Association between stroke and physical activities in Shanghai Community Elderly Cohort

Objective·To compare the physical activities of stroke population and non-stroke population based on the baseline survey of the elderly population cohort in Shanghai communities, and explore the participation in different types of physical activities of stroke population.Methods·The subjects were screened from Shanghai Community Elderly Cohort constructed from February to August, 2019 according to the admission criteria. The subjects were divided into non-stroke group and stroke group according to whether they had reported a history of stroke by themselves, and the two groups were matched 2 to 1 by controlling age and sex with propensity score matching. The baseline characteristics of the two groups were collected, and the physical activities related to sports, transportation and housework in the last week were investigated with the International Physical Activity Questionnaire (IPAQ). Pittsburgh Sleep Quality Index (PSQI) was used to evaluate the sleep quality of the subjects. Generalized Anxiety Disorder (GAD-7) and Patient Health Questionnaire-9 (PHQ-9) were used to evaluate anxiety and depression of the subjects, respectively. The above characteristics were compared between the stroke group and non-stroke group, and the participation of different types of physical activities were compared between the two groups by multivariate Logistic regression model.Results·Among the 17 948 people included, there were 993 (5.5%) in the stroke group and 16 955 (94.5%) in the non-stroke group. After propensity score matching, there were 1 984 people (66.7%) in the non-stroke group and 992 people (33.3%) in the stroke group. There were significant differences in education level, pre-retirement occupation, waist circumference, body mass index, sleep status, anxiety symptoms, depression symptoms and disease history between the two groups (all P<0.05). In terms of physical activities, the female stroke group had shorter daily moderate exercises time, fewer riding and walking days in one week, and shorter daily riding time, compared with the non-stroke people, with statistical significance (all P<0.05). Compared with the non-stroke people, the weekly housework days and daily housework time in the male and female stroke groups were lower than those in the non-stroke group, while the daily sedentary time was longer, with statistical significance (all P=0.000). In terms of physical activity level, the proportions of men and women in the stroke group who reached medium or high level were lower than those in the non-stroke group, and the differences were statistically significant (all P=0.000). After adjusting for gender, age, occupation, anxiety symptoms, history of hyperlipidemia, history of atrial fibrillation, history of chronic gastritis and history of hip fracture by multivariate Logistic regression model, the level of vigorous exercise participation in the stroke group was lower, the proportions of no housework in the last week and sedentary time greater than 180 min per day were higher, and the proportion at medium and high activity levels was lower (all P<0.05).Conclusion·The frequency and duration of housework participation and the physical activity level of elderly people with a history of stroke in Shanghai communities are at a lower level than those without stroke, and they also have a longer sedentary time

The Mitochondrial Calcium Uniporter Matches Energetic Supply with Cardiac Workload during Stress and Modulates Permeability Transition

Cardiac contractility is mediated by a variable flux in intracellular calcium (Ca2+), thought to be integrated into mitochondria via the mitochondrial calcium uniporter (MCU) channel to match energetic demand. Here, we examine a conditional, cardiomyocyte-specific, mutant mouse lacking Mcu, the pore-forming subunit of the MCU channel, in adulthood. Mcu−/− mice display no overt baseline phenotype and are protected against mCa2+ overload in an in vivo myocardial ischemia-reperfusion injury model by preventing the activation of the mitochondrial permeability transition pore, decreasing infarct size, and preserving cardiac function. In addition, we find that Mcu−/− mice lack contractile responsiveness to acute β-adrenergic receptor stimulation and in parallel are unable to activate mitochondrial dehydrogenases and display reduced bioenergetic reserve capacity. These results support the hypothesis that MCU may be dispensable for homeostatic cardiac function but required to modulate Ca2+-dependent metabolism during acute stress

G protein-coupled receptor kinase 2 promotes cardiac hypertrophy

<div><p>The increase in protein activity and upregulation of G-protein coupled receptor kinase 2 (GRK2) is a hallmark of cardiac stress and heart failure. Inhibition of GRK2 improved cardiac function and survival and diminished cardiac remodeling in various animal heart failure models. The aim of the present study was to investigate the effects of GRK2 on cardiac hypertrophy and dissect potential molecular mechanisms. In mice we observed increased GRK2 mRNA and protein levels following transverse aortic constriction (TAC). Conditional GRK2 knockout mice showed attenuated hypertrophic response with preserved ventricular geometry 6 weeks after TAC operation compared to wild-type animals. In isolated neonatal rat ventricular cardiac myocytes stimulation with angiotensin II and phenylephrine enhanced GRK2 expression leading to enhanced signaling via protein kinase B (PKB or Akt), consecutively inhibiting glycogen synthase kinase 3 beta (GSK3β), such promoting nuclear accumulation and activation of nuclear factor of activated T-cells (NFAT). Cardiac myocyte hypertrophy induced by in vitro GRK2 overexpression increased the cytosolic interaction of GRK2 and phosphoinositide 3-kinase γ (PI3Kγ). Moreover, inhibition of PI3Kγ as well as GRK2 knock down prevented Akt activation resulting in halted NFAT activity and reduced cardiac myocyte hypertrophy. Our data show that enhanced GRK2 expression triggers cardiac hypertrophy by GRK2-PI3Kγ mediated Akt phosphorylation and subsequent inactivation of GSK3β, resulting in enhanced NFAT activity.</p></div

G-protein coupled receptor kinase 2 (GRK2) regulates proteinkinase B (Akt) phosphorylation.

<p>(A-E) G-protein coupled receptor (GPCR) promoted Akt phosphorylation depends on GRK2. (A) Representative Western Blots of Akt phosphorylation at residues Ser 473 (p_SAKT) and Thr 308 (p_TAKT), total Akt expression, GRK2 expression and GAPDH under respective conditions. Samples shown on each lane are blotted, antibody stained and developed on the same western blot membrane. Vertical dotted line indicates 2 excluded samples with non ANG II/PE GPCR stimulation. Quantification of p_SAKT under PE (B; n = 4) and ANG II (C; n = 3) and p_TAKT under PE (D; n = 3) and ANG II (E; n = 4) stimulation and total Akt under respective treatment conditions, values normalized to control scrambled siRNA (Scr), * P < 0.05. (F) GRK2 overexpression promotes full Akt phosphorylation. Representative Western Blots and quantification of p_SAKT (n = 7), p_TAKT (n = 7) and total Akt (n = 4) expression after transfection with an adenovirus harboring GRK2 (AdGRK2) or β-galactosidase/LacZ as control (AdLacZ), GAPDH served as loading control. Values normalized to untreated control (-). * P < 0.05, *** P < 0.001.</p

G-protein coupled receptor kinase 2 (GRK2) expression is regulated by G-protein coupled receptor (GPCR) stimulation <i>in vitro</i>.

<p>(A) representative Western Blots of GRK2 expression in neonatal rat ventricular myocytes (NRVMs) after stimulation with Phenylephrine (PE), or Angiotensin II (ANG II), GAPDH as loading control. (B) Quantification of GRK2 protein expression after PE stimulation normalized to GAPDH and unstimulated (-) NRVM as control. Samples shown on each lane are blotted, antibody stained and developed on the same western blot membrane. Vertical dotted line indicates 2 excluded samples with non ANG II/PE GPCR stimulation. n = 10, * P < 0.05. (C) Quantification of GRK2 protein expression after ANG II stimulation normalized to GAPDH, n = 11, *** P < 0.001. (D) GRK2 knockdown by siRNA. Treatment of NRVM with siRNA against GRK2 (siGRK2, n = 6) and scrambled siRNA as control (Scr, n = 4), ** P < 0.01. (E) Adenoviral GRK2 overexpression. Representative Western Blots of GRK2 protein expression in untreated NRVMS, NRVMs treated with a control adenovirus harboring β-galactosidase (AdLacZ) and NRVMs after treatment with an adenovirus expressing GRK2 (AdGRK2), MOI = multiplicity of infection. (F) Quantification of GRK2 protein expression following AdLacZ or AdGRK2 transfection normalized to GAPDH as loading control and normalized to untreated cells (-), n = 2.</p

Characterizing the cellular and molecular variabilities of peripheral immune cells in healthy recipients of BBIBP-CorV inactivated SARS-CoV-2 vaccine by single-cell RNA sequencing

AbstractOver 3 billion doses of inactivated vaccines for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been administered globally. However, our understanding of the immune cell functional transcription and T cell receptor (TCR)/B cell receptor (BCR) repertoire dynamics following inactivated SARS-CoV-2 vaccination remains poorly understood. Here, we performed single-cell RNA and TCR/BCR sequencing on peripheral blood mononuclear cells at four time points after immunization with the inactivated SARS-CoV-2 vaccine BBIBP-CorV. Our analysis revealed an enrichment of monocytes, central memory CD4+ T cells, type 2 helper T cells and memory B cells following vaccination. Single-cell TCR-seq and RNA-seq comminating analysis identified a clonal expansion of CD4+ T cells (but not CD8+ T cells) following a booster vaccination that corresponded to a decrease in the TCR diversity of central memory CD4+ T cells and type 2 helper T cells. Importantly, these TCR repertoire changes and CD4+ T cell differentiation were correlated with the biased VJ gene usage of BCR and the antibody-producing function of B cells post-vaccination. Finally, we compared the functional transcription and repertoire dynamics in immune cells elicited by vaccination and SARS-CoV-2 infection to explore the immune responses under different stimuli. Our data provide novel molecular and cellular evidence for the CD4+ T cell-dependent antibody response induced by inactivated vaccine BBIBP-CorV. This information is urgently needed to develop new prevention and control strategies for SARS-CoV-2 infection. (ClinicalTrials.gov Identifier: NCT04871932).Trial registration: ClinicalTrials.gov identifier: NCT04871932.