How does patient-centered hospital culture affect clinical physicians' medical professional attitudes and behaviours in chinese public hospitals: a cross-sectional study?

BACKGROUND: An increasing number of studies on physicians' professionalism have been done since the 2002 publication of Medical Professionalism in the New Millennium: A Physician Charter. The Charter proposed three fundamental principles and ten responsibilities. However, most studies were done in developed countries, and few have been done in China. Additionally, few studies have examined the effect of patient-centered hospital culture (PCHC) on physicians' professionalism. We aimed to investigate physicians' medical professionalism in public hospitals in China, and to assess mediating effect of professional attitudes in the relationship of PCHC with professional behaviours. METHODS: Self-administered questionnaires including professional attitudes (20 items) and behaviours (10 items) survey and PCHC scale (22 items) were given to clinical physicians in five public hospitals, China. The mediating effect of professional attitudes in the relationship of PCHC with professional behaviours was tested. RESULT: 232 valid questionnaires were collected. More than 90% (208) respondents agreed with 15 of 20 specific statements on medical professionalism. As for the responsibility of improving quality of care, 54 (23%) respondents disagreed with reporting of incompetent colleagues and as for the responsibility of maintaining professional competence, 49 (21%) disagreed with recertification. More than 185 (83%) respondents reported that they sometimes, usually, or always showed the four positive behaviours on the questionnaire, and 173 (77%) reported that they never showed the six negative behaviours. Mediating effect analysis revealed that two dimensions of PCHC (i.e. value/institution culture and behaviour/material culture) had a significant positive impact on physicians' professional behaviour, and professional attitude played a complete mediation role between them, but another dimension of PCHC (i.e. negative evaluation of hospital) directly affected professional behaviour without influencing professional attitude. CONCLUSION: Chinese physicians showed positive professional attitudes and behaviours. Different dimensions of PCHC affected physicians' attitudes and behaviours in different ways.

Enhanced Spectral Response of ZnO-Nanorod-Array-Based Ultraviolet Photodetectors by Alloying Non-Isovalent Cu-O with CuAlO2 P-Type Layer.

CuAlO2 was synthesized by a hydrothermal method, in which the Cu-O dimers were incorporated by simply altering the ratio of the reactants and the temperature. The incorporation process increases the grain size in CuAlO2, and modulates the work function and binding energies for CuAlO2 due to the partial substitution of Cu+ 3d10 with Cu2+ 3d9 orbitals in the valence band maximum by alloying non-isovalent Cu-O with a CuAlO2 host. Based on the ZnO nanorod arrays (NRs) ultraviolet photodetector, CuAlO2/Cu-O fabricated by the low-cost drop-coating method was used as the p-type hole transport layer. The incorporation of the Cu-O clusters into CuAlO2 lattice to enhance the conductivity of CuAlO2 is an effective way for improving ZnO NRs/CuAlO2 device performance. The photodetectors exhibit significant diode behavior, with a rectification ratio approaching 30 at ±1 V, and a dark saturation current density 0.81 mA cm-2. The responsivity of the ZnO-NRs-based UV photodetector increases from 13.2 to 91.3 mA/W at 0 V bias, with an increase in the detectivity from 2.35 × 1010 to 1.71 × 1011 Jones. Furthermore, the ZnO NRs/[CuAlO2/Cu-O] photodetector exhibits a maximum responsivity of 5002 mA/W at 1.5 V bias under 375 nm UV illumination.

Revisiting the relationship between turgor pressure and plant cell growth.

Growth is central to plant morphogenesis. Plant cells are encased in rigid cell walls, and they must overcome physical confinement to grow to specific sizes and shapes. Cell wall tension and turgor pressure are the main mechanical components impacting plant cell growth. Cell wall mechanics has been the focus of most plant biomechanical studies, and turgor pressure was often considered as a constant and largely passive component. Nevertheless, it is increasingly accepted that turgor pressure plays a significant role in plant growth. Numerous theoretical and experimental studies suggest that turgor pressure can be both spatially inhomogeneous and actively modulated during morphogenesis. Here, we revisit the pressure-growth relationship by reviewing recent advances in investigating the interactions between cellular/tissular pressure and growth.

Mechanosensing, from forces to structures.

Sessile plants evolve diverse structures in response to complex environmental cues. These factors, in essence, involve mechanical stimuli, which must be sensed and coordinated properly by the plants to ensure effective growth and development. While we have accumulated substantial knowledge on plant mechanobiology, how plants translate mechanical information into three-dimensional structures is still an open question. In this review, we summarize our current understanding of plant mechanosensing at different levels, particularly using Arabidopsis as a model plant system. We also attempt to abstract the mechanosensing process and link the gaps from mechanical cues to the generation of complex plant structures. Here we review the recent advancements on mechanical response and transduction in plant morphogenesis, and we also raise several questions that interest us in different sections.

Looking beyond the gene network - metabolic and mechanical cell drivers of leaf morphogenesis.

The above-ground organs in plants display a rich diversity, yet they grow to characteristic sizes and shapes. Organ morphogenesis progresses through a sequence of key events, which are robustly executed spatiotemporally as an emerging property of intrinsic molecular networks while adapting to various environmental cues. This Review focuses on the multiscale control of leaf morphogenesis. Beyond the list of known genetic determinants underlying leaf growth and shape, we focus instead on the emerging novel mechanisms of metabolic and biomechanical regulations that coordinate plant cell growth non-cell-autonomously. This reveals how metabolism and mechanics are not solely passive outcomes of genetic regulation but play instructive roles in leaf morphogenesis. Such an integrative view also extends to fluctuating environmental cues and evolutionary adaptation. This synthesis calls for a more balanced view on morphogenesis, where shapes are considered from the standpoints of geometry, genetics, energy and mechanics, and as emerging properties of the cellular expression of these different properties.

Non-autonomous stomatal control by pavement cell turgor via the K+ channel subunit AtKC1.

Stomata optimize land plants' photosynthetic requirements and limit water vapor loss. So far, all of the molecular and electrical components identified as regulating stomatal aperture are produced, and operate, directly within the guard cells. However, a completely autonomous function of guard cells is inconsistent with anatomical and biophysical observations hinting at mechanical contributions of epidermal origins. Here, potassium (K+) assays, membrane potential measurements, microindentation, and plasmolysis experiments provide evidence that disruption of the Arabidopsis thaliana K+ channel subunit gene AtKC1 reduces pavement cell turgor, due to decreased K+ accumulation, without affecting guard cell turgor. This results in an impaired back pressure of pavement cells onto guard cells, leading to larger stomatal apertures. Poorly rectifying membrane conductances to K+ were consistently observed in pavement cells. This plasmalemma property is likely to play an essential role in K+ shuttling within the epidermis. Functional complementation reveals that restoration of the wild-type stomatal functioning requires the expression of the transgenic AtKC1 at least in the pavement cells and trichomes. Altogether, the data suggest that AtKC1 activity contributes to the building of the back pressure that pavement cells exert onto guard cells by tuning K+ distribution throughout the leaf epidermis.

NanoIndentation, an ImageJ Plugin for the Quantification of Cell Mechanics.

Growth and morphogenesis in plants depend on cell wall mechanics and on turgor pressure. Nanoindentation methods, such as atomic force microscopy (AFM), enable measurements of mechanical properties of a tissue at subcellular resolution, while confocal microscopy of tissues expressing fluorescent reporters indicates cell identity. Associating mechanical data with specific cells is essential to reveal the links between cell identity and cell mechanics. Here we describe an image analysis protocol that allows us to segment AFM scans containing information on tissue topography and/or mechanics, to stitch several scans in order to reconstitute an entire region of the tissue investigated, to segment the scans and label cells, and to associate labeled cells to the projection of confocal images. Thus all mechanical data can be mapped to the corresponding cells and to their identity. This protocol is implemented using NanoIndentation, a plugin that we are developing in the Fiji distribution of ImageJ.

Roadmap for the multiscale coupling of biochemical and mechanical signals during development.

The way in which interactions between mechanics and biochemistry lead to the emergence of complex cell and tissue organization is an old question that has recently attracted renewed interest from biologists, physicists, mathematicians and computer scientists. Rapid advances in optical physics, microscopy and computational image analysis have greatly enhanced our ability to observe and quantify spatiotemporal patterns of signalling, force generation, deformation, and flow in living cells and tissues. Powerful new tools for genetic, biophysical and optogenetic manipulation are allowing us to perturb the underlying machinery that generates these patterns in increasingly sophisticated ways. Rapid advances in theory and computing have made it possible to construct predictive models that describe how cell and tissue organization and dynamics emerge from the local coupling of biochemistry and mechanics. Together, these advances have opened up a wealth of new opportunities to explore how mechanochemical patterning shapes organismal development. In this roadmap, we present a series of forward-looking case studies on mechanochemical patterning in development, written by scientists working at the interface between the physical and biological sciences, and covering a wide range of spatial and temporal scales, organisms, and modes of development. Together, these contributions highlight the many ways in which the dynamic coupling of mechanics and biochemistry shapes biological dynamics: from mechanoenzymes that sense force to tune their activity and motor output, to collectives of cells in tissues that flow and redistribute biochemical signals during development.

Cellular Heterogeneity in Pressure and Growth Emerges from Tissue Topology and Geometry.

Cell-to-cell heterogeneity prevails in many systems, as exemplified by cell growth, although the origin and function of such heterogeneity are often unclear. In plants, growth is physically controlled by cell wall mechanics and cell hydrostatic pressure, alias turgor pressure. Whereas cell wall heterogeneity has received extensive attention, the spatial variation of turgor pressure is often overlooked. Here, combining atomic force microscopy and a physical model of pressurized cells, we show that turgor pressure is heterogeneous in the Arabidopsis shoot apical meristem, a population of stem cells that generates all plant aerial organs. In contrast with cell wall mechanical properties that appear to vary stochastically between neighboring cells, turgor pressure anticorrelates with cell size and cell neighbor number (local topology), in agreement with the prediction by our model of tissue expansion, which couples cell wall mechanics and tissue hydraulics. Additionally, our model predicts two types of correlations between pressure and cellular growth rate, where high pressure may lead to faster- or slower-than-average growth, depending on cell wall extensibility, yield threshold, osmotic pressure, and hydraulic conductivity. The meristem exhibits one of these two regimes, depending on conditions, suggesting that, in this tissue, water conductivity may contribute to growth control. Our results unravel cell pressure as a source of patterned heterogeneity and illustrate links between local topology, cell mechanical state, and cell growth, with potential roles in tissue homeostasis.

Visualizing Protein Associations in Living Arabidopsis Embryo.

Protein-protein interactions (PPI) are essential for a plethora of biological processes. These interactions can be visualized and quantified with spatial resolution using Förster resonance energy transfer (FRET) measured by fluorescence lifetime imaging microscopy (FLIM) technology. Currently, FRET-FLIM is routinely used in cell biology, and it has become a powerful tool to map protein interactions in native environments. However, implementing this technology in living multicellular organism remains challenging, especially when dealing with developing plant embryos where tissues are confined in multiple cell layers preventing direct imaging. In this chapter, we describe a step-by-step protocol for studying PPI using FRET-FLIM of the two transcription factors SCARECROW and SHORTROOT in Arabidopsis embryos. We provide a detailed description from embryo isolation to data analysis and representation.

Xyloglucans and Microtubules Synergistically Maintain Meristem Geometry and Phyllotaxis.

The shoot apical meristem (SAM) gives rise to all aerial plant organs. Cell walls are thought to play a central role in this process, translating molecular regulation into dynamic changes in growth rate and direction, although their precise role in morphogenesis during organ formation is poorly understood. Here, we investigated the role of xyloglucans (XyGs), a major, yet functionally poorly characterized, wall component in the SAM of Arabidopsis (Arabidopsis thaliana). Using immunolabeling, biochemical analysis, genetic approaches, microindentation, laser ablation, and live imaging, we showed that XyGs are important for meristem shape and phyllotaxis. No difference in the Young's modulus (i.e. an indicator of wall stiffness) of the cell walls was observed when XyGs were perturbed. Mutations in enzymes required for XyG synthesis also affect other cell wall components such as cellulose content and pectin methylation status. Interestingly, control of cortical microtubule dynamics by the severing enzyme KATANIN became vital when XyGs were perturbed or absent. This suggests that the cytoskeleton plays an active role in compensating for altered cell wall composition.

Use of Atomic Force Microscopy to Measure Mechanical Properties and Turgor Pressure of Plant Cells and Plant Tissues.

We present here the use of atomic force microscopy to indent plant tissues and recover its mechanical properties. Using two different microscopes in indentation mode, we show how to measure an elastic modulus and use it to evaluate cell wall mechanical properties. In addition, we also explain how to evaluate turgor pressure. The main advantages of atomic force microscopy are that it is non-invasive, relatively rapid (5~20 min), and that virtually any type of living plant tissue that is superficially flat can be analyzed without the need for treatment. The resolution can be very good, depending on the tip size and on the number of measurements per unit area. One limitation of this method is that it only gives direct access to the superficial cell layer.

Emergence of robust patterns from local rules during plant development.

The formation of spatial and temporal patterns is an essential component of organismal development. Patterns can be observed on every level from subcellular to organismal and may emerge from local rules that correspond to the interactions between molecules, cells, or tissues. The emergence of robust patterns may seem in contradiction with the prominent heterogeneity at subcellular and cellular scales, however it has become increasingly clear that heterogeneity can be instrumental for pattern formation. Here we review recent examples in plant development, involving genetic regulation, cell arrangement, growth and signal gradient. We discuss how patterns emerge from local rules, whether heterogeneity is stochastic or can be patterned, and whether stochastic noise is amplified or requires filtering for robust patterns to be achieved. We also stress the importance of modelling in investigating such questions.

Optimizing FRET-FLIM Labeling Conditions to Detect Nuclear Protein Interactions at Native Expression Levels in Living Arabidopsis Roots.

Protein complex formation has been extensively studied using Förster resonance energy transfer (FRET) measured by Fluorescence Lifetime Imaging Microscopy (FLIM). However, implementing this technology to detect protein interactions in living multicellular organism at single-cell resolution and under native condition is still difficult to achieve. Here we describe the optimization of the labeling conditions to detect FRET-FLIM in living plants. This study exemplifies optimization procedure involving the identification of the optimal position for the labels either at the N or C terminal region and the selection of the bright and suitable, fluorescent proteins as donor and acceptor labels for the FRET study. With an effective optimization strategy, we were able to detect the interaction between the stem cell regulators SHORT-ROOT and SCARECROW at endogenous expression levels in the root pole of living Arabidopsis embryos and developing lateral roots by FRET-FLIM. Using this approach we show that the spatial profile of interaction between two transcription factors can be highly modulated in reoccurring and structurally resembling organs, thus providing new information on the dynamic redistribution of nuclear protein complex configurations in different developmental stages. In principle, our optimization procedure for transcription factor complexes is applicable to any biological system.

A tension-adhesion feedback loop in plant epidermis.

Mechanical forces have emerged as coordinating signals for most cell functions. Yet, because forces are invisible, mapping tensile stress patterns in tissues remains a major challenge in all kingdoms. Here we take advantage of the adhesion defects in the Arabidopsis mutant quasimodo1 (qua1) to deduce stress patterns in tissues. By reducing the water potential and epidermal tension in planta, we rescued the adhesion defects in qua1, formally associating gaping and tensile stress patterns in the mutant. Using suboptimal water potential conditions, we revealed the relative contributions of shape- and growth-derived stress in prescribing maximal tension directions in aerial tissues. Consistently, the tension patterns deduced from the gaping patterns in qua1 matched the pattern of cortical microtubules, which are thought to align with maximal tension, in wild-type organs. Conversely, loss of epidermis continuity in the qua1 mutant hampered supracellular microtubule alignments, revealing that coordination through tensile stress requires cell-cell adhesion.

The parts of a plant that protrude from the ground are constantly shaken by the wind, applying forces to the plant that it must be able to resist. Indeed, mechanical forces are crucial for the development, growth and life of all organisms and can trigger certain behaviours or the production of particular molecules: for example, forces that bend a plant trigger gene activity that ultimately makes the stem more rigid. Mechanical forces can also originate from inside the organism. For example, the epidermal cells that cover the surface of a plant are placed under tension by the cells in the underlying layers of the plant as they grow and expand. The exact pattern of forces in the plant epidermis was not known because they cannot be directly seen, although scientists have tried to map them using theoretical and computational modeling. A mutant form of the Arabidopsis plant is unable to produce some of the molecules that allow epidermal cells to adhere to each other. Verger et al. placed the mutants in different growth conditions that lowered the pressure inside the plant, and consequently reduced the tension on the epidermal cells. This partly restored the ability of epidermal cells to adhere to each other, although gaps remained between cells in regions of the plant that have been predicted to be under high levels of tension. Verger et al. could therefore use the patterns of the gaps to map the forces across the epidermis, opening the path for the study of the role of these forces in plant development. Further experiments showed that cell adhesion defects prevent the epidermal cells from coordinating how they respond to mechanical forces. There is therefore a feedback loop in the plant epidermis: cell-cell connections transmit tension across the epidermis, and, in turn, tension is perceived by the cells to alter the strength of those connections. The results presented by Verger et al. suggest that plants use tension to monitor the adhesion in the cell layer that forms an interface with the environment. Other organisms may use similar processes; this theory is supported by the fact that sheets of animal cells use proteins that are involved in both cell-cell adhesion and the detection of tension. The next challenge is to analyse how tension in the epidermis affects developmental processes and how a plant responds to its environment.

In vivo FRET-FLIM reveals cell-type-specific protein interactions in Arabidopsis roots.

During multicellular development, specification of distinct cell fates is often regulated by the same transcription factors operating differently in distinct cis-regulatory modules, either through different protein complexes, conformational modification of protein complexes, or combinations of both. Direct visualization of different transcription factor complex states guiding specific gene expression programs has been challenging. Here we use in vivo FRET-FLIM (Förster resonance energy transfer measured by fluorescence lifetime microscopy) to reveal spatial partitioning of protein interactions in relation to specification of cell fate. We show that, in Arabidopsis roots, three fully functional fluorescently tagged cell fate regulators establish cell-type-specific interactions at endogenous expression levels and can form higher order complexes. We reveal that cell-type-specific in vivo FRET-FLIM distributions reflect conformational changes of these complexes to differentially regulate target genes and specify distinct cell fates.

Analysis of a Plant Transcriptional Regulatory Network Using Transient Expression Systems.

In plant biology, transient expression systems have become valuable approaches used routinely to rapidly study protein expression, subcellular localization, protein-protein interactions, and transcriptional activity prior to in vivo studies. When studying transcriptional regulation, luciferase reporter assays offer a sensitive readout for assaying promoter behavior in response to different regulators or environmental contexts and to confirm and assess the functional relevance of predicted binding sites in target promoters. This chapter aims to provide detailed methods for using luciferase reporter system as a rapid, efficient, and versatile assay to analyze transcriptional regulation of target genes by transcriptional regulators. We describe a series of optimized transient expression systems consisting of Arabidopsis thaliana protoplasts, infiltrated Nicotiana benthamiana leaves, and human HeLa cells to study the transcriptional regulations of two well-characterized transcriptional regulators SCARECROW (SCR) and SHORT-ROOT (SHR) on one of their targets, CYCLIN D6 (CYCD6).Here, we illustrate similarities and differences in outcomes when using different systems. The plant-based systems revealed that the SCR-SHR complex enhances CYCD6 transcription, while analysis in HeLa cells showed that the complex is not sufficient to strongly induce CYCD6 transcription, suggesting that additional, plant-specific regulators are required for full activation. These results highlight the importance of the system and suggest that including heterologous systems, such as HeLa cells, can provide a more comprehensive analysis of a complex gene regulatory network.