Long intergenic noncoding RNA p21 (lincRNA-p21) is considered a target of wild-type p53, but little is known about its regulation by mutant p53 and its functions during the progression of head and neck squamous cell carcinoma (HNSCC).RNAscope was used to detect the expression and distribution of lincRNA-p21. Chromatin immunoprecipitation and electrophoretic mobility shift assays were performed to analyze the transcriptional regulation of lincRNA-p21 in HNSCC cells. The biological functions of lincRNA-p21 were investigated in vitro and in vivo. RNA immunoprecipitation and pull-down assays were used to detect the direct binding of lincRNA-p21.Lower lincRNA-p21 expression was observed in HNSCC tissues and indicated worse prognosis. Both wild and mutant type p53 transcriptionally regulated lincRNA-p21, but nuclear transcription factor Y subunit alpha (NF-YA) was essential for mutant p53 in the regulation of lincRNA-p21. Ectopic expression of lincRNA-p21 significantly inhibited cell proliferation capacity in vitro and in vivo and vice versa. Moreover, the overexpression of lincRNA-p21 induced G1 arrest and apoptosis. Knockdown NF-YA expression reversed tumor suppressor activation of lincRNA-p21 in mutant p53 cells, not wild-type p53 cells. A negative correlation was observed between lincRNA-p21 and the phosphorylation of signal transducer and activator of transcription 3 (p-STAT3) in HNSCC tissues. High lincRNA-p21 expression inhibited Janus kinase 2 (JAK2)/STAT3 signal activation and vice versa. Further, we observed direct binding to STAT3 by lincRNA-p21 in HNSCC cells, which suppressed STAT3-induced oncogenic potential.Our results revealed the transcriptional regulation of lincRNA-p21 by the mutant p53/NF-YA complex in HNSCC. LincRNA-p21 acted as a tumor suppressor in HNSCC progression, which was attributed to direct binding to STAT3 and blocking of JAK2/STAT3 signaling.The online version of this article (10.1186/s12943-019-0993-3) contains supplementary material, which is available to authorized users. Head and neck squamous cell carcinoma (HNSCC) is an aggressive malignancy with high local recurrence and lymph node metastasis that affects 600,000 new patients worldwide per year . DespiteTP53 gene [TP53 gene can not only result in loss of wild-type p53 function or exert a dominant-negative effect over the remaining wild-type allele but also lead to a gain in oncogenic properties that promote tumor growth [Accumulating studies show that long noncoding RNAs (lncRNAs) participate in diverse physiological and pathological processes, such as cell cycle, apoptosis and metabolism –6, as weP53 gene , 19. Mutr growth . As a trIn this study, we demonstrated that lincRNA-p21 is transcriptionally regulated by the mutant p53/nuclear transcription factor Y subunit alpha (NF-YA) complex. Low lincRNA-p21 expression promoted aggressive progression in HNSCC in vitro and in vivo. Meanwhile, lincRNA-p21 inhibited Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling by binding to STAT3 and suppressing its transcriptional activation, which is a novel mechanism of lincRNA-p21. Our findings provide insight into how the p53/lincRNA-p21/STAT3 axis contributes to HNSCC development and indicate that lincRNA-p21 may serve as a novel therapeutic target for HNSCC.We obtained 70 HNSCC tissues and 9 normal oral mucosal tissues from patients who had undergone surgery between 2007 and 2008 and who were diagnosed by pathological examination. No local or systemic treatment was conducted in these patients before surgery. The tissues were embedded into a tissue microarray. Fresh tumor specimens were collected at surgery and stored at − 80 °C until use. This study was approved by the Ethics Committee of the Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine.The RNAscope probe targeting lincRNA-p21 was designed and synthesized by Advanced Cell Diagnostics company, and detection of lincRNA-p21 expression was performed on an HNSCC tissue array using an RNAscope 2.5 High Definition (HD)-BROWN Assay kit according to the manufacturer’s instructions . The images were acquired with a Pannoramic MIDI Viewer . The signals were visually scored based on the average number of dots per cell using the following criteria: 0 (no staining or < 1 dot/10 cells), 1 (1–3 dots/cell), 2 (4–9 dots/cell. None or very few dot clusters), 3 (10–15 dots/cell and < 10% of the dots presented in clusters), 4 (> 15 dots/cell and > 10% of the dots presented in clusters).Fluorescence in situ hybridization (FISH) was strictly performed according to the manufacture’s protocol . Briefly, target-specific probe was designed, synthesized and incubated overnight at 4 °C. 18S RNA and U6 were used as control to indicate the cytoplasm and nucleus.Immunohistochemistry (IHC) was performed as described in our previous study , with pr2 at 37 °C.The HNSCC cell lines HN4, HN6 and HN30 were kindly provided by Professor Mao Li, University of Maryland and were verified by STR genotyping, and HEK293T, Cal27, SCC25, Detroit562, MCF7 and MDA-MB-231 cell lines were purchased from Type Culture Collection of Chinese Academy of Sciences . Normal oral primary keratinocytes were cultured from gingival tissues after tooth extraction from healthy patients. An informed consent was signed by the patients. SCC25 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM)/F12, and the other cell lines were maintained in DMEM  supplemented with 10% fetal bovine serum, 1% glutamine, and 1% penicillin-streptomycin. Cells were cultured in a standard humidified atmosphere of 5% COCytoplasmic and nuclear RNA was extracted using an RNA Purification Kit  according to the manufacturer’s instructions. Real-time PCR (qRT-PCR) was performed using a StepOnePlus Real-time PCR system  as previously described  and follpcDNA3.1 vector containing the human lincRNA-p21 sequence was synthesized by OBiO Technology . Plasmids encoding wild-type p53 or mutant p53s  and lentivirus, including for lincRNA-p21 and si-lincRNA-p21, were synthesized by Gene Pharma Co., Suzhou, China. The lincRNA-p21 promoter reporter gene was cloned into the vector pGL4.0-basic . Small interfering RNA (siRNA) was synthesized by Biotend Biotechnology, Shanghai, China. The siRNA#3 sequence of lincRNA-21 was packaged into a pGLV-h1-GFP-puro vector for in vivo experiments. Cells were transfected with siRNAs or plasmids using Lipofectamine™ 3000  according to the manufacturer’s instructions. Treatments were administered 24 h after transfection. The sequences are provided in Additional file The chromatin immunoprecipitation (ChIP) assay was strictly performed according to the protocol of a SimpleChIP Enzymatic Chromatin IP kit  according to the manufacturer’s instructions as described in our previous study , 23. TheLuciferase assays were performed as described in our previous study . BrieflyElectrophoretic mobility shift assay (EMSA) was performed as described previously  and accoWestern blotting was performed as described in our previous study . DOX, MGImmunoprecipitation was operated by incubation with anti-p53 or anti-STAT3 monoclonal antibody with cell lysate at 4 °C rotation overnight. The reaction mix was further incubated with Protein A/G agarose beads for 4 h. The precipitated proteins were detected by immunoblotting with the corresponding antibodies. Anti-GAPDH antibody was used for non-specific control.Cells were plated on coverslips, and incubated at 4 °C overnight with antibodies specific for p-STAT3 (1:100), NF-YA (1:500) and p53 (1:1000). Then, cells were incubated with Cy3 conjugate goat anti-rabbit IgG  or FITC conjugate goat anti-mouse IgG (SA00003–1)  and then stained with 4′, 6-diamidino-2-phenylindole (DAPI). Labeled cells were visualized on an Axio Vert. A1 microscope .4 cells/200 μl) were added into the upper portion of a migration (uncoated insert) or invasion (matrigel-coated insert) chamber, with 500 μl DMEM containing 10% FBS added into the lower chamber. After crystal violet staining, the crossed cells were dissolved in 33% of acetic acid. OD 570 nm absorbance was measured.HN6 and Cal27 cells were transfected with siRNA or plasmids of lincRNA-p21 for 24 h, and then seeded in the plates. As described in our previous study , cell prThe treated HN6 and Cal27 cells were incubated with 50 μM EdU  100 μl per well for 2 h. After fixation with paraformaldehyde and washes, the cells were treated with 100 μl of 1 × Apollo reaction for 30 min. Then, the DNA of cells were stained with DAPI for 5 min and visualized under Axio Vert. A1 fluorescence microscope.Flow cytometry was performed using previously described methods , 25. HN6As described in our previous study , TUNEL (RNA Immunoprecipitation (RIP) assay was performed according to the manufacturer’s instructions of Millipore Magna RIP Kit . HN6 cells were transfected with vector or lincRNA-p21 plasmid for 24 h and harvested. The cell lysates were incubated with RIP buffer containing magnetic beads conjugated with control IgG, STAT3, phosphorylated STAT3 antibodies overnight at 4 °C. The samples were then incubated with proteinase K to isolate immunoprecipitated RNA. The isolated RNA was reverse transcribed to cDNA and then analyzed by qRT-PCR. qRT-PCR was performed with SYBR Green PCR  and primers targeting lincRNA-p21 truncations . LincRNA-p21 RNA was transcribed in vitro using a DNA template that included the T7 promoter according to the instructions for a Ribo™ RNAmax-T7 Transcription Kit . Then, lincRNA-p21 was end-labeled with desthiobiotin using a Pierce RNA 3′ End Desthiobiotinylation Kit . HN6 cells were treated with lincRNA-p21 plasmid for 48 h and then harvested. Biotinylated lincRNA-p21 was captured with streptavidin magnetic beads and then mixed with HN6 cell extract. The protein was eluted from the RNA-protein complex and detected by western blot using STAT3 antibody.6) stably transduced with lentivirus. The animals were divided into 4 groups: (a) si-Scramble; (b) si-lincRNA-p21; (c) vector; (d) lincRNA-p21. The tumor sizes and animal weights were monitored once a week. The tumor volumes were calculated using the following formula: (length×width2)/2. Mice were sacrificed, and the tumor tissues were excised after 6 weeks.BALB/c nude mice  were purchased from the Shanghai Laboratory Animal Center  and were bred in SPF facilities at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine. The Laboratory Animal Care and Use Committees of the hospital approved all experimental procedures. The tumor xenograft model was established with Cal27 cells as described in our previous study . The CalP < 0.05 was considered statistically significant . All values are expressed as the means ± standard error.Statistical analyses were performed using SPSS 13.0 software . GraphPad Prism version 6  was used to plot the data. Student’s t-test and one-way analysis of variance (ANOVA) were performed to assess the significance of differences. P < 0.0001, Fig. P = 0.0168, Fig. P = 0.009, Fig. P = 0.03, Fig. To investigate the clinical relevance of lincRNA-p21 expression in HNSCC, we first examined 70 HNSCC tissues and 9 normal oral mucosa tissues using an RNAscope assay. RNAscope® is a novel in situ hybridization (ISH) approach for detection of target RNA with a unique probe design strategy that allows simultaneous target-specific signal amplification and background suppression to achieve single-molecule visualization while preserving tissue morphology . We obseWe next conducted qRT-PCR to examine lincRNA-p21 expression in HNSCC tissues and cell lines. LincRNA-p21 was poorly expressed in tumor tissues from 73 patients, but it was highly expressed in matched adjacent non-tumor tissues  and MDA-MB-231 (mutant TP53), which indicated that regulation of lincRNA-p21 by p53 may be a common mechanism in different cancers , which reduced the mRNA expression levels of lincRNA-p21  and HN30 (wt-p53) cells after co-transfection with lincRNA-p21 plasmids and si-NF-YA were analyzed to observe that knockdown NF-YA expression could reverse the inhibition of proliferation capacity of lincRNA-p21 overexpression in Cal27 cells, not HN30 cells . According to our abovementioned results, p53 can transcriptionally activate lincRNA-p21 expression. Thus, we wondered whether p53 can regulate the expression of STAT3 through lincRNA-p21. As shown in Fig. Abnormal activation of the JAK2/STAT3 pathway plays critical roles in tumor cell proliferation, invasion, survival, angiogenesis and immunosuppression . OverexpUsing the UCSC database, we identified the location and sequence of human lincRNA-p21 Fig. a. Eight In this study, we first investigated the difference between wild-type and mutant p53 in regulating lincRNA-p21 expression in HNSCC. Moreover, high lincRNA-p21 expression exerted tumor suppressor activity by inhibiting JAK2/STAT3 signaling activation, which suggest that lincRNA-p21 was deeply involved in aggressive progression in HNSCC.LincRNA-p21 is a relatively novel lncRNA that plays a significant role in the initiation and development of multiple cancers. Previous studies have reported that lincRNA-p21 was significantly downregulated in hepatocellular carcinoma , colorecTP53 missense mutation mainly occurs in its DNA-binding domain [p53 is a sequence-specific DNA-binding protein that interacts with promoter regions of target genes and regulates transcription. Accumulating evidences indicate that p53 not only transcribes messenger RNAs but also noncoding RNAs, including microRNAs and long noncoding RNAs. Liu et al.  reportedg domain . Mutatiog domain . Even ing domain . Howeverg domain . We chosg domain . We propLncRNAs impact cellular functions through various mechanisms, such as interaction with chromatin remodeling proteins, genomic DNA, mRNA or proteins in the nucleus or cytoplasm . The basIn conclusion, our findings showed that mutant p53 bound to the promoter of lincRNA-p21 and activated the transcription of lincRNA-p21 depending on the co-factor NF-YA. We further demonstrated that lincRNA-p21 acts as a tumor suppressor via direct binding to STAT3 and inhibition of its transcriptional activation. Overall, the p53/NF-YA/lincRNA-p21/STAT3 axis is a novel mechanism that drives malignant progression and thus may be an attractive therapeutic target in HNSCC.Additional file 1:Table S1. The primers and sequences for qPCR, siRNA, ChIP and RIP assays. (XLSX 12 kb)Additional file 2Figure S1. The expression of lincRNA-p21 was detected by qPCR in breast cancer cells transfected with si-p53 for 24 h. Figure S2. (a) Transfection efficiency was detected after siRNA using qPCR in HN6 and Cal27 cells. (b)Transfection efficiency was detected after expression plasmid transfection using qPCR in HN6, HN30 and Cal27 cells. Figure S3. Cell viability (a) and colony formation ability (b) of HN6 and Cal27 cells after transfection with ASO-lincRNA-p21 or scrambled were determined using the CCK8 and colony formation assay. Figure S4. Cell viability (a) and colony formation ability (b) of HN30 cells after transfection with lincRNA-p21 or vector were detected using the CCK8 and colony formation assay. Figure S5. AKT and ERK1/2 signaling were analysed using western blot after si-lincRNA-p21 transfection for 48 h. Figure S6. The transfection efficiency was confirmed in fresh xenograft tissue using qPCR. Figure S7. LincRNA-p21 regulates cell cycle and apoptosis related protein in HNSCC cells. (a) The cell cycle regulation related proteins were detected in HN6 and Cal27 cells after si-lincRNA-p21 for 48 h. (b) PARP, Caspase-3 and its active forms were detected in HN6 and Cal27 cells after si-lincRNA-p21 for 48 h. Figure S8. Migration (a) and invasion (b) assays were performed with si-lincRNA-p21 or scrambled transfected HN6 and Cal27 cells using Transwell inserts. Figure S9. LincRNA-p21 reducing STAT3 expression is independent on ubiquitination degradation. Expression of STAT3 and Ubiquitin protein was detected after transfection for 48 h and then stimulation with 0.5 μM MG132 for 24 h in HN6 and Cal27 cells. Figure S10. The staining score of p-STAT3 in in the xenograft tumour tissues. Figure S11. IC50 was calculated using cryptotanshinone (a STAT3 inhibitor) at indicated concentrations for 72 h in HN6 and Cal27 cells. (DOCX 1296 kb)


The primary goal of the Helsinki VideoMEG Project is to enable magnetoencephalography (MEG) practitioners to record and analyze the video of the subject during an MEG experiment jointly with the MEG data. The project provides:•Hardware assembly instructions and software for setting up video and audio recordings of the participant synchronized to MEG data acquisition.•Basic software tools for analyzing video and audio together with the MEG data.The resulting setup allows reliable recording of video and audio from the subject in various real-world usage scenarios. The Helsinki VideoMEG Project allowed successful establishment of video-MEG facilities in four different MEG laboratories in Finland, Sweden and the United States. Magnetoencephalography (MEG) is a non-invasive functional brain imaging method that monitors neuronal activity by measuring the associated magnetic fields . In clinMEG is in many respects similar to the closely related technique of electroencephalography (EEG): the two techniques share the underlying sources of the signal, and rely on similar mappings from the activity of these sources to the signals measured by the sensors, yielding similar temporal and spatial resolutions . In clinDespite the potential benefits, video-MEG recordings have gained little traction with MEG practitioners. One of the main impediments to wider adoption of video-MEG is the lack of practical solutions that would allow an MEG laboratory to integrate video recordings into the workflow with reasonable cost and manpower requirements. While several MEG manufacturers have advertised integrated video recording capabilities in their future products, currently there are no commercial video-MEG solutions on the market. Moreover, these advertised capabilities are only available for new MEG device installations, which makes them irrelevant to existing MEG sites.1.Practicality. The project's goal is to allow a typical MEG facility to establish a video-MEG operation in practice with reasonable monetary and manpower costs.2.Openness. All the materials  are freely available on GitHub3.Vendor- and device-neutrality. The project aims at providing a video-MEG solution that is compatible with any MEG device.The Helsinki VideoMEG Project described in this paper aims at remedying this situation by providing MEG practitioners with tools for setting up video-MEG recordings. The project is guided by three main principles:Establishing a video-MEG operation entails two relatively separate developments: (1) creating an instrumentation setup for recording video and audio of the participant during the MEG experiment in a way that is synchronized to MEG data acquisition, and (2) setting up a facility for the analysis of the resulting video, audio, and MEG data streams.The first task—establishing a video-MEG recording setup—necessarily involves acquisition and installation of video recording hardware. To put video-MEG within a reach of a typical MEG facility, the Helsinki VideoMEG Project has developed a hardware setup that only uses widely available standardized off-the-shelf components and requires no special skills for assembling. The project provides all the material  necessary for setting up video-MEG recordings.The second task, setting up a video-MEG analysis facility, presents a different set of challenges. On one hand, it does not require any hardware development, as it can be completely addressed by developing appropriate software tools. On the other hand, integration with the existing instruments poses much bigger challenge for video-MEG analysis than it does for video-MEG recordings. Currently, MEG practitioners use a whole spectrum of different tools for analyzing the data. These include open-source packages, such as FieldTrip  or MNE , that prFor recording video and audio of the participant, the Helsinki VideoMEG Project employs video recording system, which essentially constitutes a separate instrument that is independent of the MEG device. The only link between the two is the synchronization line that carries the timing information encoded as a sequence of trigger pulses. Thus, any MEG device capable of recording external trigger pulses can be used with the system.The AV computer is a standard amd64-based office PC running a 64-bit version of Linux operating system. The setup was tested with Ubuntu 14.04 LTS and Ubuntu 16.04 LTS distributions of Linux, but other distributions may work as well. The computer needs to be equipped with a parallel port interface for outputting timestamps and an IEEE 1394 (firewire) interface for connecting the cameras. The firewire link can use either a standard copper cable or an optical fiber for communicating with the cameras, requiring either a standard or an optical interface, respectively. The AV computer runs custom Qt-based software written in C++. The software monitors and records the video and audio streams, and generates the timing signals that are emitted over the computer's parallel port.The project supports firewire machine-vision cameras that implement the industry-standard protocol—IIDC 1394-Based Digital Camera Specifications, also known as the DCAMThe AV computer allows recording the audio from inside the MSR via the computer's sound card. The exact configuration of the audio recording setup depends on the site's specific requirements. One possible choice  is an optical microphone, such as Sennheiser MO 2000 . The microphone itself does not contain any magnetic parts, and the signal connection is optical, eliminating any sources of interference inside the MSR. For a low-cost alternative, an electret microphone can be used.The system uses the Unix time (POSIX time) in milliseconds as timestamps. The recording software timestamps every audio buffer and video frame as soon as they are available. The timestamps are also sent to the parallel port at regular intervals (by default every 10 s). They are represented as sequences of constant-amplitude pulses, where bits are encoded as delays between subsequent pulses’ rising edges . Values Typically the parallel port output is connected to an MEG trigger channel, which is recorded synchronously with other MEG data. Thus a timestamped MEG sample is available every 10 s. In postprocessing, the timestamps for MEG samples between the timing pulses can then be computed by linear interpolation.The video is recorded as individual JPEG-encoded frames; no intraframe coding is currently performed. Thus, individual frames can be easily retrieved independently of each other at the expense of increased disk usage. The JPEG frames are stored consecutively in a single file per camera, interleaved with the timestamps for each frame.The audio is recorded as buffers. Buffer size may depend on the particular sound card and drivers used; a typical buffer size is 1024 samples, corresponding to about 23 ms at a typical sampling rate of 44,100 Hz. Similar to video, audio buffers are stored into a single file, interleaved with their corresponding timestamps.The exact specification of the audio and video file formats is available from the project's website.The Helsinki VideoMEG Project approaches the task of analyzing video-MEG recordings in two ways described below.For the users implementing their own analysis pipeline in MATLAB or Python programming environments, the project offers a complete set of functions for integrating audio and video streams into the analysis, namely routines for loading the audio and video data, and extracting and interpolating timestamps.Additionally it provides utility programs, written in Python, for various ancillary tasks—e.g. exporting video and audio to standard AVI format. The project also offers several documented examples of simple analysis pipelines written in Python.At the current stage, the selection of GUI-based tools for interactive review of the video-MEG data is quite limited. Elekta Oy has demonstrated an experimental version of it's Graph software that allows review of video and audio jointly with the MEG data from the company's VectorView and Triux MEG devices . This soThe developers of the FieldTrip software  have addNotwithstanding the aforementioned options, the lack of a practical GUI-based video-MEG data analysis tool currently constitutes the main hindrance to the project's progress and the main focus of the future development roadmap.We performed a test measurement to assess the accuracy of synchronization between the audio and video streams and the MEG data. The test system consisted of a Dell Precision 490 office PC  running the Ubuntu Linux 14.04 operating system. The motherboard integrated sound system based on the Sigmatel STAC9200 chip was used for audio. For video, two Allied Vision Stingray F033 cameras  were connected to the system via optical firewire in a daisy-chained configuration.The Elekta TRIUX system  in the BioMag laboratory of Helsinki University Central Hospital was used as the MEG device. The MEG was set up to generate trigger pulses once per second. The video cameras were pointed at the trigger interface unit which has LEDs indicating trigger onset. A function generator was set up to output a 1-kHz sinusoidal pulse with a 100-ms duration on the rising edge of the trigger signal. Using an oscilloscope, we verified that the delay between the trigger rising edge and the sinusoid onset was below 1 ms. The output of the function generator was then connected to the line input of the soundcard. Thus the rising edge of the pulse served as a reference event used for testing the synchronization between the three data streams—MEG, video and audio. Each rising edge (which was recorded in the MEG data as a trigger) coincided with the onset of the LED flash (which was recorded on video) and onset of the sinusoidal pulse (which was record on audio). Data were acquired continuously for 20 min.Next, the data were synchronized using the VideoMEG Python utilities. After synchronization, tone onsets were localized from the recorded audio data by a correlation with a 1-kHz complex exponential and thresholding. LED onsets were detected from the video data by thresholding: the frame where the intensity of the LED first reached 50% of its maximum was denoted as the LED onset frame. Since the LED may be on for only a part of the video camera's frame acquisition interval, the on/off transition is not necessarily instantaneous in the video.If the synchronization between video, audio, and MEG traces were perfect, the reference event—rising edge of the trigger pulse—would be assigned the same time in all the three data streams. The extent of the discrepancies between the timings assigned to the same event in different data streams characterize the accuracy of synchronization.We have presented the architecture of a low-cost video recording system whose output can be accurately synchronized with the rest of the MEG data stream. The total cost of the system hardware depends on the number and type of cameras and on whether an optical microphone is used; a basic system can be put together for approximately 5000 €.On our setup based on a standard office PC, the system was verified to have relatively short audio and video latencies. Critically, audio and video jitter were small: <1 ms for audio and 1 frame for video in this hardware setup. The fixed audio delay of approximately 16 ms should be low enough for clinical applications. However, for applications demanding maximally accurate synchronization of the external audio stream (e.g. neuroscience experiments), we recommend compensating for the fixed delay. This is possible as long as the jitter is negligible. The delay and jitter might depend on the particular hardware used, so we recommend validating individual setups before deployment.Below we outline the rationale for some of the project's design decisions.To synchronize the AV computer to the MEG, the project relies on binary timing signals from the AV computer recorded by the MEG devices’ trigger channel. This approach offers the advantages of being simple and reliable, and, importantly, independent of the internals of the MEG acquisition system. The only requirement for the MEG device is to be able to accurately register external trigger pulses, which in practice is satisfied by any modern MEG system.Encoding the absolute time in a sequence of pulses every 10 s  significantly increases the robustness of the synchronization scheme. It allows synchronization in the presence of partially missing or corrupt data and prevents accidental missynchronization of unrelated data streams. The redundancy of such an encoding, complemented by the redundancy provided by parity bits, allows reliable automatic detection of the timing information in the MEG data without the need for the user to explicitly specify the timing channel.To emit the timing signal, the AV computer requires a binary output facility that can produce an arbitrary sequence of pulses and allow accurate enough control of their timing. Whereas there is a multitude of options available for solving this problem—such as digital-to-analog converters, digital input/output cards, etc.—by far the cheapest, simplest and easiest solution employs parallel  port. Although the LPT technology is obsolete, parallel ports are still widely available for desktop computers as add-on PCI cards.•Such cameras offer much longer product life cycle than most of the alternatives. That means that the cameras, associated materials , and technical support will be available for many years into the future.•The cameras rely on open and stable IIDC interface for communicating with the host computer. This allows to avoid a vendor/model lock-in, as the camera can be easily replaced with another one from a wide selection of models by different vendors that support the interface.•Being designed for integration into third-party systems, machine-vision cameras offer the user an extensive control over the camera configuration both physical  and programmatic . This level of control is critical to the project's ability to accommodate a wide range of requirements posed by different clinical and research use cases—such as the demand for recordings in low levels of visible light or different experiment-specific requirements for video quality and frame rate.•Some firewire-based machine vision cameras  allow the use of optical fiber for data connection to the host computer instead of the standard copper firewire cable. The cameras still require a conductive cable for supplying the power, however the power supply can be completely independent from the host computer. This greatly simplifies the practicalities of installing the camera inside the MSR. Using an optical fiber instead of a copper cable may also reduce the amount of electromagnetic interference introduced by the setup, however, this was not systematically tested.When selecting a type of camera to be used for recording the patient, we have considered a number of possible alternatives, such as consumer web-cameras, surveillance cameras, and industrial machine-vision cameras. We have finally decided to adopt firewire-based machine vision camera a number of reasons:For the initial clinical validation we constructed a prototype that employs two black-and-white Stingray F-033 cameras at resolution 640 × 480 and frame rates of either 30 or 60 fps. This choice of a relatively low-end camera model was motivated by the intent to minimize the costs and avoid potential performance problems. In line with previous reports , our expThe project employs a simple custom format for storing video and audio data. The audio is stored uncompressed and video—as a sequence of JPEG-compressed individual frames. In the initial stages of the project, we prefer a custom format over commonly used ones  because it considerably simplifies the development of video recording and analysis tools. The main drawbacks of using a simple custom data format are: (1)unavailability of the data for analysis by numerous third-party tools, such as video editors, and (2) the increase in storage requirements (due to a very simple compression scheme).The address the first issue, the project provides conversion utilities that allow export of the video and audio data to a widely-used AVI format.For the default camera resolution currently provided by the project, the second shortcoming is tolerable, if not negligible. The amount of storage space required by video and audio is comparable to that required by MEG data, thus most MEG sites should be able to handle the additional storage demands effected by the video without any significant modifications to their data storage facilities. This situation is, however, bound to change as the project embraces higher-resolution cameras. Hence, transitioning to a more sophisticated video storage format is one of the projects near-term goals.With the exception of a comprehensive GUI-based analysis tool, the Helsinki VideoMEG Project provides a complete set of instruments for integration of video and audio recordings of the subject into the MEG laboratory workflow.


This study aimed to investigate temporal trends in the incidence rates (IRs) of vision impairment (VI) and explore its associated factors among middle-aged and older Chinese. Data were obtained from China Health and Retirement Longitudinal Study  and Chinese Longitudinal Healthy Longevity Survey . There were 20,467 participants with a mean age of 57.4 years from CHARLS and 21,210 participants with a mean age of 90.0 years from CLHLS. Distance and near vision were self-reported in CHARLS and performance-based vision was assessed in CLHLS. Crude and age-adjusted IRs per 1,000 person years (PY) of distance vision impairment (DVI), near vision impairment (NVI) and both distance and near vision impairment (DNVI) in CHARLS and VI in CLHLS were calculated for each wave of surveys. Risk factor adjusted temporal trends in the incidence were examined using regression models. Age-standardised IRs for DVI, NVI and DNVI significantly decreased from 79.4 to 41.2, 95.1 to 48.0 and 40.0 to 21.3 (P < 0.001) from 2013 to 2015, respectively. Male gender, higher education, no ADL disability, no depression, no multimorbidity were associated with lower IRs of DVI, NVI or DNVI. Younger age was associated with lower IRs of DVI and DNVI but higher IRs of NVI. Age-standardised IR of VI significantly decreased from 131.6 to 71.9 (P = 0.010) from 2000 to 2014, and higher education and living in the South were associated with lower IR of VI. Future studies may further investigate the causality of such phenomenon.


Plow教授惠赠）溶液中4 ℃孵育过夜，然后在带辣根过氧化物酶标签的羊抗鼠二抗溶液（1:4 000稀释，Jackson ImmunoResearch，PA，USA）中室温孵育1 h。最后采用Western Blotting Luminol Reagent（Santa Cruz Biotechnology, CA, USA）发光试剂盒检测，X线胶片曝光、显影、定影。以β-actin作为内参蛋白，使用β-actin鼠单抗（1:3 000, Sigma-Aldrich, MO, USA）作为一抗。1.4以上述ENO1鼠单抗（1:100稀释，Edward F. Plow教授惠赠）为捕获抗体，包被96孔板，4 ℃过夜后，以2%BSA封闭96孔板4 h，之后每孔加入50 L待测血浆孵育1 h。以ENO1兔抗人ENO1多克隆抗体（1:4 000稀释，AVIVA Systems Biology，San Diego，CA）为检测抗体，孵育1 h。然后加入带辣根过氧化物酶标签的羊抗兔二抗溶液（1:4 000稀释，Jackson ImmunoResearch，PA，USA），孵育30 min。加入显色液，避光反应30 min，最后由酶标仪（Bio-Rad Laboratory, CA, USA）于450 nm/570 nm双波长设定下读取光密度（optical density, OD）值。由于目前没有商业化的ENO1标准蛋白，故采用肺癌细胞系A549的总蛋白用1× PBS梯度稀释，稀释比依次为1:20、1:40、1:80、1:160、1:320、1:640，以此绘制标准曲线，进行板间差校正。1.5Mann-Whitney Test比较两组人群血浆ENO1蛋白水平。P < 0.05为差异具有统计学意义。采用SPSS 11.5软件（SPSS INC, IL, USA）进行统计学分析。利用非参数检验中的22.1ENO1蛋白在16例肺鳞癌患者的肿瘤组织及其配对正常肺组织中的表达情况如2.2P=0.031）和肺部良性疾患者（P=0.019）。血浆中ENO1蛋白水平在肺部良性疾病患者和健康体检者之间没有明显差异（P=0.904）。肺腺癌患者血浆中ENO1蛋白水平明显高于肺鳞癌患者（P=0.023）。采用双抗体夹心ELISA方法检测ENO1蛋白在三组人群（即健康体检者、肺部良性疾病患者和非小细胞肺癌患者）血浆中的水平。结果如3-11，Altenberg等综合分析了肿瘤的糖酵解通路中代谢酶相关基因的表达情况，发现在24种肿瘤中糖酵解通路增强，肺癌是其中之一。糖酵解通路中涉及10个代谢酶，其中丙酮酸激酶（pyruvate kinase, PK）是糖酵解通路中的限速酶，其亚型PKM2除了发挥催化能量代谢的作用，还具有与肿瘤发生发展相关的生物学功能, 14，并且肿瘤患者血浆中PKM2的水平显著升高-17，提示其可能作为血浆诊断标志物。因此，糖酵解通路中其它代谢酶可能也与肿瘤有着紧密的联系。糖酵解途径是机体获取能量的一种方式，在供氧不足的生理状态下，它为机体提供能量。肿瘤细胞普遍存在糖酵解途径的增强，作为肺癌相关抗原, 7，ENO1极可能是潜在的肺癌标志物。本研究结果显示，ENO1蛋白在肺鳞癌患者肿瘤组织中的表达高于其配对正常肺组织，此结果与其它针对临床肿瘤样本的研究结果基本一致, 7, 18，尽管与Chang等的发现相反。烯醇化酶是糖酵解途径中的代谢酶，该家族包含三个成员，分别是ENO1、ENO2和NSE。NSE作为肺癌相关血清学蛋白标志物，已被用于临床上指导小细胞肺癌患者的疗效监测、复发预测和预后评估。我们实验室前期建立的肺癌相关分泌/释放蛋白数据库中包含与NSE同家族的ENO1, 21，从而进入外周血。如糖酵解途径中大多数的代谢酶，包括ENO1，均可借助外来体途径（exosome pathway）被分泌到细胞外。结果显示，较之健康人群和肺部良性疾病人群，非小细胞肺癌患者血浆中ENO1蛋白水平明显升高，因此ENO1可能作为肺癌相关血浆蛋白标志物。同时需要指出的是，因为目前尚无商业化的ENO1标准蛋白，所以我们无法绘制标准曲线来获得具体的蛋白浓度值。尽管ENO1可通过特定途径进入外周血，但其在外周血中的浓度很低，采用自行构建的双夹心ELISA体系检测的信号值偏低（OD值< 0.1）。不过，鉴于A549细胞内的ENO1蛋白的OD值最大值仅在0.3左右，所得血浆ENO1蛋白OD值亦为合理范围。由于目前尚未见采用ELISA方法检测外周血血浆中ENO1蛋白的报道，我们自行建立的ENO1蛋白ELISA检测体系及其实验结果具有一定的意义和参考价值。ENO1位于细胞质，它可能通过肿瘤细胞坏死和细胞更新（cell turnover）或者非经典的分泌途径被释放到细胞外，定位于细胞质的ENO1可促进肿瘤细胞的生长和运动，定位于细胞核的ENO1可抑制肿瘤细胞的生长。但我们结果提示ENO1可能在肺癌的发生发展过程中更多地发挥着促进的作用。另外，我们还发现非小细胞肺癌患者血浆中ENO1蛋白水平与肺癌的组织病理类型相关，即肺腺癌患者血浆中ENO1蛋白水平显著高于肺鳞癌患者，推测ENO1作为潜在的肺腺癌血浆蛋白标志物可能更具有临床应用价值；因而有必要继续深入研究。ENO1对肿瘤进展具有双向作用，如定位于细胞膜的ENO1可促进肿瘤细胞的侵袭和转移


Social engagement technologies have the potential to benefit health and quality of life in older adults with and without mild cognitive impairment (MCI). However, technologies are rarely designed to accommodate the interests, capabilities, and limitations of these populations. In the current study, we focused on examining the potential of video chat to socially engage older adults with and without MCI by providing opportunities to link people with shared interests. Eight cognitively normal older adults (Mage: 73.3 years) and five with MCI (Mage: 70.0 years) completed a four-week experiential field trial using a novel online video chat platform called OneClick. System Usability Scale scores at both pre- and post-assessment revealed that OneClick was easy to use for older adults with and without MCI, however individuals with MCI experienced more technical issues and required additional assistance to use the system. Pre- to post- comparisons of questionnaire data revealed positive changes for the Quality of Life, Friendship/Social Isolation, and Loneliness scales in both groups. Of the 13 participants, five cognitively normal and four individuals with MCI reported that they would be interested in continuing to use the video chat system at home to connect with family and friends or to discuss topics of mutual interests. Overall, all participants enjoyed using the video chat system as a means for social engagement and showed trends for social health and quality of life benefits. This field trial illustrates the potential for video chat to provide social engagement opportunities for older adults with and without cognitive impairment.


To report a qualitative study of themes Registered Nurses raised spontaneously about their work environment, in a cross‐sectional survey study when responding to the Essentials of Magnetism II (EOMII) scale.Qualitative descriptive survey.. Of the 247 nurses who completed the EOMII scale, 30% (N = 75) provided comments. Inductive content analysis was used to analyse the textual information generated.At the end of the EOMII scale, a free form text section was included asking nurses to add comments about their ward/work environmentThree key themes emerged: “nurses need nurses to nurse”; working as a team and workplace environment. Participants described issues they were facing which comprised high turnover rates, inadequate staffing levels, increasing workload and high stress levels. Particular attention was drawn to the role of the ward manager in promoting a positive work environment, good teamwork and quality patient care. Findings from this study have provided better understanding of the challenges experienced by nurses in their work environment, particularly in terms of constraints on their ability to provide high quality patient care.This study highlights the ward manager as a key determinant in nurses’ decisions to leave or to remain in the job, and makes evident the impact of the ward manager’s behaviour on staff morale.Findings from this paper should be an impetus for nurse leaders and policy makers to involve ward nurses in decision‐making and policies for practice.1The Essentials of Magnetism II (EOMII) scale was developed in the United States by Schmalenberg and Kramer , based oThe EOMII is a 58‐item four‐point, rating scale based on the process element of the Donabedian's  Structur2The EOMII scale was distributed to 438 eligible Registered Nurses providing direct adult patient care in 29 wards in two NHS hospitals between May–October 2012. The EOMII scale was used in this study to address questions about the factor structure of the scale and to examine the associations, if any, between the factors and nurse‐assessed care quality. In our earlier study  who completed the EOMII scale wrote comments about their work environment. It is worth considering the potential effect of non‐response bias. It was possible that nurses who made comments about their work environment were more likely to be unhappy about the condition of their work environment and those least likely to comment were happy or indifferent about their work conditions. Although the response rate was low, it was the strength of feeling expressed by a small number of respondents around issues in their work environment that prompted the formal analysis of these data.3.5N = 70), with ages ranging from 20–≥60 years. Fifty‐five were Staff Nurses, 19 were Sisters and one was a Charge Nurse. Table There was an even split of respondents across the two hospitals in the study; 37 from Hospital A and 38 from Hospital B. Most respondents were females .3.7It was presumed that the 75 participants who wrote comments about their work environment would have responded to the stimulus provided by the items of the eight attributes measured by the EOMII scale. Examination of the comments revealed that data were not organized in a pre‐defined manner. Their comments went beyond the scope of the EOMII scale, addressing issues such as staffing numbers, increasing workload, high stress levels and work engagement.Inductive content analysis was used to analyse the data and it is a highly flexible method, which is applicable to a wide variety of different kinds of unstructured textual information Bryman, . The manward manager formed a dominant theme, with ward manager supportive and ward manager never giving positive feedback forming subthemes ]. One participant was specific about the rate of staff turnover stating: “…Has been difficult recently – 4 Ward Managers within the last 12 months” [P1070(RP/SR:5y9m)]. Another participant attributed poor skill mix to retention problems: “…Poor skill mix with retention problems…” [P1071(RP/SN:3y9m)].Participants also stated that inadequate staffing led to each nurse being assigned a large number of patients which had negative impact on the quality of care that staff, patients and relatives experienced: “…14 for one nurse is too much. And if you are the nurse in charge you have to look after everyone i.e. patients, staff and even relatives” [P2072(EM/SR:18y)]. According to participants, staff shortage placed increased pressure on nurses, leading to low staff morale: “…when we are short staffed – this puts pressure on everyone and reduces staff morale” [P2112(GS/SN:5y)]. In addition to staffing issues, participants further highlighted that their well‐being and ability to function effectively were being compromised, as described below.4.1.2Participants highlighted their experiences of increased workloads, which in turn led to being stretched to their limits: “…staff stretched to limits, work load very high and staff here work to the limits…” [P2048(RP/SN:5y)]. As a result of high workload, which is a direct consequence of staff shortage, participants found it difficult to provide good quality care to their patients: “We…have to deal with a high work load and we are short staff – the work is hard and we try to help our patients to the best of our ability” [P1036(GS/SN:6y)]; and “…Due to the nature of the ward often I feel there is too much going on to give really good care to patients” [P2100(GM/SN:5m)]. Participants’ inability to provide high quality care to their patients was seen as more of a challenge when nurses were tasked with the dual duty of coordinating the whole ward and simultaneously looking after patients: “Working as a nurse in charge performing to coordinate the whole ward and at the same time looking after 6–10 in‐patients enabling delay and hinders provision of quality patient care” [P2108(GS/SR:19y)].Apart from the busy nature of the nursing work, extensive attention was drawn to issues around increasing work demands attributed to age‐related comorbidity, rising complexity and acuity of the patients. Specifically, participants stressed the complexity of care needs related to those patients who were generally older, with mental health issues. According to the participants, the care of this category of patients was being compromised due to inadequate staffing and increased workload: “This ward is an elderly acute medical ward, with patients who have dementia/confused. The staffing levels could be better, as you don't feel like a nurse” [P1026(EM/SN:25y6m)]. In arguing for the increased work demands, especially regarding older patients and those with acute mental/health issues, one participant expressed belief that the Trust increased workload to save money: “Staff are given too many jobs to save money” . The increasing demand for documentation which takes participants away from bedside care prevents them from delivering high quality of care to their patients: “Very busy ward with not enough staff to meet the needs of our patients …an ever increasing amount of paperwork. Registered Nurses need to be “at the bedside”…” [P1110(GS/SN:37y)].Participants voiced concerns over their physical and psychological well‐being. Nurses perceived their work environment as being stressful and were worried about their inability to give quality care to patient as a result of staff shortage, increasing workload and pressure, leading to exhaustion, burnout and stress: “Due to staffing levels and pressure I feel we are not able to give the quality of care we would all want to give. Staff feel burn out and stressed” [P2049(RP/SN:4y10m)].In addition to individual staffing issues highlighted above, participants also indicated that teamwork significantly influenced the quality of their work environment.4.2Participants identified teamwork as a source of support in their work environment. Many participants placed emphasis on teamwork being demonstrated in staff members’ professional relationship with one another. They described teamwork as one of the facilitating aspects of their work environment that they considered essential to improving their work experiences, as well as supporting them in providing quality patient care. Professional relationships linked to teamwork included ward manager support and support from other members of staff. They also highlighted specific inhibiting factors in teamwork, such as the absence of collaborative doctor–nurse relationships and negative attitudes of some nursing staff. These factors are elaborated under three subthemes.4.2.1Many of the participants made positive comments about their ward managers. They described the supportive role of their ward managers as facilitating nursing practice. Supportive behaviours of the ward manager were identified as “understanding,” “helping to improve confidence,” “approachable,” “accessible,” “pleasant,” “good” and “very good.” Typical responses were as follows: “…Ward manager is very good and approachable…” [P2015(OT/SR:18y6m)], “…an experienced ward manager. She is very accessible and pleasant” [P1074(RP/SN:27yr)] and “…So far it's getting better here due to good management of our new manager” [P1015(OT/SN:14y)]. Participants’ descriptions of their excellent or committed team, as one which is enhanced by their ward managers: “…Good staff who all practice to their best. An excellent team and ward manager Xxx very supportive” [P1012(OT/SR:40y)], “… We have a committed team run by 2 supportive managers…” [P1104(OT/SN:22y4m)]; and “N Ward is a fantastic place to work due to management of the ward…” [P2067(CD/SN:11m)].Participants stated that due to the supportive role of their ward managers, their confidence improved. For example, one participant emphasized the understanding nature of the ward manager: “I have got a wonderful ward manager who has helped me to improve in myself and my confidence because of her understanding nature” [P2057(GM/SN:3m)]. Participants described the supportive role of the ward manager as contributing to enhanced teamwork and better work environment. They indicated that at times when there was tension between staff and the ward got busy and stressful, the supportive role of the ward manager helped to alleviate the unfavourable work situation, thereby enhancing patient care: “…busy ward where it can sometimes be stressful to work. Well supporting ward manager…” [P1045(RP/SR:4y3m)] and “Although at times, there is tension between staff. We work well as a team and provide excellent care to patients. Ward manager supports the ward” [P2042(HM/SN:1y9m)].N = 12) were complimentary; however, two participants emphasized the lack of interpersonal relationship and the managers’ inability to give encouragement or constructive feedback, leading to staff members feeling unappreciated and undervalued. The participants also revealed that those ward managers were instrumental to their resignation from the ward: “…The manager never gives positive feedback or encouragement so left feeling unappreciated & devalued” [P2111(GS/SR:11y11m)]. The above comment was echoed by another participant who revealed that the attitude of the ward manager led to his resignation. As the participant stated, the ward manager's attitude destroyed the potential for the ward to shine. However, this participant stated that staff nurses were supportive towards each other: “Staff nurses support each other very well but the ward manage…her attitude towards staff is combative and usually aggressive. This has destroyed any potential for the ward to shine. Hence I have recently left…” [P2135(MW/SN:1y)].Out of the 14 participants that commented about their ward managers, most  to the managers at board of hospital director level who are outside the wards , as they have different perceptions about the two levels of management. One participant identified the lack of support as stemming from the Trust rather than the actions of the ward manager and colleagues: “I enjoy working with my colleagues and our Ward manager as we work as a team. However, I feel we do not get the right support and back up from the Trust” [P1091(RP/SN:3y10m)]. The lack of appreciation or acknowledgement for their team efforts from management made them feel undervalued: “The ward team works well together, but this is not always acknowledged by senior mgt. A thank you from management can go a long way” [P1019(CD/CN:18y)]; and “Our involvement as nurse is not appreciate enough…” [P1064(GY/SN:4y3m)].The presence of blame culture in the hospital was revealed as a factor that could inhibit participants from putting their best performance to team efforts: “…but feel there is a culture of blame…” [P2125(GM/SN:6y)] and “A hospital with blame culture, not good…” [P2072(EM/SR:18y)]. In addition to management, the success of working as a team rests in the degree of support received from the nurses’ immediate colleagues.4.2.2Many of the participants indicated that the level of support received from other members of staff through teamwork increased their sense of belonging and strengthened their relationship with colleagues: “…staffs are competent, friendly, enthusiastic with a positive attitude towards patients care and with colleagues. Team work is always present among colleagues” [P2094(GS/SN:17y1m)]. The ethos of collegial support was echoed by other participants from their comments such as: “…my colleagues are really friendly, supportive…” [P2103(ST/SN:7y6m)], “… fellow colleagues working so well as a team and their support” [P2067(CD/SN:11m)], “Staff nurses support each other very well…” [P2135(MW/SN:1y)]; and “Very supportive and staff work well together in a team” [P2123(GM/SN:1y)].However, one participant provided a contrasting view from those who were in praise of collegial support. This participant indicated that retention problems and increased workload, which led to an unproductive ward, were direct consequences of being “…Unsupported by other staff which increases work load and leads to unproductive ward…” [P1071(RP/SN:3y9m)]. In the same vein, some participants applauded the supportive role of their colleagues and ward managers, but nevertheless highlighted the rude behaviour of some of the staff as an inhibiting factor to effective patient care: “… The support I have had as a newly qualified nurse has been more than I could have anticipated, received from staff nurses, clinical sisters and the ward manager alike…the clinical support workers are amazing…The only negative comment I have to make is that I find some specialist nurses who work at times on the ward can be rude and treat nurses on the ward as if they have less value in contributing to that particular area of care” [P2107(GS/SN:4m)].The importance of inter‐professional relationships in the provision of high quality care was also emphasized by the participants. They indicated that although they had excellent teamwork and good collegial relationships that were enhanced by managerial support, a collaborative nurse–doctor relationship was missing. For example, a participant linked inadequate staffing to the breakdown of nurse–doctor relationships, leading to loss of important information, affecting patient care: “…Due to less nurses on the ward, the relationships between Drs & SN can be affected and important information can often be omitted” [P2113(GS/SR:7y11m)]. Another participant also supported the view that good inter‐professional relationships were important in facilitating high quality care: “The CSWs  working particularly hard on this ward they are an asset…Doctors‐nurse relationship very poor but other MDT [Multidisciplinary Team] members work really well together as a team and have a shared approach to patient care…” [P2015(OT/SR:18y6m)]. Although participants highlighted the presence of cohesiveness among the nurses, demonstrated through hard work as a team, they also drew attention to how frustrating it was to work alongside colleagues who were evidently not interested in their jobs, or committed to the team.4.2.3Participants expressed frustration over some nurses’ poor motivation and lack of interest on the job. According to the participants, being absorbed in and being enthusiastic about their job would contribute to patient care, team cohesion, as well as improving their relationship with other members of staff. Some participants expressed disillusionment when they observed others displaying lack of interest in the job, often at the expense of good quality care to patients: “…Some staff are just frustrating to work with. Other staff feels/thinks they know everything and does not want to be told what to do, but they don't do things the way it should be done. Other staff goes to work because they have to, no interest or enthusiasm…” [P2072(EM/SR:18y)].my ward.…” [P2125(GM/SN:6y)].In contrast to the above comments, many of the participants acknowledged their wards as good working environments where they were either happy or enjoyed working: “I am happy to work here as a nurse …” [P2103(ST/SN:7y6m)]; and “I enjoyed working in my workplace since I started here” [P1102(EM/SN:11yr1m)]. In addition, some participants enjoyed their jobs and would be happy to have their family members admitted on their wards: “As a nurse, I enjoy and am proud of the level & quality of care my ward gives, that I would be happy for any family member to admitted to Some participants also commented that even though working on their wards could sometimes be stressful, they still enjoyed coming to work due to the workplace culture: “…I enjoy coming to work sometimes it is a bit stressful but we also have our good days” [P2119(GS/SN:2y)]; and “Stressful but fun and engaging + challenging…” [P1035(GS/SR:14y)]. These comments indicated that the participants were engaged with their jobs. In addition to inadequate staffing and teamwork, the workplace environment itself, in the form of pressure to create beds, health and safety issues and professional development, was identified as having impact on their work and quality of care given to patients.4.3Participants expressed worries over their workplace environment in terms of the physical work space and the need for further training and development. They were concerned that the absence of specific features such as adequate ventilation in their work environment could have an impact on health and safety of both the staff and the patients. They were also concerned over the priority the management placed over bed management.4.3.1“Clients need nurses not beds…” is a direct participant comment and captures participants’ understanding that the quality of care given by nurses is not about the physical resources. Rather, it involves how well trained or skilled the nurse is and the availability of the nurse. Participants believed that the focus of the Trust was on bed management, not on ensuring that patients were given the best care. According to the participants, bed managers continued to mount pressure on the nurses to create empty beds when it was inappropriate to discharge current patients, a situation the participants believed was detrimental to patient care. Nurses reported that they do not feel “listened to” by bed managers: “…feel we are not listened to by bed management who “bed” our query homes, even though they may not go home. When we have no beds we do actually mean we have no beds!” [P2025(GY/SN:6m)]. Other participants elaborated on how bed management and bed pressure were taking priority over patient care: “Trust seems to care about the day to day bed capacity on the wards and not how challenging it is to give good quality care along with other pressures that ward staff encounter” [P2047(RP/SR:11y)]; and “Recent financial constraints are causing problems and bed pressure are reducing patient care” [P2077(ST/SN:10y)]. This issue of improper bed management resulted in compromised patient care when, consequently, patients were admitted into inappropriate wards: “Our ward is extremely busy and high dependency, we are under constant pressure, day and night, to create empty beds, by transferring patients to other unsuitable wards, when not suitable for patients…” [P1101(GS/SN:31y)]. Furthermore, participants identified that apart from the pressure they were facing to create beds, the physical environment of their wards was creating health hazards and potentially compromising the quality of care nurses could provide to patients.4.3.2Participants highlighted that the physical condition and appearance of their wards could constitute health and safety issues. For instance, lack of ventilation in the ward was an issue: “Too hot – airless – troubled by outside noise – car/ambulance fumes…“ [P2026(GY/SN:37y10m)]; and “Hot – cramped – airless – windowless…” [P2133(GY/SN:38y2m)]. Participants also expressed their dissatisfaction with the lack of repair and maintenance of hospital equipment and furniture: “…Repairing of bed side lights are almost impossible” [P1059(GY/SN:8y6m)]; and: “…The ward itself is old and needs decorating…” [P2015(OT/SR:18y6m)]. Nevertheless, despite the stated shortcomings of the ward environment, one participant was of the view that she worked in a “Good ward and environment, also well organised” [P2003(EM/SR:20y)]. Finally, participants identified that apart from the physical environment, their work was influenced by workplace practices consisting of policies, which guide the provision of continuous professional development courses for nurses.4.3.3own time…” [P2111(GS/SR:11y11m)]; in contrast, a participant who was newly employed in the hospital nevertheless expressed satisfaction over the planned development and study days: “I am new at this trust. I am very happy about the development I am getting and all the study days planned for me” [P1003(GM/SN:8y8m)].Participants emphasized the importance of workplace practices such as professional development for the performance of particular tasks, which have an impact on patient care. They registered their dissatisfaction over the Trust's inability to meet their educational and training needs. Accessibility of these courses would have enhanced the fulfilment and performance of their roles. For example, a participant expressed the need for further training: “…The staff nurses need a lot of development…” [P2015(OT/SR:18y6m)]. Whilst one participant expressed dissatisfaction over the lack of encouragement and time for the completion of a six‐day course: “…I recently updated my skills in stoma care completing a course that lasted 6 days in total. This had to be done in my Participants in this study provided an insight into factors which affected the quality of their work environment and how these factors influenced their ability to provide quality care.5In response to the invitation to make comments about their work experiences, many nurses offered additional insight into aspects of their work environment which gave them concerns. This paper is the first to identify that despite the staffing problems that nurses faced and the resultant high workload and stress they were experiencing, nearly all the participants who commented about their ward managers, made positive comments. In describing their ward managers, most participants used positive words such as “approachable,” “accessible” and “pleasant”. Staff nurses in an American study by Schmalenberg and Kramer  identifiA study of nurses in acute hospitals in London demonstrated the importance of the ward manager in the effective functioning of the ward and a key determinant in nurses’ decisions to leave or to remain in the job  commented negatively on the behaviour of specialist nurses, noting their behaviour as rude and condescending. It could be argued that the staff nurse could be experiencing what Kramer, Brewer, and Maguire : 348 desFindings from this study indicate that many nurses working in the two hospitals faced huge challenges in their work environment with extensive emphasis placed on staffing shortage, having huge impact on their ability to provide high quality care. They also identified that financial constraints in the organization made alleviation of staff shortages very difficult. Participants in Kieft, Brouwer, Francke, and Delnoij  also indThis research provided the opportunity for nurses to express their feelings about factors inhibiting high quality care and teamwork, such as the presence of blame culture, lack of staff appreciation/acknowledgement, bed management and inadequate staff development. Comments made by the participants may imply that no one was listening to the nurses and that they have very little executive power to influence or change situations in the hospitals. A report by the King's Fund Ham,  has desc5.1It is acknowledged that the data gathered were short comments from the participants and this method of data collection did not present the researchers any opportunity to probe the participants or to clarify information given. In addition, this study was conducted in two NHS hospitals in the South East of England; findings may not be typical of all acute trusts in England, it may therefore limit generalization of the study.6Findings from this study have provided better understanding of the challenges experienced by nurses in their work environment, particularly in terms of constraints on their ability to provide high quality patient care. These constraints are currently largely beyond the resources of nurses themselves to address. The practice and policy implications of this research are for nurse leaders and policy makers to involve ward nurses in decision and policies for practice, to ensure effective delivery of nursing care. Further work on adapting the EOMII for use in England might focus on incorporating questions about structural elements of NWE. At the moment, the EOMII focuses exclusively on the “process” element of Donabedian's Structure–Process–Outcome model of the determinants of quality and it may have been the absence of attention to structural aspects that prompted so many nurses to give their comments.No conflict of interest has been declared by the authors.http://www.icmje.org/recommendations/browse/roles-and-responsibilities/defining-the-role-of-authors-and-contributors.html).All authors have agreed on the final versions and meet at least one of the following criteria (based on those recommended by the ICMJE*): (a) substantial contributions to conception and design of, or acquisition of data or analysis and interpretation of data; (b) drafting the article or revising it critically for important intellectual content. *


For data collection, the following validated instruments were used: the Instrument for Validation of Educational Content used for the validation by nurses and the Assistive Technology Assessment Questionnaire for the evaluation by deaf students. The criterion for validation was concordance higher than 80%, analyzed through the content validation index and binomial test. the final version of the video lasted seven minutes and thirty seconds, covered the steps that should be performed by a lay person to help a victim of cardiorespiratory arrest, presented animations and the narration in the Brazilian sign language. All the items obtained agreement among nurses and of among deaf students equal or superior to 80%. the video was considered to present valid content by the judges and comprehensible content by deaf students. Thus the video represents an inclusive technology for health education of deaf people about cardiopulmonary resuscitation. Recommendations from the American Heart Association highlight the relevance of teaching cardiopulmonary resuscitation (CPR) to lay people-Among the target public are the deaf people, who have no cognitive or motor impairment and only differ from the majority of the population for their auditory impairment. Although physically and mentally fit, they are in disadvantage when it comes to accessing information about identification of a CA and performance CPR, as they face communication barriers and a scarcity of available health education material in sign languageConsidering that the use of technological resources contributes to the success of health education and that communication with the deaf occurs through visual means, it is worth highlighting the feasibility of using videos as a resource for the provision of health information-Video also represent an effective option for teaching lay people regarding CPR-The construction and validation of an educational video for deaf people about CPR is relevant because it provides a didactic resource that can be used for training of a great number of people with standardized instructions and also corroborates with a self-directed and flexible learning process, because it provides the possibility of autonomy for the learners to watch the video in any moment they prefer and as many times as they deem it necessaryMoreover, it is worth noting that nursing has a relevant role in health education, because nurses carry out their professional activities in the various types of health services. They make up the professional category with the greatest number of professionals in the health area, and because they have education as an intrinsic trait of their professional practice. Thus, studies about the construction and validation of educational technologies about CPR are relevant to nursing because they inform about a resource option that can be used in educational interventions.Thus, this study aimed to create and validate an educational video for teaching deaf students about cardiopulmonary resuscitation.Methodological study consisting in the creation of an educational video, validation by judges, and evaluation by deaf students representatives of the target audience.Recommendations for the construction of audiovisual materials were followed, from pre-production (storyboard planning and design), production and post-production,,-The selection of the content about safety of the scene, correct identification of CA, call for help, and realization of the CPR followed the recommendations directed to lay people of the Brazilian Society of Cardiology, the American Heart Association, and the Asian and European Councils of ResuscitationThe storyboard with the scenes and drawings of the animations was built by a design company hired for this purpose under the supervision of the researcher. The choice for the development of video with digital animations instead of scenes recorded with actors was due to the fact that animations allow aesthetic improvement of the drawings, besides requiring less time for production and having a more attractive appearance than the recordings of scenes with actors.2.P (1-P)/e2. In this equation, Za (confidence level) was stipulated in 95%, P (proportion of experts agreeing with the item) was set at 85% and “e” (expected difference) was 15%, so that the calculated sample was 22 participants.After construction, the storyboard was submitted to content validation by judges with expertise in Basic Life Support (BLS). The sample calculation was performed with the equation for finite populations n = ZaIn order to recruit participants for this stage, the snowball technique was used: electronic contacts of teachers of undergraduate nursing courses and specialization in urgency and emergency and intensive therapy were sought in the websites of the state and federal public institutions located in Fortaleza - CE. These teachers were contacted via e-mail, and they, in turn, indicated other teachers from the states of Pernambuco, Rio de Janeiro, São Paulo and Minas Gerais who had an eligible profile to integrate the sample of this stage of the study.The inclusion criteria used were: to be a nurse with experience of CPR care and to have teaching or research experience in this theme. The exclusion criterion was incomplete completion of the collection instrument. Thus, 45 professionals were contacted via e-mail (via Google form), in which they received the Informed Consent Term, the storyboard, and the Instrument for Validation of Educational Content (IVEC), which consisted of an instrument with 18 questions  built and validated by the research group of assistive technologies for the health promotion of people with disabilities of the Federal University of Ceará. Thus, the 22 professionals who first sent a response were included in the study.It should be noted that the adjustments made, suggested by the judges, were related to the reformulation of some sentences to improve the chance of the contents to be understood by people without training in the health area. After completing the validation of the storyboard, the video was produced by the same design company that built the storyboard. Three characters were created: a victim affected by CA and a couple of lay rescuers. The images were first drawn in pencil, then vectorized in Corel Draw, converted into a Photoshop object and animated in After Effects.The content of the video was presented in the order of steps that should be followed to correctly provide help to the CA victim: observation of safety of the scene; identification of the problem; call for help from the Mobile Emergency Service (SAMU); and CPR without interruptions until the arrival of health professionals, with emphasis on the need of relay between rescuers who perform the compressions every two minutes.Since the objective was that the deaf students understood the content, in the construction of the video the whole narration was made in Brazilian Sign Language (LIBRAS). Thus, the narration was recorded in Libras in a studio by an interpreter of the Institute of Education of Deaf people of Ceará. It should be noted that the text of the narration was on the storyboard that had been previously validated during the stage of validation of content by judges. Synchronization of the components of the video  was performed in the Premiere program, and the interpreter was exposed in the lower right corner, in an area that corresponded to 1/6 of the screen.As the video is a technology aimed at teaching and demonstrating information, its preparation focused on the reference of Gagné’s instructional events. This reference includes components necessary for the content of an instruction to promote cognitive activation, mental processing, and long-term memory storage. These events focus on attracting the attention of student, informing objectives, stimulating memories and previous knowledge, presenting visual stimuli  and exposing content in organized blocks, with increasing order of complexityIn the construction of the video, the first event (catching the attention of the student) was contemplated by the presentation, at the beginning of the video, of questions about the need to know how to act in cases of cardiorespiratory arrest. The second event (presentation of the objectives) was shown before the presentation of the objective, with emphasis on the fact that a correct attitude may make a huge difference in saving lives. The third event (stimulus to memories of previous knowledge) was observed in front of the presentation of questions about what the viewer knew about the theme.The fourth event of Gagné  was present in the form of schemes/flowcharts and arrows in the illustration of the narrated content. The fifth event, related to the information presented in blocks, was also contemplated, since the content was organized and exposed in three blocks . The video’s compliance with Gagné’s instructional events is summarized in At the end of the preparation, the video was watched by deaf students from the Institute of Education of Deaf people of Ceará, in the city of Fortaleza/CE, for them to offer opinions about the clarity and understanding of the educational technology. The Institute of Education of Deaf people of Ceará, which is the only public institution exclusive dedicated for the education of deaf students in Ceará, has primary and secondary school classes and had, during the data collection period, approximately 70 students enrolled in each shift , totaling about 210 students.The inclusion criteria for selection of deaf students to evaluate the material were: to be enrolled in the educational institution and not to have cognitive limitations incompatible with the evaluation of the video. It should be noted that it was possible to check the existence of cognitive limitation based on information provided by the schools, because they were adapted to the teaching of deaf students and had, in the registers of enrollment, medical evaluation reports with details and attestation of deafness, pathologies, as well as motor and cognitive limitations. These records were consulted to confirm the absence of cognitive limitations in the students who participated in this study. The incomplete completion of the instrument was the exclusion criterion used.Recruitment occurred at the break between classes, in a day, shift and schedule defined by the head office of the institution. At that moment, students were approached in the courtyard and, with the help of a Libras interpreter, they were invited to go to the audiovisual room of the school reserved for evaluation. In view of this recruitment strategy, there was no calculation to define the number of participants that would be part of the sample of this stage of the study; this number was defined by the number of students who agreed and volunteered to participate. Thus, 16 students agreed to participate in the study and signed the ICF. The video was presented on a 32-inch television, with deaf students accommodated in school desks.After watching the video, the participants completed the Assistive Technology Assessment QuestionnaireThe researcher read the instrument, which was simultaneously translated by the Libras interpreter. One question at a time was read and translated; after the reading (of the researcher) and the translation into Libras (of the interpreter) of each question, it was expected that all the deaf students answered of the question explained, so that the next question was read and interpreted. Thus, all deaf students simultaneously filled out each of the questions on the instrument.The analysis of the data was carried out in the R software, version 3.1.1. In order to consider the validation, the Content Validation Index (CVI) was used in three ways: judges’ agreement on each item with the I - CVI ; proportion of items in which there was agreement of each judge, with the S-CVI/AVE ; and the mean S-CVI/AVE, which is the S-CVI . The binomial test, with significance level of 5%, was used to verify if the agreement ratio was statistically equal to or greater than 0.8The research was developed as established by Resolution 466/12 and was approved by the Ethics Committee of the Federal University of Ceará .The final version of the video had a duration of 7 minutes and 30 seconds and was composed by the opening, questions about the need to know how to act correctly in cases of cardiorespiratory arrest, presentation of the objective, questions regarding the prior knowledge of the spectator, information about the importance of safety in the scene, and three blocks of content, namely: information regarding the correct way to identify the problem; calling for help, highlighting the correct number for calling the SAMU; and performance of CPR, as observed in Thus, the educational video (available for access at: https://www.youtube.com/watch?v=V6_CnIn6TOo&t=1s) has accessibility in Libras to contribute, as a technological resource, to the teaching-learning of deaf students about the correct provision of assistance to victims of CA in an out-of-hospital setting.The judges who validated the content were all graduates of nursing course, four (18.2%) had a master’s degree, five (22.7%) had PhD degree, and the others were specialists. Regarding the professional activity, all had previous experience in urgency and emergency care or intensive care, and at the time of data collection, 15 (68.2%) were teaching in higher education institutions and/or specialization courses, and they provided content related to CPR. All had already participated in courses on CPR and 17 (77.3%) had already taught these types of courses. Moreover, 18 (81.8%) had published scientific articles on CPR in periodicals or in annals of events.In the validation of storyboard content presented in the video, there was a minimum 86% of agreement on the objectives, structure and presentation. Among the items referring to relevance, the lowest agreement was 77% in the item about adequacy of the size of the material. The five judges who disagreed with this item reported that the time programmed for the video (approximately seven minutes) was too long and did not present suggestions of adjustments/modifications, however, as the binomial test of such item was not significant, such agreement was considered statistically equal to or greater than 80%, so that the item was considered valid and there was no change.There was 86% agreement of the judges in the item “the information has accurate content”. The three judges who disagreed presented as justification the need of the educational material to show/teach how to check the carotid pulse in the identification of CA. It is worth noting that it was impossible to meet this suggestion because the protocols of the American Heart Association recommend that the presence of pulse is an action indicated for health professionals  and that lay rescuers should detect a CA if the individual is unconscious and in apnea.Furthermore, there was unanimity of agreement among the judges in the items concerning the fact that the video favors a reflection on the theme, deals with a modern theme, and collaborates with the area of knowledge .Regarding the S-CVI/AVE of the content validation, 12 judges disagreed with two items (agreement with 88.8% of the items and S-CVI/AVE = 0.88); six judges disagreed with one item (agreement of 94.4% and S-CVI/AVE = 0.94); and four judges agreed with all items (S-CVI-AVE = 1). Thus, the minimum S-CVI/AVE was 0.88 and the S-CVI of the content validation was 0.91.Among the deaf students who evaluated the video, 11 (68.7%) were women, 14 (87.5%) were single, none had children and all were in high school.According to the evaluation of these students, the video was considered comprehensible and there was a minimum agreement of 87% in the item “the acquisition of new concepts facilitates the search for information”. The deaf students who disagreed with this item argued that the search for information would only be facilitated if the viewer had control over the video projection technology so that they could pause and review some parts where necessary.In addition, there was a minimum agreement of 93% regarding the relevance and efficacy and unanimous agreement on clarity and interactivity, as observed in Regarding the S-CVI/AVE of the evaluation by the deaf students, four disagreed with one of the 14 items, so that there was agreement in 93.7% of the items and the S-CVI/AVE of each one was 0.93. The other 12 deaf students agreed on all items (S-CVI/AVE = 1). Thus, the S-CVI of the evaluation of deaf students was 0.98.The content that subsidized the construction of the video contemplated the safety of the scene, correct identification of the problem, call for help, and correct way to perform the CPR. These contents were relevant to be addressed in the video, because, once performed correctly, contribute to reduce the probability of death.,,Regarding the first information presented by the video (safety of the scene), the American Heart Association and the European Resuscitation Society emphasize the need to verify such safety before any approach to the victimOther relevant steps presented in the sequence of the content of the video concern the correct way to identify a CA and the request for help. When considering the incompatibility of life with the CA, fast assistance of specialized professionals becomes of major importance. However, the access of health professionals to the victim depends directly on the identification of the grievance, so that the mobile medical service be called right away-As soon as an out-of-hospital CA is identified, the task to call the mobile emergency service-The content used in the construction of the educational video also included, in particular, the correct realization of CPR, which explains the position of the rescuer, the point of the victim’s chest where the compressions must applied, the rate and depth that characterize high quality compressionsThe correct realization of these conducts is related to the effectiveness of the CPR and the probability of return of spontaneous circulation, as it is pointed out in the results of a North American multicenter study carried out with 8719 participants, where there was a correlation of correct rate and depth with the lowest mortalityDespite the rigorous construction based on scientific literature on the subject, the validation of the content of the video was important as well as its evaluation by deaf students, for scientific anchoring and attributing credibility to this educational technology.,-Regarding the identification of CA, some judges disagreed with the content because they thought the video needed to inform the need to check the carotid pulse to identify a CA. However, as individuals who do not respond and do not breathe have a high chance of being in CA, and because confirming the presence of a palpable central pulse is pertinent to health professionals, it is recommended that the population be oriented to treat as a CA all cases where there is no responsiveness or breathing-In the view of the nurses who participated in the validation, the video presented sufficient and necessary information. This result corroborates Brazilian methodological researches on the validation of a booklet on first aid at school and a video for parents of children using clean intermittent catheters, whose results showed that the evaluators agreed with the pertinence and adequacy of the content presentedThese Brazilian findings corroborate a study carried out in Venezuela regarding the construction and validation of an assistive technology for deaf people on oral health, which showed similar agreement among the evaluators-In the validation of the content, there was agreement that the video had clear information, with a language compatible with the understanding of the public, corroborating the evaluation of the deaf students, in which there was agreement on the comprehension and attractiveness of the video. A Brazilian study of validation of a printed material on prevention of metabolic syndrome, and a research conducted in New York about a video animation aimed at the teaching about the human genome found similar agreement regarding the understanding of the technologiesAs the health area uses specific terms, it is necessary to evaluate the language used in teaching materials for the population so that the educational technologies do not become of little usefulness and human inputs and materials do not be wasted in the dissemination and distribution of unclear or confusing materials. The incompatibility about the clarity of educational technologies in health is presented in a study whose results showed that 80% of the virtual contents of the American Academy of Orthopedic Surgery used in patient health education were incomprehensible to the target audience. Similar findings were presented in a study in Germany, whose results showed that educational materials on ophthalmology from 32 hospitals were not understood by the population for which they were createdIn this context, it is necessary that educational technologies be evaluated by representatives of the public who will use them, so that confusing and little comprehensible parts are adjusted and become compatible with popular understanding. Thus, the agreement of nurses and deaf students regarding the clarity of the video on CPR corroborates with the greater probability of the technology to enable the multiplication of information and contribute more effectively with the training of lay people on the subject.According to the evaluation of deaf students, the educational video stimulates interest and learning. Similar findings were observed in a research with schoolchildren in Indonesia on a video animation about metabolismThe positive evaluation of deaf students is relevant because, although the video was considered long by five judges (who disagreed about the adequacy of its size), the agreement of deaf students about the clarity of the content and the stimulation to learn confirmed that the duration of the video was necessary to properly expose the content, in a manner compatible with the understanding.The longer of time needed in the exposition of the information is due to the synchrony of the video directed to deaf students needs to respect the speed of capture of visual stimuli of this public, because the deaf spectator needs first observe the narration in Libras for then leave the visual field of the Libras interpreter to observe the demonstrations and animations. Thus, scenes and demonstrations need to be presented/displayed slower to allow the deaf student to see both the narration in Libras and the demonstrations/animations. This aspect makes sign language videos longer than videos with audio narration only.-The educational video on CPR was considered adequate and easy to use by the deaf students who evaluated it. Similar results were found in a study that evaluated electronic technology for teaching sign language in Bogotá and in a Brazilian study that included assistive technology for the blind people about breastfeedingIn this context, it is necessary to stress the need to empower the population to use educational technologies, especially deaf students, in order to stimulate autonomy, so that the search for information is facilitated. The findings also point to the relevance of investigating the opinion of the target population about the use of technologies so that the final version available is as compatible as possible with their autonomy. This corroborates the active participation of the population in the processes of teaching-learning about prevention, recovery and rehabilitation in health.The agreement among deaf students was influenced not only by the narration of the video in Libras, but also by the use of the Gagné’s theoretical reference in its construction. It is worth noting that the events related to the presentation of the content in blocks (of increasing complexity) and to the exhibition of visual stimuli  converge to benefit the understanding of the deaf students, due to the exacerbation of their visual perception. Thus, in methodological researches, it is pointed out the necessity of using references that fit and contemplate specific aspects of the target audience to which the technologies are intended.As a limitation out of the study is that the research was conducted in one Brazilian state and the participants were students. It is possible that its findings are distinct from the other states or from the reality of deaf people who are not students.It is worth noting that the construction and validation of the educational video about CPR adapted for use with deaf students contribute to the advancement of scientific knowledge in view of the availability of a didactic resource constructed with basis on an educational reference  and technical and scientific rigor, attesting to the validity of the content of the material as well as its comprehension by deaf students. The video can be used in the teaching-research-outreach activities tripod and enable the access to information about the CPR for deaf students.Considering that the role of health educators is inherent to the professional practice of nursing and that this category works in the assistance to CA and therefore has the expertise to carry out educational interventions about CPR, the construction and validation of the video about this topic is very relevant. Furthermore, this methodological study provides a replicable method to be used in nursing science for the construction and validation of other assistive technologies, with accessibility in Libras, in future studies.It is also worth noting that the availability of the educational video in Libras, with valid content and comprehensible for deaf students, not only makes it possible to use them during educational interventions, but also for self-instruction to build knowledge even in the absence of health professionals/instructors. In addition, the material contributes to the multiplication of information about CA, a theme little disseminated in Libras, for a public who, once trained to act correctly, may intervene and contribute to reducing mortality.An educational video on cardiopulmonary resuscitation was developed for deaf students. Its final version lasted 7 minutes and 30 seconds. It dealt with the sequence of behaviors to be adopted by a lay person for the correct provision of help for a victim of CA, and had its narration presented in audio and in Libras, in which the interpreter occupied about 1/6 of the screen.The validation of content had over 80% of agreement in all items, regarding objectives, presentation and relevance, and the S-CVI was 0.91. In the evaluation by deaf students, an agreement greater than 80% was also observed, regarding interactivity, objectives, relevance, effectiveness and clarity, and the SCV-I was 0.98. By means of the binomial and CVI tests, in the forms of I-CVI, S-CVI/AVE and S-CVI, the validity of the video regarding the content and appearance and its approval by the deaf students was demonstrated.The video can enable deaf students to access content on CPR and it is, therefore, a viable technological resource to be used by nursing and other health professionals in health education. It is necessary to investigate the effectiveness of using the video in the knowledge and ability of deaf students about CPR.