ABSTRACT
SUMMARY OBJECTIVE: The purpose of this study was to compare arterial stiffness and ultrasound indices in patients with and without chronic obstructive pulmonary disease. METHODS: In our retrospective study, 83 chronic obstructive pulmonary disease patients were assigned to the chronic obstructive pulmonary disease group and 80 healthy controls were enrolled. Pearson's correlation analysis software was used to analyze the correlation between arterial stiffness (including brachial ankle pulse wave velocity and ankle-brachial blood pressure index) and ultrasound index (including resistance index, pulsatility index, and intima-media thickness) at the carotid artery in chronic obstructive pulmonary disease patients. RESULTS: The ultrasound resistance index and pulsatility index level of chronic obstructive pulmonary disease group were lower than those of control group (t=6.326, 8.321, p<0.001). Compared with the control group, the chronic obstructive pulmonary disease group had higher intima-media thickness, total plaque area, and number of plaques (t=4.574, 7.493, 5.093, p<0.001). The arterial stiffness and ankle-brachial blood pressure index level in the chronic obstructive pulmonary disease group were higher than those in the control group (t=6.392, 5.109, p<0.001). Moreover, arterial stiffness in patients with chronic obstructive pulmonary disease was negatively correlated with the ankle-brachial blood pressure index, resistance index, and pulsatility index levels (p<0.05), while it is positively correlated with intima-media thickness, total plaque area, and number of plaques (p<0.05). CONCLUSION: Our results indicated that patients with chronic obstructive pulmonary disease have stiffer arteries compared with healthy control subjects; the ultrasound index could be used as an auxiliary indicator for clinical prediction of arterial stiffness, which is helpful to improve the accuracy of prediction and thus better guide clinical interventions in high-risk groups of chronic obstructive pulmonary disease in time.
ABSTRACT
Abstract Autophagy plays an important role in maintaining cell homeostasis through degradation of denatured proteins and other biological macromolecules. In recent years, many researchers focus on mechanism of autophagy in apicomplexan parasites, but little was known about this process in avian coccidia. In our present study. The cloning, sequencing and characterization of autophagy-related gene (Etatg8) were investigated by quantitative real-time PCR (RT-qPCR), western blotting (WB), indirect immunofluorescence assays (IFAs) and transmission electron microscopy (TEM), respectively. The results have shown 375-bp ORF of Etatg8, encoding a protein of 124 amino acids in E. tenella, the protein structure and properties are similar to other apicomplexan parasites. RT-qPCR revealed Etatg8 gene expression during four developmental stages in E. tenella, but their transcriptional levels were significantly higher at the unsporulated oocysts stage. WB and IFA showed that EtATG8 was lipidated to bind the autophagosome membrane under starvation or rapamycin conditions, and aggregated in the cytoplasm of sporozoites and merozoites, however, the process of autophagosome membrane production can be inhibited by 3-methyladenine. In conclusion, we found that E. tenella has a conserved autophagy mechanism like other apicomplexan parasites, and EtATG8 can be used as a marker for future research on autophagy targeting avian coccidia.
Resumo A autofagia desempenha um papel importante na manutenção da homeostase celular através da degradação de proteínas desnaturadas e outras macromoléculas biológicas. Nos últimos anos, muitos pesquisadores se concentraram no mecanismo da autofagia em parasitas apicomplexos, mas pouco se sabe sobre esse processo na coccidia aviária. No presente estudo, a clonagem, sequenciamento e caracterização de gene relacionado à autofagia Etatg8 foram investigados pela PCR quantitativa em tempo real (RT-qPCR), mancha ocidental (WB), ensaios indiretos de imunofluorescência (IFAs) e microscopia eletrônica de transmissão (TEM), respectivamente. Os resultados mostraram que o gene Etatg8 de E. tenella possui uma ORF de 375 bp, codificando uma proteína de 124 aminoácidos com estrutura e propriedades semelhantes à de outros apicomplexos. RT-qPCR revelou que Etatg8 é expresso durante os quatro estágios de desenvolvimento de E. tenella. Entretanto, seus níveis transcricionais foram significativamente mais elevados na fase de oocisto não esporulados. Os ensaios de manchas ocidental (WB) e de imunofluorescência (IFA) mostraram que a proteína EtATG8 foi lipidada para ligar-se à membrana do autofagossomo sob condições de deficiência nutritiva (em presença de rapamicina) e se agregar no citoplasma de esporozoítas e merozoítas. No entanto, o processo de produção de membrana do autofagossomo pode ser inibido por um inibidor de autofagia (3-meetiladeninatiladenina, 3-MA). Em conclusão, foi demonstrado que E. tenella tem um mecanismo de autofagia conservado, semelhante ao de outros parasitas apicomplexos, e que EtATG8 pode ser usado como um marcador para futuras pesquisas sobre autofagia direcionada à coccidiose aviária.