ABSTRACT Purpose To study effects of Rehmannia glutinosa polysaccharides (RGP) on bone tissue structure and skeletal muscle atrophy in rats with disuse. Methods A rat model of disuse osteoporosis combined with muscle atrophy was established by removing the bilateral ovaries of rats and fixing their hind limbs for a long time. Forty SD rats were administered intragastrically for 12 weeks. The bone histomorphometry parameters and the level of oxidative stress were measured. In addition, the changes of muscle atrophy F-box (MAFbx), muscle RING-finger protein-1 (MuRF1), forkhead box O1 (FOXO1) mRNA expression in skeletal muscle of rats were observed. Results RGP significantly increased the percentage of fluorescence perimeter and bone mineralization deposition rate of the second lumbar vertebrae of rats. It also significantly increased the wet weight ratio and muscle fiber cross-sectional area of the gastrocnemius muscle of rats. At the same time, RGP significantly increased the levels of super oxide dismutase (SOD) and catalase (CAT) in the skeletal muscle of rats, and reduced the content of malondialdehyde (MDA). Rehmannia glutinosa polysaccharides also significantly reduced the expression levels of FOXO1, MAFbx and MuRF1 mRNA in rat skeletal muscle. Conclusions RGP could improve the bone structure of osteoporotic rats. It could also improve muscle that atrophy may be related to the inhibition of FOXO1-mediated ubiquitin-proteasome pathway.

Exercise-based training decreases hospitalizations in heart failure patients but such patients have exercise intolerance. The objectives of the study were to evaluate the effect of 12 weeks of Tai Chi exercise and lower limb muscles' functional electrical stimulation in older chronic heart failure adults. A total of 1,084 older adults with chronic systolic heart failure were included in a non-randomized clinical trial (n=271 per group). The control group did not receive any kind of intervention, one group received functional electrical stimulation of lower limb muscles (FES group), another group practiced Tai Chi exercise (TCE group), and another received functional electrical stimulation of lower limb muscles and practiced Tai Chi exercise (FES & TCE group). Quality of life and cardiorespiratory functions of all patients were evaluated. Compared to the control group, only FES group had increased Kansas City Cardiomyopathy Questionnaire (KCCQ) score (P<0.0001, q=9.06), only the TCE group had decreased heart rate (P<0.0001, q=5.72), and decreased peak oxygen consumption was reported in the TCE group (P<0.0001, q=9.15) and FES & TCE group (P<0.0001, q=10.69). FES of lower limb muscles and Tai Chi exercise can recover the quality of life and cardiorespiratory functions of older chronic heart failure adults (trial registration: Research Registry 4474, January 1, 2015).

Tumor necrosis factor-alpha (TNF-α) plays an important role in autoimmune diseases. Previous studies have investigated the association of TNF-α-238G/A (rs361525) and -308G/A (rs1800629) polymorphisms with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). However, no agreed conclusion had been made. Therefore, this meta-analysis was conducted to assess the associations of TNF-α-238G/A and -308G/A polymorphisms with RA and SLE risk. A systematic search was conducted in commonly used databases. Meta-analysis was performed by STATA12.0. A total of 43 studies were included. In the overall population, the TNF-α-238A allele was observed to be a protective factor for RA (A vs G: OR=0.75, 95%CI=0.57–0.99, P=0.040) and the TNF-α-308A allele was found to be a risk factor for SLE (A vs G: OR=1.78, 95%CI=1.45–2.19, P<0.001). However, no evidence of association was found between TNF-α-238 G/A polymorphism and SLE nor between -308G/A and RA. In the subgroup analysis, TNF-α-308A allele played a pathogenic role for RA in Latin Americans (A vs G: OR=1.46, 95%CI=1.15–1.84, P=0.002) and for SLE in Latin Americans (A vs G: OR=2.12, 95%CI=1.32–3.41, P=0.002) and Europeans (A vs G: OR=2.03, 95%CI=1.56–2.63, P<0.001), while it played a protective role for RA in Asians (A vs G: OR=0.54, 95%CI=0.32–0.90, P=0.017). No significant association was found between TNF-α-308G/A and SLE susceptibility in Africans and Asians. This meta-analysis demonstrated that TNF-α-238A was associated with decreased risk of RA rather than SLE, while -308G/A polymorphism was associated with SLE rather than RA. Stratification analysis indicated that different ethnicities would have different risk alleles.

OBJECTIVES: The pedunculopontine nucleus (PPN) is considered a promising new target for neurostimulation in Parkinson’s disease (PD) patients with postural instability and gait disturbance that is refractory to other treatment modalities. However, the PPN is typically difficult to visualize with magnetic resonance imaging (MRI) at clinical field strengths, which greatly limits the PPN as a viable surgical target for deep brain stimulation (DBS). Thus, the aim of this study is to directly visualize the PPN based on 7.0T ultrahigh-field MRI. METHODS: Five PD patients were enrolled and scanned using the MP2RAGE sequence on a 7.0T ultrahigh-field MRI scanner. Then, the MP2RAGE sequences were imported into a commercially available navigation system. The coordinates of the directly localized PPN poles were recorded in the navigation system relative to the anterior commissure-posterior commissure plane. RESULTS: Our results indicated that the PPN presented intermediate signal intensity in the 7.0T ultrahigh-field MR images in comparison with the surrounding structure, such as the hypo-intensity of the periaqueductal gray and the hyperintensity of the neighboring white matter tracts, in PD patients. The mean coordinates for the rostral and caudal poles of PPN were 6.50 mm and 7.20 mm lateral, 1.58 mm and 2.21 mm posterior, and 8.89 mm and 13.83 mm relative to the posterior commissure. CONCLUSION: Our findings provide, for the first time, direct visualization of the PPN using the MP2RAGE sequence on a 7.0T ultrahigh-field MRI, which may improve the accuracy of stereotactic targeting of the PPN and improve the outcomes in patients undergoing DBS.

Background: Bloodstream infections (BSIs) due to Enterococcus faecium (E. faecium), particularly those due to vancomycin-resistant enterococcus (VRE), are still a therapeutic challenge. Aim: To evaluate mortality from BSI due to E. faecium and VRE in central Taiwan. Materials and Methods: We retrospectively analyzed cases of significant E. faecium BSI in the Changhua Christian Hospital System between January 1, 2010 and December 31, 2013. Results: Of the 76 cases, 28 patients (36.8%) were admitted to intensive care units (ICUs) at the onset of BSI, 10 (13.2%) cases were associated with polymicrobial bacteremia, and 29 (38.2%) cases were associated with entry via the biliary tract. VRE was observed in 18 (23.7%) cases. The 30-day mortality rate was 13.1% (10/76). Multivariate logistic regression analysis showed that bacteremia of non-biliary tract origin (OR = 8.43, 95% confidence interval (95% CI) = 1.32-54.00, p = 0.002) and ICU admission (OR = 4.2, 95% CI = 1.7-10.0, p = 0.002) were significant risk factors for 30-day mortality, whereas appropriate antimicrobial therapy was a protective factor for 30-day mortality (OR = 0.33, 95% CI = 0.14-0.79, p = 0.013). Conclusions: Our results underscore the need to assist patients admitted to ICUs with E. faecium BSIs with a non-biliary tract origin. We emphasize the use of appropriate antimicrobial therapy for E. faecium BSI with the aim to rescue more patients with these infections.

Antecedentes: Las infecciones del torrente sanguíneo por Enterococcus faecium, particularmente aquellas causadas por enterococos resistentes a vancomicina (ERV), representan aún un desafío para los tratamientos. Este estudio está orientado a la evaluación de la mortalidad debido a la infección del torrente sanguíneo (ITS) por E. faecium y por enterococos resistentes a vancomicina (ERV) en Taiwán central. Materiales y Métodos: Analizamos de forma retrospectiva casos de ITS causadas por E. faecium genuinas en el Sistema del Hospital Changhua Christian, entre los días 1 de enero de 2010 y 31 de diciembre de 2013. Resultados: De los 76 casos analizados, 28 pacientes fueron ingresados a las Unidades de Cuidados Intensivos (UCI) al comienzo de una ITS (36,8%), 10 casos fueron asociados a bacteriemia polimicrobiana (13,2%), y 29 casos tuvieron como puerta de entrada la vía biliar. En 18 casos se pudieron observar ERV (23,7%). La mortalidad a 30 días fue de 13,1% (10/76). El análisis multivariado mediante regresión logística mostró que la bacteriemia de origen no biliar (OR = 8,43, 95% intervalo de confianza (95% CI) = 1,32-54,00; p = 0,002), y el ingreso a la UCI (OR = 4,2; 95% CI = 1,7-10,0; p = 0,002), fueron factores de riesgo significativos para el rango de mortalidad de 30 días, así como un tratamiento de antimicrobiano apropiado constituye un factor protector en contra la mortalidad (OR = 0,33; 95% CI = 0,14-0,79; p = 0,013). Conclusiones: Nuestros resultados destacan la necesidad de asistir a los pacientes ingresados a la UCI con ITS por E. faecium con origen no biliar. Hacemos énfasis a la aplicación de una antibioterapia adecuada para sacar adelante a un mayor número de pacientes con este tipo de infecciones.

La fibrilación auricular (FA) es la arritmia más frecuente en la población adulta. Es sabido que la FA se asocia con un aumento del riesgo de presentar muchos eventos adversos, entre los que se incluyen accidente cerebrovascular, insuficiencia cardíaca, muerte por todas las causas, así como muerte súbita cardíaca. Más recientemente también ha habido un creciente interés en la asociación de FA con deterioro cognitivo y demencia. En esta revisión narrativa describimos la actual evidencia a favor de la relación entre la FA y la declinación cognitiva, el deterioro cognitivo o la demencia en pacientes sin antecedentes de accidente cerebrovascular. Aun cuando no queda muy claro cuáles son los mecanismos exactos que explican la asociación entre la FA y la declinación cognitiva, los mecanismos propuestos incluyen isquemia silente, estado pro inflamatorio, microsangrados cerebrales e hipoperfusión cerebral. Se necesitan más estudios para dilucidar los mecanismos de base que faciliten el descubrimiento de alguna estrategia de prevención.

Atrial fibrillation (AF) is the most common arrhythmia in the adult population. AF is widely known to be associated with an increased risk of many adverse outcomes including stroke, heart failure, all-cause death, as well as sudden cardiac death. More recently, there has also been growing interest in the association of AF with cognitive impairment and dementia. In this narrative review, we describe the current evidence that supports the relationship of AF to cognitive decline, cognitive impairment, or dementia in patients without a history of stroke. The exact mechanisms underlying the association of AF with cognitive decline are unclear; however, proposed mechanisms include silent ischemia, pro-inflammatory state, cerebral microbleeds, and cerebral hypoperfusion. Further research is needed to elucidate the underlying mechanisms to facilitate discovery of prevention strategies.

FUNDAMENTO: O programa de biogênese mitocondrial no coração parece apresentar remodelação adaptativa após estresse biomecânico e oxidativo. Os mecanismos adaptativos que protegem o metabolismo do miocárdio durante a hipóxia são coordenados, em parte, pelo óxido nítrico (NO). OBJETIVO: Observar a biogênese mitocondrial e expressão do óxido nítrico sintase (NOS) em corações de cardiopatia congênita com cianose; discutir a resposta mitocondrial à hipóxia crônica do miocárdio. MÉTODOS: Foram investigados 20 pacientes com defeitos cardíacos cianóticos (n = 10) ou acianóticos (n = 10). Foram estudadas amostras do miocárdio na via de saída ventricular direita, tomadas durante a operação. A análise morfométrica de mitocôndrias foi realizada por microscopia eletrônica de transmissão. A relação mtDNA/nDNA foi determinada com PCR em tempo real. Os níveis de transcrição da subunidade I da citocromo c oxidase (COXI), coativador-1α do receptor γ ativado por proliferador de peroxissoma (PGC-1α), o fator respiratório nuclear 1 (NRF1), e fator de transcrição mitocondrial A (Tfam) foram detectados por reação em cadeia da polimerase via transcriptase reversa (RT-PCR) ativado por fluorescência em tempo real. Os níveis proteicos de COXI e nNOS, iNOS e eNOS foram medidos por técnica de Western Blot. RESULTADOS: A densidade volumétrica mitocondrial (Vv) e a densidade numérica (Nv) foram significativamente elevadas em pacientes com cianose, em comparação com a cardiopatia congênita acianótica. MtDNA elevada e suprarregulação dos níveis de COXI, PGC-1 α, NRF1 e Tfam mRNA foram observadas em pacientes cianóticos. Os níveis de proteína de COXI e eNOS foram significativamente maiores no miocárdio de pacientes cianóticos que nos de acianóticos. Os níveis de transcrição do PGC-1α se correlacionam com os níveis de eNOS. CONCLUSÃO: A biogênese mitocondrial é ativada no miocárdio da via de saída ventricular na cardiopatia congênita com cianose, que poderia ser a resposta adaptativa à hipóxia crônica e possivelmente envolve suprarregulação da eNOS. (Arq Bras Cardiol. 2012; [online].ahead print, PP.0-0)

BACKGROUND: Mitochondrial biogenesis program in heart appears to exhibit adaptive remodeling following biomechanical and oxidative stress. The adaptive mechanisms that protect myocardium metabolism during hypoxia are coordinated in part by nitric oxide (NO). OBJECTIVE: To observe mitochondrial biogenesis and nitric oxide synthase (NOS) expression in hearts of congenital heart disease with cyanosis, discuss mitochondrial response to chronic hypoxia in myocardium. METHODS: 20 patients with cyanotic (n=10) or acyanotic cardiac defects (n=10) were investigated. Samples from the right ventricular outflow tract myocardium taken during operation were studied. Morphometric analysis of mitochondria was performed with transmission electron microscope. Relative mtDNA/nDNA ratio was determined with real-time PCR. Cytochrome c oxidase subunit I (COXI), peroxisome-proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (Tfam) transcript levels were detected by real-time fluorescent RT-PCR. COXI and nNOS, iNOS and eNOS protein levels were measured with western blot. RESULTS: Mitochondrial volume density (Vv) and numerical density (Nv) were significantly elevated in patients with cyanotic compared to acyanotic congenital heart disease. Elevated mtDNA and up-regulated COXI, PGC-1α, NRF1 and Tfam mRNA levels were observed in cyanotic patients. Protein levels of COXI and eNOS were significantly higher in the myocardium of cyanotic than of acyanotic patients. PGC-1α transcript levels correlated with the levels of eNOS. Conclusion: Mitochondrial biogenesis is activated in right ventricular outflow tract myocardium in congenital heart disease with cyanosis, which could be the adaptive response to chronic hypoxia and possibly involves eNOS up-regulation. (Arq Bras Cardiol. 2012; [online].ahead print, PP.0-0)

Objective: To investigate the course of tubular cell senescence and expressions of p53, p21, and Rb during the late phase of ischemia/reperfusion (IRI) in the kidney, and assess the effects of the p53-Rb pathway on tubular cell senescence. Methods: Experimental models of unilateral renal IRI were used in p53(+/+) and p53(-/-) mice. Histological changes at the tubular level, progress of cell senescence, and the expression of Rb, p21, and/or p53 proteins in tubular cells were studied at different moments in time after IRI. Results: Chronic tubulointerstitial fibrosis was much more severe and widely distributed in IRI kidneys of p53(+/+) mice in later stages than in earlier stages. Senescent tubular cells were significantly increased at 3 and 6 months after IRI. In contrast, in contralateral kidneys of p53(+/+) mice and in both kidneys of p53(-/-) mice, almost no senescent cells were observed at 1 and 3 months after IRI, and only a few senescent cells were detected in IRI kidneys of p53(-/-) mice at 6 months. In mice of both genotypes, cell senescence was correlated with the expression levels of p53, p21, and Rb proteins. Conclusion: The IRI accelerated tubular cell senescence is presumed to be one of the mechanisms of the "long-term effect" of IRI. Furthermore, the activation of p53-Rb signaling pathway may play a vital role in tubular cell senescence induced by IRI.

Objective: To investigate the functions of Fibroblast Growth Factor Receptor-2 (FGFR2) at different stages of cell differentiation. The engineered murine embryonic stem (ES) cells with conditional knockout of FGFR2 were developed depending on Cre-loxP. Methods: Cre-loxP system was used in a conditional targeting vector. The competent AM-1 bacteria, which expressed Cre-recombinase, was used to confirm the Cre-mediated deletion of the floxed exons 7 and 8 of FGFR2. The targeting vector was electroporated into the ES cells, and the transfected ES cells were screened with G418 and Ganciclovir. Finally, the ES clones with correct targeting events were identified by Southern Blot and PCR. Results: The targeting vector with conditional knockout of murine FGFR2 was successfully constructed and confirmed by PCR and digestion analysis in bacteria. 86 ES clones were collected by selective culture with G418 and Ganciclovir. Four of the 86 ES clones were found containing the targeting gene sequence in genomic DNA proved by Southern Blot with a 5'-end flank probe. Two of the four ES clones had the correct targeting events that included the insertion of the targeting gene sequence in genomic DNA and were checked by Southern Blot with a 3'-end flanking probe. Finally, the insertion of loxP (loxP3) between exons 8 and 9 in genomic DNA was identified in one of the two ES clones by Southern Blot and PCR. Conclusion: FGFR2 conditional knockout depending on Cre-loxP can be successfully used in ES cells.