RESUMO
This study aimed to evaluate 12 genes (18S, GAPDH, B2M, ACTB, ALAS1, GUSB, HPRT1, PBGD, PPIA, PUM1, RPL29, and TBP) for their reliability and stability as reference sequences for real-time quantitative PCR (RT-qPCR) in bone marrow-derived mesenchymal stem cells (BMSCs) isolated from patients with avascular necrosis of the femoral head (ANFH). BMSCs were isolated from 20 ANFH patients divided into four groups according to etiology, and four donors with femoral neck fractures. Total RNA was isolated from BMSCs and reverse transcribed into complementary DNA, which served as a template for RT-qPCR. Three commonly used programs were then used to analyze the results. Reference gene expression varied within each group, between specific groups, and among all five groups. Based on comparisons of all five groups, two of the programs used suggested that HPRT1 was the most stable reference gene, while 18S and ACTB were the most variable. Among the 12 candidate reference genes, HPRT1 exhibited the greatest reliability, followed by PPIA. Thus, these sequences could be used as references for the normalization of RT-qPCR results.
Assuntos
Necrose da Cabeça do Fêmur/genética , Células-Tronco Mesenquimais/metabolismo , Biossíntese de Proteínas/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , Osso e Ossos/metabolismo , Osso e Ossos/patologia , Feminino , Necrose da Cabeça do Fêmur/metabolismo , Necrose da Cabeça do Fêmur/patologia , Regulação da Expressão Gênica/genética , Humanos , Masculino , Reação em Cadeia da Polimerase em Tempo Real/métodos , Padrões de ReferênciaRESUMO
The aim of this study was to investigate the impacts of positive acceleration (+Gz) on the gastric mucosal tissues in cases of acute gastric mucosal injury and to explore the role of oxygen free radicals. Thirty Sprague Dawley rats were randomly divided into the absolute ethanol control group (A group), absolute ethanol +5Gz group (B group), absolute ethanol +10Gz group (C group). Following centrifugation, the gastric tissues of each group were studied for the presence of gastric mucosal injuries and morphological changes. The concentrations of malondialdehyde (MDA) and superoxide dismutase (SOD) contents were simultaneously investigated. Degree of gastric mucosal injuries were as follows: C group (visually 49.080 ± 10.254, under light microscopy 9.400 ± 2.011) > B group (visually 23.654 ± 9.678, under light microscopy 5.000 ± 1.054) > A group (visually 11.410 ± 3.742, under light microscopy 3.800 ± 1.399). The gastric mucosal MDA content (0.376 ± 0.084 vs 0.235 ± 0.044) was significantly higher in the C group than in the A group, whereas the SOD content (8.852 ± 1.001 vs 10.694 ± 0.965) was lower than that in the A group. However, the MDA and SOD contents did not change much in the B group. Our results suggest that the +Gz exposure might aggravate the acute gastric mucosal injury, and changes in MDA and SOD contents in the gastric tissues indicated that the oxygen free radicals play an important role in this regard.
Assuntos
Mucosa Gástrica/lesões , Hipergravidade/efeitos adversos , Malondialdeído/análise , Superóxido Dismutase/análise , Doença Aguda , Animais , Etanol/toxicidade , Mucosa Gástrica/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Superóxido Dismutase/efeitos dos fármacosRESUMO
We aimed to investigate the role of 4 single nucleotide polymorphisms of the xeroderma pigmentosum complementation group F (XPF) gene (rs3136038, rs1799798, rs1800067, and rs2276466) in glioma, and the roles of gene-gene interactions in the risk of developing this type of cancer. We collected samples from 225 glioma cases and 262 controls and genotyped the rs3136038, rs1799798, rs1800067, and rs2276466 polymorphisms using a 384-well plate format with the Sequenom MassARRAY platform. Individuals carrying the rs1800067 GG genotype were more likely to have an increased risk of glioma when compared with carriers of the A/A genotype in a co-dominant model, with an odds ratio (OR) [95% confidence interval (CI)] of 2.85 (1.14-7.76). However, we did not find an association with increased risk of glioma for the polymorphisms rs3136038, rs1799798, and rs2276466 in XPF. The combination genotype of the rs1800067 G allele and the rs2276466 G allele was associated with a moderate risk of glioma (OR = 1.71, 95%CI = 1.02-2.87). Our study suggests that the rs1800067 genetic variant of XPF functions in the development of glioma.