RESUMEN

OBJECTIVE: To determine changes in mitochondrial DNA (mtDNA) copy number in peripheral blood in Rett syndrome caused by methyl-CpG-binding protein-2 (MECP2) variants and explore the mechanism of mitochondrial dysfunction in Rett syndrome. STUDY DESIGN: Female patients who were diagnosed with Rett syndrome and had an MECP2 variant (n = 142) were recruited in this study, along with the same number of age- and sex-matched healthy controls. MtDNA copy number was quantified by real-time quantitative polymerase chain reaction with TaqMan probes. The differences in mtDNA copy number between the Rett syndrome group and the control group were analyzed using the independent-samples t test. Linear regression, biserial correlation analysis, and one-way ANOVA were applied for the correlations between mtDNA copy number and age, clinical severity, variant types, functional domains, and hot-spot variants. RESULTS: MtDNA copy number was found to be significantly increased in the patients with Rett syndrome with MECP2 gene variants compared with the control subjects. Age, clinical severity, variant types, functional domains, and hot-spot variants were not related to mtDNA copy number in patients with Rett syndrome. CONCLUSIONS: MtDNA copy number is increased significantly in patients with Rett syndrome, suggesting that changes in mitochondrial function in Rett syndrome trigger a compensatory increase in mtDNA copy number and providing new possibilities for treating Rett syndrome, such as mitochondria-targeted therapies.

Asunto(s)

Variaciones en el Número de Copia de ADN , ADN Mitocondrial , Proteína 2 de Unión a Metil-CpG/genética , Mitocondrias/genética , Síndrome de Rett/genética , Adolescente , Adulto , Estudios de Casos y Controles , Niño , Preescolar , Estudios Transversales , Femenino , Marcadores Genéticos , Humanos , Lactante , Modelos Lineales , Gravedad del Paciente , Síndrome de Rett/fisiopatología , Adulto Joven

RESUMEN

The aim of this study was to make a comparison between the tissue fusion technique and conventional methods for sealing bowel anastomosis. Eighteen female domestic pigs (Suidae, Sus) were used in our study. Tissue-fused anastomoses (LigaSure groups) were made in 13 animals (5 anastomoses per animal), which were subdivided into 4 groups according to different manufacturing settings: "LigaSure-L-1" and "LigaSure-L-2", with low energy output level with 1 or 2 device-activated tissue sealing times, and "LigaSure-M" and "LigaSure-H", with medium or high energy output level. As controls, automatically stapled (GIA group) and hand-sewn (suture group) anastomoses were utilized in 3 and 2 animals, respectively. There was no statistical difference in the overall leakage rate between the GIA group (6.7%) and the LigaSure groups (15%) (P=1.000). There was less proliferating epithelium covering the anastomosis gap in the LigaSure groups compared with the other two groups. The gap between the two extremities of muscular layers of the anastomosis in the LigaSure groups was filled with collagen fibers. More proliferating cell nuclear antigen (PCNA)-positive cells were found in the anastomoses of the LigaSure groups compared with the other two groups (P=0.010). Our results showed that the tissue fusion technology was a feasible and safe method for anastomoses.

Asunto(s)

Anastomosis Quirúrgica/métodos , Intestino Delgado/cirugía , Técnicas de Sutura , Ingeniería de Tejidos , Animales , Femenino , Modelos Animales , Reproducibilidad de los Resultados , Porcinos

RESUMEN

The aim of this study was to make a comparison between the tissue fusion technique and conventional methods for sealing bowel anastomosis. Eighteen female domestic pigs (Suidae, Sus) were used in our study. Tissue-fused anastomoses (LigaSure groups) were made in 13 animals (5 anastomoses per animal), which were subdivided into 4 groups according to different manufacturing settings: "LigaSure-L-1" and "LigaSure-L-2", with low energy output level with 1 or 2 device-activated tissue sealing times, and "LigaSure-M" and "LigaSure-H", with medium or high energy output level. As controls, automatically stapled (GIA group) and hand-sewn (suture group) anastomoses were utilized in 3 and 2 animals, respectively. There was no statistical difference in the overall leakage rate between the GIA group (6.7%) and the LigaSure groups (15%) (P=1.000). There was less proliferating epithelium covering the anastomosis gap in the LigaSure groups compared with the other two groups. The gap between the two extremities of muscular layers of the anastomosis in the LigaSure groups was filled with collagen fibers. More proliferating cell nuclear antigen (PCNA)-positive cells were found in the anastomoses of the LigaSure groups compared with the other two groups (P=0.010). Our results showed that the tissue fusion technology was a feasible and safe method for anastomoses.

Asunto(s)

Animales , Femenino , Anastomosis Quirúrgica/métodos , Técnicas de Sutura , Ingeniería de Tejidos , Intestino Delgado/cirugía , Porcinos , Reproducibilidad de los Resultados , Modelos Animales

RESUMEN

ABSTRACT Cyclin-dependent kinase-like 5 (CDKL5) is a protein kinase that is homologous to mitogen-activated protein kinases (MAPKs) and cyclin-dependent kinases (CDKs). Mutations in the CDKL5 gene cause X-linked infantile spasms and phenotypes that overlap with that of Rett syndrome, which is a neurodevelopmental disorder caused primarily by mutations in the methyl CpG binding protein 2 gene (MECP2). Previous studies in transfected cell lines showed that CDKL5 interacts with MeCP2 and DNA (cytosine-5)-methyltransferase 1 (Dnmt1). However, little is known about the relationships of CDKL5 with interacting proteins in primary neuronal cultures. In this study, we investigated the expression patterns of CDKL5, MeCP2 and Dnmt1, and their interaction in cultured rat cortical neurons. Using real-time PCR analysis, we found that CDKL5, MeCP2 and Dnmt1 have similar expression patterns at the mRNA level. In contrast, the expression patterns of those proteins at the protein level are different and could be inversely correlated, as shown by western blotting. Using co-immunoprecipitation, we further demonstrated that CDKL5 interacts with MeCP2 and Dnmt1 in primary rat cortical neurons. These data suggest that a functional link exists among CDKL5, MeCP2 and Dnmt1 during neuronal development and may provide further insight into the pathogenesis of Rett syndrome.

RESUMEN

Intramuscular fat (IMF) content plays an important role in meat quality. Many genes involved in lipid and energy metabolism were identified as candidate genes for IMF deposition, since genetic polymorphisms within these genes were associated with IMF content. However, there is less information on the expression levels of these genes in the muscle tissue. This study aimed at investigating the expression levels of sterol regulating element binding protein-1c (SREBP-1c), diacylglycerol acyltransferase (DGAT-1), heart-fatty acids binding protein (H-FABP), leptin receptor (LEPR) and melanocortin 4 receptor (MC4R) genes and proteins in two divergent Banna mini-pig inbred lines (BMIL). A similar growth performance was found in both the fat and the lean BMIL. The fat meat and IMF content in the fat BMIL were significantly higher than in the lean BMIL, but the lean meat content was lower. The serum triacylglycerol (TAG) and free fatty acid (FFA) contents were significantly higher in the fat than in the lean BMIL. The expression levels of SREBP-1c, DGAT-1 and H-FABP genes and proteins in fat BMIL were increased compared to the lean BMIL. However, the expression levels of LEPR and MC4R genes and proteins were lower.