Part one examined the perspectives on the selection and development processes of senior academy male players (n = 8), whereas part two explored the perceptions of male coaches (n = 7). A total of three focus groups were used. Three main themes were confirmed by players and coaches: (a) task constraints, (b) performer constraints, and (c) environmental constraints. Specifically, although athletes and coaches believed that performer constraints were highly impactful on players' career in RU, there were inconsistencies surrounding the task and environmental constraints. Despite an indication that three common themes impacted an players path, this preliminary study shows an imbalance in the understanding of some of the key factors perceived to be important for talent progression in the present rugby academy. More research using similar qualitative methods is recommended to better understand the differences in opinions between players and coaches. Meanwhile, practitioners should consider implementing objective and holistic strategies to improve the talent pathway in English RU academies.The pathway towards senior professional status in sport is affected by a multitude of factors. An abductive examination of the talent identification and development processes at an English Premiership rugby union (RU) club was undertaken for the present study.  Becoming a senior professional athlete is the ambition of many youth participating in organised youth sports. Several authors have attempted to provide guidelines to optimise the Talent Identification (TID) and Talent Development (TD) paths to support this journey e.g., . Althougwide and emergent  and narrow and focused  framework. The English system is managed by fourteen regional academies aligned to the respective professional RU clubs (The increasing popularity of Rugby Union (RU) has led to large financial investment in TID and TD systems from World Rugby Union  and natiRU clubs , 8. A reRU clubs , which iRU clubs  found thRU clubs . DespiteRU clubs . In partactors on the talent stage, are those acting as “selectors”  and “selected” . Notably, the few qualitative studies exploring the TID and TD systems have focused on the coaches' perspective  rarely feature in the literature to understand how organisational approaches actually affect TID and TD, despite their ability to offer valuable insights into the transition to the professional level  were also present during the focus groups to assist with recording equipment.Institutional research ethics board approval was granted by (institution name blinded for review process) prior to data collection. Participants reviewed a letter of information and provided informed consent form prior data collection. The data collection was comprised of two parts in order to collect and analyse the data from both players and coaches' separately. All group discussions were organised by the lead author on three different days across two consecutive weeks in February 2020. Each focus group lasted approximately 60-minutes and was held in a meeting room at the club training ground, which had a comfortable setting that encouraged open discussion and interaction. Holding the focus groups at the club's facility was convenient and allowed participants to feel in a familiar setting and in a space where they were more comfortable to share information. Each focus group conversation was recorded by two video cameras (Sony HDR-CX240E Handycam) and two microphones (7RYMS RimoMic Lite LN Mini) in order to collect details of the discussions, and facilitate subsequent anonymised transcription of dialogue. While data collection was set by the first author, the conversation was facilitated by the first author for PG1 and PG2, and by the fifth author for the CG. Other authors  to provide results that are consistent with the data collected . Rigor wworthy topic. The use of two independent focus groups for the players and a separate one for the coaches provided both similarities and contrasting perspectives on the selection and development processes in one of the few English Premiership RU academies, which ensured that the study met the criteria for rich rigor. The criterion of sincerity was encompassed throughout all the steps of the research process, whereby each author avoided bias during data collection and analysis. Credibility in the present study was met through the accuracy of the reported data and the reflections from all participants, whilst following and adapting widely used thematic guidelines  to validate previous findings. Since this piece of work received both organisational and institutional ethical approval from their respective administrative and ethics board, adequate ethical procedures were always followed. Finally, to ensure meaningful coherence, the research group believed this study achieved its stated goals and interconnected each stage of the research process so as to accomplish the intended outcomes. These considerations are based on the criteria suggested when analysing qualitative data  sport participation history, (b) activity type, and (c) game exposure. From a “[…] But the biggest thing is making sure you do as many sports as possible because there's so many different skills that are transferable to the game of rugby […]. So, I think it's good to do as many sports as possible, just to increase your variety of skillsets.”Moreover, a player from PG2 reported his diverse sporting background while focusing more specifically on RU:“So, rugby is probably the main sport that I've really done. Done football, done kickboxing, done most sports, golf, but rugby's the main one”.activity type performed , was seen to have an advantageous effect on progression throughout a professional academy. As an example, it was reported that a larger accumulation of hours towards RU-specific activities was considered important for a player from PG1:It also emerged that the “I think mine was just definitely dedicating more time to it [training] than anyone else, really, when I was in those school kind of years. I was always kind of semi obsessed with going and getting passing and kicking done. In all my spare time it was just me and a couple of mates going to the local rugby club and kick for hours and hours. And it was our way of socialising as well through summers and stuff. So yeah, I think for me it was just kind of dedicating that time to go and develop my skills”.Similarly, a player from PG2 reinforced the importance of engaging in additional RU-specific activities:“So, you just find yourself doing extra gym sessions, extra kicking, like tackling with coaches and asking for more time and more feedback and stuff than like your peers around you who are not doing that”.A player from PG1 reported that training similar in the form of deliberate play was what made the difference in his career:“For me it was probably more enjoyment. When I was at school, I played it with my friends and we all went like training after school, which would be quite fun and we'd go back home. Then weekends, we'd play a game and then we'd go out for food or something afterwards and it was kind of like a fun activity to do […]. But for me, I think it was just making sure I kept enjoying it and didn't like overdo it and take it too seriously and ruin it for myself”.game exposure was believed important for growth and development from a participant of PG2:The fact that the RU academy players had a large amount of “So, I was playing like a Saturday, Sunday, sometimes on a Wednesday as well for the school. And when I look back on it and think you might say, ‘It was too much’. But I actually think it actually helped me a lot, because if you're tired and you physically don't feel as strong going into a game, it encourages you to challenge your skillset and do something in a different way”.anthropometric viewpoint, a player from PG2 reported that being tall was a prerequisite to be a successful rugby player:Performer constraints consisted of four categories: (a) anthropometric, (b) physiological, (c) psychological and psychosocial, and (d) technical-tactical. From an “[…] you’ve got to be people that are tall [to play professional rugby…]”.physiological requirement important for the different game-positions in RU, a player from PG1 reported:Regarding “[…] it's specific. It's not like football where, ‘Oh you'll pass, you'll shoot.’ Like the front row have just got to be big strong brutes, if you be a strong brute, you can , that's what you've got to bring. If you're a 9, you've got to bring your quick kicking game and your fast passing game. If you're a back, you've got to bring just speed in everything, over the top everything, just fast get in people's faces and stuff. I think you've got to bring that in your position and when you spend time with your mentor, you just learn off him”.psychological and psychosocial characteristics, such as cohesiveness with the rest of the team, in order to progress and set apart from less successful academy players. For example, a player from PG2 reported:Players also require “It's like the things that we're describing are sort of like the skeleton of the ideal player. But I feel like each individual person in that skeleton has got their own like flare or specific thing that they're good at. So, I feel like as a team we sort of fit into that skeleton by bringing our own like individual attributes. So, I feel like that's the best thing about being here, everyone has those individual attributes that just fit into that ideal player. So, I think there's no like specific, ‘He's the ideal player’. Everyone's got their own ability to become that ideal player, it's just fitting into that skeleton. […] It's more an ideal team”.psychological and psychosocial characteristics  had an important role in the TD process throughout the academy. These helped both in creating a successful teamwork environment and gaining trustworthiness. An example was provided from a player in PG1:Moreover, results from the players' focus groups showed that several other “If you're working hard and the guys around you know you're working hard they can trust you. And then if everyone's doing that together, then you'll work together well and it just makes a better team”.psychological and psychosocial characteristic both for the TID and TD process. For example, a player from PG mentioned:The communication and the capacity to be resilient when asking for individual feedback was recognised by players to be a fundamental “So, I think selection is a massive part of what we do in terms of like speaking to the coach and stuff, that's where you can get on their backs and say, ‘What do I need to do better to play? What can I improve? Where are my opportunities?’ that kind of thing”.psychological and psychosocial characteristic that differentiated standards of players. As an example, a player from PG1 stated:Mindset was also considered a “I think that's the difference between a really good club player and sort of the senior academy and making the first team. It's just that mentality”.psychological and psychosocial characteristics in RU, was reported by a participant from PG2 when they stated that several other behavioural characteristics were considered beneficial to distinguish level of players :Another example confirming the importance of “[…] And I think one thing that sets boys apart, so, from that jump where you go from academy to senior academy, you kind of see the boys that want it more. They go out on the field, they're more aggressive, they don't want to lose. So, when you've got some people that are just kind of there, they're good rugby players, they're talented, they're not going to make it because they don't have that extra little bit of grit”.technical-tactical skills, a player from PG1 stated that practising basic RU drills was the prerequisite to be successful during the TID and TD process:Regarding “If you don't have your basics [skills] then you don't have any chance, really”.The connection between these four categories, which underscores the impact of a multitude of factors on the TID and TD processes in RU and the existing individual differences among players and positions, was explained by a player from PG1:“I think around the table [focus group participants], like *player's name* sort of like brings a lot of physicality and work rate to games because he's like a very fit number 8. Like *player's name* brings a lot of speed like sort of around rucks and stuff like that and, like he says, with his kicking game. And *player's name* is just sort of like a wall in defence and like a strong runner. And personally, me, I like ball carrying, stuff like that is what I like to pride myself on”.Another quote that supported the previous statement reported that every player has their own strengths relevant for the TID process. However, participants reported that for each player, there could be multiple important factors that could have influenced their personal progression across a professional academy, which was suggested by a player from PG1:“But it's just everyone has their own individual thing that they're good at. There could be like two or three things, but I've just named one for each of us that I've seen in them. But there could be three or four, there isn't just one thing that you pride yourself on so there could be like a whole lot of various things”.national outlook, a statement from a player from PG2 proposed how he may have played rugby due to his national sport tradition:Environmental constraints consisted of three categories: (a) national, (b) socio-economic, and (c) family. From a “But probably because rugby is the main sport in Wales as well, so everybody does rugby […] Because rugby's much bigger in Wales for kids than football […]”.socio-economic perspective, in a sentence from a player in PG1 it was evident the clear impact of the sport orientation that different type of schools  could have on a RU player pathway:From a “[…] Yeah well, not really [rugby] at state school, it was just football, everyone played football. Yeah, there was no rugby. It was literally the only sport you did; it was the only sport anyone did. And then, obviously, when I went to [private] school, it was just school rugby, nothing else. Unless it was cricket and so on but I hate cricket”.Family was considered from PG2 one of the most important factors affecting progression in RU and in general in sports:“Probably, a huge thing for I guess everyone round the country [England] would be family input because if you're raised up in a household of football then you're more than likely to be going into football and playing football more often. So, I was quite lucky in that sense because my family are massively into rugby and sort of like a wide range of different sports. So, they were open to me to play whatever sport I wanted to play and support me in whatever I wanted to do. And it turned out to be rugby and they were very happy about that. So, I think family is a huge thing for sportsmen to start off their career”.A useful connection among these categories was highlighted throughout various statements. More specifically, a player from PG1 suggested:“I went to a state school until Year 7 and then private school [in England] because all my brothers went there so I just followed the family. And then that's where I picked up rugby and started enjoying it”.sport participation history perspective, one coach highlighted the importance of a multi-sport background:Task constraints consisted of two categories: (a) sport participation history, and (b) game exposure. From a “Just through my background and what I like to see in a player. If I see someone that plays [standoff or scrumhalf] in rugby, who's naturally a good footballer as an identified space from a young age, then you're like, ‘Yeah, we can work with this kid’. A lot of fly-halves that are at the standoff position in rugby have come through a football background. They will start with football and then be encouraged to see things and then move into rugby, and when you see them from an early age, they've usually got a good skillset, they've come from a background where they've been encouraged to work. And then it's just trying to give them more fine detail about the tactical stuff. And if they can take that on board, then usually you've got a bit of hope for them”.game exposure and developmental opportunities was explained by another coach:Moreover, the connection between “We do look at the scale of how much rugby they're playing week to week to keep it fair, when you're comparing them [for selection]”.anthropometric and long-term change upon TD outcomes. For instance, one coach reported that those players who did not mature physically in the time of 3 years would not progress across the academy:Performer consisted of four main categories: (a) anthropometric, (b) physiological, (c) psychological and psychosocial, and (d) technical-tactical. The CG highlighted the implications of “There are some kids who, through their athletic, their size, they haven't grown in a year or two or three, and they just won't fit into it naturally and we can't see any growth in them sometimes, physically, which is upsetting for them sometimes, but that's the way the modern game is going”.physiological viewpoint, coaches reported that players' physical attributes have a key role during TID and TD processes. In fact, an example of statement confirming this concept is:From a “Really just looking if a player's got an X factor. We talk about physical attributes. They might be really tall, you might have a guy who is really fast, they're not always the best rugby player but we might just have a look at someone athletically”.psychological and psychosocial area was also connected to TID and TD. In particular, coaches were unanimous on the topic of work ethic. As an example, one coach suggested:The “Just within that, once they come a bit older, it's also seeing a work ethic in them. So, when you're doing any hard work, and regardless if they're X factor or they're not, when you see a kid working, then it gives you a bit of hope that they'll learn and want to do better and they'd be competitive and they've got a work ethic to try and improve and get better […]. So, when you see the people graft and work hard, then you've probably got an eye for them as well when they stand out in bits and pieces that are not a glamorous part of the game. You realise that they've got an edge to them, and that usually stands out for me, which helps put them in a good place moving forward, as well”.psychological and psychosocial characteristics in young RU players, another coach specifically reported that the commitment to work hard was essential for successful players:Confirming the importance of this “So, […] is about having people who are really committed to working hard”.psychological and psychosocial characteristics believed pivotal for a player to be selected and be able to progress across the academy was their capacity to ask for feedback and engage with training putting in practice information provided by coaches:Another “[…] during the sessions, making sure that… little things like they're listening when we're talking to them as a group, making sure that they're paying attention, they're listening and maybe asking good questions and feedback around what we're trying to deliver to them. And for us, being able to see them trying to implement any points we're trying to work on with them. So, if we're looking at a certain skill, whatever it might be, seeing them actually trying to work on that and trying improve that when we then put them back into the game or the drill or whatever it might be; then seeing if they can, as the weeks go by, slowly take their individual points off everything and just keep adding to their game”.psychological and psychosocial deficiencies seemed to be less successful during the selection process, whereby a coach reported that:On this basis, players with “We also take into account […] learning difficulties and things like that. Because we've had kids who we don't necessarily think are listening in the summer [which is the period when U15 first selection take place], but then we've had a chat with them and we've realised that they're really dyslexic or they're slow in processing things. I think the most important thing is knowing your [player] that you're working with to boost the selection process”.Technical-tactical factors were also an important element for TD. For instance, one of the coaches commented:“We spend a lot of time on the 14 s, 15 s stage that we're at, the basics if you like, so being able to catch, pass well, manipulate defenders, footwork around the contact area, that sort of thing. Which hopefully then later in the years […] but how then we put that into, when the players get to more of the senior levels of the academy or into more of the first team area, whether they're still holding onto those points and hopefully adding to them as they go through, I would say”.socio-economic factors, which references the type of school, is highlighted by the following statement of a coach:Environmental constraints consisted of two categories: (a) socio-economic and (b) culture. An example of “For example, when you compare a state school kid to a private school kid, the difference is how much rugby they play is very different. But that state school kid, you might be able to look at him and go, ‘Well, he's got more potential if we put a bit more rugby into him.’ So, we take that into account, and we look at where they go to school […]”.culture also appears to influence the TD process. For instance, one coach reported:The club's “I think the whole culture of *club's name* adds to it [referred to the TD path] massively […]. But it's that whole culture of there's a friendship and quality there, so you feel comfortable, and you feel comfortable in whatever you do, which adds to the strength of what we're trying to achieve in the bigger picture”.culture for young players to flourish:Furthermore, another coach suggested the importance of creating a developmentally appropriate “the environments we have for them are challenging but actually are fun environments, such as the coaching groups that we have within the pathway”.Additionally, another coach provided a similar sentiment to reiterate the importance of club culture and quality of coaches in developing young RU players:“[…] it's the people that they have here. It's not as such just bringing in quantity of anyone with a rugby background and interest; it's making sure it's the quality that comes in […]”.Interestingly, a statement from a coach outlined the connection among the high-order themes emerged:“We've just offered some academy players contracts, and a couple of them have been given contracts just because of the size of them, and we think that they're going to grow. We're hoping that they're going to grow, being in a professional environment. So, they've been at school every day, and they've not had a chance to go to gym every day. They're restricted to how much training they can do, and hopefully by bringing them into this environment, they will grow physically as well as athletically and they'll fit into that” .constraints-led approach” and are understood as boundaries that shape the path  and academy coaches (part two) from an English Premiership RU club. Findings revealed that despite inconsistencies in some of the factors compounding categories and sub-categories, both players and coaches perceived task, performer, and environmental constraints to be important aspects of players progression. Previous studies indicated that, although not always analysed in unison, these three areas were the most researched in different countries during the TID and TD processes in RU . These cthe path  and emerthe path . Broadlythe path . Task cothe path .task constraints were expressed as important aspects across both cohorts. In fact, all participants suggested that sport participation history and game exposure were characteristics that would discriminate players progressions across a RU talent path since, as found in previous studies, these aspects facilitated skill acquisition, skill transfer, and were decisive in reaching high performance status  along with (b) players' maturation status, should be carefully monitored during TID and TD assessments in order to observe progression and avoid potential bias.ce in RU , 62, it ce in RU . Importace in RU . This inPosition-specific physiological traits were considered important for players' progression from both players and coaches. While players outlined that, despite the position individual characteristics of a player, a multitude of physical aspects  contribute to players’ success, coaches indicated that physicality was the most important attribute (even more than RU skills) to distinguish young talents in RU. These findings are in line with recent research on regional English , 13, 64 hard work  was the most cited by both cohorts, indicating that, in line with previous findings in sports  were believed important from both groups of participants. Interviewed players believed that one of the reasons for an initial successful identification and development in RU was the nation of provenience, since an elevated popularity of RU in the area of origin could offer more chance of initiation and continuous engagement. This concept is in agreement with Winn et al. , were often considered “better schools” and were more oriented to either form of rugby training than state schools placed in poorer areas of the country –75. PresFamily support was seen as an important factor from both player groups. Specifically, they suggested that both parents and brothers provided the right motivation, attitude, and economic support to engage in RU activities. This result was in line with TID and TD literature in sport , wherebyculture to all-age players during their path towards the senior professional status to improve motivation in their athletes.According to coaches, the culture surrounding the academy environment of a Premiership RU club is a fundamental aspect for players' growth. The interviewed members of staff mentioned that a challenging, positive, and supportive full-time RU academy environment was the key for optimal TD. However, in the coaches' opinion, these environments could only be created by clubs who recruit high-quality coaches. In fact, the importance of the appropriateness of personnel in elite sport academies has been already reported by several authors investigating the TID and TD processes. For example, in order to potentiate the athletic development of an athlete, Lloyd et al.  reportedA limitation of this study is the number of participants recruited. A similar number of individuals has previously been recommended for focus groups , yet a hThis is the first published study that has analysed the perceptions of senior academy players and academy coaches on the TID and TD processes in an English Premiership RU club. These preliminary findings demonstrate that task, performer, and environmental constraints were the parameters considered important during the players journey towards senior professional status. Despite results aligning to previous holistic findings , and add


Augmented reality (AR) integrates computer-generated content and real-world scenarios. Artificial intelligence's continuous development has allowed AR to be integrated into medicine. Neurosurgery has progressively introduced image-guided technologies. Integration of AR into the operating room has permitted a new perception of neurosurgical diseases, not only for neurosurgical planning, patient positioning, and incision design but also for intraoperative maneuvering and identification of critical neurovascular structures and tumor boundaries. Implementing AR, virtual reality, and mixed reality has introduced neurosurgeons into a new era of artificial interfaces. Meningiomas are the most frequent primary benign tumors commonly related to paramount neurovascular structures and bone landmarks. Integration of preoperative 3D reconstructions used for surgical planning into AR can now be inserted into the microsurgical field, injecting information into head-up displays and microscopes with integrated head-up displays, aiming to guide neurosurgeons intraoperatively to prevent potential injuries. This manuscript aims to provide a mini-review of the usage of AR for intracranial meningioma resection. Augmented reality (AR) has been defined as adding virtual components within the real world . An AR sIn the past few years, AR has been demonstrated to be helpful for resecting both brain  and skulAll steps for setting up AR for meningioma resection do not change compared to other brain lesions. The information injected depends on the capacity of the surgical planning software to integrate not only vessels but also diffusion tensor imaging or other remarkable neuroimaging data. The HUDs of the microscope are used for AR support. Injection of information for AR interface requires advanced processing from 3D reconstruction software. Checking the calibration of AR is performed by centering the microscope above the divot of the registration array. Thus, the optical outline and the AR visualization of the reference array can be adjusted, and the 3D reconstructed objects can also be visualized  . All opeThe integration of AR for tumor resection, including meningiomas resection, and mainly skull-base located meningiomas, has been pivotal for developing the technique . In 2010The field also advanced toward minimally invasive surgeries. Jean et al. reported a video article using AR templates for a minimally invasive transorbital approach for intradural tumors, including meningiomas . This puOncologic neurosurgery is a highly complex field that has evolved by integrating various technologies to provide better patient outcomes. The first prototype of AR was designed in the 1960s, but only in the 1990s and 2000's that the interest in this technology increased, particularly in the medical field , 18. In Intraoperative AR in micro neurosurgery depends on integrating the neuroimaging processing software into the hardware of both the microscope and the neuronavigation system. Currently, few systems with integrated HUDs allow this integration and adequate projection of images. The cost of these devices is considerably high, which undoubtedly poses a clear access limitation, even more so without demonstrating their actual usefulness. Regarding the operating room setup, the times can initially be increased up to ~30 min in the first cases, and progressively reduced to ~12 min. In order to improve workflow, the same surgical team must be trained in its implementation. The quality of the projected images will depend on the quality of their acquisition. To maximize the benefits of AR, we always suggest supplementing with other imaging modalities. We have always performed image fusion with CT and time-of-fly (TOF) magnetic resonance imaging for skull base lesions, achieving adequate vascular reconstructions. During the AR projection, the surgeon must adapt to the image overlay. However, the system allows easy removal of the projection. Thus, the surgeon can determine when the projected image will be most helpful.The position of the microscope is one of the most relevant references for proper navigation, and the microscope optics determines the depth of the focus point . This is a pivotal aspect, given the adjustments required during surgery that may modify the projected image. The images of AR are projected from the neuronavigation system and have the same limitations as this technology. We also recommend looking for submillimeter precision during neuronavigation registration as much as possible to improve AR accuracy, as the information of regular volumetric CT/MR images and 3D segmentation reconstructions will be integrated into one single microscopical surgical view . It is iNeurosurgeons face the challenge of learning, planning, and performing increasingly complex surgical procedures with a narrow window for error . AR has Despite being a benign tumor in most cases, meningiomas remain a neurosurgical challenge, given their close relationships to crucial neurovascular structures, especially for those in the cranial base . MultiplThe literature and evidence remain scarce, and intraoperative AR indications remain in research.Depuration of indications and real pros and cons are necessary to understand the benefits of its use. For now, it is mandatory to understand the basic principles of AR, introduce technology cautiously, given the progressive learning curve, and be aware of the limitations and possible errors associated with new technologies. At some point, the advancements in artificial intelligence may focus on AR into real benefits for a safer procedure. In the meantime, research is necessary, and a comparison of this technology with other image-guided tools is also necessary. AR and other technologies will never replace knowledge of microsurgical anatomy, nor the surgical skills required for adequate resection  . Still, Augmented reality (AR) integration in neurosurgery has shown promise in enhancing surgeons' awareness of critical neurovascular structures, facilitating tumor dissection, and improving overall surgical outcomes. While the literature on intraoperative AR remains limited, ongoing research has shown, along with other emerging technologies, that it can serve as a valuable tool to guide surgeons on an individual, case-by-case basis. There will always be a need to improve meningioma surgical results, and as advancements in artificial intelligence continue, integrating AR and mixed reality scenarios into neurosurgical procedures holds the promise of safer and more effective meningioma resections.DG: Conceptualization, Formal analysis, Methodology, Supervision, Writing—original draft, Writing—review & editing. EO-R: Conceptualization, Data curation, Investigation, Methodology, Writing—original draft, Writing—review & editing. AR-S: Conceptualization, Investigation, Methodology, Writing—original draft, Writing—review & editing. LF: Formal analysis, Software, Visualization, Writing—original draft, Writing—review & editing. MV-O: Investigation, Software, Writing—original draft, Writing—review & editing. JR: Conceptualization, Data curation, Investigation, Writing—review & editing. JM: Investigation, Supervision, Validation, Writing—review & editing. FH: Validation, Writing—original draft, Writing—review & editing.


Hepatic fibrosis is a common consequence of chronic liver diseases without approved antifibrotic therapies. Long noncoding RNAs (lncRNAs) play an important role in various pathophysiological processes. However, the functions of certain lncRNAs involved in mediating the antifibrotic role remain largely unclear.4) or bile duct ligation (BDL) to investigate the function of lnc-Helf in liver fibrosis.The RNA level of lnc-High Expressed in Liver Fibrosis (Helf) was detected in both mouse and human fibrotic livers. Furthermore, lnc-Helf-silenced mice were treated with carbon tetrachloride  and activated hepatic stellate cells (HSCs). In vivo studies showed that silencing lnc-Helf by AAV8 vector alleviates CClOur results unveil a lnc-Helf/PTBP1/PIK3R5/AKT feedforward, amplifying signaling that exacerbates the process of hepatic inflammation and fibrosis, thus providing a possible therapeutic strategy for hepatic fibrosis.The online version contains supplementary material available at 10.1186/s11658-023-00492-3. Hepatic fibrosis is a common consequence of chronic liver diseases caused by various etiologies, such as chronic viral hepatitis B and C, cholestasis, or alcohol abuse , 2. If nFibrosis is characterized by the excessive deposition of extracellular matrix (ECM) components , 2. Acti4) or bile duct ligation (BDL)-induced proinflammatory M1 macrophage polarization and pyroptosis [The enormous transcripts transcribed from the mammalian genome are non-coding RNAs (ncRNAs) . Althougroptosis . Moreoveroptosis . In addiroptosis . DespiteIn this study, we identified a novel lnc-High Expressed in Liver Fibrosis (Helf) that is increased in mouse and human fibrotic livers and systemically investigated its regulatory and functional role in hepatic inflammation and fibrosis.Study population analysis was conducted as previously reported . BrieflyThree pathologists blinded to the study protocol scored liver fibrosis according to the meta-analysis of histological data in viral hepatitis (METAVIR) fibrosis staging system. The clinical characteristics were summarized in Additional file 4, 60 mice were randomly separated into six groups: AAV8-negative control (NC), NC + CCl4, AAV-lnc-Helf-shRNA-1#, AAV-lnc-Helf-shRNA-1# + CCl4, AAV-lnc-Helf-shRNA-2#, and AAV-lnc-Helf-shRNA-2# + CCl4 (n = 10). The adeno-associated virus was injected 2 weeks after the first injection of CCl4 via the tail vein (2 × 1011 pfu/mouse). NC + CCl4 group, lnc-Helf-sh1# + CCl4, and lnc-Helf-sh2# + CCl4 group mice were treated with CCl4 (0.2 ml/kg) twice per week for additional 6 weeks after the virus was injected. AAV8-NC, AAV-lnc-Helf-shRNA-1# and AAV-lnc-Helf-shRNA-2# group mice were injected with an equivalent amount of olive oil. For hepatic fibrosis model induced by BDL, 90 Balb/c mice were randomly separated into six groups: AAV8-NC, NC + BDL, lnc-Helf-sh1#, lnc-Helf-sh1# + BDL, lnc-Helf-sh2#, and lnc-Helf-sh2# + BDL (n = 15). Mice were injected with AAV8-NC or AAV8-lnc-Helf-shRNAs virus (2 × 1011 pfu/mouse) via tail vein 2 days before sham operation or bile duct ligate operation. A total of 21 days after the operation, serum and liver specimens were collected for further analyses.All animal work was conducted according to the guidelines approved by the Animal Care and Use Committee of Tianjin Medical University. Eight-week-old Balb/c male mice were purchased from Beijing HFK bioscience . For hepatic fibrosis model induced by CCl4, AAV-lnc-Helf-shRNA-1# + CCl4, and AAV-lnc-Helf-shRNA-2# + CCl4 (n = 3) was qualified by Agilent 2100 bioanalyzer  and subsequently screened on the BGIseq500 platform . The threshold was fold change > 2 and padj < 0.05. The sequencing data have been deposited in NCBI GEO database: GSE182485 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE182485).RNA harvested from liver tissues of mice treated with NC + CClMouse primary HSCs were isolated from 40-week-old male Balb/c mice by in situ ethylene glycol tetra-acetic acid (EGTA)/Hank’s balanced salt solution (HBSS) solution, pronase, collagenase perfusion, further digestion, and Histodenz gradient centrifugation as previously reported , 19. Mor7 cells were lysed. RNA-bound beads were mixed to the lysates and incubated for 1 h at 4 °C. Followed by washing with wash buffer, we added elution buffer to the beads, mixed and incubated at 37 °C with agitation for 30 min. Supernatant was removed for downstream analysis .For biotinylated RNA pull-down assay, lnc-Helf was transcribed in vitro with T7 RNA polymerase . Then, T4 RNA ligase was used to attach a single biotinylated nucleotide to the 3′ terminus of an RNA strand at 16 °C for 16 h . The labeled RNA was bound to the Streptavidin Magnetic Beads at 25 °C incubating for 30 min. The mixture was washed with 20 mM Tris (pH 7.5) two times. Liver single cell suspensions of mice were prepared and total of 2 × 10RIP was performed using the Magna RIP RNA-Binding Protein Immunoprecipitation Kit  according to the manufacturer’s instructions. Briefly, mouse liver single cell suspensions and RAW 264.7 cells were lysed by RIP lysis buffer and stored at −80 °C. A total of 5 µg of the antibody (anti-PTBP1 and control IgG) was added to magnetic beads and incubated with rotation for 30 min. Then, the RIP lysate was centrifuged at 14,000 rpm for 10 min at 4 °C. The beads were added to the lysates and RIP immunoprecipitation buffer and incubated at 4 °C for overnight. Next, RIP buffer was briefly centrifuged to remove the supernatant. The purified RNAs were used for cDNA synthesis and evaluated by quantitative reverse transcription polymerase chain reaction (qRT–PCR).t-test (two group comparison), and p < 0.05 indicated a significant difference.Data were expressed as mean ± standard error of the mean (SEM) with at least three independent experiments. All statistical analyses were conducted using SPSS 23.0 . Statistical analysis was conducted using either one-way analysis of variance (more than two groups) or Student’s Details on other materials and methods are provided in the Additional file 4 for various time periods, and the data revealed that the level of lnc-Helf was markedly increased with persistent injury  staining, Masson staining, Sirius red staining, and immunohistochemistry (IHC) for α-SMA, COL1α1, and TGFβ, and the data revealed that lnc-Helf silencing alleviated hepatic fibrosis induced by CCl4   and proliferation  in the livers  assay to identify the proteins binding with lnc-Helf Fig. B. Moreov4-treated lnc-Helf-shRNAs mice compared with CCl4-treated AAV8-NC mice  The fold change of lnc-Helf was shown according to the microarray data.  The nucleotide sequence of mouse lnc-Helf and human lnc-HELF. (D) The comparison between human and mouse lnc-Helf sequences. (E) The nuclei or cytoplasm of primary HSCs was isolated, and qRT–PCR analysis detected the expression of lnc-Helf, Neat1, and Gapdh. (F) The intracellular localization of lnc-Helf in primary HSCs was measured by RNA-FISH assays; scale bar is 10 μm. (G) qRT–PCR analysis of lnc-Helf in AML12 cells, LX-2 cells, RAW264.7 cells, BMMs, and HUVECs (H) qRT–PCR was used to assess the expression of lnc-Helf and Acta2 in primary HSCs cultured at day 2 treated with 10 ng/ml TGF-β for 24 h. (I–L) The correlations of lnc-HELF, ACTA2, COL1α1, ALT, and AST were assessed by Pearson correlation analysis, n = 34. *p < 0.05, unpaired Student’s t test (H). Fig. S2, related to Fig. S2. Mice were divided into six groups: AAV8-NC, NC + CCl4, lnc-Helf-sh1#, lnc-Helf-sh1# + CCl4, lnc-Helf-sh2#, and lnc-Helf-sh2# + CCl4. Mice were injected with AAV8-lnc-Helf-shRNAs or AAV8-NC virus 2 weeks after the first injection of CCl4 via tail vein. After CCl4 treatment for 8 weeks, (A–D) the volcano map and GO analysis of differentially expressed mRNAs in lnc-Helf-sh1# + CCl4 mice and lnc-Helf-sh2# + CCl4 mice are shown, compared with NC + CCl4 mice. Fig. S3, related to Fig. S2. Mice were divided into six groups: AAV8-NC, NC + CCl4, lnc-Helf-sh1#, lnc-Helf-sh1# + CCl4, lnc-Helf-sh2# and lnc-Helf-sh2# + CCl4. Mice were injected with AAV8-lnc-Helf-shRNAs or AAV8-NC virus 2 weeks after the first injection of CCl4 via tail vein. After CCl4 treatment for 8 weeks (A), IHC for COL1α1 and TGFβ are shown; scale bar is 100 μm for 40× and 400 μm for 10× magnifications . (B) qRT–PCR was used to assess the expression of Acta2, Col1α1, Mmp2, Timp1, and Tgfβ1.  Serum ALT and AST was examined. Data are presented as mean ± SEM.*/#p < 0.05. *p < 0.05 for AAV8-NC. #p < 0.05 for NC + CCl4, one-way ANOVA (B-D). Fig. S4, related to Fig. S2. Mice were divided into six groups: AAV8-NC, NC + CCl4, lnc-Helf-sh1#, lnc-Helf-sh1# + CCl4, lnc-Helf-sh2#, and lnc-Helf-sh2# + CCl4. Mice were injected with AAV8-lnc-Helf-shRNAs or AAV8-NC virus 2 weeks after the first injection of CCl4 via tail vein. After CCl4 treatment for 8 weeks, (A) western blot was used to determine the expression of CD11b, TNF-α, IL-1β, and MCP1. GAPDH was used as an internal control.  qRT–PCR was used to assess the RNA level of proinflammatory genes  and proliferation-related genes  in livers of each group. (D) IHC for CD11b, F4/80, TNF-α, IL-1β, LY6C, and PCNA; scale bar is 100 μm for 40× and 400 μm for 10× magnifications. Data are presented as mean ± SEM. */#p < 0.05. *p < 0.05 for AAV8-NC. #p < 0.05 for NC + CCl4, one-way ANOVA (B and C). Fig. S5, related to Fig. S3. Mice were divided into six groups: AAV8-NC, NC + BDL, lnc-Helf-sh1#, lnc-Helf-sh1# + BDL, lnc-Helf-sh2# and lnc-Helf-sh2# + BDL. Mice were injected with AAV8-lnc-Helf-shRNAs or AAV8-NC virus 2 days before sham operation or bile duct ligate operation via tail vein. After 21 days of operation, (A) qRT–PCR was used to examine the RNA level of lnc-Helf in livers of each group.  Serum AST and ALT was examined. (D) qRT–PCR was used to assess the RNA level of Pcna, Ki67, Cyclin D1, and Bcl-2 in livers of each group. Data are presented as mean ± SEM. */#p < 0.05. *p < 0.05 for AAV8-NC. #p < 0.05 for NC + BDL, one-way ANOVA (A-D). Fig. S6, related to Fig. S4. (A) Quantitative analysis of western blot of Fig. S4B. (B) Primary HSCs at day 2 were infected with lenti-control and lnc-Helf-shRNA or lenti-lnc-Helf for 48 h, following the treatment with 10 ng/ml TGF-β1. The expression and location of COL1α1 was assessed by confocal microscopy. Scale bar is 20 μm. (C) Quantitative analysis of western blot of Fig. S4F. (D) Primary HSCs at day 2 were infected with lenti-control and lenti-lnc-Helf for 48 h, following the treatment with 10 ng/ml TGF-β1. The expression and location of COL1α1 was assessed by confocal microscopy. Scale bar is 20 μm. (E–G) LX-2 cells were infected with lenti-control or lenti-lnc-HELF for 48 h, following the treatment with TGF-β1 for 24 h. qRT–PCR was used to assess the expression of lnc-HELF, ACTA2, COL1α1, COL1α2, TIMP1, TGF-β1, PCNA, KI67, and CYCLIN D1 (E); western blot was used to determine the expression of the protein level of α-SMA, MMP2, CYCLIN D1, CYCLIN B1, and PCNA (F). GAPDH was used as an internal control. Cell proliferation was detected by CCK8 (G). (H) Primary HSCs at day 3 were infected with lenti-control and lenti-lnc-Helf for 96 h, cell proliferation was detected by CCK8. Data are presented as mean ± SEM. */#p < 0.05. *p < 0.05 for LV-control. #p < 0.05 for LV-control + TGF-β1, one-way ANOVA  and unpaired Student’s t test (G and H). Fig. S7, related to Fig. S5. (A) Mouse primary HMs transfected with siRNA for 24 h, qRT–PCR analysis was used to detect the RNA level of Mrc1, Arg1, Cd163, and Il10. (B) HMs were transfected with pcDNA3.1-lnc-Helf or pcDNA3.1 for 48 h, qRT–PCR was used to assess the expression of Mrc1, Arg1, Cd163, and Il10. (C) RAW264.7 cells were transfected with pcDNA3.1 or pcDNA3.1-lnc-Helf for 48 h, qRT–PCR was used to assess the expression of lnc-Helf, Tnf-α, Mcp-1, Il-1β, Il-6, Pcna, Ki67, Cyclin D1, Cyclin E1, Mrc1, Arg1, and Il10. (D) RAW264.7 cells were transfected with pcDNA3.1 or pcDNA3.1-lnc-Helf for 48 h following treatment with 20 ng/ml IFN-γ for 24 h. Mature supernatant IL-1β level was detected by ELISA. Data are presented as mean ± SEM. */#p < 0.05. *p < 0.05 for pcDNA3.1. #p < 0.05 for pcDNA3.1 + IFN-γ, unpaired Student’s t test (A–C) and one-way ANOVA (D). Fig. S8, related to Fig. S6. (A) Mice were divided into six groups: AAV8-NC, NC + BDL, lnc-Helf-sh1#, lnc-Helf-sh1# + BDL, lnc-Helf-sh2#, and lnc-Helf-sh2# + BDL. Western blot was used to determine the protein level of phos-AKT (Thr308), phos-c-Raf1 (Ser259), phos-GSK-3β (Ser9), and AKT in liver tissues of each group. (B) LX-2 cells were infected with lenti-control or lenti-lnc-HELF for 48 h following treatment with 10 ng/ml TGF-β1 for 24 h. Western blot was used to determine the protein level of phos-AKT (Thr308) and AKT. GAPDH was used as an internal control. (C) RAW264.7 cells treated with or without the AKT inhibitor MK2206 were transfected with pcDNA3.1 or pcDNA3.1-lnc-Helf for 72 h. qRT–PCR was used to detect the expression of lnc-Helf, Tnf-α, Mcp-1, Il-1β, Il-6, Pcna, Cyclin D1, and Cyclin E1. Data are presented as mean ± SEM. */#p < 0.05. *p < 0.05 for pcDNA3.1 + DMSO. #p < 0.05 for pcDNA3.1-lnc-Helf + DMSO, one-way ANOVA (C). Fig. S9, related to Fig. S7. The expression and location of α-SMA in lnc-Helf-increased HSCs simultaneously transfected with siPTBP1 was assessed by confocal microscopy. Scale bar is 20 μm. Fig. S10, related to Fig. S8.  Prediction of the interaction probabilities between lnc-Helf and PTBP1 using the RPIseq and prediction of lncRNA–protein interactions database. (C) qRT–PCR was used to detect the RNA level of PTBP1 in livers of healthy people (n = 6) and fibrotic patients (n = 28).  The correlation of PTBP1, lnc-HELF, and PIK3R5 was assessed by Pearson correlation analysis, n = 34. (F–I) Mice were divided into six groups: AAV8-NC, NC + CCl4/BDL, lnc-Helf-sh1#, lnc-Helf-sh1# + CCl4/BDL, lnc-Helf-sh2#, and lnc-Helf-sh2# + CCl4/BDL. qRT–PCR was used to detect the RNA level of Pik3r5 and Ptbp1 in livers of each group. Data are presented as mean ± SEM. */#p < 0.05. *p < 0.05 for AAV8-NC. #p < 0.05 for NC + CCl4/BDL, one-way ANOVA (F–I). Fig. S11, related to Fig. S8.  Primary HSCs at day 2 were infected with lenti-control or lnc-Helf-shRNA or lenti-lnc-Helf for 48 h, following the treatment with TGF-β1 for 24 h. qRT–PCR was used to detect the RNA level of Pik3r5. (C) LX-2 cells were infected with lenti-control or lenti-lnc-HELF for 48 h, following the treatment of TGF-β1 for 24 h. qRT–PCR was used to assess the expression of PIK3R5. (D) Mouse primary HMs were transfected with lnc-Helf siRNA for 24 h following treatment of 20 ng/ml IFN-γ for 24 h. qRT–PCR was used to assess the expression of Pik3r5.  Mouse primary HMs and RAW264.7 cells were transfected with pcDNA3.1 or pcDNA3.1-lnc-Helf for 48 h, qRT–PCR was used to assess the expression of Pik3r5. (G) qRT–PCR was used to assess the expression of Pik3r5 in lnc-Helf-overexpressed RAW264.7 cells simultaneously transfected with siPTBP1. Data are presented as mean ± SEM. */#p < 0.05. *p < 0.05 for LV-control or si-NC or pcDNA3.1or pcDNA3.1 + si-NC. #p < 0.05 for LV-control + TGF-β1/IFN-γ or pcDNA3.1-lnc-Helf + si-NC, one-way ANOVA  and unpaired Student’s t test (E and F). Fig. S12. Schematic diagram shows the function and mechanism of lnc-Helf in the progression of hepatic inflammation and fibrosis. Upon liver injury, increased lnc-Helf binds with PTBP1 to promote its interaction with PIK3R5 mRNA, resulting in increased stability and activating the AKT pathway, thus promotes HSCs and HMs activation and proliferation, which augments hepatic inflammation and fibrosis. Table S1. Clinical characteristics of patients. Table S2. The results of mass spectrometry from proteins pulled down by the sense and antisense of lnc-Helf. Table S3. Cloning primers for lnc-Helf. Table S4 siRNA sequences. Table S5 qRT–PCR primers. Table S6 RACE primers for lnc-Helf.Additional file 2: The results of mass spectrometry from proteins pulled down by the sense and antisense of lnc-Helf.


SLC1A1, SLC25A12, and ATP2B2) and three related long non-coding RNAs  from this pathway in 39 patients with refractory epilepsy and 71 healthy controls. Expression of all genes except for lnc-SLC25A12 was higher in total epileptic cases compared with controls  for differentiation of epileptic cases from controls. Moreover, lnc-MTR-1 and lnc-SLC25A12 were sensitive markers for such purpose . The highest value belonged to LINC01231 with the value of 0.76. Taken together, this study demonstrates dysregulation of calcium-signaling related genes in epileptic patients and suggests these genes as potential biomarkers for epilepsy.Calcium signaling is a metabolic pathway that is essential in neurons development and can be involved in the pathobiology of epilepsy. We assessed expression of three mRNA coding gene ( One third of patients with epilepsy do not respond to anti-seizure drugs and are known to have refractory epilepsy2. This condition affects quality of life and has adverse results in daily routines3.Brain tissue is involved in many neurological disorders and one of these chronic disorders is epilepsy. Epilepsy is defined as a condition that patient has long-lasting generating seizures4. Intrinsic burst firing, mediated by inward calcium ion is considered as the initiator of epileptic activity5. Calcium signaling is regulated by long non-coding RNAs (lncRNAs). Several calcium-binding proteins are subjected to regulation by these transcripts6.Calcium signaling is a metabolic pathway that is essential in neurons development. Calcium acts as an important second messenger and contributes to the correct synapse activities. So, it is obvious that distraction in this signaling pathway may result in neurodevelopmental disordersATP2B2 gene codes a protein that removes bivalent calcium ions from cells and plays a critical role in calcium homeostasis. This gene is regulated by an lncRNA, namely lnc-MTR-17. Solute Carrier Family 1 Member 1 (SLC1A1) is a glutamate transporter that is related with a number of signaling pathways including Transport of inorganic cations/anions. Mutations in this gene might be involved in the pathogenesis of schizophrenia8. Most notably, rare variants in this gene have been found to be associated with a kind of sensory epilepsy9.Several of proteins and non-coding RNAs are involved in calcium signaling. For instance, SLC25A12 codes for a mitochondrial aspartate/glutamate transporter which is principally expressed in neurons10. This protein is involved in the exchange of aspartate for glutamate and protons in the mitochondria11. Dysfunction of this protein leads to defects in the transport of aspartate into the cytoplasm, reduction of N-acetyl glutamate production in the cytoplasm and subsequent defects in the production of myelin11. SLC25A12 variants are involved in the pathogenesis of epilepsy13 possibly due to accumulation of glutamate which results in cellular injury14.15. Since calcium signaling has an efficient role in the neurodevelopmental disorders through regulation of normal neuronal activity, the related genes may play a role in the epilepsy as well. Thus, we analyzed expression of SLC1A1, SLC25A12, and ATP2B2 genes that encode calcium signaling proteins that play a part in storing, regulation, and transmission of calcium in neurons. We also assessed expression of three lncRNAs that are related with these genes, namely LINC01231:1, lnc-SLC25A12-8:1 and lnc-MTR-1:1T, respectively, as predicted by the LNCipedia16 and RNAcentral17 databases. The regulatory network constructed by lncRNA-mRNA interactions may affect epileptogenesis and can be considered as a putative target for therapeutic interventions, particularly in refractory epilepsy.The role of calcium signaling related genes has been discussed in the context of brain disorders such as autism spectrum disorder18. Diagnosis was based on electroencephalogram and brain magnetic resonance imaging. Informed consent forms were signed by all participants. The study protocol was permitted by ethical committee of Shahid Beheshti University of Medical Sciences.In total, 40 patients with refractory seizures and 71 healthy persons  were enrolled in the current study. Patients were classified according to the definition of International League Against Epilepsy (ILAE), “failure of adequate trials of two tolerated, properly selected and used antiepileptic drug plans (whether as monotherapy or in combination) to accomplish continued seizure freedom”In this assay, we measured expression of six genes (coding and non-coding) which are shown in Table SLC1A1, SLC25A12, ATP2B2 and their related lncRNAs, namely LINC01231, lnc-SLC25A12, and lnc-MTR-1 were measured in the peripheral blood obtained from 39 patients with refractory epilepsy and 71 healthy controls. Expression levels were assessed using the comparative − delta Ct method. The normal/gaussian distribution of the values was evaluated using the Shapiro–Wilk test. Unpaired t test or non-parametric test (Mann–Whitney U test) was used to identify differentially expressed genes between two groups. Effects of main factors on gene expression were determined using the two-way ANOVA and Tukey post hoc tests. Correlations between expression levels of genes were measured using the Spearman’s rank correlation coefficient.Statistical analysis was accomplished in GraphPad Prism version 9.0 . Expression levels of three genes encoding ion channels and transporters, namely All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent forms were obtained from all study participants. The study protocol was approved by the ethical committee of Shahid Beheshti University of Medical Sciences. All methods were performed in accordance with the relevant guidelines and regulations.The study included 39 patients with refractory epilepsy . Moreover, we included 71 healthy persons as controls .Expression levels of SLC1A1, SLC25A12, ATP2B2 and their related lncRNAs were significantly different between patients with epilepsy and healthy subjects  had a significant effect on expression levels of SLC1A1, SLC25A12, LINC01231 and lnc-MTR-1. On the other hand, gender factor and interaction between gender and group had no effect on expression levels of any of genes  for differentiation of epileptic cases from controls. Moreover, lnc-MTR-1 and lnc-SLC25A12 were sensitive markers for such purpose . The highest value belonged to LINC01231 with the value of 0.76  are associated with autism spectrum disorder26. In spite of up-regulation of this gene in epileptic cases, expression of the related lncRNA with this gene (lnc-SLC25A12) was not different between cases and controls. This finding might suggest that the role of this mRNA coding gene in epilepsy is independent from the related lncRNA.In line with our findings, mutations in We also noticed that the most robust correlations were between lnc-MTR-1/ATP2B2, ATP2B2/lnc-SLC25A12 and lnc-MTR-1/lnc-SLC25A12 pairs among epileptic cases. In fact, the strongest correlation was detected between ATP2B2 and its related lncRNA, i.e. lnc-MTR-1. However, the correlations between other mRNA coding genes and their related lncRNAs were not so strong.Remarkably, SLC25A12, lnc-MTR-1 and lnc-SLC25A12 were found to be sensitive markers for differentiation of epileptic cases from controls. However, the specificity of these transcripts were not appropriate. Taken together, this study demonstrates dysregulation of calcium-signaling related genes in epileptic patients and suggests these genes as potential biomarkers for epilepsy.Supplementary Information.


Atrioventricular block (AVB) is a serious complication following coronary artery bypass grafting (CABG) surgery, and its high-grade form may necessitate the implantation of a permanent pacemaker (PPM). AVB is associated with increased morbidity and mortality rates. This study aims to estimate the incidence of AVB and subsequent PPM implantation after isolated CABG surgery.I2 statistic was reported to evaluate the heterogeneity of studies.We searched electronic databases of PubMed, Embase, and Scopus from inception to 18 November 2022. Clinical trials and observational studies reporting the incidence of post-CABG AVB or subsequent PPM implantation in adult patients were included. The total incidence for all included outcomes was calculated using the inverse variance method, and the A total of 28 studies met the inclusion criteria. Four studies  reported AVB without specifying its type; one (cohort) reported different degrees of AVB, 20  reported complete heart block (CHB) (or AVB requiring temporary pacing), and nine  reported the number of PPM inserted due to AVB. The pooled incidence of AVB, CHB (or AVB requiring temporary pacing), and PPM due to AVB was 1.16%, 1.73%, and 0.58%, respectively. Meta-regression analysis revealed that age, gender, diabetes, hypertension, hyperlipidemia, or smoking were not significantly associated with AVB, CHB, or PPM implantation.This study highlights the incidence of AVB and the need for PPM implantation following CABG surgery. The findings emphasize the importance of postoperative monitoring and surveillance to improve patient outcomes.https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42022377181, identifier PROSPERO CRD42022377181. It is recommended when a severe blockage in one of the major coronary arteries cannot be relieved by percutaneous coronary intervention , 2. ApprPatients with high-grade AVB have a higher risk for syncope, congestive heart failure, ventricular tachycardia, asystole, or sudden cardiac death –17. HighDespite the importance of this issue, few studies have investigated the incidence of AVB after CABG, and to our knowledge, no systematic review or meta-analysis has been performed on this subject. The present systematic review and meta-analysis aims to estimate the incidence of AVB and subsequent PPM implantation following CABG and to review associated risk factors. It helps to better understand the significance of AVB as a complication of CABG and to build the foundation for preventive measures.2.2.1.This systematic review was conducted according to the Preferred Reporting Items for Systematic Review and Meta-analysis (PRISMA) statements . The stu2.2.The inclusion and exclusion criteria were as follows: randomized clinical trials and observational studies reporting the incidence of AVB or PPM implantation due to AVB after isolated CABG surgery in adult patients (18 years or older) were included. Animal studies, studies not in English, conference abstracts, review articles, research letters, or studies whose full texts were not available were excluded. After the initial search and removing duplicates, two authors screened the titles and abstracts to identify relevant studies. The full texts of potentially relevant studies were independently reviewed following inclusion and exclusion criteria, and relevant data were extracted into a pre-defined Microsoft Excel spreadsheet . The way of consensus resolved any conflict in this step. The following variables were extracted from each study: first author, publication year, study design, sample size and the major cardiovascular risk factors , mean follow-up time, number of AVBs, complete heart blocks (CHB) (or AVBs requiring TP) and PPMs inserted due to AVB. We also extracted the list of risk factors reported to have a statistically significant relationship with the review outcomes from the included studies.It is worth mentioning that some studies also reported pacemaker implantations for indications other than post-CABG AVB, such as sinus bradycardia, bundle branch block, atrial fibrillation, and cardiac arrest. To maintain the focus and integrity of our analysis, we only entered data related to pacemakers that were specifically indicated for post-CABG high-grade AVB. Furthermore, while some included studies did not exclude patients with pre-existing AVB, and some did not explain whether they did, all of the studies clearly stated that they were reporting on newly developed AVBs after CABG. Due to the limited availability of studies that exclude patients with pre-existing AVB, we deemed it appropriate to include all relevant studies that reported post-CABG AVB.2.3.Two reviewers independently evaluated the quality of each study, and disagreements were resolved with the help of a third reviewer. The quality of observational studies was assessed following the Newcastle-Ottawa Quality Assessment Scale . The sca2.4.The primary outcomes were the incidence of AVB, CHB (or AVB requiring TP), and PPM implantation. The secondary outcome was to determine if major cardiovascular risk factors are associated with the occurrence of primary outcomes.AVB is a significant complication after CABG surgery that may range from transient and self-resolving to persistent and requiring medical intervention such as pacemaker implantation. High-grade AVB or AVB requiring TP increase morbidity and mortality , 22. UndThe frequency of desired outcomes and methods used to identify them in each study are shown in 2.5.τ2) was calculated through the restricted maximum-likelihood estimator (REML)  format for continuous variables and percentage (number) for categorical variables. The total incidence for all included outcomes was calculated using data from the longest follow-up of each study and through Freeman-Tukey double arcsine transformation and back transformation in the inverse variance method . Betweenr (REML) . We alsoFurthermore, a meta-regression analysis was conducted to assess the relationship between desired outcomes and risk factors such as age, gender, diabetes, hypertension, hyperlipidemia, and smoking. We assessed publication bias using funnel plots and Egger's test. However, there is no universally accepted definition of what constitutes a positive result in a proportional meta-analysis. The presumption that positive results are more frequently published is not always valid for proportional studies , 50. R P3.3.1.The initial search of electronic databases returned 1,228 documents. After removing duplicates and initial screening, the full text of 135 articles was evaluated for inclusion and exclusion criteria. Finally, reviewers extracted the data of the thirty included articles into pre-defined spreadsheets. The PRISMA flowchart of the study is presented in 3.2.The baseline demographic features and comorbidities of patients have been presented in 3.3.τ2 = 0.003, I2 = 88.7%; p-values of 0.425, 0.273, 0.727, and 0.611, respectively).Five studies  reported AVBs following CABG. The pooled incidence of AVB was 1.16% [95% CI , 3.4.τ2 = 0.007, I2 = 96.0%; p-values of 0.972, 0.110, 0.366, 0.929, 0.569 and 0.619, respectively).Twenty-one studies  reported the incidence of post-CABG CHB or AVB requiring TP, resulting in an aggregate incidence of 1.73% [95% CI , 3.5.τ2 = 0.002, I2 = 73%; p-values of 0.997, 0.947, 0.784, 0.858, and 0.978, respectively).The pooled estimate for post-CABG pacing was calculated using data from nine studies  and equaled 0.58% [95% CI , 3.6.p-values of Egger's test for AVB, CHB, and permanent pacing were 0.1821, 0.8425, and 0.0623, respectively.Funnel plots were used to evaluate publication bias for all three outcomes . The plo4.To our knowledge, this is the first systematic review and meta-analysis of the incidence of postoperative AVB and subsequent pacing requirements in patients undergoing isolated CABG surgery. The current meta-analysis found that after isolated CABG, the incidence of AVB in 4,116 patients was 1.16%, the incidence of CHB or AVB requiring pacemaker implantation in 10,707 patients was 1.73%, and the percentage of PPM implantation due to advanced AVB in 6,851 patients was 0.58%. Based on these results, 0.33 or one-third of patients with CHB or AVB requiring TP will need PPM implantation. In the meta-regression analysis, age, gender, diabetes, hypertension, hyperlipidemia, or smoking had no significant association with AVB, CHB, or PPM implantation. However, these variables were reported for study populations, and we were not able to access the individual data required for a more robust analysis. In this review, we considered studies that reported AVB without further explanation as high-grade AVB due to the similarity between their incidence rates and those that reported high-grade AVB requiring TP.The results of this study show that the incidence of high-grade AVB and the subsequent need for PPM implantation is a serious complication after CABG surgery. Therefore, it is crucial to take preventive measures, especially in high-risk patients. For this purpose, the risk factors should be further investigated, and then clinical guidelines should be designed to identify high-risk patients.4.1.I2 of 96.0%. The overall heterogeneity for AVB and PPM implantation was lower . Since the diagnosis method was serial electrocardiograms in most studies, it does not seem to be the source of variability. Instead, two outlier studies in the CHB group  who needed pacing after releasing the ACC, only five (10%) required it upon discharge from the operating room . Accordi4.2.Literature has reported many risk factors associated with post-CABG AVB and the subsequent need for pacing. Here we review the risk factors in the studies we included in our meta-analysis. These consisted of old age, gender, LVD, perioperative MI, number of vessels bypassed, longer ACC time, perioperative use of an intra-aortic balloon, preoperative arrhythmia, and the type of cardioplegia .One of the most frequently mentioned risk factors was age. In four studies, old age was associated with a higher incidence of AVB after CABG or after releasing the ACC. Four studies had a significant association between LVD and post-CABG AVB or pacing. In a large cohort study, Pianta et al. found that perioperative MI increased post-CABG AVB requiring TP, but the same was not true for patients with previous MI. In the same study, post-CABG AVB was more frequent in patients with preoperative atrial fibrillation . In otheThe type of cardioplegia can be a risk factor since different types of cardioplegia methods have been related to increased postoperative conduction disorders. A study by Cook et al. suggested that crystalloid cardioplegia increased post-CABG conduction defects compared with blood cardioplegia . Gozal eThe other factor that should be further investigated is the effect of medications. Pianta et al. found that the incidence of high-grade AVB after CABG was similar between patients using beta-blockers and those without . In a coBecause only one study  specific4.3.Preventing post-CABG AVB and its related complications is crucial to avoid adverse consequences. Several studies suggest the incidence of AVB after CABG surgery is associated with increased mortality. Pianta et al. indicated AVB requiring TP prolonged hospital stay and increased mortality after CABG . In a st4.4.There are several restrictions on how broadly applicable our findings can be. First, we only searched three online databases and included full-text English journal articles, which may have resulted in selection bias. Second, five studies reported post-CABG AVB without further explanation of its type, which limited analysis of the incidence of different types of AVB. However, we analyzed them separately to prevent the pooling of heterogeneous data. Third, studies did not separately report characteristics in patients with and without AVB, except in one . Consequ4.5.Our study provides important insights into the incidence of post-CABG AVB. The pooled prevalence rates calculated for the incidence of CHB (or AVB requiring TP) and PPM implantation can serve as reference points for clinicians. These incidence rates highlight the need for increased postoperative surveillance and monitoring to reduce the risk of high-grade AVB sequelae. The identification of high-risk patients should be prioritized. Risk factors such as age, preoperative arrhythmia, LVD, perioperative MI, and the use of certain medications should be considered when assessing the patients.As for future research, we recommend conducting large-scale prospective cohort studies investigating the incidence of post-CABG AVB, its risk factors, and its long-term prognosis. These studies would provide a more comprehensive understanding of the multiple factors influencing post-CABG AVB and facilitate the development of risk prediction models. Moreover, including other patient-centered outcomes, such as quality of life measures and healthcare resource utilization, would contribute to a more holistic assessment of the impact of post-CABG AVB. The STROBE guidelines should be used for complete and transparent reporting.1.Determine whether the study focuses on developing new-onset AVB or worsening pre-existing AVB.2.Identify the type of AVB of interest .3.Determine the time frame for defining post-CABG AVB .4.Define the criteria for determining the presence of AVB. It could include identifying new electrocardiographic findings consistent with the type of AVB of interest.5.•Pre-existing AVB.•pre-existing pacemakers or implantable cardioverter-defibrillators.•Previous cardiac surgeries.•Severe comorbidities that may affect the study outcomes, such as mortality.•Use of medication that can exacerbate AVB.•Inadequate medical records.Provide clear exclusion criteria. It could include:One of the most significant limitations of this review was the nonuniform definitions and reporting across different studies. It underscores the importance of a uniform definition and diagnostic criteria for post-CABG AVB, which should be concise, clear, and clinically relevant. This consistency will allow for more accurate comparisons and assessment of the incidence and outcomes of post-CABG AVB across different studies. The following are our recommendations in this regard:5.The current study was designed to determine the overall incidence of AVB, CHB, and subsequent PPM implantation following the CABG procedure and identify its associated risk factors. We calculated a pooled prevalence of 1.16% for postoperative AVB incidence, 1.73% for CHB or AVB requiring TP, and 0.58% for PPM implantation. These findings underscore the significance of AVB as a serious complication of CABG, emphasizing the need for postoperative monitoring and surveillance to ensure satisfactory patient outcomes. As for further research, we recommend conducting large-scale prospective cohorts with primary outcomes focusing on the incidence, risk factors, and prognosis of post-CABG AVB to yield more precise and comprehensive results. Using a uniform definition and diagnostic criteria for post-CABG AVB and adhering to the reporting standards will allow for more accurate comparisons and assessment of the incidence and outcomes of post-CABG AVB across different studies.


The mean age of onset was 59.5 years. The severity of this disease was evenly distributed between mild (147 patients) and severe (149 patients) forms. There was a positive, statistically significant, medium correlation between the severity of the disease and its time of progression. Moreover, hypothyroidism was present in 70 patients (22.9%) and classic signs of concomitant lichen planopilaris were observed in just 30 patients (9.8%), while other forms of lichen planus were uncommon. The estimated prevalence in our population is 0.15% and the incidence is 15.47 new cases per 100,000 inhabitants; (4) Conclusions: The time of progression was positively correlated with the severity of FFA. However, the presence of clinical signs, such as inflammatory trichoscopic signs, was not associated with the progression of this condition. Frontal fibrosing alopecia (FFA) is a primary scarring and irreversible type of alopecia first described in a group of postmenopausal women by Kossard in 1994 . This coInterestingly, FFA was initially described as a lichen planopilaris (LPP) variation by Kossard himself in 1997 , since tThe FFA pathogenesis still remains unknown, although there seems to be hormonal factors involved, as the majority of patients affected are women. Actually, most affected women are postmenopausal, suggesting that ovary hormones could play a protective role in the hair follicles of the affected zones ,9. AccorThe clinical course of FFA is variable but usually entails an insidious progression which may stabilize after several years . In relaDespite its increasing incidence, the pathogenesis of FFA remains unknown and its treatment still represents a challenge. In this context, the aim of this study was to describe the epidemiology, clinical and trichoscopic features, diagnostic findings and comorbidities of a series of 306 patients with FFA.Studied population:A retrospective, observational single-center study was conducted in the Hospital General Universitario in Ciudad Real, Spain. In this study, patients with a diagnosis of FFA between 2010 and 2021 were included. All diagnoses were made following well-established diagnostic criteria which included clinical, trichoscopic and even histopathological aspects, as previously described . Whe. Whe1]. Whether AFF is a variant of LPP or a distinctive entity is yet to be elucidated. Both pathologies share primary histological findings, although clinically LPP often produces alopecic plaques in the vertex and parietal scalp rather than a homogeneous recession on the frontotemporal hairline ,7. HistoRegarding treatments for FFA, there are no standardized guidelines for these patients, since its etiopathogenesis remains to be clarified. The goals of treatment of this condition are to reduce clinical signs of inflammation, stop the progression of alopecia and appease clinical symptoms. Potent topical corticosteroids and calcineurin inhibitors reduce the inflammation signs and ameliorate symptoms, but do not seem to interfere with the progression of alopecia ,44.Among systemic therapies, 5-alpha reductase inhibitors appear to be the most effective treatment, as they seem to achieve the highest stabilization rates ,44,45. OOur study has some limitations, the primary one being the observational and retrospective design, as it limits the collection of representative data. In addition, it is still necessary to evaluate the potential efficacy of the diverse treatments which are in used and their correlation with FFA progression. Despite these limitations, the single-center nature of the study provides a highly homogeneous assessment from the investigators involved and these results could be considered representative of this locality.In conclusion, this is probably the largest retrospective single-center study analyzing a cohort of FFA patients reported in the literature. FFA is a scarring type of alopecia affecting mainly postmenopausal women, inducing a recession of the frontotemporal hairline. We found a slightly increased number of premenopausal women in our study compared to previous ones (16%). Its pathogenesis still remains unknown, although there is evidence that hormonal factors, genetic susceptibility, autoimmunity and some exogen factors may play an important role. The grade of severity of the alopecia is correlated with the time of progression of the disease. We found eyebrow alopecia was associated with milder forms of FFA, although we did not find an association between the grade of severity and the presence of facial papules or hair body alopecia, as other studies suggest. In our study, the presence or absence of inflammatory trichoscopic signs did not seem to relate to the progression and therefore the severity of FFA, which might change our therapeutic approach for this entity. In addition, to our knowledge, this is the first study conducted in this particular population that shows the incidence, prevalence and main clinical features associated with this entity. Moreover, it is important to conduct more retrospective or prospective clinical studies to develop new quantitative diagnostic criteria to establish and grade the diagnosis of FFA. Finally, histological and molecular studies are still needed to elucidate the biological processes that could be involved in the genesis, progression or response to new treatments of this pathological condition.


It is the most practical goal of limb rehabilitation for stroke patients to make the upper limb, trunk, and lower limb joints link together as a whole and restore the ability to self-care. However, many previous studies focused on the single joint or single muscle group movement of stroke patients and did not integrate self-care ability training into the whole process of rehabilitation, which lacks accuracy, integrity, and systematization.n = 80) and a control group (n = 80) by the medical district. The control group received the routine physical rehabilitation intervention. The experimental group adopted the physical rehabilitation program based on self-care ability led by the nurses specializing in stroke rehabilitation to carry out the multi-joint coordinated exercise based on the control group. The training time and frequency were the same in both groups . The primary outcome was myodynamia. Secondary outcomes were the modified Barthel Index (MBI) and Stroke Specific Quality of Life Scale (SS-QOL). The primary and secondary outcomes were assessed before the intervention and at 1 and 3 months of intervention. In this study, the TREND checklist was followed for non-randomized controlled trials.A quasi-experimental study was conducted in a tertiary hospital. Eligible patients were recruited according to the inclusion criteria and exclusion criteria and then divided into an experimental group (P < 0.05), and the myodynamia of lower limbs recovered faster than that of upper limbs. In the control group, the myodynamia of the affected limb was not significantly improved (P > 0.05), with only a small increase in MBI and SS-QOL scores (P < 0.05).A total of 160 participants completed the study. The physical rehabilitation program based on self-care ability was better than the routine rehabilitation program. With the prolongation of intervention time, all outcomes improved gradually in the experimental group (The physical rehabilitation program based on self-care ability after stroke was beneficial for acute ischemic stroke patients and improved the patient's myodynamia, quality of life, and self-care ability within the third month. Stroke has become the leading cause of death and disability worldwide , 2. AcutSelf-care ability assessment is not only the basis of treatment, nursing, and rehabilitation but also one of the most important indicators to evaluate the effect of rehabilitation for AIS patients , 10. StuThe improvement of motor function and self-care ability in AIS patients is a slow and long-term process . HoweverThe physical rehabilitation training of AIS patients is a gradual and complex process, requiring the coordination of multiple joints and muscle groups , 19. MorOverall, focusing on the five levels of self-care ability, this study constructed and implemented a physical rehabilitation program based on the self-care ability of AIS patients and carried out a 3-month intervention. The starting point of rehabilitation training was determined by rehabilitation therapists and neurology physicians according to the patient's self-care ability. The continuous rehabilitation plan, which included admission and discharge, was implemented by nurses specializing in stroke rehabilitation. This study developed a systematic, accurate, and continuous limb rehabilitation program for AIS patients, aiming to improve the patient's myodynamia, self-care ability, and quality of life and provide a reference for the rehabilitation of stroke patients.Patients were included if they were (1) diagnosed with AIS according to the Chinese guidelines for the diagnosis and treatment of AIS 2018 (Patients were excluded if they (1) had evidence of intracranial hemorrhage or other cerebral diseases ; (2) had severe complications such as liver or renal dysfunction; (3) had evidence of massive ischemic stroke  ; (4) werThis study was approved by the Medical Ethics Committee of Shaanxi Provincial People's Hospital . All the participants provided written informed consent.All the stroke physical rehabilitation therapists involved in this study obtained health professional and technical certificates approved and issued by the National Health and Family Planning Commission and the Ministry of Human Resources and Social Security of the People's Republic of China. They have been engaged in stroke limb rehabilitation for more than 5 years.n = 80) and control group (n = 80). The purpose of this study was to compare the effects of physical rehabilitation programs based on self-care ability led by nurses specializing in stroke rehabilitation with routine physical rehabilitation intervention. All participants were informed of the purpose of the study and agreed to participate in the study. The TREND checklist was followed for non-randomized controlled trials  During the hospitalization, rehabilitation therapists and doctors assessed the patient's condition, determined the level of self-care ability, and the starting point of rehabilitation according to the patient's MBI score and actual physical condition. Twenty-four hours after the patient's vital signs were stabilized, the rehabilitation training plan was implemented by the nurses specializing in stroke rehabilitation. (2) The WeChat group of physical rehabilitation for patients in the experimental group was established, and the group members included members of the multidisciplinary rehabilitation team, patients, and their caregivers. Nurses specialized in stroke rehabilitation pushed videos of the physical rehabilitation training to the group. (3) Before discharge, patients were informed to do physical rehabilitation training at home every day after discharge, and caregivers were informed to record the process of the patient's physical rehabilitation training in the form of video and feedback in the WeChat group every 2 weeks. (4) The patients entered the WeChat group upon discharge, and nurses specializing in stroke rehabilitation again explained and demonstrated to them and their caregivers the movement essentials and precautions of the rehabilitation training. (5) After discharge, the caregivers filmed the process of the patients' physical rehabilitation training at home in a video and provided feedback in the WeChat group once every 2 weeks. In the WeChat group, doctors and nurses specializing in stroke rehabilitation and rehabilitation therapists continued to follow up on the patient's training process and provided targeted guidance for the content of the feedback. For patients who did not provide feedback in time, nurses specializing in stroke rehabilitation reminded and urged patients to carry out physical rehabilitation at home via WeChat, and guided patients online to carry out physical rehabilitation training.In order to improve the rehabilitation compliance of patients after discharge, the following measures were mainly adopted in this study. First of all, it not only provided patients with professional guidance on physical rehabilitation but also regularly arranged training for caregivers. During the period of hospitalization, the nurses specializing in stroke rehabilitation carried out rehabilitation training education for the caregivers in the teaching classroom on Monday, Wednesday, and Friday. Second, when the nurse implemented rehabilitation training for patients in the ward, she also explained training methods and matters needing attention for caregivers. After assisting the patients with rehabilitation training, the nurse instructed the patients to repeat the same training. Before the patients were discharged, the nurse explained and demonstrated the method of physical rehabilitation training for the caregivers again. When patients were discharged from the hospital, the Department of Neurology arranged online rehabilitation courses free of charge for caregivers every 2 weeks and pushed rehabilitation training videos in the WeChat group. The nurses specializing in stroke rehabilitation urged and guided caregivers to consolidate rehabilitation actions when urging patients to provide feedback on WeChat every 2 weeks. In addition, different rewards were given for the consecutive times of feedback.Routine drugs and rehabilitation therapy were given to patients in the Department of Neurology every day. Within 24 h after the patient's condition stabilized, nurses in the Department of Neurology provided rehabilitation training guidance for them and their caregivers, mainly with passive and active joint activity training in bed with the single joint or single muscle group and walking training assisted by walking AIDS. After discharge, patients were also urged to perform routine limb rehabilitation training. The training time, intensity, frequency, caregiver training, and compliance improvement methods of patients in the control group were the same as those in the experimental group.The primary outcomes were the six levels of the myodynamia assessment method commonly used in clinical practice. Level 0 myodynamia is complete paralysis of the limb, and muscle contractions cannot be seen or measured by the eye. The myodynamia of level 1 is that the muscle contractions can be seen by the eye, but no movement can be formed. Level 2 myodynamia is the ability of the limbs to move in parallel on the bed, but it cannot resist gravity and be lifted off the bed surface. Level 3 myodynamia is that the limbs can resist gravity and lift off the bed surface but cannot resist resistance. Level 4 myodynamia is the ability of the limb to resist only partial resistance. Level 5 myodynamia indicates that the patient has normal myodynamia.The secondary outcomes were the MBI and SS-QOL scales. The self-care ability was evaluated by the MBI, comprising 10 items including eating, dressing, bathing, and grooming , with a Myodynamia, MBI, and SS-QOL were assessed at baseline (t0), 1 month (t1), and 3 months (t2) of intervention. We also recorded sociodemographic and clinical data. Patients directly reported any unintentional injuries and serious adverse events during hospitalization or follow-up.t-test, chi-square, or Fisher's exact test. A non-parametric test was used when the data did not conform to normal distribution. The Mann–Whitney U-test in rank-sum test was used to compare grade data. The repeated measurement data of outcomes were analyzed using GEE or analysis of variance of repeated measurement data. Data analysis was performed using SPSS V.25.0 .Descriptive statistics were applied, with results provided as numbers and percentages for categorical variables and means with SD for continuous variables. When the data conform to the normal distribution, between-group data were compared using the independent P > 0.05). These results suggested that the groups were comparable at baseline  and t1 . However, the myodynamia of affected limbs in the experimental group was better than that in the control group at t2, and the difference between the groups was statistically significant (P < 0.01). In the experimental group, only the difference in myodynamia at 3-time points before and after the intervention was statistically significant (P < 0.01). By pairwise comparison, myodynamia was gradually improved with the extension of the intervention time . The MBI score of patients in the control group was only significantly higher at t2 than that at t0 . However, the MBI scores in the experimental group were higher than those in the control group , and the SS-QOL scores of the two groups at t2 were higher than those that at t1 (P < 0.01). Moreover, there was an interaction effect between time and group . With the extension of intervention time, the SS-QOL scores of the two groups gradually increased, and the increased range of the scores of the experimental group was greater than that of the control group (Repeated measurement analysis of variance by Greenhouse–Geisser corrected showed that the SS-QOL scores of the experimental group were higher than those of the control group after intervention (ol group .This quasi-experimental study found that a physical rehabilitation program based on self-care ability was superior to a routine physical program. In the experimental group, myodynamia, self-care ability, and quality of life of patients were significantly improved, and myodynamia of the affected lower limb recovered faster than that of the upper limb.Dyskinesia in stroke patients was mainly manifested in abnormal movement patterns . SpasticMyodynamia of lower limbs recovered faster than that of upper limbs, which may be related to the following factors. The lower limb sensorimotor area of the brain received a dual blood supply from the middle cerebral artery and the anterior cerebral artery, while the upper limb sensorimotor area received a single blood supply . The uppThe quality of life of AIS patients depends on the function of their affected limbs and their self-care ability . It has Therefore, it was necessary to construct a physical rehabilitation program for AIS patients based on self-care ability. We should not only attach great importance to the rehabilitation of stroke patients but also provide more systematic and accurate physical rehabilitation strategies. In addition, the hospital should also reasonably allocate medical resources and strengthen the training of nurses specializing in stroke rehabilitation so as to improve patients' self-care ability and quality of life.This study had several limitations. First, it is a single-center study with a limited sample size, and the applicability and generalizability of the findings will be limited. Further multicenter studies with larger sample sizes are needed to support our conclusions. Second, considering the cooperation and compliance of patients at follow-up, our study only evaluated the effects of physical rehabilitation programs on myodynamia, quality of life, and self-care ability. Third, most of our data were collected from rehabilitation scales and lacked sufficient quantitative and objective measures, which may introduce some bias to the evaluation. Finally, this rehabilitation method can cause any potential adverse reactions such as falls, bed falls, or pressure sores. However, before the implementation of the study plan, we have developed the relevant contingency plans. During the implementation process, we also modified and improved the contingency plan continuously according to the actual situation. In future studies, we will apply objective measures, increase interventions, and increase the sample size to carry out multicenter clinical trials to identify risk factors for AIS patients, as well as improve and refine the follow-up system, enhance communication with patients, and increase outcome indicators.The physical rehabilitation program for AIS patients based on self-care ability may be beneficial, as it improved the patient's myodynamia, quality of life, and self-care ability within the third month. Due to the limited time, this study was only conducted in a tertiary hospital in China for 3 months. It is necessary to conduct independently replicated studies with large samples to validate our findings.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by Medical Ethics Committee of Shaanxi Provincial People's Hospital. The patients/participants provided their written informed consent to participate in this study.RH and Y-PZ designed the study, oversaw data safety and monitoring, and revised the manuscript. YL performed the analysis and drafted the manuscript. QW and X-LL were site principal investigators and oversaw site recruitment and site data collection. All authors contributed to the manuscript's initial draft, read, and approved the final manuscript.