days of gestation for primary culture. The cortical neurons were incubated with different concentrations (0.1, 1, and 10 micro;mol/L) of maslinic acid prior to OGD. The cell damage and viability were evaluated for lactate dehydrogenase (LDH) leakage and using MTT assay, respectively, and the expression of Bax protein was detected using Western blotting.

RESULTSOGD significantly increased LDH release rate and decreased the viability of the cells. After preconditioning with maslinic acid (1 and 10 micro;mol/L), LDH leakage rate was decreased and cell the viability increased in cells exposed to OGD. Western blotting showed that Bax expression in the cells decreased as maslinic acid concentrations increased.

CONCLUSIONPretreatment with maslinic acid can protect cultured embryonic rat cortical neurons against OGD-induced injury possibly in relation to decreased expression of Bax.

Animals ; Apoptosis ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; cytology ; drug effects ; Embryo, Mammalian ; Glucose ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Neurons ; drug effects ; metabolism ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley ; Triterpenes ; pharmacology ; bcl-2-Associated X Protein ; metabolism

Objective To isolate the glioblastoma multiforme stem cells (GBM-SCs) from GBM specimens and to investigate the biological role ofmiR-153 in GBM-SCs so as to explore the application of gene therapy of GBMs.Methods CD133+ cells were separated using magnetic cell sorting technique (MACS) after primary culture.Immunofluorescence staining was employed to detect the CD133,nestin,glial fibrillary acidic protein (GFAP),microtubule-associated protein 2 (MAP2) expressions; real time-PCR was used to analyze miR-153 mRNA expression in CD 133-cells and CD 133+ cells.Lipofectamine RNAiMAX was used to transfect MiR-153 mimic (miR-153 group) and scrambled control oligonucleotides (NC group) into GBM-SCs; 7 d after that,sphere formation assay was performed to determine the self-renewal ability of GBM-SCs.Real time-PCR and immunofluorescence were carried out to examine the CD133,nestin,GFAP,MAP2 mRNA and protein expressions.At last,the proliferation ability of miR-153 treated GBM-SCs and NC cells was determined by CCK-8 at 24,48,72,96 and 120 h after the transfection and the apoptosis ratio was detected by flow cytometry 3 d after the transfection.Results GBM-SCs isolated from GBM specimens could express stem cell markers CD133 and nestin; after differentiation,the cells could express astrocyte marker GFAP and neuron marker MAP2; miR-153 expression in CD133+ cells was signficantly down-regulated as compared with that in CD133-cells (P＜0.05).Seven d after transfection,the number of spheres in the NC group was significantly larger than that in the miR-153 group (P＜0.05); real time-PCR indicated that the mRNA expressions of CD 133 and nestin in the miR-153 group were significantly decreased,and the GFAP and MAP2 mRNA expressions were statistically increased as compared with those in the NC group (P＜0.05).The results detected by immunofluorescence were in accordance with those by real time-PCR; 48,72,96h after the transfection,cell viability in cells from miR-153 group was statistically significant lower than that in the NC group (P＜0.05); flow cytometry showed that the apoptosis rate of cells from the miR-153 group (9.4 1％±1.98％) was significantlyhigher than that in the NC group (4.28％±0.31％,P＜0.05).Conclusion Reactivation of miR-153 expression suggests novel therapeutic strategy for GBM-SCs.

OBJECTIVE: To investigate the effects of overexpression of long noncoding RNA (lncRNA) MEG3 on the proliferation and invasion of glioblastoma U251 cells by suppressing the expression of hypoxia inducible factor 1 METHODS: The expression of lncRNA MEG3 and HIF1 RESULTS: The expression of MEG3 was significantly lower and HIF1 CONCLUSIONS MEG3 overexpression inhibits the proliferation and invasion of U251 cells through suppressing the expression of HIF1
Apoptosis ; Cell Line, Tumor ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Gene Expression Regulation, Neoplastic ; Glioblastoma/genetics* ; Humans ; MicroRNAs ; RNA, Long Noncoding/genetics*

OBJECTIVETo study the serotypes, antimicrobial resistance, virulence genes and molecular characteristics of Vibrio parahaemolyticus isolated from outbreaks and sporadic cases in Guangdong, 2013.

METHODS36 Vibrio parahaemolyticus strains isolated from outbreaks and 43 strains from sporadic cases were sero-typed and tested for antimicrobial resistance. PCR was used to detect for tdh(thermostable direct hemolysin gene), trh (tdh(-) related hemolysin gene), GS-PCR and orf8 genes. All the samples were analyzed by pulsed-field gel electrophoresis (PFGE).

RESULTS36 isolates from outbreaks were all identified as O3 : K6, and among the 43 sporadic isolates, O3 : K6 (23, 53.49%) was the dominant serotype. Vibrio parahaemolyticus isolates showed high resistance rate to ampicillin (96.20%) and cefalotin (40.50%), but were high sensitive to cotrimoxazole (100%) and chloramphenicol (100%). 83.33% (30/36) outbreak isolates were resistant to multi-drugs but only 37.21% (16/43) of the sporadic isolates showed so. Results from virulence gene detection suggested that all the 36 outbreak isolates belonged to tdh(+) trh(-) strains, while 86.05% (37/43) of the sporadic isolates were tdh(+)trh(-) and 11.63% (5/43)were tdh(-)trh(+) . Only one tdh(+)trh(+) strain was found. All the outbreak isolates contained GS-PCR and/or orf8 genes, whereas among the sporadic isolates only 51.16% (22/43) of them carrying the similar genes. Results from PFGE analysis suggested that 79 isolates were discriminated into 3 clusters and 32 different PFGE patterns with the similarity value between 59.8% and 100.0%. Outbreak isolates seemed to gather in the same cluster, while the sporadic isolates spreading in all the three clusters.

CONCLUSIONO3 : K6 was the dominant serotype in Vibrio parahaemolyticus strains isolated in Guangdong, 2013. These strains showed high sensitivity to most antibiotics, but with multi-drug resistance. Positive rate of tdh gene was high, and most O3 : K6 strains contained GS-PCR and/or orf8 genes. PFGE analysis revealed genetic diversity was within the Vibrio parahaemolyticus strains in Guangdong.

Bacterial Toxins ; China ; epidemiology ; Disease Outbreaks ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Genetic Variation ; Hemolysin Proteins ; Humans ; Polymerase Chain Reaction ; Serotyping ; Vibrio Infections ; epidemiology ; Vibrio parahaemolyticus ; genetics ; pathogenicity ; Virulence