TNFα stabilizes myocardin mRNA which upregulates myocardin expression and activity in differentiated VSMCs.

<p>(<b>A</b>) VSMCs were serum-starved for 24 h and transcription was blocked using α-amanitin (50 µM). Cells were then treated with TNFα (50 ng/ml) for 1, 3, 6 and 9 h. Graph shows the effect of TNFα on myocardin mRNA degradation. (<b>B</b>) VSMCs containing the T-REx system for myocardin over-expression were treated with Dox. Western blot analysis shows the effect of Dox on myocardin and contractile protein expression levels. (<b>C</b>) RT-qPCR analysis showing the effect of myocardin over-expression on MRTF-A and B. (<b>D</b>) Light microscope images showing the effect of Dox on VSMC morphologies. (<b>E,F,G</b>) VSMCs were serum-starved for 24 h and treated with TNFα (50 ng/ml) and Dox for 24 h. Representative western blots and RT-qPCR analysis show the effect of TNFα on myocardin and contractile marker protein and mRNA expression levels compared with those control without Dox (C). (<b>H</b>) VSMCs without the T-REx system to over-express myocardin were treated with TNFα and Dox to determine non-specific effects of Dox on myocardin. RT-qPCR analysis shows the effect of TNFα and Dox on myocardin mRNA expression levels compared with the control (C). (<b>I</b>) Primary rat aortic VSMCs (passage 2) were serum-starved for 24 h and treated with TNFα (50 ng/ml) for 24 h. Representative western blots show the effect of TNFα on myocardin and contractile marker protein expression levels. (<b>J,K</b>) Western blot and RT-qPCR analyses showing the concentration- and time-dependent effects of TNFα on myocardin and contractile marker expression in VSMCs treated with Dox. qPCR results were normalized using GAPDH (n = 7, *p<0.05).</p

TNFα down-regulates endogenous myocardin expression and activity.

<p>VSMCs were serum-starved for 24 h and treated with TNFα (50 ng/ml) for 24 h. (<b>A</b>) Western blot analysis showing the effects of TNFα on myocardin and contractile marker protein expression levels compared with the control (C). (<b>B</b>) RT-qPCR analysis showing the effect of TNFα on myocardin and contractile marker mRNA expression levels compared to control (C). (<b>C,D</b>) Western blot and RT-qPCR analysis showing the concentration dependent effects of TNFα on myocardin expression and activity. qPCR results were normalized using GAPDH (n = 8, *p<0.05).</p

Model of TNFα-mediated myocardin mRNA stability.

<p>(<b>A</b>) The effect of TNFα in de-differentiated VSMCs. (<b>B</b>) The effect of TNFα in differentiated VSMCs.</p

TNFα decreases contractility of VSMCs.

<p>VSMCs were serum-starved for 24 h, subcultured into collagen gel matrix and treated with TNFα with or without QNZ for 24 h. (<b>A</b>) Representative images of collagen gels showing the change in area of the gel. (<b>B</b>) The cross-sectional area was analyzed using ImageJ and plotted on a graph to show the changes in the collagen area (n = 3, *p<0.05).</p

Inhibition of NF-κB blocks TNFα-induced inhibition of myocardin expression and activity.

<p>VSMCs were serum-starved for 24 h and treated with TNFα (50 ng/ml). (<b>A</b>) Western blot analysis showing the effect of NF-κB inhibitor TPCK (1 µM) on TNFα-induced inhibition of myocardin and contractile marker expression. (<b>B,C</b>) Western blot and RT-qPCR analyses showing the effects of NF-κB inhibitor QNZ (1 µM) on TNFα-induced inhibition of myocardin expression and activity compared with the control (C). (<b>D,E</b>) VSMCs were treated with TNFα for different time points and the p65 subunit was stained using anti-p65 antibody and AF488-conjugated secondary antibody. The nuclear localization of the p65 subunit was determined using LSC based on AF488 florescence and propidium iodide (PI) nuclear staining. (<b>F</b>) VSMCs were treated with TNFα for different time points and cellular and nuclear proteins were extracted separately. Western blot analysis was performed to show localization of the p65 subunit in the cytoplasm and nuclei of VSMCs. qPCR results were normalized using GAPDH (n = 4, *p<0.05).</p

TNFα increases proliferation of VSMCs.

<p>VSMCs were serum-starved for 24 h, treated with the indicated drugs for 24 h and labeled with BrdU for 60 min. Cells were then fixed with 80% ethanol and BrdU incorporation was detected using anti-BrdU antibody and AF488-conjugated secondary antibody using LSC. (<b>A</b>) Cells were treated with TNFα with and without QNZ. Graph shows the percentage of BrdU positive cells of the total scanned cells. (<b>B</b>) Cells were treated with TNFα and/or Dox to induce myocardin expression. Graph shows the percentage of BrdU positive cells of the total number of cells (n = 6, *p<0.05).</p

The colours and contours of compassion: A systematic review of the perspectives of compassion among ethnically diverse patients and healthcare providers

<div><p>Objective</p><p>To identify and describe the perspectives, experiences, importance, and impact of compassionate care among ethnically diverse population groups.</p><p>Methods</p><p>A systematic search of peer-reviewed research focused on compassionate care in ethnically diverse populations published between 1946 and 2017 was conducted.</p><p>Results</p><p>A total of 2296 abstracts were retrieved, out of which 23 articles met the inclusion criteria. Synthesis of the literature identified the perspectives, facilitators and barriers of compassion in healthcare within ethnic groups. Compassion was described as being comprised of healthcare provider (HCP) virtues (honesty, kindness, helpful, non-judgment) and actions (smile, touch, care, support, flexibility) aimed at relieving the suffering of patients. The importance and impact of providing compassion to ethnically diverse patients was also identified which included overcoming cultural differences, alleviating distress at end-of-life, promoting patient dignity and improving patient care. This review also identified the need for more contextual studies directly exploring the topic of compassion from the perspectives of individuals within diverse ethnic groups, rather than superimposing a pre-defined, enculturated and researcher-based definition of compassion.</p><p>Conclusions</p><p>This review synthesizes the current evidence related to perceptions of compassion in healthcare among diverse ethnic groups and the role that compassion can play in bridging ethno-cultural differences and associated challenges, along with identifying gaps in literature related to compassionate care within diverse ethnic groups. Establishing an evidence base grounded in the direct accounts of members of diverse ethnic communities can enhance culturally sensitive compassionate care and improve compassion related health outcomes among diverse ethnic groups.</p></div

Data extraction and evaluation of studies.

<p>Data extraction and evaluation of studies.</p

Search and selection process.

<p>Search and selection process.</p

Ovid MEDLINE(R) (1946 to 2017) search strategy.

<p>Ovid MEDLINE(R) (1946 to 2017) search strategy.</p