Effects of Fragmented Liuzijue Breathing Exercise on Sleep Quality, Anxiety, and Attention in College Students

Effects of Fragmented Liuzijue Breathing Exercise on Sleep Quality, Anxiety, and Attention in College Students | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effects of Fragmented Liuzijue Breathing Exercise on Sleep Quality, Anxiety, and Attention in College Students Yuanyuan Wei, Ahmad Taha Khalaf, Junyi Su, Yang Zhan, Shiyu Luo, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9357142/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 6 You are reading this latest preprint version Abstract Objective Health issues caused by prolonged sitting among college students are increasingly prominent, and fragmented exercise can provide an effective intervention. This study explored the application value of the traditional health-preserving exercise "Liuzijue" in fragmented exercise for college students through 10-minute training sessions during breaks conducted from April 2025 to May 2025. Methods A total of 34 college students (16 males and 18 females) were enrolled. After 6 weeks of Liuzijue breathing regulation training, serum brain-derived neurotrophic factor (BDNF) levels were measured. Anxiety status was assessed using psychological scales such as the Self-rating Anxiety Scale (SAS), and sleep status was evaluated using the Pittsburgh Sleep Quality Index (PSQI). Selective attention was assessed via the Stroop Color-Word Test (SCWT). Results After intervention, serum BDNF levels(22.6 ± 3.2 pg/ml) in the experimental group were significantly higher than those before intervention ༈15.2 ± 2.5 pg/ml༉and in the control group༈16.5 ± 2.8 pg/ml༉ (P < 0.01). SAS scores༈35.6 ± 4.2 points༉ and PSQI scores ༈4.8 ± 1.2 points༉in the experimental group were significantly lower than those before intervention ༈52.3 ± 5.8 points、11.2 ± 2.5 points༉and in the control group ༈50.9 ± 5.9 points、10.5 ± 2.2 points༉(P < 0.01). In the SCWT, the experimental group showed significantly shorter naming time༈59.6 ± 7.3 seconds༉ and fewer errors ༈2.1 ± 1.1 times༉in the interference task (conflicting color words), with significantly reduced time interference values༈24.9 ± 5.2 seconds༉ and error interference values༈0.9 ± 0.8 times༉ (P < 0.01), and these indices were significantly better than those in the control group༈76.5 ± 8.1 seconds、4.9 ± 1.2 times、41.7 ± 5.9 seconds、3.5 ± 1.3 times༉ (P < 0.01). However, there were no significant changes in indices of neutral tasks. No statistically significant differences were observed in all indices of the control group before and after intervention (P > 0.05). Conclusion This study confirms that Liuzijue breathing regulation can alleviate anxiety, improve sleep quality, and enhance attention by upregulating BDNF expression, providing research evidence for its application in anxiety prevention and treatment. Fragmented exercise Liuzijue College students Sleep quality Anxiety Introduction With the acceleration of social rhythms and the popularization of digital learning models, contemporary college students face prolonged sitting time and insufficient physical activity, leading to increasingly prominent physical and mental health risks [ 1 ]. Prolonged sedentary behavior not only causes physiological problems such as decreased physical fitness and metabolic disorders but also easily triggers psychological distress, including anxiety and stress accumulation [ 2 ], which in turn impair sleep quality and cognitive functions, especially attention and executive functions crucial for academic performance [ 3 ]. Sleep disorders and inattention have become common issues among college students, negatively affecting their quality of life, academic achievement, and long-term development. Against this background, fragmented exercise has attracted growing attention as a flexible health intervention that adapts to students' schedules. Breaking the constraints of time and space in traditional exercise, it can be easily integrated into fragmented scenarios such as class breaks and study intervals, providing a feasible solution to address insufficient physical activity among college students [ 4 ]. As a traditional health-preserving practice, Liuzijue emphasizes simplicity, ease of learning, and guidance on breathing regulation and mindfulness, making it uniquely suitable for fragmented settings [ 5 ]. Previous studies have shown that Liuzijue can improve autonomic nervous function and relieve psychological stress by regulating breathing rhythm [ 6 , 7 ]. However, its specific effects on key indicators such as sleep quality and attention when implemented as fragmented exercise among college students, as well as potential mechanisms (e.g., association with brain-derived neurotrophic factor, BDNF), remain to be further explored. Based on this, this study aimed to investigate the effects of fragmented Liuzijue breathing training on serum BDNF levels, anxiety, sleep quality, and attention in college students, and to reveal its potential mechanisms. The findings provide scientific evidence and practical guidance for using traditional health-promoting practices to improve college students’ physical and mental health and academic performance. Additionally, by analyzing the compatibility between Liuzijue’s characteristics and college students’ physical and mental needs, this study designed a simplified exercise protocol suitable for class breaks, exploring its positive effects on physical function, psychological stress relief, and learning efficiency, offering a theoretical basis and practical model for promoting health-related physical activity in colleges and universities. Materials and Methods 1.1 Study Subjects This study was approved by the Ethics Committee of Xinjiang Medical University (Ethics Approval No.: IACUC-20240514-182) and conducted from April to May 2025. A total of 34 undergraduate students aged 19–21 years were recruited from a university. Subjects were initially screened using the SAS, with exclusions for severe physical or mental illnesses, recent significant traumatic experiences, use of anti-anxiety or sedative-hypnotic drugs, and abnormal visual acuity (including uncorrected color blindness). Eligible subjects were randomly assigned to the experimental group (n = 18) and control group (n = 16) using a random number table. There were no significant differences in age, gender ratio, baseline SAS/PSQI scores, serum BDNF levels, or daily exercise frequency between the two groups (P > 0.05), indicating comparability. The study strictly adhered to ethical principles, ensuring subjects’ right to informed consent, voluntary participation, and privacy protection. Subjects could withdraw from the survey at any time, unconditionally and without affecting their relevant rights and interests. 1.2 Grouping and Intervention 1.2.1 Grouping Method Stratified random sampling was used, with baseline SAS scores (< 50 points, 50–60 points), gender, and daily exercise frequency as stratification factors. Within each stratum, subjects were assigned to the experimental or control group using random numbers generated by SPSS 26.0. No significant differences in general characteristics were observed between groups (P > 0.05). 1.2.2 Experimental Intervention Experimental Group: Received 6 weeks of Liuzijue breathing regulation training guided by professional instructors. Training was conducted twice daily (10:00 and 15:00 during class breaks), 10 minutes per session (including 2 minutes of warm-up, 6 minutes of core Liuzijue practice, and 2 minutes of relaxation), 5 days a week (10 sessions/week) for 6 weeks. Warm-up: 2 minutes of simple stretching and joint mobilization to relax the body. Core Liuzijue Practice: Performed according to the protocol in *Fitness Qigong·Liuzijue* edited by the Fitness Qigong Management Center of the General Administration of Sport of China [ 8 ]. The six characters ("Xu", "He", "Hu", "Si", "Chui", "Xi") were practiced in sequence. Each character was pronounced with a 6-second exhalation, followed by a natural inhalation, repeated 6 times as one set, for a total of 2 sets. Emphasis was placed on accurate pronunciation, deep, even breathing, and coordination with appropriate body movements (e.g., raising the hands and turning the head during "Xu" to enhance qi regulation). Relaxation: 2 minutes of progressive muscle relaxation and mindfulness meditation to calm the mind and consolidate training effects. Control Group: Maintained routine study and daily activities without additional intervention during the experimental period. 1.3 Outcome Measures and Methods 1.3.1 Serum BDNF Determination Fasting venous blood (2 ml) was collected at baseline (1 week before intervention) and endpoint (1 week after intervention). Serum BDNF levels were measured using ELISA kits (R&D Systems, USA) and a microplate reader (Bio-Tek ELx808, USA) at 450 nm absorbance. 1.3.2 Sleep Quality Assessment The Pittsburgh Sleep Quality Index (PSQI) was used to evaluate sleep quality over the past month [ 9 ]. The scale includes seven dimensions (e.g., sleep latency, sleep duration) with a total score ranging from 0 to 21; lower scores indicate better sleep quality. The PSQI has good reliability and validity among college students (Cronbach’s α = 0.82) [ 10 ]. 1.3.3 Anxiety Assessment The Self-rating Anxiety Scale (SAS) consists of 20 items covering physiological, emotional, cognitive, and behavioral dimensions, using a 4-point Likert scale [ 11 ]. Total scores were categorized using a cutoff of 50 points: 70 (severe anxiety). The SAS has stable reliability and validity in adults (Cronbach’s α = 0.83–0.85) [ 12 ]. 1.3.4 Attention and Executive Function Assessment The Stroop Color-Word Test (SCWT) was used to evaluate inhibitory control and interference sensitivity [ 13 ], a classic tool for executive cognitive function with good cross-population applicability (test-retest reliability: 0.76–0.84). The test included three tasks: (1) neutral color-word reading; (2) color block naming; (3) conflicting color-word naming. The experimental program was designed using E-Prime 3.0, with stimuli presented as 640×480-pixel images and subjects seated 38 cm from the screen. All participants completed cognitive assessments at baseline and at the endpoint in a quiet, well-lit environment. The instructions were: “Quickly and accurately judge the font color and press the corresponding key” (F = red, D = yellow, J = blue, K = green; color labels were placed on the keys). A red “+” fixation point was presented for 500 ms, followed by the stimulus, which remained on the screen for a maximum of 3000 ms or until a response was made. Participants completed 32 practice trials (8–12 per task, covering all colors), and a minimum accuracy of 90% was required to proceed to formal testing. The formal assessment consisted of 36 randomized trials per task. Reaction time (ms) and error counts were recorded automatically. Each assessment session lasted approximately 20 minutes. 1.4 Data Collection and Statistical Analysis Data were collected by two trained researchers, with double data entry and verification to ensure accuracy. SPSS 26.0 software was used for statistical analysis. After confirming normality and homogeneity of variance, a repeated-measures analysis of variance (ANOVA) was used to test group time interaction effects. If significant, paired t-tests (intra-group) and independent samples t-tests (inter-group, Bonferroni correction) were performed. Categorical data were compared using the χ2 test. P < 0.05 was considered statistically significant. Results 2.1 General Characteristics of Subjects No significant differences in age, gender ratio, baseline SAS/PSQI scores, or serum BDNF levels were observed between the two groups (P > 0.05), confirming comparability (Table 2.1 ). Table 2.1 Comparison of general conditions between the two groups of subjects (n, ‾X ± s) Indicators Experimental group(n = 18) Control group(n = 16) t/χ² P Value Age (years) 20.1 ± 0.8 19.9 ± 0.9 t = 0.63 0.53 Gender(Male/Female) 7(38.9%)/ 11(61.1%) 9(56.3%)/ 7(43.7%) χ²=0.41 0.52 Baseline SAS score 52.3 ± 5.8 51.8 ± 6.1 t = 0.35 0.73 Baseline PSQI score 11.2 ± 2.5 10.8 ± 2.3 t = 0.48 0.63 Baseline BDNF (pg/ml) 15.2 ± 2.5 14.8 ± 2.3 t = 0.42 0.68 2.2 Changes in Serum BDNF Levels At baseline, serum BDNF levels were comparable between groups (P > 0.05). After intervention, BDNF levels in the experimental group were significantly higher than baseline (P < 0.001) and the control group (P 0.05) (Table 2.2 ). Table 2.2 Comparison of serum BDNF levels between the two groups (n, X ± s, pg/ml) Group n Baseline BDNF (pg/ml) Post-intervention BDNF (pg/ml) Intra-group Comparison (t/P) Inter-group Comparison (t/P) Experimental group 18 15.2 ± 2.5 22.6 ± 3.2 t = 7.82 P < 0.001 t = 5.91 P 0.05 - 2.3 Changes in Self-Rating Anxiety Scale (SAS) Scores Before the intervention, there was no statistically significant difference in SAS scores between the two groups of students (P > 0.05). After the experiment, the SAS scores of students in the experimental group were significantly lower than before (P < 0.01) and significantly lower than those in the control group (P 0.05). Detailed data are presented in Table 2.3 Table 2.3 Changes in Self-Rating Anxiety Scale Scores of the Two Groups of Subjects (n, ‾X ± s) Group n Pre-Interv SAS score Post-Interv SAS score Intra-group Comp. (Pre vs.Post) Inter-group Comp. (Post) Exp.Grp. 18 52.3 ± 5.8 35.6 ± 4.2 t = 8.15 P < 0.001 t = 6.22 P 0.05 - 2.4 Changes in Pittsburgh Sleep Quality Index (PSQI) Scores Before the intervention, there was no statistically significant difference in PSQI scores between the two groups of students (P > 0.05). After the experiment, the PSQI scores of students in the experimental group were significantly lower than those before the experiment (P < 0.01) and significantly lower than those in the control group (P 0.05). Detailed data are presented in Table 2.4 . Table 2.4 Comparison of PSQI between the Two Groups of Subjects (n, ‾X ± s) Group n Pre-Interv PSQI Score Post-Interv PSQI Score Intra-group Comp. (Pre vs.Post) Inter-group Comp. (Post) Exp.Grp. 18 11.2 ± 2.5 4.8 ± 1.2 t = 10.32 P < 0.001 t = 8.95 P 0.05 - 2.5 Results of the Stroop Color-Word Test Before the intervention, there were no statistically significant differences in neutral tasks (color naming, word reading) or interference tasks (conflicting color-word naming) between the two groups (all p > 0.05). After the intervention, the experimental group showed a significant reduction in response time and error rates for conflicting color-word naming in the interference task, with marked decreases in time-interference and error-interference values (all p < 0.01). Inter-group comparison revealed that the indicators of the interference task in the experimental group after intervention were significantly superior to those in the control group (all p < 0.01). Detailed data are presented in Table 2.5 . Table 2.5 Comparison of Stroop Test Results between the Two Groups of Subjects (n,‾ X ± s) Indicators Group Before Interv After Interv Intra-group Comp. (Pre vs. Post) Inter-group Comp. (Post) Neutral Task -Color Word Reading Time (s) Exp.Grp. 32.5 ± 4.2 31.8 ± 3.9 t = 0.42, P > 0.05 t = 0.38, P > 0.05 Ctrl.Grp. 33.1 ± 4.5 32.9 ± 4.1 t = 0.28, P > 0.05 - Neutral Task - Color Block Naming Time (s) Exp.Grp. 35.2 ± 5.1 34.7 ± 4.8 t = 0.35, P > 0.05 t = 0.25, P > 0.05 Ctrl.Grp. 34.9 ± 5.3 34.6 ± 4.9 t = 0.21, P > 0.05 - Interf. Task- Conflicting Color-Word Naming Time (s) Exp.Grp. 78.3 ± 8.6 59.6 ± 7.3 t = 8.25,P < 0.001 t = 6.12, P 0.05 - Interf. Task - Conflicting Color-Word Error Count (times) Exp.Grp. 5.2 ± 1.8 2.1 ± 1.1 t = 5.32, P < 0.01 t = 4.98,P 0.05 - Interf.Effect- Time Interf. Value (s) Exp.Grp. 43.1 ± 6.5 24.9 ± 5.2 t = 7.89 P < 0.001 t = 6.05,P 0.05 - Interf.Effect- Error Interf. Value Exp.Grp. 3.8 ± 1.5 0.9 ± 0.8 t = 6.11 P < 0.01 t = 5.87 P 0.05 - Discussion After six weeks of fragmented Liuzijue exercise intervention, which was conducted 10 times a week for five consecutive days, the subjects showed varying degrees of improvement in anxiety, sleep quality, and attention. 3.1 Improvement of Sleep Quality and Anxiety by Liuzijue All subjects in this study exhibited mild anxiety (SAS > 50 points) accompanied by decreased serum brain-derived neurotrophic factor (BDNF) levels. Following the 6-week fragmented intervention, the experimental group showed a significant reduction in SAS scores and a marked elevation in BDNF levels compared with both the control group and their pre-intervention status. BDNF, a neurotrophic factor, primarily regulates neuronal synaptic plasticity and plays a crucial role in brain regions closely associated with emotional processing, such as the hippocampus and prefrontal cortex. Current research has demonstrated a negative correlation between BDNF levels and anxiety [ 11 , 12 ]. Meanwhile, numerous studies have indicated a bidirectional association between anxiety and sleep [ 13 , 14 ]. Compared with the control group and the pre-intervention baseline, subjects receiving the fragmented Liuzijue intervention showed a significant decrease in Pittsburgh Sleep Quality Index (PSQI) scores, suggesting that the fragmented Liuzijue intervention exerts a beneficial effect on sleep quality among college students. From a physiological perspective, the slow breathing pattern of Liuzijue, characterized by prolonged exhalation on specific syllables such as "He" and "Xi", may stimulate the vagus nerve [ 15 ], thereby inhibiting the overactivation of the hypothalamic-pituitary-adrenal (HPA) axis and reducing cortisol secretion. Existing evidence indicates that elevated cortisol levels are a key contributor to sleep-onset difficulties and sleep fragmentation [ 16 ]. Additionally, the alleviation of anxiety can reduce nocturnal rumination, thereby improving sleep continuity and forming a positive cycle of "anxiety reduction-sleep improvement". This finding is consistent with the research conclusions of Guo Shuai et al., who reported that traditional health-preserving exercises improve sleep through regulating emotional pathways [ 17 , 18 ]. During the 6-week intervention period, no significant changes were observed in anxiety and sleep indicators in the control group. 3.2 Improvement of Cognitive Function by Liuzijue and Its Neural Mechanisms The present study found that the experimental group showed significant improvements on the conflict task of the Stroop Color-Word Test (SCWT), as evidenced by shorter response times and reduced error rates in naming conflicting color words, as well as significant decreases in time and error interference. In contrast, no significant changes were observed in neutral tasks (color-word reading and color-block naming). These results indicate that Liuzijue breathing regulation does not enhance basic perceptual or reading abilities but specifically strengthens inhibitory control within executive functions, i.e., the ability to inhibit the dominant response of "automatically reading word meanings" and focus on the task of "naming font colors". This suggests that Liuzijue may simultaneously improve attention allocation and task-switching abilities, functions that primarily rely on the neural regulatory network of the prefrontal cortex [ 19 , 20 ]. A large body of research has confirmed that BDNF can promote synaptic plasticity in the prefrontal cortex and enhance the efficiency of neural circuit connections, particularly by playing a central role in regulating circuits related to inhibitory control and attention [ 21 , 22 ]. The current study demonstrated that short-term fragmented intervention effectively upregulates BDNF expression, which may enhance inhibitory control by improving prefrontal function. Furthermore, the "breathing regulation" of Liuzijue may reduce anxiety's interference with prefrontal function by balancing the autonomic nervous system (e.g., reducing sympathetic nervous system activity). This is consistent with the decreased SAS scores observed in the experimental group, forming a synergistic effect between emotional regulation and cognitive enhancement: on the one hand, the optimization of prefrontal neural circuits by BDNF directly enhances inhibitory control and attention; on the other hand, the autonomic nervous regulation induced by breathing exercises alleviates anxiety, removing obstacles to the normal functioning of the prefrontal cortex. Together, these two mechanisms contribute to improved cognitive performance. Although direct evidence linking Liuzijue to prefrontal function is currently lacking, several studies have reported similar findings. For instance, Birdee et al. [ 23 ] found that yoga-style slow exhalation can significantly reduce psychological stress. A recent survey by Balban et al. [ 24 ] showed that, compared with mindfulness meditation, breathing exercises, especially cyclic exhalation-focused breathing, are more effective at improving mood and reducing respiratory rate. 3.3 Research Significance of Liuzijue in Modern Neuroscience Originating in the Huangting Scripture, Liuzijue is a traditional health-preserving exercise that focuses on achieving "harmonious cultivation of body and spirit" through regulated breathing and focused attention [ 25 , 26 ]. The present study confirmed that fragmented Liuzijue intervention can upregulate BDNF expression, providing a potential molecular basis for the TCM theory that "qi is the mother of spirit. This interdisciplinary research enriches the scientific connotation of traditional health-preserving theories. Compared with aerobic exercise, Liuzijue features gentle, slow movements that avoid disrupting students' inter-class state. Its emphasis on prolonged exhalation may directly affect cerebral blood supply [ 27 ], which may be one of the reasons for the short-term and efficient intervention effects observed in this study. Future research could further explore this mechanism by incorporating neuroimaging techniques such as functional magnetic resonance imaging (fMRI) or electroencephalography (EEG) [28, 29]. 3.4 Research Limitations and Future Directions This study has several limitations. First, the small sample size and single-center design may limit the generalizability of the results; future studies should expand the sample size to more than 50 subjects per group. Second, in terms of mechanism research, only BDNF was measured as an indicator. Future studies should investigate additional related pathways, such as neurotransmitters like serotonin and gamma-aminobutyric acid (GABA). The lack of neuroimaging data on brain regions such as the prefrontal cortex also prevents direct confirmation of improvements in brain function. Third, long-term effects are lacking: the 6-week intervention only reflects short-term outcomes, and the sustained impact of Liuzijue, including the duration of effect maintenance after ceasing practice and the dose-response relationship, remains unclear. To address these issues, future research should increase the sample size, extend the study duration, and incorporate diffusion tensor imaging (DTI) to examine the effects of Liuzijue on prefrontal-limbic system connectivity. Declarations Ethics approval and consent to participate: The study adhered to the Declaration of Helsinki. The University Committee of Xinjiang Medical University approved the study. Informed consent to participate was obtained from all participants in the study. Consent for publication: Consent for publication was obtained from all participants involved in the study. Competing Interests: No conflicts of interest Clinical trial number not applicable. Funding: This study was supported by the Higher Education Talent Training Quality and Teaching Reform Project of Sichuan Province (2024–2026; No. 2024016) and by the Higher Education Scientific Research Planning Project entitled Study on the Interactive Effects of Exercise and Diet on Health-Related Physical Fitness of Overweight and Obese College Students (No. 23TY0213). Author Contribution Y.W. and A.T.K. contributed equally to this work. Y.W., A.T.K., and J.S. conceived and designed the study. Y.W., Y.Z., and S.L. collected the data. A.T.K., J.S., and K.J. performed data analysis and interpretation. T.J. supervised the study and provided critical revisions. All authors reviewed and approved the final manuscript. 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Diagram of the Interior of Huangting's Body Showing the Balancing and Suppression of Internal Organs[M].Ming Dynasty: Orthodox Taoist Canon, Dongxuan Section Spiritual Chart Category. ZOU P A.The Liuzijue Breath Technique of the Supreme Deity[M].Song Dynasty:Folk Hand - copied Edition. Pontus Söderström, Anders Eklund, Nina Karalija, et al.Respiratory influence on cerebral blood flow and blood volume - A 4D flow MRI study [J]..J. Cereb. Blood Flow Metab.,45(8),271678X251316395. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Reviews received at journal 18 May, 2026 Reviewers agreed at journal 18 May, 2026 Reviewers invited by journal 09 Apr, 2026 Editor assigned by journal 09 Apr, 2026 Submission checks completed at journal 09 Apr, 2026 First submitted to journal 08 Apr, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-9357142","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":620494903,"identity":"0ab9a515-35e1-4d82-8064-4147669cee66","order_by":0,"name":"Yuanyuan Wei","email":"","orcid":"","institution":"Chengdu University","correspondingAuthor":false,"prefix":"","firstName":"Yuanyuan","middleName":"","lastName":"Wei","suffix":""},{"id":620494904,"identity":"47f08478-2402-4afc-9d56-139ca1f09704","order_by":1,"name":"Ahmad Taha Khalaf","email":"","orcid":"","institution":"Anhui University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Ahmad","middleName":"Taha","lastName":"Khalaf","suffix":""},{"id":620494905,"identity":"42787134-0050-4128-a85c-453b0428d436","order_by":2,"name":"Junyi Su","email":"","orcid":"","institution":"The University of Newcastle","correspondingAuthor":false,"prefix":"","firstName":"Junyi","middleName":"","lastName":"Su","suffix":""},{"id":620494906,"identity":"dfb2835a-6087-4411-9149-bbc6f598041f","order_by":3,"name":"Yang Zhan","email":"","orcid":"","institution":"Chengdu University","correspondingAuthor":false,"prefix":"","firstName":"Yang","middleName":"","lastName":"Zhan","suffix":""},{"id":620494907,"identity":"674eb40f-d9e3-47df-92d5-caacf22a261c","order_by":4,"name":"Shiyu Luo","email":"","orcid":"","institution":"Chengdu University","correspondingAuthor":false,"prefix":"","firstName":"Shiyu","middleName":"","lastName":"Luo","suffix":""},{"id":620494908,"identity":"d423f188-2b68-4527-a9d4-dca785c5b8ce","order_by":5,"name":"Kunrong Jing","email":"","orcid":"","institution":"Chengdu University","correspondingAuthor":false,"prefix":"","firstName":"Kunrong","middleName":"","lastName":"Jing","suffix":""},{"id":620494909,"identity":"79cf68a3-6189-4e14-b074-4d31d55dfbfa","order_by":6,"name":"Tao Jiang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1ElEQVRIiWNgGAWjYBACNvnDBx98/GMjx9jffIA4LXwSbMmGMxvSjJlnHEsgToucBI+ZNG/D4UT2hhwDIh0m3WAgzbvjcAJvw5mPN94w2MnpNhDSInMgwXDumfQ8yebezZZzGJKNzQ4Q0sKQcCDhDZt1sWHD2W3SPAwHErcR1pLYcICHjTlx/4GcZ0RqkUhmbORtc05sbMhhI1ILzzFmxhln0owZZxwztpxjQIRf5Nv7v//4UAGOyoc33lTYyRHUggIkeIiMGmQtpOoYBaNgFIyCEQEAsvtGIryyxmUAAAAASUVORK5CYII=","orcid":"","institution":"Xinjiang Medical University","correspondingAuthor":true,"prefix":"","firstName":"Tao","middleName":"","lastName":"Jiang","suffix":""}],"badges":[],"createdAt":"2026-04-08 12:41:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9357142/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9357142/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":107483675,"identity":"ee3c2464-a8f9-46da-9e0c-ae6b0fb1ad76","added_by":"auto","created_at":"2026-04-22 02:28:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":428433,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9357142/v1/7666885d-45dd-41d5-88bc-8f82b07c29b9.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Effects of Fragmented Liuzijue Breathing Exercise on Sleep Quality, Anxiety, and Attention in College Students","fulltext":[{"header":"Introduction","content":"\u003cp\u003eWith the acceleration of social rhythms and the popularization of digital learning models, contemporary college students face prolonged sitting time and insufficient physical activity, leading to increasingly prominent physical and mental health risks [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Prolonged sedentary behavior not only causes physiological problems such as decreased physical fitness and metabolic disorders but also easily triggers psychological distress, including anxiety and stress accumulation [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e], which in turn impair sleep quality and cognitive functions, especially attention and executive functions crucial for academic performance [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Sleep disorders and inattention have become common issues among college students, negatively affecting their quality of life, academic achievement, and long-term development. Against this background, fragmented exercise has attracted growing attention as a flexible health intervention that adapts to students' schedules. Breaking the constraints of time and space in traditional exercise, it can be easily integrated into fragmented scenarios such as class breaks and study intervals, providing a feasible solution to address insufficient physical activity among college students [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. As a traditional health-preserving practice, Liuzijue emphasizes simplicity, ease of learning, and guidance on breathing regulation and mindfulness, making it uniquely suitable for fragmented settings [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Previous studies have shown that Liuzijue can improve autonomic nervous function and relieve psychological stress by regulating breathing rhythm [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. However, its specific effects on key indicators such as sleep quality and attention when implemented as fragmented exercise among college students, as well as potential mechanisms (e.g., association with brain-derived neurotrophic factor, BDNF), remain to be further explored.\u003c/p\u003e \u003cp\u003eBased on this, this study aimed to investigate the effects of fragmented Liuzijue breathing training on serum BDNF levels, anxiety, sleep quality, and attention in college students, and to reveal its potential mechanisms. The findings provide scientific evidence and practical guidance for using traditional health-promoting practices to improve college students\u0026rsquo; physical and mental health and academic performance. Additionally, by analyzing the compatibility between Liuzijue\u0026rsquo;s characteristics and college students\u0026rsquo; physical and mental needs, this study designed a simplified exercise protocol suitable for class breaks, exploring its positive effects on physical function, psychological stress relief, and learning efficiency, offering a theoretical basis and practical model for promoting health-related physical activity in colleges and universities.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e1.1 Study Subjects\u003c/h2\u003e \u003cp\u003e This study was approved by the Ethics Committee of Xinjiang Medical University (Ethics Approval No.: IACUC-20240514-182) and conducted from April to May 2025. A total of 34 undergraduate students aged 19\u0026ndash;21 years were recruited from a university. Subjects were initially screened using the SAS, with exclusions for severe physical or mental illnesses, recent significant traumatic experiences, use of anti-anxiety or sedative-hypnotic drugs, and abnormal visual acuity (including uncorrected color blindness). Eligible subjects were randomly assigned to the experimental group (n\u0026thinsp;=\u0026thinsp;18) and control group (n\u0026thinsp;=\u0026thinsp;16) using a random number table. There were no significant differences in age, gender ratio, baseline SAS/PSQI scores, serum BDNF levels, or daily exercise frequency between the two groups (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05), indicating comparability. The study strictly adhered to ethical principles, ensuring subjects\u0026rsquo; right to informed consent, voluntary participation, and privacy protection. Subjects could withdraw from the survey at any time, unconditionally and without affecting their relevant rights and interests.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e1.2 Grouping and Intervention\u003c/h2\u003e \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e \u003ch2\u003e1.2.1 Grouping Method\u003c/h2\u003e \u003cp\u003eStratified random sampling was used, with baseline SAS scores (\u0026lt;\u0026thinsp;50 points, 50\u0026ndash;60 points), gender, and daily exercise frequency as stratification factors. Within each stratum, subjects were assigned to the experimental or control group using random numbers generated by SPSS 26.0. No significant differences in general characteristics were observed between groups (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e1.2.2 Experimental Intervention\u003c/h2\u003e \u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003eExperimental Group: Received 6 weeks of Liuzijue breathing regulation training guided by professional instructors. Training was conducted twice daily (10:00 and 15:00 during class breaks), 10 minutes per session (including 2 minutes of warm-up, 6 minutes of core Liuzijue practice, and 2 minutes of relaxation), 5 days a week (10 sessions/week) for 6 weeks.\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003eWarm-up: 2 minutes of simple stretching and joint mobilization to relax the body.\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003eCore Liuzijue Practice: Performed according to the protocol in *Fitness Qigong\u0026middot;Liuzijue* edited by the Fitness Qigong Management Center of the General Administration of Sport of China [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. The six characters (\"Xu\", \"He\", \"Hu\", \"Si\", \"Chui\", \"Xi\") were practiced in sequence. Each character was pronounced with a 6-second exhalation, followed by a natural inhalation, repeated 6 times as one set, for a total of 2 sets. Emphasis was placed on accurate pronunciation, deep, even breathing, and coordination with appropriate body movements (e.g., raising the hands and turning the head during \"Xu\" to enhance qi regulation).\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003eRelaxation: 2 minutes of progressive muscle relaxation and mindfulness meditation to calm the mind and consolidate training effects.\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003eControl Group: Maintained routine study and daily activities without additional intervention during the experimental period.\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e1.3 Outcome Measures and Methods\u003c/h2\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e1.3.1 Serum BDNF Determination\u003c/h2\u003e \u003cp\u003eFasting venous blood (2 ml) was collected at baseline (1 week before intervention) and endpoint (1 week after intervention). Serum BDNF levels were measured using ELISA kits (R\u0026amp;D Systems, USA) and a microplate reader (Bio-Tek ELx808, USA) at 450 nm absorbance.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e1.3.2 Sleep Quality Assessment\u003c/h2\u003e \u003cp\u003eThe Pittsburgh Sleep Quality Index (PSQI) was used to evaluate sleep quality over the past month [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. The scale includes seven dimensions (e.g., sleep latency, sleep duration) with a total score ranging from 0 to 21; lower scores indicate better sleep quality. The PSQI has good reliability and validity among college students (Cronbach\u0026rsquo;s α\u0026thinsp;=\u0026thinsp;0.82) [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e1.3.3 Anxiety Assessment\u003c/h2\u003e \u003cp\u003eThe Self-rating Anxiety Scale (SAS) consists of 20 items covering physiological, emotional, cognitive, and behavioral dimensions, using a 4-point Likert scale [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Total scores were categorized using a cutoff of 50 points: \u0026lt;50 (normal), 50\u0026ndash;60 (mild anxiety), 61\u0026ndash;70 (moderate anxiety), and \u0026gt;\u0026thinsp;70 (severe anxiety). The SAS has stable reliability and validity in adults (Cronbach\u0026rsquo;s α\u0026thinsp;=\u0026thinsp;0.83\u0026ndash;0.85) [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e \u003ch2\u003e1.3.4 Attention and Executive Function Assessment\u003c/h2\u003e \u003cp\u003eThe Stroop Color-Word Test (SCWT) was used to evaluate inhibitory control and interference sensitivity [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], a classic tool for executive cognitive function with good cross-population applicability (test-retest reliability: 0.76\u0026ndash;0.84). The test included three tasks: (1) neutral color-word reading; (2) color block naming; (3) conflicting color-word naming. The experimental program was designed using E-Prime 3.0, with stimuli presented as 640\u0026times;480-pixel images and subjects seated 38 cm from the screen.\u003c/p\u003e \u003cp\u003e All participants completed cognitive assessments at baseline and at the endpoint in a quiet, well-lit environment. The instructions were: \u0026ldquo;Quickly and accurately judge the font color and press the corresponding key\u0026rdquo; (F\u0026thinsp;=\u0026thinsp;red, D\u0026thinsp;=\u0026thinsp;yellow, J\u0026thinsp;=\u0026thinsp;blue, K\u0026thinsp;=\u0026thinsp;green; color labels were placed on the keys). A red \u0026ldquo;+\u0026rdquo; fixation point was presented for 500 ms, followed by the stimulus, which remained on the screen for a maximum of 3000 ms or until a response was made. Participants completed 32 practice trials (8\u0026ndash;12 per task, covering all colors), and a minimum accuracy of 90% was required to proceed to formal testing. The formal assessment consisted of 36 randomized trials per task. Reaction time (ms) and error counts were recorded automatically. Each assessment session lasted approximately 20 minutes.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e1.4 Data Collection and Statistical Analysis\u003c/h2\u003e \u003cp\u003eData were collected by two trained researchers, with double data entry and verification to ensure accuracy. SPSS 26.0 software was used for statistical analysis. After confirming normality and homogeneity of variance, a repeated-measures analysis of variance (ANOVA) was used to test group time interaction effects. If significant, paired t-tests (intra-group) and independent samples t-tests (inter-group, Bonferroni correction) were performed. Categorical data were compared using the χ2 test. P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e2.1 General Characteristics of Subjects\u003c/h2\u003e \u003cp\u003eNo significant differences in age, gender ratio, baseline SAS/PSQI scores, or serum BDNF levels were observed between the two groups (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05), confirming comparability (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e2.1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2.1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of general conditions between the two groups of subjects (n, \u0026oline;X\u0026thinsp;\u0026plusmn;\u0026thinsp;s)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIndicators\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eExperimental group(n\u0026thinsp;=\u0026thinsp;18)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl group(n\u0026thinsp;=\u0026thinsp;16)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003et/χ\u0026sup2;\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eP Value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge (years)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.53\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGender(Male/Female)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7(38.9%)/ 11(61.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9(56.3%)/ 7(43.7%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eχ\u0026sup2;=0.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.52\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBaseline SAS score\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e52.3\u0026thinsp;\u0026plusmn;\u0026thinsp;5.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e51.8\u0026thinsp;\u0026plusmn;\u0026thinsp;6.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBaseline PSQI score\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.63\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBaseline BDNF (pg/ml)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.68\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Changes in Serum BDNF Levels\u003c/h2\u003e \u003cp\u003eAt baseline, serum BDNF levels were comparable between groups (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05). After intervention, BDNF levels in the experimental group were significantly higher than baseline (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and the control group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001). No significant change was observed in the control group (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2.2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2.2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of serum BDNF levels between the two groups (n, X\u0026thinsp;\u0026plusmn;\u0026thinsp;s, pg/ml)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBaseline BDNF\u003c/p\u003e \u003cp\u003e(pg/ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePost-intervention BDNF\u003c/p\u003e \u003cp\u003e(pg/ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eIntra-group Comparison (t/P)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eInter-group Comparison (t/P)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExperimental group\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e15.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e22.6\u0026thinsp;\u0026plusmn;\u0026thinsp;3.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;7.82 P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;5.91 P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl group\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e14.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e16.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;1.24, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Changes in Self-Rating Anxiety Scale (SAS) Scores\u003c/h2\u003e \u003cp\u003eBefore the intervention, there was no statistically significant difference in SAS scores between the two groups of students (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05). After the experiment, the SAS scores of students in the experimental group were significantly lower than before (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) and significantly lower than those in the control group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01). There was no statistically significant difference in SAS scores of students in the control group before and after the experiment (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Detailed data are presented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e2.3\u003c/span\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2.3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eChanges in Self-Rating Anxiety Scale Scores of the Two Groups of Subjects (n, \u0026oline;X\u0026thinsp;\u0026plusmn;\u0026thinsp;s)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePre-Interv\u003c/p\u003e \u003cp\u003eSAS score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePost-Interv\u003c/p\u003e \u003cp\u003eSAS score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eIntra-group Comp. (Pre vs.Post)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eInter-group Comp. (Post)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExp.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e52.3\u0026thinsp;\u0026plusmn;\u0026thinsp;5.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e35.6\u0026thinsp;\u0026plusmn;\u0026thinsp;4.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;8.15 P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;6.22 P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCtrl.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e51.8\u0026thinsp;\u0026plusmn;\u0026thinsp;6.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e50.9\u0026thinsp;\u0026plusmn;\u0026thinsp;5.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.38 P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Changes in Pittsburgh Sleep Quality Index (PSQI) Scores\u003c/h2\u003e \u003cp\u003eBefore the intervention, there was no statistically significant difference in PSQI scores between the two groups of students (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05). After the experiment, the PSQI scores of students in the experimental group were significantly lower than those before the experiment (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) and significantly lower than those in the control group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01). There was no statistically significant difference in PSQI scores of students in the control group before and after the experiment (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Detailed data are presented in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e2.4\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2.4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of PSQI between the Two Groups of Subjects (n, \u0026oline;X\u0026thinsp;\u0026plusmn;\u0026thinsp;s)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePre-Interv PSQI Score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePost-Interv PSQI Score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eIntra-group Comp. (Pre vs.Post)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eInter-group Comp. (Post)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExp.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e11.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e4.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;10.32 P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;8.95 P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCtrl.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e10.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e10.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.36 P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Results of the Stroop Color-Word Test\u003c/h2\u003e \u003cp\u003eBefore the intervention, there were no statistically significant differences in neutral tasks (color naming, word reading) or interference tasks (conflicting color-word naming) between the two groups (all p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). After the intervention, the experimental group showed a significant reduction in response time and error rates for conflicting color-word naming in the interference task, with marked decreases in time-interference and error-interference values (all p\u0026thinsp;\u0026lt;\u0026thinsp;0.01). Inter-group comparison revealed that the indicators of the interference task in the experimental group after intervention were significantly superior to those in the control group (all p\u0026thinsp;\u0026lt;\u0026thinsp;0.01). Detailed data are presented in Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e2.5\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2.5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of Stroop Test Results between the Two Groups of Subjects (n,\u0026oline; X\u0026thinsp;\u0026plusmn;\u0026thinsp;s)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIndicators\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBefore Interv\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAfter Interv\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eIntra-group Comp. (Pre vs. Post)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eInter-group Comp. (Post)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNeutral Task -Color Word Reading Time (s)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eExp.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e32.5\u0026thinsp;\u0026plusmn;\u0026thinsp;4.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e31.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.42, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.38, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCtrl.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e33.1\u0026thinsp;\u0026plusmn;\u0026thinsp;4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e32.9\u0026thinsp;\u0026plusmn;\u0026thinsp;4.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.28, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNeutral Task - Color Block Naming Time (s)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eExp.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e35.2\u0026thinsp;\u0026plusmn;\u0026thinsp;5.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e34.7\u0026thinsp;\u0026plusmn;\u0026thinsp;4.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.35, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.25, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCtrl.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e34.9\u0026thinsp;\u0026plusmn;\u0026thinsp;5.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e34.6\u0026thinsp;\u0026plusmn;\u0026thinsp;4.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.21, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eInterf. Task- Conflicting Color-Word Naming Time (s)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eExp.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e78.3\u0026thinsp;\u0026plusmn;\u0026thinsp;8.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e59.6\u0026thinsp;\u0026plusmn;\u0026thinsp;7.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;8.25,P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;6.12, P\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCtrl.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e77.9\u0026thinsp;\u0026plusmn;\u0026thinsp;8.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e76.5\u0026thinsp;\u0026plusmn;\u0026thinsp;8.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.41, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eInterf. Task - Conflicting Color-Word Error Count (times)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eExp.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e5.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e2.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;5.32, P\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;4.98,P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCtrl.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e5.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e4.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.31, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eInterf.Effect- Time Interf.\u003c/p\u003e \u003cp\u003eValue (s)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eExp.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e43.1\u0026thinsp;\u0026plusmn;\u0026thinsp;6.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e24.9\u0026thinsp;\u0026plusmn;\u0026thinsp;5.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;7.89 P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;6.05,P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCtrl.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e42.8\u0026thinsp;\u0026plusmn;\u0026thinsp;6.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e41.7\u0026thinsp;\u0026plusmn;\u0026thinsp;5.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.45, P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eInterf.Effect- Error Interf. Value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eExp.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e3.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e0.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;6.11 P\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;5.87 P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCtrl.Grp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e3.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e3.5\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003et\u0026thinsp;=\u0026thinsp;0.29 P\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eAfter six weeks of fragmented Liuzijue exercise intervention, which was conducted 10 times a week for five consecutive days, the subjects showed varying degrees of improvement in anxiety, sleep quality, and attention.\u003c/p\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Improvement of Sleep Quality and Anxiety by Liuzijue\u003c/h2\u003e \u003cp\u003eAll subjects in this study exhibited mild anxiety (SAS\u0026thinsp;\u0026gt;\u0026thinsp;50 points) accompanied by decreased serum brain-derived neurotrophic factor (BDNF) levels. Following the 6-week fragmented intervention, the experimental group showed a significant reduction in SAS scores and a marked elevation in BDNF levels compared with both the control group and their pre-intervention status. BDNF, a neurotrophic factor, primarily regulates neuronal synaptic plasticity and plays a crucial role in brain regions closely associated with emotional processing, such as the hippocampus and prefrontal cortex. Current research has demonstrated a negative correlation between BDNF levels and anxiety [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Meanwhile, numerous studies have indicated a bidirectional association between anxiety and sleep [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Compared with the control group and the pre-intervention baseline, subjects receiving the fragmented Liuzijue intervention showed a significant decrease in Pittsburgh Sleep Quality Index (PSQI) scores, suggesting that the fragmented Liuzijue intervention exerts a beneficial effect on sleep quality among college students.\u003c/p\u003e \u003cp\u003eFrom a physiological perspective, the slow breathing pattern of Liuzijue, characterized by prolonged exhalation on specific syllables such as \"He\" and \"Xi\", may stimulate the vagus nerve [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e], thereby inhibiting the overactivation of the hypothalamic-pituitary-adrenal (HPA) axis and reducing cortisol secretion. Existing evidence indicates that elevated cortisol levels are a key contributor to sleep-onset difficulties and sleep fragmentation [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Additionally, the alleviation of anxiety can reduce nocturnal rumination, thereby improving sleep continuity and forming a positive cycle of \"anxiety reduction-sleep improvement\". This finding is consistent with the research conclusions of Guo Shuai et al., who reported that traditional health-preserving exercises improve sleep through regulating emotional pathways [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. During the 6-week intervention period, no significant changes were observed in anxiety and sleep indicators in the control group.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Improvement of Cognitive Function by Liuzijue and Its Neural Mechanisms\u003c/h2\u003e \u003cp\u003eThe present study found that the experimental group showed significant improvements on the conflict task of the Stroop Color-Word Test (SCWT), as evidenced by shorter response times and reduced error rates in naming conflicting color words, as well as significant decreases in time and error interference. In contrast, no significant changes were observed in neutral tasks (color-word reading and color-block naming). These results indicate that Liuzijue breathing regulation does not enhance basic perceptual or reading abilities but specifically strengthens inhibitory control within executive functions, i.e., the ability to inhibit the dominant response of \"automatically reading word meanings\" and focus on the task of \"naming font colors\". This suggests that Liuzijue may simultaneously improve attention allocation and task-switching abilities, functions that primarily rely on the neural regulatory network of the prefrontal cortex [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eA large body of research has confirmed that BDNF can promote synaptic plasticity in the prefrontal cortex and enhance the efficiency of neural circuit connections, particularly by playing a central role in regulating circuits related to inhibitory control and attention [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. The current study demonstrated that short-term fragmented intervention effectively upregulates BDNF expression, which may enhance inhibitory control by improving prefrontal function. Furthermore, the \"breathing regulation\" of Liuzijue may reduce anxiety's interference with prefrontal function by balancing the autonomic nervous system (e.g., reducing sympathetic nervous system activity). This is consistent with the decreased SAS scores observed in the experimental group, forming a synergistic effect between emotional regulation and cognitive enhancement: on the one hand, the optimization of prefrontal neural circuits by BDNF directly enhances inhibitory control and attention; on the other hand, the autonomic nervous regulation induced by breathing exercises alleviates anxiety, removing obstacles to the normal functioning of the prefrontal cortex. Together, these two mechanisms contribute to improved cognitive performance.\u003c/p\u003e \u003cp\u003eAlthough direct evidence linking Liuzijue to prefrontal function is currently lacking, several studies have reported similar findings. For instance, Birdee et al. [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] found that yoga-style slow exhalation can significantly reduce psychological stress. A recent survey by Balban et al. [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] showed that, compared with mindfulness meditation, breathing exercises, especially cyclic exhalation-focused breathing, are more effective at improving mood and reducing respiratory rate.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Research Significance of Liuzijue in Modern Neuroscience\u003c/h2\u003e \u003cp\u003eOriginating in the Huangting Scripture, Liuzijue is a traditional health-preserving exercise that focuses on achieving \"harmonious cultivation of body and spirit\" through regulated breathing and focused attention [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. The present study confirmed that fragmented Liuzijue intervention can upregulate BDNF expression, providing a potential molecular basis for the TCM theory that \"qi is the mother of spirit. This interdisciplinary research enriches the scientific connotation of traditional health-preserving theories.\u003c/p\u003e \u003cp\u003eCompared with aerobic exercise, Liuzijue features gentle, slow movements that avoid disrupting students' inter-class state. Its emphasis on prolonged exhalation may directly affect cerebral blood supply [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], which may be one of the reasons for the short-term and efficient intervention effects observed in this study. Future research could further explore this mechanism by incorporating neuroimaging techniques such as functional magnetic resonance imaging (fMRI) or electroencephalography (EEG) [28, 29].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec23\" class=\"Section2\"\u003e \u003ch2\u003e3.4 Research Limitations and Future Directions\u003c/h2\u003e \u003cp\u003eThis study has several limitations. First, the small sample size and single-center design may limit the generalizability of the results; future studies should expand the sample size to more than 50 subjects per group. Second, in terms of mechanism research, only BDNF was measured as an indicator. Future studies should investigate additional related pathways, such as neurotransmitters like serotonin and gamma-aminobutyric acid (GABA). The lack of neuroimaging data on brain regions such as the prefrontal cortex also prevents direct confirmation of improvements in brain function. Third, long-term effects are lacking: the 6-week intervention only reflects short-term outcomes, and the sustained impact of Liuzijue, including the duration of effect maintenance after ceasing practice and the dose-response relationship, remains unclear.\u003c/p\u003e \u003cp\u003eTo address these issues, future research should increase the sample size, extend the study duration, and incorporate diffusion tensor imaging (DTI) to examine the effects of Liuzijue on prefrontal-limbic system connectivity.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003cstrong\u003eEthics approval and consent to participate:\u003c/strong\u003e \u003cp\u003e The study adhered to the Declaration of Helsinki. The University Committee of Xinjiang Medical University approved the study. Informed consent to participate was obtained from all participants in the study.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eConsent for publication:\u003c/strong\u003e \u003cp\u003eConsent for publication was obtained from all participants involved in the study.\u003c/p\u003e \u003c/p\u003e\u003cp\u003e \u003ch2\u003eCompeting Interests:\u003c/h2\u003e \u003cp\u003eNo conflicts of interest\u003c/p\u003e \u003c/p\u003e\u003cp\u003e \u003ch2\u003eClinical trial number\u003c/h2\u003e \u003cp\u003enot applicable.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding:\u003c/h2\u003e \u003cp\u003eThis study was supported by the Higher Education Talent Training Quality and Teaching Reform Project of Sichuan Province (2024\u0026ndash;2026; No. 2024016) and by the Higher Education Scientific Research Planning Project entitled Study on the Interactive Effects of Exercise and Diet on Health-Related Physical Fitness of Overweight and Obese College Students (No. 23TY0213).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eY.W. and A.T.K. contributed equally to this work. Y.W., A.T.K., and J.S. conceived and designed the study. Y.W., Y.Z., and S.L. collected the data. A.T.K., J.S., and K.J. performed data analysis and interpretation. T.J. supervised the study and provided critical revisions. All authors reviewed and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eAvailable from the corresponding author upon reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eGAO X L, ZHANG J H, YANG Y, et al. Meta-analysis of sedentary behavior/screen time and mental health among college students [J]. Chin J Epidemiol, 2023, 44(3): 477\u0026ndash;485.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJiang, L., Cao, et al.(2020). Association of Sedentary Behavior With Anxiety, Depression, and Suicide Ideation in College Students[J]. Front. 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Brief structured respiration practices enhance mood and reduce physiological arousal[J].Cell Rep Med, 4(1), 100895\u0026ndash;100895.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHU Y. Diagram of the Interior of Huangting's Body Showing the Balancing and Suppression of Internal Organs[M].Ming Dynasty: Orthodox Taoist Canon, Dongxuan Section Spiritual Chart Category.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZOU P A.The Liuzijue Breath Technique of the Supreme Deity[M].Song Dynasty:Folk Hand - copied Edition.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePontus S\u0026ouml;derstr\u0026ouml;m, Anders Eklund, Nina Karalija, et al.Respiratory influence on cerebral blood flow and blood volume - A 4D flow MRI study [J]..J. Cereb. Blood Flow Metab.,45(8),271678X251316395.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"sport-sciences-for-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ssfh","sideBox":"Learn more about [Sport Sciences for Health](http://link.springer.com/journal/11332)","snPcode":"11332","submissionUrl":"https://submission.nature.com/new-submission/11332/3","title":"Sport Sciences for Health","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Fragmented exercise, Liuzijue, College students, Sleep quality, Anxiety","lastPublishedDoi":"10.21203/rs.3.rs-9357142/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9357142/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eObjective\u003c/h2\u003e \u003cp\u003eHealth issues caused by prolonged sitting among college students are increasingly prominent, and fragmented exercise can provide an effective intervention. This study explored the application value of the traditional health-preserving exercise \"Liuzijue\" in fragmented exercise for college students through 10-minute training sessions during breaks conducted from April 2025 to May 2025.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eA total of 34 college students (16 males and 18 females) were enrolled. After 6 weeks of Liuzijue breathing regulation training, serum brain-derived neurotrophic factor (BDNF) levels were measured. Anxiety status was assessed using psychological scales such as the Self-rating Anxiety Scale (SAS), and sleep status was evaluated using the Pittsburgh Sleep Quality Index (PSQI). Selective attention was assessed via the Stroop Color-Word Test (SCWT).\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eAfter intervention, serum BDNF levels(22.6\u0026thinsp;\u0026plusmn;\u0026thinsp;3.2 pg/ml) in the experimental group were significantly higher than those before intervention ༈15.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5 pg/ml༉and in the control group༈16.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8 pg/ml༉ (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01). SAS scores༈35.6\u0026thinsp;\u0026plusmn;\u0026thinsp;4.2 points༉ and PSQI scores ༈4.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2 points༉in the experimental group were significantly lower than those before intervention ༈52.3\u0026thinsp;\u0026plusmn;\u0026thinsp;5.8 points、11.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5 points༉and in the control group ༈50.9\u0026thinsp;\u0026plusmn;\u0026thinsp;5.9 points、10.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.2 points༉(P\u0026thinsp;\u0026lt;\u0026thinsp;0.01). In the SCWT, the experimental group showed significantly shorter naming time༈59.6\u0026thinsp;\u0026plusmn;\u0026thinsp;7.3 seconds༉ and fewer errors ༈2.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1 times༉in the interference task (conflicting color words), with significantly reduced time interference values༈24.9\u0026thinsp;\u0026plusmn;\u0026thinsp;5.2 seconds༉ and error interference values༈0.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8 times༉ (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01), and these indices were significantly better than those in the control group༈76.5\u0026thinsp;\u0026plusmn;\u0026thinsp;8.1 seconds、4.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2 times、41.7\u0026thinsp;\u0026plusmn;\u0026thinsp;5.9 seconds、3.5\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3 times༉ (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01). However, there were no significant changes in indices of neutral tasks. No statistically significant differences were observed in all indices of the control group before and after intervention (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThis study confirms that Liuzijue breathing regulation can alleviate anxiety, improve sleep quality, and enhance attention by upregulating BDNF expression, providing research evidence for its application in anxiety prevention and treatment.\u003c/p\u003e","manuscriptTitle":"Effects of Fragmented Liuzijue Breathing Exercise on Sleep Quality, Anxiety, and Attention in College Students","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-17 20:50:19","doi":"10.21203/rs.3.rs-9357142/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorInvitedReview","content":"","date":"2026-05-18T11:12:41+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"281899808921855207154310478411627564274","date":"2026-05-18T10:19:12+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-04-09T17:14:15+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-04-09T04:12:40+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-04-09T04:11:41+00:00","index":"","fulltext":""},{"type":"submitted","content":"Sport Sciences for Health","date":"2026-04-08T12:27:27+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"sport-sciences-for-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ssfh","sideBox":"Learn more about [Sport Sciences for Health](http://link.springer.com/journal/11332)","snPcode":"11332","submissionUrl":"https://submission.nature.com/new-submission/11332/3","title":"Sport Sciences for Health","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"551ed7c3-b9d1-4d52-b38a-e4784a2300ee","owner":[],"postedDate":"April 17th, 2026","published":true,"recentEditorialEvents":[{"type":"editorInvitedReview","content":"","date":"2026-05-18T11:12:41+00:00","index":36,"fulltext":""},{"type":"reviewerAgreed","content":"281899808921855207154310478411627564274","date":"2026-05-18T10:19:12+00:00","index":35,"fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-04-17T20:50:19+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-17 20:50:19","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9357142","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9357142","identity":"rs-9357142","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}