RESUMO
Bacillus anthracis, B. thuringiensis and B. cereus are members of the B. cereus group. They share high genetic similarity. Whereas plcR (Phospholipase C regulator) usually encodes a functional pleiotropic activator protein in B. cereus and B. thuringiensis isolates, a characteristic nonsense mutation is found in all B. anthracis strains investigated, making the gene dysfunctional. To study the function of PlcR in B. anthracis, we used the B. cereus CMCC63301 genome as a template and constructed a recombinant expression plasmid pBE2A-plcR, and introduced it into the B. anthracis vaccine strain A16R, and then analyzed the activity of the hemolysin and sphingomyelinase. The results showed that transformation of B. anthracis with plasmid pBE2A-plcR carrying the native B. cereus plcR gene active the expression of sphingomyelinase gene, but did not activate expression of hemolysin genes of B. anthracis A16R.
Assuntos
Bacillus anthracis/genética , Bacillus cereus/genética , Proteínas de Bactérias/metabolismo , Vacinas Bacterianas/genética , Regulação Bacteriana da Expressão Gênica , Transativadores/metabolismo , Bacillus anthracis/enzimologia , Bacillus anthracis/crescimento & desenvolvimento , Bacillus anthracis/metabolismo , Bacillus cereus/metabolismo , Proteínas de Bactérias/genética , Vacinas Bacterianas/metabolismo , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/metabolismo , Esfingomielina Fosfodiesterase/genética , Esfingomielina Fosfodiesterase/metabolismo , Transativadores/genéticaRESUMO
Arsenic trioxide has shown remarkable biological activity against bladder cancer in some clinical studies. However, the mechanism of its action is unknown. Our aim was to find the relationship between miRNAs and arsenic trioxide treatment by using T24 human bladder carcinoma cells. By performing microRNA microarray and quantitative real-time PCR after ATO treatment, we found that expression levels of several miRNAs, in particular, miRNA-19a, were significantly decreased in T24 cell line. Furthermore, cell proliferation assay, flow cytometry analysis, prediction of miRNA targets, Western blot analysis, and luciferase reporter assay were performed to determine the role of mir-19a in affecting the biological behaviors of T24 cells. Several miRNAs were up-regulated or down-regulated in T24 cells treated with arsenic trioxide compared to their controls. If only changes above two folds were considered, two miRNAs were identified, miRNA-19a was down-regulated, while miRNA-222* was up-regulated. Among them, knockdown of miRNA-19a by anti-miRNA-19a transfection showed a positive therapeutic effect in bladder cancer cells by inhibiting cell growth and inducing cell apoptosis targeting PTEN through the PTEN/Akt pathway. Besides this, a synergy effect was detected between knockdown of miRNA-19a and arsenic trioxide. Arsenic trioxide altered miRNA expression profile in T24 cells. It seems miRNA-19a plays a critical role in the mechanism of arsenic trioxide treatment in bladder cancer. The synergy effect between miRNA-19a and arsenic trioxide that advocates targeting the mir-19a may represent a potential approach to enhance the efficacy and safety of ATO to treat bladder cancer by a decrease in dose.
Assuntos
Antineoplásicos/farmacologia , Arsenicais/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , MicroRNAs/fisiologia , Óxidos/farmacologia , PTEN Fosfo-Hidrolase/genética , Neoplasias da Bexiga Urinária/tratamento farmacológico , Neoplasias da Bexiga Urinária/genética , Apoptose/efeitos dos fármacos , Trióxido de Arsênio , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Western Blotting , Proliferação de Células/efeitos dos fármacos , Perfilação da Expressão Gênica , Humanos , Luciferases/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , PTEN Fosfo-Hidrolase/metabolismo , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais Cultivadas , Neoplasias da Bexiga Urinária/patologiaRESUMO
BACKGROUND: Diabetes is a common chronic disease. Given the increasing incidence of diabetes, more individuals are affected by diabetic optic neuropathy (DON), which results in decreased vision. Whether DON leads to abnormalities of other visual systems, including the eye, the visual cortex, and other brain regions, remains unknown. AIM: To investigate the local characteristics of spontaneous brain activity using regional homogeneity (ReHo) in patients with DON. METHODS: We matched 22 patients with DON with 22 healthy controls (HCs). All subjects underwent resting-state functional magnetic resonance imaging. The ReHo technique was used to record spontaneous changes in brain activity. Receiver operating characteristic (ROC) curves were applied to differentiate between ReHo values for patients with DON and HCs. We also assessed the correlation between Hospital Anxiety and Depression Scale scores and ReHo values in DON patients using Pearson correlation analysis. RESULTS: ReHo values of the right middle frontal gyrus (RMFG), left anterior cingulate (LAC), and superior frontal gyrus (SFG)/left frontal superior orbital gyrus (LFSO) were significantly lower in DON patients compared to HCs. Among these, the greatest difference was observed in the RMFG. The result of the ROC curves suggest that ReHo values in altered brain regions may help diagnose DON, and the RMFG and LAC ReHo values are more clinically relevant than SFG/LFSO. We also found that anxiety and depression scores of the DON group were extremely negatively correlated with the LAC ReHo values (r = -0.9336, P < 0.0001 and r = -0.8453, P < 0.0001, respectively). CONCLUSION: Three different brain regions show ReHo changes in DON patients, and these changes could serve as diagnostic and/or prognostic biomarkers to further guide the prevention and treatment of DON patients.
RESUMO
A measurement method of 85Kr using an internal gas proportional counter (IGPC) is presented in this study. The operation conditions of the IGPC were determined and optimized, including the operating voltage, pressure, sample volume, interference from other gas components such as nitrogen or air, and mitigation of the memory effect. The IGPC was calibrated using certified standards, and the detection efficiency was approximately 58% for typical samples. A lower limit of detection of approximately 0.11 MBq/m3(Kr) was achieved after counting for 5â¯h with 1â¯mL pure Kr, corresponding to the atmospheric activity concentration of 0.18 Bq/m3 (air). It was shown that the IGPC could be used effectively for measuring 85Kr.