Milvus milvus) broods across an elevational gradient toward the upper range margin and we GPS‐tagged nestlings to assess their start of dispersal. While considering timing of breeding and breeding density across elevation, this allowed disentangling effects of elevational food gradients from co‐varying environmental gradients on the age at departure from the natal home range. We found an effect of food supplementation on age at departure, but no elevational gradient in the effect of food supplementation. Similarly, we found an effect of breeding density on departure age without an underlying elevational gradient. Supplementary‐fed juveniles and females in high breeding densities departed at younger age than control juveniles and males in low breeding densities. We only found an elevational gradient in the timing of breeding. Late hatched juveniles, and thus individuals at high elevation, departed at earlier age compared to early hatched juveniles. We conclude that favorable natal food conditions, allow for a young departure age of juvenile red kites. We show that the elevational delay in breeding is compensated by premature departure resulting in an elevational gradient in departure age. Thus, elevational differences in dispersal behaviour likely arise due to climatic factors affecting timing of breeding. However, the results also suggest that spatial differences in food availability and breeding density affect dispersal behavior and that their large‐scale gradients within the distributional range might result in differential natal dispersal patterns.Attributes of natal habitat often affect early stages of natal dispersal. Thus, environmental gradients at mountain slopes are expected to result in gradients of dispersal behavior and to drive elevational differences in dispersal distances and settlement behavior. However, covariation of environmental factors across elevational gradients complicates the identification of mechanisms underlying the elevational patterns in dispersal behavior. Assuming a decreasing food availability with elevation, we conducted a food supplementation experiment of red kite ( Milvus milvus) along an elevational gradient toward the upper range margin. We found an effect of food supplementation and of breeding density on age at departure, as well as an elevational gradient in timing of breeding. We show that the elevational delay in breeding is compensated by premature departure resulting in an elevational gradient in departure age and therefore conclude that elevational differences in dispersal behavior likely arise due to climatic factors affecting timing of breeding.By conducting a food supplementation experiment, we aimed at disentangling food availability and co‐varying elevational gradients affecting the onset of dispersal in juvenile red kites ( We tracked large samples of juvenile red kites from fledging onwards, enabling identification of the age at departure across an elevational gradient. We disentangled the effect of potential elevational gradients in food availability from the effect of other elevational gradients on the age at departure by using a food supplementation experiment across elevations. We controlled for effects of gradients in hatching date and breeding density as well as for the potential confounding effects of hatching rank, sex, and potential correlations with body mass at fledging using statistical models. We assume that availability and accessibility of food decreases towards to high margin of the elevational gradient due to associated changes in habitat structure and seasonal weather conditions  and managed forests (26.95%). The agricultural parts are dominated by crops and dairy farming, as well as meat production, resulting in large areas with meadows .The red kite is a medium‐sized diurnal raptor species with a pan‐European distribution and an opportunistic feeding behavior Carter, . In Swits StatA,  and, thus StatA, . While as StatA, . The stus StatA, . The low2.22 squares. We repeatedly visited territories until a nest was located, and we then monitored nests by observations to confirm incubation and hatching of the nestlings. If the growth of brown covert feathers was observed (nestling age 15–20 days), we climbed the nest to ring and measure nestlings. Based on wing length measurements, we approximated age using a generic growth curve by Aebischer (3D DEM (© swisstopo 2017). During the 2 years, a total of 239 nestlings were equipped with solar powered GPS‐GSM‐UHF transmitters (Ecotone SKUA/CREX type) using backpack style diagonal‐loop harnesses  when they reached at least 670 g in body weight in order to fit the harness properly, but were not yet at risk of escaping capture due to impending fledging .Red kite territories were identified by a standardized territory survey in 2.5 x 2.5 kmebischer  in orderKenward, , 2001 wiAt tagging, we measured body mass, tarsus length, and the length of the eighth primary feather. This allowed us to determine the exact hatching date based on a study area‐specific primary feather growth curve Nägeli, . In addi2.3ad libitum supplementation setting. We assessed whether the food supplementation was accepted by observing the platform and nest shortly after a feeding event using either binoculars or video cameras.In order to disentangle elevational gradients of food availability from other environmental gradients across elevations, we conducted a supplementary feeding experiment. We selected nests in a stratified design along the elevational gradient . Every other day, we placed five dead day‐old chickens per adult and per nestling <10 days, and 10 chickens per nestling >10 days of age on wooden platforms located 20–200 m away from the target nests until the nestlings were close to fledging, that is, approximately 52 days old. Thus, food availability during nestling development, but not in the post‐fledging period was experimentally increased. The provided food exceeds the known daily food requirements of red kite nestlings  showed that departures from the natal home range exceeding 2 days are abrupt large‐scale movements and returns to the 2‐km radius for an entire day occur rarely  a radius surrounding the nest, and (b) a duration in which the individual continuously had to stay outside the radius. This spatio‐temporal threshold assumes that once the individual stays outside the radius longer than the temporal threshold, it can be considered as having departed from the parental home range. We used a 2 km radius surrounding the nest as spatial threshold. This value was based on known breeding home range sizes of adult male breeding birds in our study area Baucks, . Daily fNon‐breeding red kites tend to form social roosts to spend the night. If social roosts occur near the natal home range, independent individuals show higher probability to incidentally return to the natal home range during roosting. The distance to the closest communal roost was therefore considered as a control variable in order to account for confounding effects of roosts located close to natal nest. The roosts were identified based on night‐time positions of tagged non‐breeders (including juveniles) in the study area with an external buffer of 10 km. We calculated the number of revisits in a radius of 50 m around each position for each year from July to September using the R‐package “recurse” . In addition, individuals that did not depart from the natal home range before starting migration were excluded (n = 17) as start of migration is likely mediated by a different set of factors than the departure to dispersal. This resulted in a total of 158 juveniles which were included in further analyses.If juveniles died or if their transmitters stopped working before departure from the natal home range, they were excluded from the analysis . There was considerable variation in the age at departure among individuals ranging from 61 to 115 days  and 108 individuals from 72 broods without food supplementation . The median age at departure was 83 days  might affect competitiveness of individuals Emlen, , timing Patrick Scherler: Conceptualization ; data curation ; formal analysis (lead); funding acquisition (supporting); investigation ; methodology ; project administration ; resources ; software ; supervision ; validation (lead); visualization (lead); writing – original draft (lead); writing – review and editing . Stephanie Witczak: Data curation ; formal analysis (supporting); funding acquisition (supporting); investigation ; methodology ; project administration ; resources ; writing – original draft (supporting); writing – review and editing . Adrian Aebischer: Conceptualization (supporting); data curation (supporting); investigation (supporting); methodology (supporting); project administration (supporting); writing – original draft (supporting); writing – review and editing (supporting). Valentijn van Bergen: Data curation ; investigation ; project administration ; resources ; validation ; writing – review and editing . Benedetta Catitti: Conceptualization (supporting); data curation ; investigation (supporting); project administration ; resources ; validation ; writing – review and editing . Martin U. Grüebler: Conceptualization ; data curation (supporting); formal analysis (supporting); funding acquisition (lead); investigation ; methodology ; project administration ; resources ; supervision (lead); validation (supporting); visualization (supporting); writing – original draft (supporting); writing – review and editing .This work was funded by the Swiss National Science Foundation (Grant 31003A_169668 to M. U. Grüebler).We declare that none of the authors is subject to any conflict of interest.Appendix S1Click here for additional data file.


The dorsal skinfold chamber is one of the most important in vivo models for repetitive longitudinal assessment of microcirculation and inflammation. This study aimed to refine this model by introducing a new lightweight chamber made from polyetheretherketone (PEEK). Body weight, burrowing activity, distress, faecal corticosterone metabolites and the tilting angle of the chambers were analysed in mice carrying either a standard titanium chamber or a PEEK chamber. Data was obtained before chamber preparation and over a postoperative period of three weeks. In the early postoperative phase, reduced body weight and increased faecal corticosterone metabolites were found in mice with titanium chambers. Chamber tilting and tilting-related complications were reduced in mice with PEEK chambers. The distress score was significantly increased in both groups after chamber preparation, but only returned to preoperative values in mice with PEEK chambers. In summary, we have shown that light chambers reduce animal distress and may extend the maximum dorsal skinfold chamber observation time. Chambers made of PEEK are particularly suitable for this purpose: They are autoclavable, sufficiently stable to withstand rodent bites, inexpensive, and widely available through 3D printing. Researchers investigated inflammation4, thrombogenesis7, wound healing9, angiogenesis, biomaterials10 and tumor vascularisation13 using the dorsal skinfold chamber16. Repetitive intravital visualisation of the microvascular dynamics is the major advantage of the model . Weight and skin stretching may lead to restricted breathing, immobilization, and pain. The severity of any dorsal skinfold chamber experiment is considered at least moderate. Therefore, the reputation of the model as a standard in microvascular research has been contested in the context of 3R 17. Weight, the chamber's high center-of-gravity, and overstretching of the skin (in particular at the fixation screws) lead to lateral tilting of the chamber in the second week after dorsal skinfold chamber preparation. Tilting comprises perfusion and causes animal distress . Chamber tilting of > 50° must be scored in the severity assessment and tilting of > 100° considered as an abort criterion. Therefore, experiments of up to 21 days are associated with high dropouts (20% in the third week) and low reliability . Experiments of longer than 21 days are impossible with standard chambers20.Lateral tilting of the dorsal skinfold chamber is another important limitation and strongly related to animal distress1. While 63% of the dorsal skinfold chamber studies were from German-speaking countries and mostly used titanium chambers, smaller titanium chambers are already standard in the US1. Research groups from Sweden and Asia proposed more advanced plastic chambers made from plexiglass, dacron and polyetheretherketone (PEEK)1.To overcome these limitations, the chambers were continuously revised. Schreiter et al. reviewed the developments until 201722. PEEK is a linear, semi-crystalline polymer that exhibits excellent mechanical and thermal properties. PEEK is bioinert and durable .For many years, PEEK has been used successfully in medicine as a replacement material for titanium to fabricate surgical devices, implants and prostheses26.Although many publications point out disadvantages of large titanium chambers, they are still used in most studies: Between 2017 and 2021, only four of 76 studies used non-titanium chambers  and general distress parameters . As bridging technology, the PEEK chamber may improve future in vivo research until sufficient in vitro and in virtuo models are established.This study examined animal distress of standard titanium dorsal skinfold chambers compared to a 3D printed design made of PEEK. Preparation of the chamber and intravital microscopy were comparable. PEEK chambers showed no signs of bites, chews or any other manipulation by the mice. In terms of lateral chamber tilting, animal weight loss and corticoid levels, the PEEK chamber was markedly superior.The new PEEK chamber was designed lower and lighter than titanium chambers (approx. 1.5 g instead of 3.8 g). The chamber was 3D-printed cost-effectively according to a reproducible protocol (€5.30 /chamber). The costs were lower compared to milled titanium chambers (€30–€110). The design and the low weight allowed the PEEK chamber to be secured with sutures Table . The traPEEK chambers significantly reduced lateral tilting in the third week Fig. . In the Postoperative weight loss was significantly reduced in PEEK chamber mice Fig. A. While Similar beneficial results for the PEEK chamber were found for faeces corticosterone metabolites . These sutures cause less trauma than previously used screws. After chamber implantation, the experiments run without differences to titanium chambers. In particular, there are no differences in the quality of the repetitive intravital fluorescence microscopy.17. By the third week, 50% of the titanium chambers tilt to a position of > 90°, which causes animal immobilization. In all PEEK chambers, tilting remained below 90° for three weeks. The only PEEK chamber that had a tilting of 50° in the second week remained constant in the following week. This is clearly because of the reduced weight. In contrast, titanium chambers continued tilting over time. Therefore, the stable position of PEEK chambers could extend the maximum duration of future experiments to four or five weeks.The most common complication of dorsal skinfold chamber experiments is lateral tilting of the chamber during the second week32.Besides decreased tilting in the third week, the use of PEEK chambers also reduced distress for the experimental animals in the postoperative period. Postoperative weight loss is significantly higher in titanium chamber animals and does not return to baseline values over three weeks. With PEEK chambers, on the other hand, the test animals reach their original weight as early as the second week. Consistent with our data, previous research described up to 15% postoperative weight loss for titanium chambers and decreased weight loss for non-metal dorsal skinfold chambers33. FCMs increased significantly after implantation of a titanium chamber while only a slight increase in FCMs was observed for PEEK chambers. Therefore, postoperative stress was primarily related to the titanium chamber and not to the surgery itself. In the intermediate and late phase, FCMs returned to baseline values in both groups. The design of future titanium chamber experiments should consider increased postoperative stress as a potential bias34.This difference in stress was confirmed by FCM measurement, a non-invasive measure of adrenocortical activityIn contrast to FCMs and body weight changes, mice in both groups did not differ in burrowing activity nor distress score. The distress score remained at a low level after the operation. The postoperative increase of 7/66 points in the distress score was statistically significant. However, the values remained in the lower range of the score.29. These smaller titanium chambers are sold commercially in the United States . Schreiter et al. describe the advantages of a self-designed small titanium chamber: postoperatively no recovery period was necessary, younger animals could be used and their stress was supposed to be reduced1. However, titanium chambers are not MRI compatible and screw fixation is necessary. Furthermore, titanium is difficult to process and cannot be manufactured in life science facilities.To our knowledge, this study is the first to investigate the distress of laboratory animals with dorsal skinfold chambers. Despite the lack of data on animal distress, many chamber improvements have already been published. These improvements were supposed to reduce animal distress and to enable MRI imaging. A simple development is a smaller titanium frame with an equally large observation window.17. The publication illustrated reduced tilting and supposed distress reduction, because of the lightweight Duracon material. However, quantification of tilting and distress was not performed. In addition, these early plastic chambers continued to use screw fixation35. These screws penetrate the skin and cause large wounds at the chamber basis.Innovative developments are chambers made of plastic, which have been used for years in Japan and the US. The first plastic chamber made of Duracon was described in 2003 by Ushiyama et al36. PEEK can therefore resist the bite of rodents. Furthermore, PEEK can be fabricated with additive manufacturing processes, which results in further advantages such as a high degree of geometric freedom, low production costs and the flexibility regarding the unique or single-part production37.A further weight reduction was achieved by using thermoplastic PEEK. PEEK is characterized by a high flexural modulus (3738 MPa) and tensile strength (100 MPa) compared to Duracon  and acrylic glass . and Seynhaeve et al.28. The PEEK chamber weight was as low as 1 g and 1.1 g, respectively. The chambers were fixed using sutures (Gaustad) or small screws (Seyhaeve). Mice fitted with the chambers showed a full capacity of motion, climbed, and gained weight as mice without chambers. We observed similar positive effects for PEEK chambers . In addition, we verified reduced distress using a standardized protocol. We have repetitively measured chamber tilting and found that the PEEK chambers significantly reduce tilting in the third week of the experiment. Lightweight chambers with reduced lateral tilting enable increased observation times of up to one month27. This is especially relevant for biomaterial and oncology research: dorsal skinfold chamber observation times of three to five weeks could enable to study long-term biomaterial integration 38. In oncology, longer observation times could significantly improve the model, since the growth of tumor cells already preoccupies large parts of the current maximum observation time39.The first lightweight PEEK chambers were introduced by Gaustad et alThe reduction in tilting was significant, although the measurement method had limitations, as the chamber position depends on the body position. The measurement was performed on anaesthetized animals in an upright position with all feet on the ground. However, when positioning the animals, slight deviations of the angles occurred.Another limitation of the study is that the standardized distress score does not focus on the immobilization of the test animals. However, the impediment of free movement, because of tilting and chamber weight, probably represents the main restriction for the experimental animals. Electronically recording of the animal mobility through tracking systems or recording of the time spent climbing the cage could increase the power of the stress analysis. However, using standardized distress scores enables comparisons to previous experiments.The low number (n = 6) of test animals may be considered another limitation of the study. However, the PEEK chamber was significantly superior in major outcomes, such as tilting and weight loss. Therefore, no additional experimental animals had to be included. Another limitation related to the study design is that the PEEK and the titanium chambers have different sizes. Hence, all conclusions are related to the design , but not to the material (PEEK vs. titanium). Low and lightweight titanium chambers may also decrease animal distress compared to large standard titanium chambers. Yet, we consider PEEK a more suitable material because it increases availability, enables imaging, and decreases costs. These main advantages render a light titanium group obsolete.In experiments with dorsal skinfold chambers, the animals are particularly stressed by classical titanium chambers. This setup should be revised, in the context of 3R. Despite the development of smaller and lighter chambers, most dorsal skinfold chamber experiments in recent years have continued to use titanium chambers. We have shown that lighter chambers can significantly reduce animal distress and even extend the maximum experiment duration. Chambers made of PEEK are particularly suitable for this purpose: They are autoclavable, sufficiently stable to withstand rodents, inexpensive, and widely available through 3D printing.The PEEK chamber was designed for geometric shape improvement, weight reduction, and optimization for additive manufacturing using SolidWorks . The Fused Filament Fabrication (FFF) process and the printer Minifactory ultra  were used for chamber printing. The design (*.stl file) was imported to Simplify3d . Biocompatible and steam sterilizable PEEK filament Intamsys Funmat HT  with a flexural modulus of 3738 MPa and tensile strength of 100 MPa was chosen. The material-dependent printing parameters were 230 °C chamber temperature, 190 °C bed temperature, 420 °C nozzle temperature 0.4 mm nozzle diameter, and 18 mm/s printing speed. Slicing was done according to the manufacture settings with a corresponding layer thickness of 250 µm. A brim of 3 mm gave optimal hold to the chamber on the printer bed. To ensure a plane chamber surface for skin contact side, the bottom of the chamber was placed on the printer glass bed for slicing. An extrusion multiplexer of 1.02 was set to fill production-related gaps between the filaments in the x–y direction  and the NIH Guide for the Care and Use of Laboratory Animals . Male hairless SKH1/hr mice (6–10 weeks of age and weight of 25–30 g) were used for all experiments. The animals were housed individually in a specific pathogen-free facility with a twelve-hour light–dark cycle and access to standard laboratory chow and water ad libitum.Twelve mice were randomly allocated to two experimental groups: titanium chamber and PEEK chamber. Each animal was one experimental unit and examined independently using body weight, faecal corticosterone metabolites (FCMs), burrowing activity and clinical distress scores on days 1/2, 10/11 and 20/21 after dorsal skinfold chamber preparation. Following distress and tilting measurements on day 21, mice were sacrificed Fig. .Figure 65. The significant change in the proposed model is the preparation without screws: After disinfection of the dorsal skin  and marking of the median line, a skin bilayer was stretched in the median line. Subsequently, the back of the PEEK chamber was sutured to the skin fold through the preformed holes . The preparation area was color-marked and the microsurgical preparation of the front side was performed. After completion of the preparation, the front of the chamber was placed congruently and fixed by three sutures connecting both chamber frames  injection of ketamine/xylazine (90/10 mg/kg bw) and positioned on a heating pad (37.8 °C). Microsurgery for dorsal skinfold titanium chamber implantation has been described beforemes Fig. .For chamber tilting analysis, animals were sedated in an isoflurane chamber for approximately ten seconds on days 7, 12, and 20/21. The sedated animals were positioned upright, with all feet on the floor. In the upright position, the animals were photographed from behind. In Photoshop software , the lateral tilting angle was measured in degrees of deviation from a vertical line.The body weight was measured on a scale  at 9:00–9:30 am. Percent change in body weight was determined by comparison with body weight in the preoperative phase.40.  filled with 200 ± 1 g of food pellets  was placed in the left back corner of the cages 3 h before the dark phase at 04:00–04:10 pm. Despite the implanted chambers, mice had free access to these pellets throughout the whole observation time. The weight of the food pellets (g) left in the tube was measured on the next day.42. FCMs were evaluated blinded, and the percentage of FCMs was determined by comparison to respective FCM concentrations in the pre-operative phase.After weighting, the bedding with old faeces was removed and replaced by fresh beddings. After 24 h, at least 400 mg faeces were collected per cage. The faeces were dried for 4 h at 65 °C and kept at − 20 °C until further processing. 50 mg of homogenized, dried faeces were extracted with 1 mL of 80% methanol and FCMs analyzed with a 5α-pregnane-3β, 11β, 21-triol-20-one enzyme immunoassayRepresentative intravital microscopy was performed on day 3. Mice were anesthetized and placed on a plexiglass pad with integrated heating. For the visualization of the microvascular system fluorescein isothiocyanate–labeled (FITC)-dextran  was injected into the lateral tail vein (or into the retrobulbar venous plexus if tail vein injection failed). Intravital microscopy was performed with 50-, 100- and 200-fold magnification using an Axiotech vario microscope  with a 100-W HBO mercury lamp with a blue filter  The microscopic images were recorded on DVD  using a charge-coupled video camera  for off-line evaluation.Data were graphed and analyzed with GraphPad Prism  and were presented as median and 95% confidence interval. The characteristics of data were assessed by Shapiro Wilk test. When analyzing the influence of time on the dependent variables, a Friedman Test was performed (corrections of multiple comparisons using Dunn test) in tilting angles, burrowing activity, percentage of FCMs and distress score, and a one-way repeated measure ANOVA was performed (corrections of multiple comparisons using Tukey test) in the percentage of body weight change analysis. When analyzing the influence of the chambers on the dependent variables, a Mann Whitney Rank sum test (for non-parametric data) or unpaired t test (for parametric data) was used. Differences with p < 0.05 were considered significant. Data are given as median/95% confidence interval.Supplementary Information.Supplementary Tables.Supplementary Figure S1.


In recent years, a variety of emerging online media, such as TikTok, Kuaishou, YouTube and other short video application platforms, have appeared. The problem of short video addiction has become an issue to education experts and the general public, as students’ excessive use of short video has become increasingly serious with many hidden concerns to the students’ learning effectiveness. In addition, to meet the growing demand for innovative design talents worldwide, the Taiwan government has been committed to promoting policies related to the cultivation of innovative and creative talents nowadays, particularly for innovative design profession students who often use the Internet and short videos for learning. Therefore, the study aims to use questionnaires to understand the habits and addiction of the innovative design profession students in using short videos, and to further investigate the relation of short video addiction to the students’ creative self-efficacy (CSE) and career interests. A total of 561 valid questionnaires were collected after eliminating invalid questionnaires and reliability analysis. Structural equation modeling and model validation were conducted afterwards. The results showed that short video addiction had a negative effect on CSE; CSE had a positive effect on career interests; and CSE had an indirect effect between short video addiction and career interests. According to the “2020 Communications Market Survey in Taiwan” and “2020 Taiwan Internet Report”, the use and access of the Internet of youths aged from 16 to 25 are the highest among all age groups: the Internet access rate is approaching 100% and the mobile Internet access reaching over 93%. Among the various Internet usage behaviors, instant message accounts for 14.2%, social media 13%, and audio-visual entertainment 11.1% respectively [With regard to the COVID-19 pandemic in the past two years, the resources and demand for e-learning in schools and society has greatly increased. Numerous emerging online media, such as TikTok, Kuaishou, YouTube and other audio-visual platforms are constantly appearing and being used in learning, teaching aids and life, which has a great impact on the living habits, learning styles and values of youths. As a result, it seems to be an indispensable learning channel for teenagers to use short video related application on smartphones to obtain knowledge and skills. Relevant studies have also demonstrated that the smartphones can help students not only learn anywhere but also improve peer interaction, maintain interpersonal relationship, improve learning tactics, plan after-school learning, document learning processes and write assignment reports ,4,5. HowFrom the above-mentioned studies, it has been proven that the popularity of short videos would cause the risk of addiction to college students ,13. MoreFrom the viewpoint of self-efficacy, it is seen as a belief in one’s own abilities . People CSE is defined as “the belief one has the ability to produce creative outcomes” . Social According to Bandura’s  self-effAccording to Hong  work on 1.Short Video Addiction and CSEIn the past few years, more and more studies have been conducted to investigate the relations between creativity and creative self-efficacy. Most of the studies have shown a positive correlation between creativity and creative self-efficacy ,38,39, wHypothesis 1 (H1) Short video addiction has a negative effect on CSE-ability.Hypothesis 2 (H2) Short video addiction has a negative effect on CSE-confidence.2.CSE and Career InterestsSeveral studies have demonstrated that the level of self-efficacy is highly correlated with learning motivation, and students with high self-efficacy tend to have higher learning motivation ,53,54,55Hypothesis 3 (H3) CSE-ability has a positive effect on career interests.Hypothesis 4 (H4) CSE-confidence has a positive effect on career interests.3.Short Video Addiction, CSE and Career InterestsFrom the above-mentioned studies, an interactive relationship among short video addiction, creative self-efficacy, and career interests is observed. Therefore, to understand the relation between each dimension, the hypotheses are conducted as follows:Hypothesis 5 (H5) CSE has an indirect effect on short video addiction and career interests.The current study was an empirical study and recruited college students in innovative design profession as the participants. Snowball sampling method was applied. The questionnaire link was sent to a number of innovative design-relevant departments to invite students to fill it out. The questionnaire was collected since 1 March 2022, and the collecting process was completed once 650 copies of the questionnaire were received. It was ensured that all processes conducted in the study comprising humans were in accordance with the ethical principles of the American Psychological Association. Informed consent was provided in the questionnaire, so that all participants were aware that they were participating in this study, and that the data they provided were presented anonymously.The current study recruited 650 participants (the number of returned questionnaires). The total number of invalid pieces was 89, while the number of the valid pieces (participants) was 561, with a valid return rate of 86.3%. Among 561 study participants, there were 247 male students (44%) and 314 female students (56%); 175 first-year students (31.2%), 112 second-year students (20%), 183 third-year students (32.6%), and 91 fourth-year students (16.2%). With regard to the average number of weekly viewers, there were 75 participants viewing for 1–3 days (13.4%), 124 participants viewing for 4–6 days (22.1%), and 362 participants viewing daily (64.5%). Additionally, regarding the average number of daily viewers, there were 58 participants watching less than 1 h (10.3%), 279 participants watching between 1-3 h (49.7%), 201 participants watching between 3-5 h (35.8%), and 23 participants watching for more than 5 h (4.1%). The average age of participants was 19–20 years old, with a standard deviation of 1.49 years.Short video addiction is seen as a psychiatric disorder and a complex bio-psychological phenomenon. It is defined as the inability of users to control their media use, resulting in disruption of their daily life. From the definition, the current study refers to the Short Video Addiction Scale by Ye  and Liu The current study refers to the Creative Self-Efficacy Scale of Hong  with sevAccording to the study of Cui , the expStructural equation modeling (SEM) is a statistical modeling technique that can be used to test complex causal structure . To ensu2/df value was between 1–3, RMSEA value was below 0.10 and GFI value was above 0.90 [To ensure the questionnaire was consistent and stable, item analysis was applied initially. Relevant studies pointed out it was adequate that χove 0.90 ,66,67. ASome related studies highly suggested that the reliability and validity should be above 0.70 ,69. AccoThe CR values of each dimension reached >0.70, indicating that there was consistency in the latent variables among dimensions of short video addiction, CSE-ability, CSE-confidence and career interests. The six questions of the dimension of short video addiction had high internal consistency, indicating that these six questions were sufficiently representative of the dimension of short video addiction.Additionally, the AVE of each dimension reached >0.50, demonstrating that the four dimensions were distinguished from each other, which each of the four dimensions being independent . In brief, the short video addiction, CSE-ability, CSE-confidence and career interests are sufficiently distinguished with stability and consistent reliability, as shown in 2 = 919.5, df = 205, χ2/df = 4.49, RMSEA = 0.08, GFI = 0.89, AGFI = 0.87, NFI = 0.89, NNFI = 0.90, CFI = 0.91, IFI = 0.91, RFI = 0.88, PNFI = 0.79 and PGFI = 0.72.Structural equation modeling (SEM) has become a standard tool in many scientific fields for explaining the interrelationships between variables and the soundness of theoretical models . It is np < 0.001); short video addiction had a negative effect on CSE-confidence ; CSE-ability had a positive effect on career interests ; CSE-confidence had a positive effect on career interests ; and CSE played a medicating role in short video addiction and career interests. As shown in The current study proposed five research hypotheses and constructed a theoretical model, which was validated by SEM. The results of the validated model showed that short video addiction had a negative effect on CSE-ability  short video addiction has a negative effect both on CSE-ability and CSE-confidence; (2) both CSE-ability and CSE-confidence have a positive effect on career interests; (3) CSE plays a mediating role between short video addiction and career interests.In addition, although the level of short video addictive behavior of the participants examined was low in the study, some relevant studies presented that most students still have had the frequent and prolonged short video watching behavior ,2. The cWith regard to the results of the current study, short video addiction was proven to have a negative impact on students’ CSE and career interests. Yet, due to the influence of popularity of short video, it is not feasible to thoroughly eradicate the use of short videos. Short video watching is not merely inadequate for students, but if short videos for learning are effectively used, they can enhance students’ willingness and effectiveness in learning ,82,83,84The study was conducted by quantitative analysis, and the corresponding results and conclusions were inferred by the statistical examination. Although the correlation between short video addiction and CSE was confirmed, it was insufficient to understand the process and effect of how the student watched short videos. As a result, by adopting qualitative interviews to gain profound understanding of the participants’ psychological states and actual thoughts, the results of the study can alternatively have more referred value.Based on the study, although short video addiction has a negative impact on innovative design profession students, it is necessary to observe that students are so accustomed to watching short videos nowadays. Therefore, it is suggested that future research can explore the use of short videos for learning in specific professional fields, thus transforming short videos into one of the learning tools and further exploring the impact on students’ learning effectiveness.Finally, and above all, the study focuses on the effects of short video addiction on CSE and career interests. However, short video addiction is only one of the Internet addictive behaviors. It is suggested that a comparative study can be conducted to investigate the effects of different Internet addictive behaviors on CSE or career interests in the future to enrich the relevant topics within CSE research.


The premise that the intensity of biotic interactions decreases with increasing latitudes and elevations is broadly accepted; however, whether these geographical patterns can be explained within a common theoretical framework remains unclear. Our goal was to identify the general pattern of elevational changes in trophic interactions and to explore the sources of variation among the outcomes of individual studies. Meta‐analysis of 226 effect sizes calculated from 134 publications demonstrated a significant but interaction‐specific decrease in the intensity of herbivory, carnivory and parasitism with increasing elevation. Nevertheless, this decrease was not significant at high latitudes and for interactions involving endothermic organisms, for herbivore outbreaks or for herbivores living within plant tissues. Herbivory similarly declined with increases in latitude and elevation, whereas carnivory showed a fivefold stronger decrease with elevation than with latitude and parasitism increased with latitude but decreased with elevation. Thus, although these gradients share a general pattern and several sources of variation in trophic interaction intensity, we discovered important dissimilarities, indicating that elevational and latitudinal changes in these interactions are partly driven by different factors. We conclude that the scope of the latitudinal biotic interaction hypothesis cannot be extended to incorporate elevational gradients. Meta‐analysis of 226 effect sizes calculated from 134 publications demonstrated a significant but interaction‐specific decrease in the intensity of herbivory, carnivory and parasitism with increasing elevation. We discovered several previously unknown sources of variation in elevational patterns, and revealed important similarities and dissimilarities between latitudinal and elevational changes in the intensity of trophic interactions. We conclude that elevational and latitudinal changes in these interactions are partly driven by different factors and that the scope of the latitudinal biotic interaction hypothesis cannot be extended to incorporate elevational gradients. The exploration of the intensity of biotic interactions in geographical gradients is of both fundamental and applied importance. Studies of these gradients can improve an understanding of how abiotic, biotic and phylogenetic factors concomitantly influence ecosystem structure and functions , which states that the intensity of biotic interactions reaches its maximum in stable and warm climates and generally decreases from low to high latitudes  the intensity of trophic interactions decreases with increasing elevation. However, the strength or even the direction of elevational changes in these interactions can differ between trophic levels because this type of variation was previously detected in latitudinal gradients: herbivory and carnivory decreased from low to high latitudes, whereas parasitism increased  and classified into herbivory, carnivory and parasitism, as in previously conducted the meta‐analysis of latitudinal changes in trophic interactions  the data were collected from natural ecosystems, (2) the data were collected from at least two study sites with elevation differences of at least 100 m within the same mountain region and (3) the magnitude of the effect could be calculated from the data or statistics presented in the publication or provided by the authors. From multiyear studies, we extracted the combined result for all years if it was presented in the publication. If the data collected in different years were not combined by the authors, then we selected the year with the highest average value of the character under study. If the study employed some manipulations , then we selected a control treatment.Whenever possible, herbivores and carnivores were divided into ectotherms  and endotherms . All parasites were ectotherms; therefore, they were excluded from this comparison. We classified host plants and prey into natural, permanently inhabiting the study areas, and standardised, that is introduced to all study sites by the researchers .The herbivory level was considered to be background unless the authors explicitly mentioned an outbreak. Herbivory was divided into folivory  and granivory (i.e. seed predation). Folivory was divided into mammalian grazing and invertebrate folivory, whereas granivory was divided into pre‐ and post‐dispersal seed predation. Invertebrate herbivores were divided into exophagous (defoliators and sap‐feeders) and endophagous  feeders; the latter group also included pre‐dispersal seed feeders.We divided studies of herbivory into those reporting damage of all plant species in a site or of several dominant species (community‐wide herbivory hereafter) and those reporting damage of a certain plant species or genus (species‐specific herbivory hereafter). Host plants were classified as woody or herbaceous, and woody plants were classified as evergreen  or deciduous.Elevational gradients were classified as those located entirely below or above the tree line and those crossing the tree line. We calculated the elevational span of a gradient as the difference in elevation between the highest and lowest sites. The geographical coordinates of study areas (to the nearest degree of latitude and longitude) were extracted from publications or searched on the internet based on information provided in the publication. The attribution of elevational gradients to climate zones  was based on climate and vegetation at the foot of the mountain and was performed in the same way as in the meta‐analysis of latitudinal patterns . We estimated the variation in effect sizes (ES) within groups by calculating the heterogeneity index (Qt). To compare ES among different groups of studies, we calculated the between‐group heterogeneity (QB) using a random effects model, and we tested QB against the χ2 distribution with the number of groups minus one degree of freedom  following an equation given by Lajeunesse , thereby justifying the combination of ES calculated by these methods in our analyses.We used three approaches for the ES calculation. When a study reported the data from two or three sites, we calculated Hedge's jeunesse  and calcn  were considered as proof of robustness of the analysis against the insignificant results  that satisfied our criteria, and we calculated 226 ES  and climate zones Figure . Of 226 zr = −0.21, CI95 from −0.30 to −0.12), and this pattern was robust against unpublished studies . We did not find publication bias in our data .The intensity of trophic interactions generally decreased with increasing elevation , mostly due to different responses of trophic interactions to elevation . The decreases with increasing elevation were stronger for carnivory and parasitism than for herbivory  was more than fourfold greater  than the decrease in herbivory (zr = −0.12). The elevational patterns in carnivory and parasitism combined did not show statistically significant heterogeneity , whereas the patterns in herbivory were highly variable .The overall effect of elevation showed high heterogeneity , although this difference was not statistically significant . The changes in trophic interactions became stronger with an increase in the elevational span of the individual gradient  and did not cross  the tree line.The association between the intensity of trophic interactions and elevation was stronger in gradients crossing the tree line than in gradients located either below or above the tree line Figure . This dizr = −0.01). This lack of an elevational decrease was mostly due to herbivory (zr = 0. 04), whereas carnivory and parasitism still tended to decrease with elevation (zr = −0.38). Meta‐regression of zr against latitude was not significant within tropical and temperate zones combined , whereas zr increased in the polar zone (i.e. became less negative) towards the pole . The latter pattern was mostly due to an elevational gradient in herbivory, which weakened substantially with increasing latitude  within the polar zone.The elevational decrease in the intensity of trophic interactions was significant in tropical and temperate climate zones, but it was not significant in the polar zone Figure , where nQB = 7.41, p = 0.008). Studies using standardised prey or plants yielded a nearly threefold greater magnitude of ES compared to studies that measured carnivory and herbivory on naturally occurring prey or plants .Within both herbivory and carnivory, the elevational decrease was significant only for ectothermic consumers Figure . Within zr = −0. 08 and zr = −0.04, respectively; QB = 0.03, df = 1, p = 0.86). Within invertebrate herbivores, the damage imposed by exophagous species  decreased with increasing elevation, while herbivory imposed by endophagous species  did not change with elevation .Invertebrate folivory decreased with elevation, whereas mammalian grazing and seed predation did not show significant elevational changes Figure ; pre‐ anQB = 0.10, df = 1, p = 0.76), and the elevational decrease tended to be greater in herbaceous than in woody plants  due to the tendency of folivory to increase with elevation in evergreen woody species  or by invertebrates feeding within plant tissues .The general lack of an elevational decrease in herbivory caused by internally feeding invertebrates is in line with the hypothesis  to noise  ratio. Furthermore, the decrease in the intensity of all studied interactions is stronger in gradients crossing the tree line (i.e. spanning more than one vegetation zone) than in gradients located entirely within a single vegetation zone either below or above the tree line Figure . These rOnly a few studies  between the equator and about 50° N/S, followed by a strong poleward decrease in the lapse rate at higher latitudes  than for latitudinal gradients , so the difference in the overall effect outlined above is due to the different responses of the top consumers to environmental changes in these two geographical gradients. Changes in carnivory are almost fourfold stronger with increasing elevation than with increasing latitude (zr = −0.58 and zr = −0.15 respectively). Even greater differences are observed in parasitism, which significantly increases with increasing latitude (zr = 0.18) but significantly decreases with increasing elevation (zr = −0.48). Importantly, the elevational decrease in parasitism is consistent with the meta‐analysis by Péré et al.  yield a stronger decrease with increases in either latitude or elevation than herbivory and carnivory measured on naturally occurring local prey. This difference can be explained by the evolved local adaptations of native prey or plant in different sites along both gradients, whereas standardised models did not participate in evolution  statistical testing for generality of patterns uncovered by case studies and narrative reviews; (2) discovery of previously unknown sources of variation in elevational patterns in biotic interactions, for example dramatic difference between trophic levels and geographic variation in strength of elevational decrease and (3) discovery of important similarities and dissimilarities between latitudinal and elevational changes in the intensity of trophic interactions. Our analysis of elevational changes in trophic interactions revealed considerable limitations for the prediction of a general decrease in their intensity with increased elevation. While carnivory and parasitism consistently decline from low to high elevation, for herbivory this prediction is unequivocally fulfilled only for ectothermic, openly living folivores at their background population densities in elevational gradients that cross tree lines in mountains located outside the polar zone. The differences between the outcomes of the two meta‐analyses conducted using similar methodologies (Zvereva & Kozlov, ELZ and MVK formulated goals and designed methodology, ELZ extracted data for meta‐analysis, conducted meta‐analysis and wrote the first draft of the manuscript, MVK participated in the writing of later drafts.


Morin displayed a remarkable ROS inhibitory ability and protected keratinocytes against PMs by downregulating matrix metalloproteinase-1 (MMP-1) expression via suppressing p-ERK and p-p38 in the MAPK pathway. Moreover, water solubility of liposomal morin (LM) prepared by the thin film hydration method was significantly better than free form of morin due to particle size reduction of LM. Our results also demonstrated that deformable liposomal vesicles were achieved for increasing dermal absorption. Additionally, LM  was able to effectively reduce generation of ROS, inactivate p-ERK, p-p38 and MMP-1 in HaCaT cells exposed to PM. In conclusion, our findings suggested that LM would be a bright candidate for various topical anti-aging and anti-pollution products.Recently, a global market for anti-aging skin care using botanicals has been noticeably developing. Morin, 3,5,7,2′,4′-pentahydroxyflavone, is a polyphenol with many pharmacological properties including antioxidant, anti-inflammation and photoprotection. However, poor aqueous solubility of morin restricts its application in pharmaceuticals. The present study aimed to encapsulate morin into liposomal vesicles to improve its water solubility and skin penetration, and further investigated its ROS inhibition and anti-aging activity in HaCaT keratinocytes induced by particulate matters (PMs). Our data presented that morin was a strong DPPH Morus alba L.), almond (Prunus dulcis Mill.) and sweet chestnut (Castanea sativa Mill.) [Staphylococcus aureus and to inhibit influenza virus in a rat model [Despite COVID-19 pandemic crisis, global anti-aging market size is expected to keep booming to reach USD 83.2 billion by 2027, equivalent to a growth rate of 6.8% . Similarly, personal care products using botanical extracts were projected to see a rise of 10–11% annually from 2015 to 2019 and are expected to explode in the coming years [a Mill.) . Morin dAmong various strategies introduced to overcome the limitations of poor soluble drugs and simultaneously enhance dermal absorption for topical application, liposomal vesicles stand out from others because of their biocompatibility and safety profile . LiposomRecently, air pollution has been emerging as a global major concern since more than 99% of the entire population are living in areas where levels of pollutants do not meet the standard for healthy air WHO, 2017). Air pollutants consist of gaseous molecules and solid particles of different chemical compositions, sizes, and shapes, also known as particulate matters . Previou. Air polThe present study aimed to prepare deformable forms of liposomal morin with various ratios of compositions, measure their physicochemical features including aqueous solubility, intermolecular bonding, particle size and morphology, and skin absorption; and further investigate the in vitro inhibitory capacity of morin and its liposomal formulation on ROS generation, the underlying mechanism of skin anti-aging property on HaCaT keratinocytes induced by PM.® E145V)) was purchased from Cosphatec GmbH . Pluronic F-68 non-ionic surfactant was ordered from Sigma . Tween-80 (edge activator) was purchased from Shimakyu’s Pure Chemicals . HaCaT keratinocyte cell line was acquired from the I.Z.S.L.E.R. . Primary antibodies including anti-phospho-ERK 1/2  and anti-phospho-p38 (Thr180/Tyr182) were obtained from EMD Millipore Corporation . GAPDH antibodies were obtained from Santa Cruz Biotechnology  and MMP-1 antibodies were purchased from Proteintech Group, . Fetal Bovine Serum (FBS) and Gibco Dulbecco’s Modified Eagle’s Medium (DMEM) for cell culture were supplied by Thermo Fisher Scientific .Morin hydrate with 90% of purity was purchased from Tokyo Chemical Industry Co., Ltd. . Lecinolws-50 . The system was set at 1 mL/min for flow rate, detecting wavelength was at 355 nm, injection volume was 10 µL and 8 min of running time. Morin was dissolved in ethanol and diluted to a series of nine concentrations ranging from 0.01 to 100 µg/mL in order to determine the calibration curve. Absorption peak of morin was observed at 6.1 min. The calibration curve of morin displayed an excellent linear with R2 = 0.9999.Identification and quantification analysis of morin in this research were acquired using HPLC. The HPLC instrument  is equipped with an L-2130 pump, an L-2200 autosampler and an L-2420 (UV–vis) detector connecting with a Mightysil RP-18 GP column . The mobile phase consists of acetonitrile and monopotassium phosphate  prior to being ten-fold diluted for HPLC analysis. The above-mentioned standard curve was employed to determine the concentrations of samples and then calculate the aqueous solubility of morin in free form and in liposomal formulations.Measuring the solubility of morin and its liposomal formulation is the primary step towards preparing an appropriate concentration for further experiments. Specifically, 1 mg of free morin and liposomal morin that contains equal amounts of morin were dissolved in water (at room temperature) under a vigorous vortex  for uniform mixing. Solutions were subsequently filtered through a 0.45 µm syringe filter (13 mm Acrodiscw/v) in distilled water was made ready to drop onto dried vesicles (200 µL was used for each copper grid). The resulting samples were stored in the empty plastic capsules until being placed in a transmission electron microscope  for their morphological observation.TEM is the most frequently used imaging technique for characterizing morphology of nano-scaled particles such as liposomes. In this study, we prepared samples by the negative-staining method due to its simplicity and availability. Each sample of liposomal morin formulations was completely dissolved in de-ionized distilled water to concentration 100 µg/mL. Then 200 µL of prepared solution was slowly dripped onto a 200-mesh copper grid and left to dry. At the same time, phosphotungstic acid  0.5% .• , a stable free radical at room temperature, is a simple screening test to investigate the antioxidant property of morin and liposomal morin in this study. First of all, morin was prepared in de-ionized distilled water and ethanol of an ascending concentration series . Liposomal morin was dissolved in water at the same concentrations. A solution of DPPH• 200 µM was made ready to use by dissolving DPPH• powder in ethanol 95%. Then 100 µL of each sample would react with 100 µL purple DPPH• solution in a 96-well plate. It was noted that the plate was incubated in darkness at room temperature. After 30 min, the optical density of each well at wavelength 517 nm (OD517) was measured by a microplate spectrophotometer . Data were collected and processed using the following formula to calculate the free radical scavenging ability of each sample.DPPH−1 which covers nearly all of the chemical group spectrums.Molecular vibration created by radiation absorbed at a specific frequency is used to identify occurring functional groups and chemical interaction of compounds. In this study, FTIR was investigated to figure fundamental chemical groups in the structure of morin, liposomes and their interaction in complexes. Samples including morin, blank liposomes and liposomal morin were handled using the compressed pellet method. Each sample was firstly blended with KBr  (ratio 1:9 in volume). The mixture was then ground into fine powder using an agate pestle and mortar. Ground powder was transferred to a dry pellet die, pressed into a thin translucent film and carefully placed in a spectrometer . Wavelength range for scanning was set at 400 to 4000 cm2-(2 cm × 2 cm)-sized pork skin was used for each FDC and placed onto the FDC with the stratum corneum facing upwards; the receptor chamber below was filled with a buffer solution containing 0.14 M NaCl, 2 mM K2HPO4, and 0.4 mM KH2PO4 (pH 7.4). A small magnetic stirring bar was placed at the bottom of the receptor chamber in order to maintain the constant compound concentration and temperature by the circulation of water at 32 °C from a water bath. The stirrer speed was set at 600 rpm during the experiment. The sample preparation included dissolving morin, liposomal morin in water comparable to 1 mg of morin per 1 mL of distilled water. Then 200 µL of testing samples were loaded onto the donor chamber in direct contact with the outer-most layer of pork skin. After 1, 2, and 4 h, testing compounds and skin samples were collected for analysis. The stratum corneum layer was removed by stripping 15–20 times with 3 M Transpore tapes. The remaining skin samples were heated at 80 °C to facilitate the separating of the viable epidermis and dermis. They were then cut into smaller pieces with a scalpel. The individual skin layer of each testing sample was soaked into methanol and placed in an ultrasonic bath sonicator for 1 h to extract the remaining morin out of the skin. The extracts were filtered with 0.45 µm pore-sized- membrane before HPLC analysis with the method mentioned earlier.According to the standard guideline of the European Cosmetic Toiletry and Perfumery Association (COLIPA), the Franz Diffusion Cell (FDC) has been employed to measure kinetics of percutaneous absorption of morin and liposomal morin. The FDC system basically consists of two primary chambers: upper donor and lower receptor chamber made with glass, separated by a membrane; in this case pork skin was used. Pork skin samples were purchased from the local traditional market. Areas of skin that did not have the intact barrier or were compromised with any wounds which could result in incorrect data were removed. A 4 cm2. To begin with, HaCaT cells were seeded in a 96-well plate, specifically, 1 × 104 cells suspended in 100 µL of culture medium per well in 24 h prior to treating. In terms of sample preparation, morin was originally dissolved in dimethyl sulfoxide (DMSO), liposomal morin was prepared in PBS, the solutions were then diluted in DMEM to desired concentrations . The cells in each well were washed with 100 µL of PBS before being exposed to morin and liposomes for the following 24 h. On the third day, the medium with the active compounds was removed from cells by washing with PBS and 150 µL of 0.5% methyl thiazolyl tetrazolium (MTT) solution was then added into each well. The plate was stored back into the incubator and left for reaction. After 3 h, MTT solution was discharged from wells. Purple formazan crystals created by the reduction of a yellow tetrazolium salt was dissolved in DMSO solution. Optical density at a wavelength of 550 nm (OD550) was determined using a microplate spectrophotometer .Cell metabolism and survival rate play a key role in screening appropriate doses of drug that do not lead to toxicity or cell death but result in therapeutic effectiveness. MTT assay is the most frequently used to measure cellular metabolic activity as an indicator of cell viability, proliferation and cytotoxicity. HaCaT keratinocytes were acquired from Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna . Culture medium included Gibco Dulbecco’s Modified Eagle Medium DMEM , 10% fetal bovine serum  and 1% penicillin–streptomycin . Cells were grown in an incubator , temperature was set at 37 °C and 5% CO4 HaCaT keratinocytes were initially seeded on the 96-well plate and cultured for 24 h with medium solution and incubating conditions mentioned above in the cell culture section. Preparing samples involved dissolving morin in DMSO and liposomal morin in PBS, then diluting to concentrations of 10 and 25 μg/mL by DMEM. Cells were washed with PBS before being treated by the sample preparations for the following 24 h. On the third day of the assay, HaCaT cells were reacted with 20 µM solution of dichlorodihydrofluorescein diacetate  for 30 min under an incubation setting. PMs  was purchased from the National Institute of Standards and Technology and suspended in PBS to the concentration of 10 mg/mL. The suspension was sonicated with ultrasound water bath for 10 min. After that, PMs was added to each well (50 µg/cm2) and kept exposed for an hour. The cell plate was then washed with PBS and measured for fluorescence absorbance at the excitation and emission wavelength of 485 and 528 nm, respectively, using a microplate reader . The experiment was triplicated and data were presented by mean value ± SD.ROS inhibition is one of the most potential bioeffects of morin and its liposomes in terms of topical application. In order to screen for their impact on the ROS generation, 1 × 105 HaCaT keratinocytes were seeded in a 6 cm dish and cultured for 24 h in the incubator set with the condition described above. PM were suspended in PBS (10 mg/mL) and sonicated for 10 min. The cells were then treated with morin or liposomal morin prepared in DMEM solution at various concentrations for 3 h. Subsequently, the HaCaT cells were induced with PM suspension (50 µg/cm2). After 2 or 6 h, the whole proteins in treated cells were extracted using RIPA Lysis Buffer  and collected into 1.5 mL tubes, centrifuged at 12,000 rpm for 10 min. The protein contents were then determined by the BCA Protein Assay Kit . An equal quantity of protein in each sample was loaded into sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for separating. Proteins were blotted onto polyvinylidene difluoride (PVDF) membranes . The membranes were then blocked for an hour and washed with tris-buffered saline (TBS) and 1% tween-20 before being incubated overnight at 4 °C with primary antibodies including p-p38 , p-ERK , MMP-1  and GAPDH  as the internal control. The membranes were washed with TBS prior to being incubated with horseradish peroxidase (HRP) conjugated secondary antibodies which were already reacted with chemiluminescence reagents  for an hour at room temperature the following day. Protein signaling bands were achieved by Touch Imager  and analyzed using ImageJ application.The anti-aging effect of morin and liposomal morin in keratinocytes were analyzed by Western blotting. Firstly, 4 × 10p < 0.05 was regarded as statistical significance.Data were shown as mean ± standard deviation (SD). For protein quantification analysis, ImageJ software  was employed. SPSS software  was used for statistical analysis. Comparison of data between multiple groups was performed using analysis of variance (ANOVA) followed by Tukey’s post-hoc test, • radicals did not exceed 40% at various concentrations since morin was water-insoluble. However, when freely dissolved in ethanol, the scavenging activity of morin at 200 µg/mL was twice as much as morin in water, reaching approximately 70%. In particular, the DPPH• removal effect of morin was concentration-dependent.Antioxidant activity is an overall ability to scavenge free radicals, the higher power of neutralizing free radicals, the better the antioxidant property of the component. As we can see in A cell viability assay was conducted to determine the non-toxic concentration range in which more than 80% of keratinocytes still alive after treatment . In thisAs shown in As shown in p < 0.05). The inhibitory effect of 25 µM on MMP-1 was small. For more details about protein expression, the percentage change was calculated and is shown in Regards to anti-aging activity, we examined the effect of morin on the suppression of MMP-1 . HaCaT kIn contrast to the water solubility, the yield rate showed an obvious proportional dependence on the lecithin portion in lipid bilayers . The totMorphological imaging of LM was taken by TEM . As we c−1 represents dimeric hydroxyl O–H stretching functional. A pair of absorptions at 1620 cm−1 is indicative of double bond alkenyl C=C stretching. Peaks between 1600 and 1500 cm−1 signify one or more aromatic ring vibrations, specifically, 1511 cm−1 was assigned to C=C–C ring bonding. Band at 1177 cm−1 suggests a phenolic C–O stretching vibration. A peak at 831 cm−1 was a C–H vibration of aromatic ring. A small sharp peak at 639 cm−1 was attributed to a bending vibration of the OH alcoholic group. P–O bending vibration peaks were demonstrated at 539, 538, 586, 558 cm−1 in liposomal morin and empty liposome preparation, which contain lecithin as a component and not seen in the infrared spectrum of morin. C–H stretch vibrations for methyl (–CH3) or methylene (–CH2–) peaking at 2922, 2923, 2925, and 2927 cm−1 are distinctive in saturated or unsaturated aliphatic compounds that were depicted in all graphs, except for morin. As we can see that the infrared spectrum of blank liposome formulations 1, 7  and morin were merged to shape the spectrum of liposomal morin formulation 1, 7 , respectively. Even though some peaks shifted to higher or lower wavelengths, no noticeable new peaks were found in the spectra of LMF1, LMF2. Therefore, morin was supposed to be successfully encapsulated in liposomes without unexpected chemical reactions between ingredients.FTIR spectra of morin, liposomal morin formulations 1 and 7, blank liposome formulations 1 and 7 are illustrated in 2 for LMF7 and LMF1, respectively. Most of the liposomal morin could overcome the stratum corneum to be deposited in viable epidermis, nearly 50 µg/cm2 of LMF1 and 60 µg/cm2 of LMF7 were quantified after 4 h of application. A dynamic and time-dependent penetration was most obviously seen in the dermis, approximately 3, 7 and 25 µg/cm2 of LMF7 were penetrated into this layer after 1, 2 and 4 h, respectively. As a result, encapsulating morin into liposomes evidently enhanced dermal absorption compared to free morin. No content of morin found in the receptor fluid (data not shown) illustrated liposomes hardly penetrated deeper than the dermal layer into systemic circulation which might cause undesirable adverse effects. LMF7 was superior to LMF1 in total dermal absorption and would be used for further bioactive investigation.• radicals, ranging from 50 to 60%. However, there was no improvement in the effectiveness of removing DPPH• radicals when increasing LMF7 dosage. This was different from the results of the above experiment in which morin worked in a dose-dependent manner.LMF7 at 25 µM showed a significant benefit to decrease oxygen-containing free radicals by nearly a half compared to the PM-induced group without treating with LMF7 . SimilarFor more details about protein expression, percentage change was calculated and is shown in Airborne PM exposure has become a noticeable risk factor related to many cutaneous disorders in the short- or long-term such as skin allergies, acne, urticaria, eczema, contact dermatitis, dermographism, seborrhea, infected skin disease, psoriasis, dyspigmentation, skin cancer and aging . Dijkhof• assay. Morin was found to be a powerful antioxidant that could strongly scavenge DPPH• radicals. We further looked at its ROS inhibitory effect in HaCaT keratinocytes exposed to PM and clarified the underlying mechanism of its action on skin antiaging. Our study proved that morin dissolved in DMSO successfully reduced the ROS level and significantly diminished the expression of MAPK proteins including p-ERK and p-p38 in HaCaT cells. Downstream signaling of p-ERK and p-p38 subsequently resulted in suppressing MMP-1 expression. MMPs play the most critical role in ECM degradation throughout the skin aging process. MMP-1, which is mainly secreted by keratinocytes, is especially responsible for breakdown and fragmentation of skin collagen fibers [In this study, we first screened the antioxidant property of free morin dissolved in DMSO by in vitro DPPHn fibers . In prevn fibers . With thHowever, low bioavailability resulted from poor aqueous solubility of morin is the biggest obstacle for its application and this is also the problematic issue for most of the other flavonoids . Active Entrapping morin into lipid carriers resulted in water solubility enhancement by nearly 300 to 500 times in this study. Liposomal formulations of other flavonoids including resveratrol, fisetin, quercetin, curcumin and epigallocatechin gallate (EGCG) were also reported to have significantly better water solubility ,30. LipoPrevious studies pointed out that dermal absorption of liposomes was inversely related to their particle size. They illustrated that liposomes with a size over 600 nm were unable to penetrate through the stratum corneum and vesicles that were smaller than 70 nm obtained the best deposition in the epidermal and dermal layer. The author also mentioned the significant role of pilosebaceous units in the skin penetration of liposomal vesicles . As a reOur data showed that liposomal morin greatly penetrated deep into the dermis while the free form of morin could not overcome the skin barrier—stratum corneum. In addition, the total dermal absorption of liposomal morin was shown to be not only size-dependent but also highly improved when the lipid outer layers of vesicles were made more flexible with appropriate ratio of lecithin and tween-80. Even though particles from formulation 7 were bigger than from formulation 1 in diameter, lower amounts of lecithin in formulation 7 could produce liposomes with more elastic and deformable lipid shells. This characteristic was thought to help LMF7 penetrate better through stratum corneum and hair follicles leading to greater dermal absorption. Moreover, a majority of liposomal vesicles deposited in epidermis and dermis without travelling to the receptor fluid which emphasized the safety property of liposomes not reaching the systemic circulation.As a result, formulation 7 would be more preferable to treating HaCaT keratinocytes in further experiments. Cytotoxicity of LMF7 was evaluated beforehand. Although the proliferation of HaCaT keratinocytes was high if treated with 100 µM of morin, a significant reduction in cell growth was seen when treated with LMF7 with concentrations over 50 μM (data not shown). This cell death could be explained by the presence of large amounts of lecithin and tween-80 in the formulations. Vater, et al. also reported that a nanoemulsion containing 5% of tween-80 led to a complete cell death of fibroblasts after 24 h exposure . TherefoTo summarize, in this study, we prepared eight formulations of liposomal morin with different ratios of morin, lecinolws-50, tween-80 and PF-68. A significant improvement in water solubility of morin loaded in liposomes was achieved. Deformable particles with a small diameter contributed to considerable skin penetration enhancement. Both morin and liposomal morin effectively protected keratinocytes against ROS generation induced by PM exposure and subsequently provided skin anti-aging and anti-pollution property by reducing MMP-1 expression via downregulating the MAPK signaling pathway. As a result, we suggest that liposomal morin can be applied in various topical products for anti-aging and anti-pollution benefits on skin.


Anorexia nervosa is a paradoxical disorder, regarded across disciplines as a body project and yet also an illness of disembodied subjectivity. This overlooks the role that material environments—including objects and spaces—play in producing embodied experiences of anorexia both within and outside treatment. To address this gap, this paper draws together two ethnographic studies of anorexia to explore the shared themes unearthed by research participants’ engagements with objects that move across boundaries between treatment spaces and everyday lives. Demonstrating how the anorexic body is at once both phenomenologically lived and socio-medically constituted, we argue that an attention to materiality is crucial to understanding lived experiences. A materialist account of anorexia extends the literature on treatment resistance in eating disorders and offers a reconceptualisation of ‘the body in treatment’, showing how  objects and spaces shape, maintain, and even ‘trigger’ anorexia. Therefore, against the background of the high rates of relapse in eating disorders, this analysis calls for consideration of how interventions can better take account of eating disordered embodiment as shaped by material environments. Anorexia nervosa is a seemingly paradoxical disorder, framed in both sociological and clinical discussions as a body project and yet also as an ethereal illness of subjectivity. Across the many disciplines that engage with anorexia—psychiatry, psychology, sociology, anthropology, history, philosophy and neuroscience, among others—reflections on meaning, subjectivity and embodiment have tended to hinge upon the emaciated body. This has been held up as the focal ‘object’ of analysis as well as assumed to be the locus of desires and practices in anorexia.Yet, such a reification of emaciation as a ‘synecdoche’ of anorexia  and the UK (Lavis:2007–2008), the authors explore the materialities of anorexia by focusing on five objects that were, each in a different way, described by participants as core to their everyday experiences of anorexia: a photo album; a chair; a television set; biscuits; and paperback memoirs of anorexia. The authors decided to examine their datasets together due to this resonance of material objects in their respective field sites. Through discussing the role of materiality in their fieldwork observations, the authors recognised that, while different material objects emerged as prominent in each field site, reflecting local structures, practices, and concerns, when brought together, these objects spoke of broader experiences of anorexia as forged at the intersections of treatment and everyday life. As such, the authors recognised that analysing their data jointly could shed new light on anorexic embodiment.Positioning this analysis of anorexic embodiment both in and out of treatment is argued to be crucial to understanding lived experiences. In Israel and the UK, where the authors conducted their fieldwork, living with a diagnosis of anorexia almost always involves living with outpatient or inpatient treatment, however desired or resisted. This is due both to the local nationalised healthcare systems, and to the position of anorexia within these systems, i.e., as the eating disorder considered most pressing, due to the significant medical complications of starvation  were part of Eli’s doctoral research. The study, which has been described in detail elsewhere Eli , focusedThe ethnography from the English eating disorders inpatient unit (2007–2008) derives from Lavis’ doctoral research. 45 patients took part in the ethnography. Forty staff members also participated in the study; however, this paper does not refer to the data collected through interviews with or observations of staff members. From both groups, informed written consent to ‘hang around’ as an ethnographer was gained, as well as additional informed written consent for each interview. The focus of this research was to explore everyday experiences of anorexia and, specifically, to interrogate the desire amongst some individuals with anorexia to maintain their existing illness. Thematic analysis  self, growing to ‘contain multitudes’ Whitman .Grace had a photo album dedicated to her life on the ward. She began to take photographs three months after she was hospitalized, keeping a detailed record of the people, spaces, and objects that marked her stay. A typical stay in inpatient care entailed weeks or months of living in the ward. Patients who showed progress were allowed to go home for weekends or to spend a weekday afternoon away; for the most part, however, inpatient stays involved continuous living in the ward’s enclosed space, with the occasional supervised excursion to the ward’s front yard, or a guided walk at night to a nearby landmark. Life on the ward was marked by the strict schedules and regimens of diet and activity prescribed by the senior clinicians, under the observation of nursing staff.Early on during Eli’s fieldwork, and following several interview hours, Grace offered to show her the photo album—the object that materialized Grace’s stories about the eating disorders ward, providing a visible and tangible illustration of what she had described. This object served as Eli’s initiation into the ward, which, at that point, she had not yet visited. Guiding Eli through the album, Grace filled in the relational gaps in her photographs: identifying roommates, best friends, and favourite members of staff, telling Eli who had since given birth, and who was no longer in touch. Grace photographed and narrated the ward with a loving inflection, and as she turned the pages—in her kitchen, years after she had been discharged—she did so with a tinge of nostalgia. “It’s fun, nice, a sweet memory”, she reflected; “but I don’t miss it anymore… I used to miss it so much”.Grace’s photographs and stories, while positive, continued to highlight experiential moments that designated the ward as a space of surveillance and resistance: there was the common room , where patients were watched for two hours after every meal; there was the bathroom, where patients were followed by nurses, who stood watch; and there was Grace’s room, with her suitcase perpetually packed. “I was always with one foot out”, she explained. This image, however, did not conflict with her story of saying lingering goodbyes, on her last day at the ward, having been discharged but feeling unable to leave.a little lab like that, that you could be inside…. A lab in the sense that it was very sterile, it was – very very exact and measured conditions, and – you knew that you, it’s not like the real world, so it eased [our burden].2The ward, as it emerged in Grace’s account, was a haven. “It was,” she said,what is she in for? For bulimia. Okay, great”. Grace and Eli then began laughing, and Grace said that clarifying what each patient was ‘in for’ was part of every new arrival. The ward may have been a haven—a space away from “the real world”—but Grace kept records that testified to the authoritatively structured, carefully surveyed grids of sociality that the ward entailed. Her photo album, then, evoked ambivalence across time: both the ambivalence she had experienced as an inpatient, years ago, and the ambivalence she experienced in the present tense, when recollecting, narrating, and making sense of her past patienthood. Leafing and talking through the album’s pages, Grace opened up the co-existing tension and wistfulness she associated with her inpatient experiences, communicating her still-ambivalent relationship with the anorexic identity forged on the ward.But the ward, as Grace later explained, was also a prison. As she went through the album, and Eli observed that Grace had parenthetically noted the diagnosis of each fellow patient she had photographed, she explained that “it was so I’d remember”. Eli asked her why, and she said, her voice somewhat tense, “I defined the disease [each patient] was in, and she also didn’t have a problem defining the disease she got, it’s not—I didn’t define it judgmentally or divisively, I wrote it in the sense of noting a fact. She got in—Eli visited the ward for the first time in the summer of 2006, several months after being introduced to the eating disorders ward, and the ambivalence therein, through Grace’s photo album. Sara, a participant with whom Eli had been in contact since her fieldwork began, entered the ward as an inpatient, and Eli visited her regularly. Through these visits, she witnessed Sara’s everyday enactments of compliance and resistance. When Eli first visited, she met Sara in the common room, where most patients congregated (with or without visitors), and where the main features were a flat-screen television, a few desktop computers, and many upholstered chairs. With the guard at the door—a clinical social work or nursing student—keeping watch, patients  were engaged in the task of sitting still. It was the two-hour surveillance window after the patients’ mid-afternoon snack, and Sara, like other patients classified as anorexic, was not allowed to move around. Although the chair might have seemed incidental to Sara’s actions, it was, in fact, a crucial actor in the scene that unfolded. In the common room, the chair was transformed from a piece of furniture into a clinical object, used to confine Sara’s rebelling body after each meal. In conversation with Eli and her aunt, observed by the watchful guard, Sara made sure to stay seated. Occasionally, however, she pretended to yawn, stretching her arms at length while keeping her torso firmly within the confines of the chair. Each furtive stretch ended with an exchange of knowing smiles; she and Eli were acknowledging the resistance embodied in each subtle, and forbidden, stretch. But Sara complied with the rules, otherwise—eating every meal, drinking water only when allowed, and honouring her clinicians’ trust when they sent her for an afternoon away with the instruction to eat an ice cream bar (which she did).Sara’s resistance also took on less obvious shapes and targets. During a later visit, when Sara and Eli were engaged in conversation, another patient announced excitedly that her sister had just texted her to say that ‘Hunger Point’ Silver,  was beinall anorexic. Through mundane objects in the ward’s common room, Sara expressed a dual resistance: small subversions of the treatment regime, embodied in shifting and stretching on a chair meant to confine her movements after meals, alongside protests against fellow patients’ performances of eating disorders, embodied in switching off the television set where they played eating disordered media. With this dual resistance, Sara was continuously engaged in maintaining a fine balance—reasserting her anorexic self, while not losing herself to the patient group, and by extension, to anorexia.Sara’s resistance was directed at more than avoiding the triggering effects of anorexic talk and pro-anorexic images. On the ward, other patients were never fully ‘other’; ward life blurred the boundaries of selfhood, creating images of oneself—past, present, and future—that had equal power to inspire and to horrify Eli . As suchRather than highlighting the so-called pathology of the anorexic body or self, the engagements with materialities discussed so far have emplaced anorexic embodiment both within and beyond the logics of everyday clinical contexts, at intersections where illness and identity are continuously negotiated, parsing out relationships between self, others, objects, and spaces. Running through the participants’ narratives and practices are the constitutive elements of biomedical power in generating and reinforcing anorexic modes of being-in-the-world, whilst also showing the affordances of objects and spaces to disrupt these. Anorexia, thus, has emerged here as not simply the manifestation of a pathologized relationship between a person and her body but as shaped by material environments.Anorexic embodiment, then, is seen to be constantly created and recreated through engagements with multiple subjects and objects. The two vignettes so far have shown that it is through the relationships among the self, clinicians, other patients, as well as the spaces and material culture of the ward, that anorexic embodiment is lived, remembered, and defined. Exploring materialities has thereby begun to offer insights into anorexic embodiment as a continual process of drawing and redrawing relationships between illness, corporeality and subjectivity; it is these intersections that will now be drawn to the fore through Lavis’ analyses.Taking up the interrelated strands of ambivalence and resistance introduced by Eli’s explorations of the material dynamics of lived experiences of eating disorders treatment, and the porous boundaries of selfhood that these illuminate, Lavis now interrogates two objects that were framed by participants as core to their experiences of treatment and  to anorexia more widely: anorexia memoirs and food—in this case, biscuits. Woven through life within the Eating Disorders Unit (EDU) and beyond it, these emerge as at once utterly mundane and yet clinical, with their material exchange between spaces transacting and disrupting multiple meanings at once. Just as the objects that Eli discussed accrete into autobiographical reference points, biscuits and memoirs draw clinical spaces and categories into the forging of anorexic embodiment in ways that speak of the continual effort it involves both with and beyond treatment.In the English Eating Disorders Unit (EDU), it was during quiet moments in the patients’ lounge, as well as the fraught mealtimes of the dining room, that participants’ complex relationships with, and embodiment of, anorexia were made, articulated, and rendered visible. In relation to the very structures of the EDU’s spaces and times, various aspects of these relationships came to the fore or receded, and one moment at which this was most apparent was during the enforced ‘rest periods’ after mealtimes.the treatment for anorexia, thereby framing anorexia primarily as both in and about the body  nothing else at all. Whilst at home for the weekend she would always walk to the same Starbucks, a mile or so from her home, and order the same latte and small pre-sealed packet of biscuits. She would then sit with these for two hours, sipping and nibbling, slowly feeling her anorexia, as she put it, “rise up around [her]”. The mechanics of this rising are intriguing; Hadia described how the biscuits—as intermingled signifiers of both the agency of choosing and paying for food herself  and the messy materiality of butter and sugar—were utterly terrifying. By sitting in Starbucks every week Hadia drew too close to food to be comfortable, thereby echoing discussion above of participants’ experiences of the threat posed by food in the EDU. Yet, this discomfort played an important role in Hadia’s anorexic embodiment, or rather perhaps, her active embodiment of anorexia. She intentionally incited her terror of food precisely because, in leading her to fear for her anorexia, it ‘triggered’ an upsurge of the illness; it made, she felt, her anorexia stronger. In her adherence to the same latte and biscuits each week, Hadia mobilised anorexia in order to hold on to it, creating a “safe space” carved through the illness. As such, her biscuits might appear at first glance to transfer her treatment regimen to the home spaces of weekend leave, adhering to the rules of the EDU and honouring her clinicians’ trust, as Eli put it. Yet, through them Hadia resisted their clinical conceptualisations of anorexia and selfhood. She felt these moments in Starbucks to release her anorexia from the grip of the EDU, thereby symbolically also shifting herself from that unwanted enclosed space to the other space offered, perhaps ambivalently, by anorexia; biscuits afforded Hadia a modality of “stepping out” of clinical conceptualisations and spaces. This echoes Kate’s reading of memoirs, which created what she felt to be an “anorexic space” within the space of the EDU, carving her own sense of everyday embodiment into the fabric of the patients’ lounge. Extending that discussion -  that food emerges here as an object through which anorexia may be maintained both goes against habitual understandings of the illness and yet is also clearly shaped by Hadia’s experiences of treatment. The biscuits were imbued with multiple meanings that did not necessarily connect to or even disrupt one another. Rather, for this moment in Starbucks they came powerfully to afford Hadia’s anorexic embodiment, recreating and renewing the “safe space” that she felt her illness to signify.On the EDU every morning all the patients had to spend fifteen minutes in the dining room for morning snacks. The second of the highly regulated six meals and 4000 calories a day, morning snacks comprised milk with chocolate, strawberry or banana Across the vignettes, anorexic embodiment has been seen to be forged through the clinic’s ordering of space, time and materiality—an ordering which becomes absorbed into, but also reconfigured by, individuals’ experiences. As such, what Hadia’s biscuits draw to the fore is that all the material engagements in this paper have been about far more than the linearity of resisting treatment. In their mutability and movement across spaces and boundaries, the objects have shown how anorexic embodiment is forged through the fraught and mobile relationships between the clinical and the mundane. In both treatment and domestic spaces, these are carved into one another in ways that problematize medicalised understandings of the relationship between the body and anorexia. In so doing, these material engagements have challenged an arguably pervasive privileging of discourse over materiality, showing how bodies and the objects and spaces they encounter develop through mutually productive processes.To date little scholarly attention has been paid to how materialities shape intimate experiences of mental ill-health. In interrogating how material objects and spaces forge, frame and disrupt what we term ‘anorexic embodiment’, this paper has opened up a consideration of how material environments relate to eating disorders and mental health experiences more broadly. We have sought to ask what theoretical and therapeutic possibilities, trajectories, and limitations are engendered by exploring mental health, illness, and treatment through a critical lens of materiality.Specifically, the objects explored in this paper have elucidated the central, but previously-overlooked, role of materiality in forging lived experiences of anorexia. Ensuing directly from participants’ narratives, objects and spaces emerged from both datasets as central to embodied practices of “being anorexic”  possibilities for material interaction prescribed by the clinic, participants thereby creatively mobilized material engagements to assert modes of being in the world. As such, reflecting on anorexia through a lens of materiality has shown that objects and the spaces they move through are a crucial part of the production of anorexic embodiment as this comes into being, is threatened, and is remade within treatment.pas de deux, an enactment of compliance with ward rules (staying seated) and simultaneous resistance , communicating and embodying ambivalence. Likewise, the exchange of memoirs and calorie counting books between patients was not only disruptive but also productive; within a clinical space, this exchange enacted alternative regimes of care defined in specifically non-clinical terms.Crucially, these analyses have also shown that this movement of objects is bidirectional. Everyday household objects—such as televisions, books, and chairs—featured centrally in ward spaces and practices, turning intimacy inside out  or craft a personal response to macro-environmental changes, such as a global pandemic, which fundamentally alter both human and non-human relationality.This analysis has thereby shown how, using the lens of objects and spaces, it becomes possible to elucidate the anorexic body as both phenomenologically lived and socio-medically constituted. In so doing, our paper has sought to extend clinical discussions of anorexia and demonstrate that moving towards a materialist account of eating disorders has profound implications for treatment.


Congenital pelvic arteriovenous malformation (AVM) is a rare vascular abnormality whereby arteries and veins are directly connected with malformed vascular plexus. Owing to its low incidence and nonspecific symptoms, the ultrasonographic characteristics of congenital pelvic AVM in males have been infrequently studied. A 30-year-old man visited our hospital complaining of progressive pain in the right lower abdomen and lumbar area since 2 months previously. Abdominal ultrasound (US) was performed at the initial examination and pelvic AVM was diagnosed, which was then confirmed by computed tomographic angiography. After right internal iliac artery embolization, the patient recovered uneventfully and remained asymptomatic during the 12-month follow-up period. Congenital pelvic AVM should thus be included in the differential diagnosis of pelvic cystic masses in males despite its low incidence, with US also being of great diagnostic value. We describe the ultrasonic features of AVM in detail and hope that this study may contribute to the ultrasonic diagnosis of congenital pelvic AVM in males. Pelvic arteriovenous malformation (AVM) is a rare vascular abnormality whereby arteries and veins are connected with a nidus of dysplastic vascular channels, and it is particularly rare in male patients . As mostThere have been some studies on congenital pelvic AVM in the last few decades despite its low incidence. However, previous studies  mainly aAgainst this background, we present a rare case of congenital pelvic AVM diagnosed by abdominal US and discuss the ultrasonic features for diagnosis. This work may contribute to the ultrasonic diagnosis of congenital pelvic AVM in the male population.A 30-year-old man visited our hospital with a complaint of progressive persistent colic in the right lower abdomen and lumbar area over a period of 2 months, with no fever, nausea, vomiting, frequency/urgency of urination, urodynia, or hematuria. The causative factor was unclear. The patient denied a history of tumor, trauma, or previous surgery. Physical examination was normal with no tenderness or rebound tenderness in the abdomen, and no palpable mass was found. Laboratory investigations  were unremarkable. As the initial imaging examination, abdominal US revealed multiple tortuous tubular structures within an irregularly shaped anechoic lesion (62 mm × 56 mm) located on the right side of the bladder. Color Doppler flow imaging (CDFI) displayed rich colorful blood flow signals as a colored mosaic pattern in the same lesion, and spectral Doppler showed continuous turbulent flow in the systolic and diastolic phases with a high-velocity low-resistance waveform . In the The patient underwent computed tomographic angiography (CTA) after the sonographic diagnosis of pelvic AVM, whereby a large pelvic AVM supplied mainly by the right internal iliac artery was found , thus coPelvic congenital AVMs in males are relatively rare. In 2002, Game et al.  have beeThe malformed vascular plexus formed by the remnant capillary network constitute the structures between the arterial and venous systems. AVM may occur in any part of the body but mainly in the brain, neck, lung, and kidney. Pelvic AVMs are relatively rare, especially in males. The presenting symptoms of pelvic AVM are nonspecific and can be either subtle or dramatic and may even be fatal . Both thDespite the fact that abdominal US is usually the initial routine imaging examination, according to previous literature reports, the diagnostic value of US remains questionable with regard to male congenital pelvic AVM. Game et al.  considerIn contrast to previous literature, we believe that, with the application of Doppler, US is also a potential tool of solid diagnostic value for congenital pelvic AVM in males. Mittal et al.  considerThe treatment options of congenital pelvic AVM in males depend on the severity of presenting symptoms, and asymptomatic or mildly symptomatic lesions are considered not to require therapy . A varieWe presented the case of a male congenital pelvic AVM first diagnosed by abdominal US. Furthermore, we described the ultrasonic features of AVM in detail and established some key points of differential diagnosis by US in pelvic cystic masses, rarely mentioned in previous literature. For pelvic cystic masses found on two-dimensional US, color Doppler and spectral Doppler are necessary for further differential diagnosis. Furthermore, congenital pelvic AVM should be included in the differential diagnosis of pelvic cystic masses in males despite its low incidence.