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 investigate the effect of overexpression of LncRNA MEG3 on proliferation,migration and cisplatin sensitivity of hepatoma cells HepG2 and LM3 and explore the underlying and mechanism.Methods The expression of MEG3 in healthy individuals and patients with hepatocellular carcinoma(HCC)was analyzed by online bioinformatics analysis,and Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect MEG3 expression in different HCC cell lines.A MEG3-overexpresing plasmid was transfected in HepG2 and LM3 cells,and the changes in cell proliferation and migration were examined using CCK8 assay and scratch assay.CCK8 assay was used to determine the inhibitory rate of cisplatin on the transfected cells.A reactive oxygen species(ROS)fluorescence probe(DCFH-DA)and malondialdehyde(MDA)kit were used to assess the changes in ROS production and MDA level in the cells.Western blotting was performed to detect the expression levels of ferroptosis-related proteins glutathione peroxidase 4(GPX4)and ferritin heavy chain 1(FTH1).Results The expression of MEG3 was significantly lower in HCC cells than in LO2 cells,which was consistent with the results of bioinformatic analysis(P<0.05).Overexpression of MEG3 in the HCC cell lines significantly suppressed cell proliferation and migration(P<0.05),increased the cell inhibition rate of cisplatin(P<0.05),enhanced cellular ROS production and increased MDA levels in the cells(P<0.05).MEG3 overexpression significantly decreased the expressions of GPX4 and FTH1 in the HCC cell lines.Conclusion The expression of MEG3 is decreased in HCC cells,and its overexpression inhibits proliferation and migration and enhances cisplatin sensitivity of HCC cells by promoting ferroptosis of the cells.

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 effect of overexpression of LncRNA MEG3 on proliferation,migration and cisplatin sensitivity of hepatoma cells HepG2 and LM3 and explore the underlying and mechanism.Methods The expression of MEG3 in healthy individuals and patients with hepatocellular carcinoma(HCC)was analyzed by online bioinformatics analysis,and Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect MEG3 expression in different HCC cell lines.A MEG3-overexpresing plasmid was transfected in HepG2 and LM3 cells,and the changes in cell proliferation and migration were examined using CCK8 assay and scratch assay.CCK8 assay was used to determine the inhibitory rate of cisplatin on the transfected cells.A reactive oxygen species(ROS)fluorescence probe(DCFH-DA)and malondialdehyde(MDA)kit were used to assess the changes in ROS production and MDA level in the cells.Western blotting was performed to detect the expression levels of ferroptosis-related proteins glutathione peroxidase 4(GPX4)and ferritin heavy chain 1(FTH1).Results The expression of MEG3 was significantly lower in HCC cells than in LO2 cells,which was consistent with the results of bioinformatic analysis(P<0.05).Overexpression of MEG3 in the HCC cell lines significantly suppressed cell proliferation and migration(P<0.05),increased the cell inhibition rate of cisplatin(P<0.05),enhanced cellular ROS production and increased MDA levels in the cells(P<0.05).MEG3 overexpression significantly decreased the expressions of GPX4 and FTH1 in the HCC cell lines.Conclusion The expression of MEG3 is decreased in HCC cells,and its overexpression inhibits proliferation and migration and enhances cisplatin sensitivity of HCC cells by promoting ferroptosis of the cells.

Objective:To understand the molecular characteristics of the mutant strain of polymerase basic protein 2 (PB2) gene of influenza A (H1N1pdm) in Guangdong province, and to explore its specific molecular sites, so as to provide scientific basis for the prevention and control of influenza virus.Methods:Throat swab samples were collected from 2 cases infected with PB2 gene variant strains for virus isolation, and 23 influenza virus strains were selected from Guangdong province for sequencing analysis. The reference sequences and vaccine strain sequences provided by GISAID were used to perform evolutionary analysis on hemagglutinin (HA) and PB2 genes. Virus strain antigen analysis and neuraminidase (NA) inhibition test were carried out. PB2 protein model was constructed and polymerase activity was analyzed.Results:H399N amino acid mutation occurred in the HA gene of PB2-D701N and PB2-A271S variant strains, both of which belonged to the branch of 6B.1A.5a.2a. They belonged to the same big branch and different small branches as the vaccine strain A/Victoria/4897/2022, and they are all vaccine-like strains. In the three-dimensional structure, the mutations of PB2-D701N and PB2-A271S change charge and hydrophobicity.Conclusions:PB2-D701 and A271 were highly conserved, and PB2 mutant strains were not the dominant strains. The PB2 mutant had high antigenicity with the vaccine. The PB2 mutant strain is sensitive to NA inhibitors. The three-dimensional model predicted that PB2-D701N mutation could enhance virulence and affect transmissibility of influenza virus, while PB2-A271S mutation could affect polymerase activity and polymerase complex synthesis of influenza virus.

Objective:To understand the etiology of a confirmed case of Coronavirus Disease 2019 (COVID-19).Methods:The pharyngeal swabs, serum and nasal swabs of a case of COVID-19 were inoculated into Vero-E6 cell tubes for virus isolation. The cytopathic effect (CPE) were observed daily. Collecting cell’s isolation when CPE was over 75%, after repeated freezing and thawing for 3 times, the supernatant was centrifugally taken, and the images of the virus were obtained by transmission electron microscopic observation, and the nucleic acid of the virus was extracted, second generation sequencing and sequence evolution analysis were used to identify and type the virus strains.Results:One strain of 2019 novel coronavirus (2019-nCoV) was successfully isolated from the nasal swab of this case of COVID-19, and one strain of herpes simplex virus type 1 (HSV-1) was also successfully isolated from the throat swab of the same case.Conclusions:COVID-19 cases have the possibility of co-infection with 2019-nCoV and HSV-1.

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*