ObjectiveTo investigate the effect of Tougu Xiaotong capsules （TGXTC） on the regulation of chondrocyte PANoptosis， delay of chondrocyte degeneration， and improvement of the symptoms in knee osteoarthritis （KOA）. MethodsIn vivo experiments： 50 male C57BL/6 mice were randomly assigned into five groups （n=10 per group）： sham operation group， model group， low-dose TGXTC group （7.2 g·kg^-1）， high-dose TGXTC group （14.4 g·kg^-1）， and diclofenac sodium group （0.05 g·kg^-1）. Except for the sham group， KOA models were established in all other groups using the modified Hulth method. Following successful model induction， the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg^-1 for 6 weeks， while the diclofenac sodium group received 0.05 g·kg^-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score； micro-computed tomography （micro-CT） was used to scan the knee， hematoxylin-eosin （HE） staining and safranin O-fast green staining were used to observe the morphology of cartilage， transmission electron microscopy （TEM） was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence （IF） co-localization assays was performed to detect the co-localization of cleaved Caspase-3， receptor-interacting protein 3 （RlPK3）， and the N-terminal domain of gasdermin D （GSDMD-N） in cartilage tissue， while western blot was employed to detect the expression levels of cleaved Caspase-3， RIPK3， and GSDMD-N. In vitro experiments： The knee cartilages of 4-week-old SD rats were isolated， and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups： the control group， the model group （pretreated with 10 mg·L^-1 lipopolysaccharide （LPS） for 24 h followed by treatment with 1 μmol·L^-1 nigericin for 4 h）， and the TGXTC treatment group （pretreated with 10 mg·L^-1 LPS for 24 h， followed by exposure to 1 μmol·L^-1 nigericin for 4 h and subsequently treated with 100 mg·L^-1 TGXTC for an additional 24 h）. The levels of reactive oxygen species （ROS）， apoptosis， necroptosis， and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection， AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels （IL-1β and IL-18） were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis， including cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3. ResultsCompared with the sham group， the Lequesne MG scores were significantly up-regulated（P<0.01） in the model group， and the pathological changes of cartilage were significantly， with joint spaces narrower， osteophyte formation increased， secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis， necroptosis， and pyroptosis， along with markedly elevated expression of cleaved Caspase-3， RlPK3， and GSDMD-N in cartilage tissue （P<0.01）. In addition， The mean fluorescence intensities of ROS， orange-red fluorescence in AO/EB staining， green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group （P<0.01） . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased （P<0.01）. The expression of PANoptosis related proteins （cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3） were also significantly upregulated（P<0.05）. Compared to the model group， the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score， an increase in joint space， a decrease in osteophyte formation， diminished cartilage damage， reduced release of ROS， and alleviation of apoptotic， necroptotic， and pyroptotic processes in chondrocytes. Additionally， mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased （P<0.05）. Furthermore， the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions （P<0.05）. Similar results were observed in chondrocytes： cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3 exhibited significant decreases as well （P<0.05）. ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.

ObjectiveTo investigate the effect of Tougu Xiaotong capsules （TGXTC） on the regulation of chondrocyte PANoptosis， delay of chondrocyte degeneration， and improvement of the symptoms in knee osteoarthritis （KOA）. MethodsIn vivo experiments： 50 male C57BL/6 mice were randomly assigned into five groups （n=10 per group）： sham operation group， model group， low-dose TGXTC group （7.2 g·kg^-1）， high-dose TGXTC group （14.4 g·kg^-1）， and diclofenac sodium group （0.05 g·kg^-1）. Except for the sham group， KOA models were established in all other groups using the modified Hulth method. Following successful model induction， the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg^-1 for 6 weeks， while the diclofenac sodium group received 0.05 g·kg^-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score； micro-computed tomography （micro-CT） was used to scan the knee， hematoxylin-eosin （HE） staining and safranin O-fast green staining were used to observe the morphology of cartilage， transmission electron microscopy （TEM） was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence （IF） co-localization assays was performed to detect the co-localization of cleaved Caspase-3， receptor-interacting protein 3 （RlPK3）， and the N-terminal domain of gasdermin D （GSDMD-N） in cartilage tissue， while western blot was employed to detect the expression levels of cleaved Caspase-3， RIPK3， and GSDMD-N. In vitro experiments： The knee cartilages of 4-week-old SD rats were isolated， and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups： the control group， the model group （pretreated with 10 mg·L^-1 lipopolysaccharide （LPS） for 24 h followed by treatment with 1 μmol·L^-1 nigericin for 4 h）， and the TGXTC treatment group （pretreated with 10 mg·L^-1 LPS for 24 h， followed by exposure to 1 μmol·L^-1 nigericin for 4 h and subsequently treated with 100 mg·L^-1 TGXTC for an additional 24 h）. The levels of reactive oxygen species （ROS）， apoptosis， necroptosis， and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection， AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels （IL-1β and IL-18） were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis， including cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3. ResultsCompared with the sham group， the Lequesne MG scores were significantly up-regulated（P<0.01） in the model group， and the pathological changes of cartilage were significantly， with joint spaces narrower， osteophyte formation increased， secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis， necroptosis， and pyroptosis， along with markedly elevated expression of cleaved Caspase-3， RlPK3， and GSDMD-N in cartilage tissue （P<0.01）. In addition， The mean fluorescence intensities of ROS， orange-red fluorescence in AO/EB staining， green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group （P<0.01） . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased （P<0.01）. The expression of PANoptosis related proteins （cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3） were also significantly upregulated（P<0.05）. Compared to the model group， the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score， an increase in joint space， a decrease in osteophyte formation， diminished cartilage damage， reduced release of ROS， and alleviation of apoptotic， necroptotic， and pyroptotic processes in chondrocytes. Additionally， mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased （P<0.05）. Furthermore， the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions （P<0.05）. Similar results were observed in chondrocytes： cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3 exhibited significant decreases as well （P<0.05）. ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.

Objective: To explore the interaction between school bullying and insomnia in relation to depression-anxiety-stress emotions among primary and secondary school students,so as to provide a basis for preventing negative emotional states in adolescents. Methods: In October 2024, a stratified cluster sampling method was used to select 3 058 students in grade 5-6 of primary, junior and senior high school in Sheyang County of Jiangsu Province. The Delaware Bullying Victimization Scale, Insomnia Severity Index, Depression-Anxiety-Stress Scale-21, and Study Condition Questionnaire were employed to investigate school bullying, insomnia, depression-anxiety-stress emotions, and academic performance. The χ 2 test and Logistic regression were used to analyze the association between school bullying and insomnia interactions and depression-anxiety-stress emotions among primary and secondary school students, multiplicative interaction analysis was conducted, and additive interaction analysis was performed using R software. Results: The detection rates of depression-anxiety-stress emotions among primary and secondary school students were 21.6%, 28.4% and 10.8%, respectively. The detection rates of physical bullying, relationship bullying, verbal bullying and cyberbullying in school bullying were 10.6%, 14.0%, 22.3%, and 6.2%, respectively. The detection rate for insomnia was 23.1%. Results from Logistic regression analysis showed that, after adjusting for relevant factors, physical, relational, verbal, and cyberbullying and insomnia were positively correlated with the detection rates of depression ( OR = 5.72- 10.93), anxiety ( OR =6.35-12.17), and stress emotions ( OR =5.97-14.52) among primary and secondary school students (all P <0.01). The multiplicative interaction between physical, relational, verbal, and cyberbullying and insomnia was positively correlated with the detection rates of depression ( OR =8.00-18.01), anxiety ( OR =11.35-17.76), and stress emotions ( OR =7.64-9.12) in primary and secondary school students (all P <0.01). Additive interactions were observed between physical, relational, verbal, and cyberbullying and insomnia in relation to the detection rates of depression, anxiety, and stress emotions among primary and secondary school students (both RERI and AP >0 and the credible interval excluded 0, SI >1 and the credible interval excluded 1). Conclusion School bullying and insomnia are associated with depression, anxiety, and stress emotions among primary and secondary school students, and they exhibit both multiplicative and additive interactions.

BACKGROUND:Rheumatoid arthritis is widely prevalent worldwide,with its high incidence and universality that considerably affects patients' quality of life.Previous studies have focused on a few immune cells or cytokines,whereas this study comprehensively provides a more complete view of the immune mechanisms in rheumatoid arthritis.OBJECTIVE:To explore the causal relationship between 731 immune cell phenotypes and rheumatoid arthritis using the Mendelian randomization method,thereby providing evidence of causality.METHODS:The 731 immune cell phenotypes used in this study were sourced from the GWAScatalog database,jointly developed by the National Human Genome Research Institute(NHGRI)and the European Bioinformatics Institute(EBI).The rheumatoid arthritis data were from the Finngen database,developed by the Finnish Institute for Molecular Medicine(FIMM).The inverse variance weighting method was employed as the primary analytical approach.Additionally,multiple analytical methods,including MR-Egger,weighted mode,simple mode,and weighted median,were concurrently utilized to complement the final results.Sensitivity analyses(Cochran's Q test,MR-Egger regression,and MR-presso analysis)were also conducted to verify the stability and feasibility of the data.RESULTS AND CONCLUSION:(1)After excluding results through heterogeneity testing,the inverse variance weighting analysis indicated that 10 absolute cell counts,15 median fluorescence intensities of surface antigen levels,1 morphological characteristic,and 9 relative cell counts had a causal relationship with the occurrence of rheumatoid arthritis.(2)According to cell classification,this study found that seven types of B cells,seven types of classical dendritic cells,six types of mature T cells,four types of monocytes,three types of myeloid cells,three types of TBNK cells(lymphocyte subset T cells,B cells and natural killer cells),and five types of Tregs had a causal association with the occurrence of rheumatoid arthritis.(3)Through comprehensive bidirectional two-sample MR analysis,we demonstrated the complex causal relationships between multiple immune phenotypes and rheumatoid arthritis,highlighting the intricate interaction patterns between the immune system and rheumatoid arthritis.These results provide new biomarkers for the early screening and diagnosis of rheumatoid arthritis in China,and help to improve the diagnostic accuracy and sensitivity.

BACKGROUND:Rheumatoid arthritis is widely prevalent worldwide,with its high incidence and universality that considerably affects patients' quality of life.Previous studies have focused on a few immune cells or cytokines,whereas this study comprehensively provides a more complete view of the immune mechanisms in rheumatoid arthritis.OBJECTIVE:To explore the causal relationship between 731 immune cell phenotypes and rheumatoid arthritis using the Mendelian randomization method,thereby providing evidence of causality.METHODS:The 731 immune cell phenotypes used in this study were sourced from the GWAScatalog database,jointly developed by the National Human Genome Research Institute(NHGRI)and the European Bioinformatics Institute(EBI).The rheumatoid arthritis data were from the Finngen database,developed by the Finnish Institute for Molecular Medicine(FIMM).The inverse variance weighting method was employed as the primary analytical approach.Additionally,multiple analytical methods,including MR-Egger,weighted mode,simple mode,and weighted median,were concurrently utilized to complement the final results.Sensitivity analyses(Cochran's Q test,MR-Egger regression,and MR-presso analysis)were also conducted to verify the stability and feasibility of the data.RESULTS AND CONCLUSION:(1)After excluding results through heterogeneity testing,the inverse variance weighting analysis indicated that 10 absolute cell counts,15 median fluorescence intensities of surface antigen levels,1 morphological characteristic,and 9 relative cell counts had a causal relationship with the occurrence of rheumatoid arthritis.(2)According to cell classification,this study found that seven types of B cells,seven types of classical dendritic cells,six types of mature T cells,four types of monocytes,three types of myeloid cells,three types of TBNK cells(lymphocyte subset T cells,B cells and natural killer cells),and five types of Tregs had a causal association with the occurrence of rheumatoid arthritis.(3)Through comprehensive bidirectional two-sample MR analysis,we demonstrated the complex causal relationships between multiple immune phenotypes and rheumatoid arthritis,highlighting the intricate interaction patterns between the immune system and rheumatoid arthritis.These results provide new biomarkers for the early screening and diagnosis of rheumatoid arthritis in China,and help to improve the diagnostic accuracy and sensitivity.

Objective:To identify the key epithelial cell characteristics that can accurately diagnose eosinophilic chronic sinusitis with nasal polyps（ECRSwNP） through nasal brush sampling and comparing with the pathological results of nasal polyp tissue sections. Methods:Ninety-one patients underwent surgery in the Ophthalmology and ENT Department of the Second People's Hospital of Longgang District, Shenzhen, from January 2022 to July 2024 were selected. The cohort comprised 58 males and 33 females（mean age: 41.4 years; range: 12.0-71.0）. The clinical characteristics of the patients, including gender, age, disease duration, smoking and drinking history, asthma history, subjective symptoms, sinus CT, and nasal endoscopy scores, were recorded. Nasal brush sampling of nasal polyps and inferior turbinate mucosa was performed before surgery to obtain cytological specimens, and nasal polyp tissues were collected during surgery. The demographic and clinical characteristics of patients with eosinophilic and non-eosinophilic nasal polyps were compared, as well as the relationship between nasal brush cytology of nasal polyps and inferior turbinate and nasal polyp histopathology. Statistical analysis was performed using SPSS 23.0 software. Results:Among the 91 patients, no significant differences were observed between ECRSwNP and NECRSwNP patients in terms of age, gender, smoking status, alcohol consumption, and disease duration. The nasal brush cell population in ECRSwNP patients was more likely to contain eosinophils（P<0.001） and less likely to contain lymphocytes and plasma cells（P<0.001）. Additionally, the ciliated cells in ECRSwNP patients exhibited larger widths（P=0.036）, shorter cilium lengths（P<0.001）, and more disordered arrangements（P<0.001） compared to NECRSwNP patients. In nasal brush cells from the inferior turbinate, ECRSwNP patients also showed shorter cilium lengths（P<0.001） and shorter cilia（P=0.024） compared to NECRSwNP patients. Conclusion:There are significant differences in obtaining epithelial cytological information from nasal polyps or inferior turbinates through nasal brush sampling between ECRSwNP and NECRSwNP patients.
Humans ; Male ; Female ; Middle Aged ; Adult ; Nasal Polyps/complications* ; Sinusitis/complications* ; Aged ; Chronic Disease ; Adolescent ; Nasal Mucosa/pathology* ; Young Adult ; Rhinitis/complications* ; Eosinophilia/pathology* ; Child ; Eosinophils/pathology* ; Rhinosinusitis

OBJECTIVES: To explore the molecular mechanism of Jiangzhi Huaban Decoction (JZHBD) for improving atherosclerosis through the "qi meridian-blood channels" pathway. METHODS: ApoE^-/- mouse models of atherosclerosis were established by high-fat diet feeding for 8 weeks, with C57BL/6 mice on a normal diet as the controls. Forty ApoE^-/- mouse models were randomized into model group, low-, medium-, and high-dose JZHBD treatment groups, and atorvastatin treatment group (n=8) for their respective treatments for 8 weeks. The changes in body weight and overall condition of the mice were monitored weekly. After the treatments, serum levels of TC, TG, HDL-C, LDL-C, TBA, ALT, and AST of the mice were measured, pathological changes in the liver and aortic root plaques were examined with HE staining, and lipid accumulation in the liver and aortic wall was assessed using Oil Red O staining. The core molecular mechanism was studied through transcriptomics, and the expressions of the key pathway proteins were confirmed using Western blotting and immunohistochemistry. RESULTS: Treatment with JZHBD significantly reduced blood lipid and total bile acid levels, improved liver function and hepatic steatosis, and decreased aortic lipid deposition and plaque area in the mouse models of atherosclerosis. Transcriptomic analysis suggested that the therapeutic mechanism of JZHBD involved reverse cholesterol transport, PPAR signaling, and the inflammatory pathways. In atherosclerotic mice, JZHBD treatment obviously up-regulated hepatic expressions of PPARγ, LXRα, ABCA1, ABCG1, and CYP7A1, down-regulated hepatic expressions of p-p65/p65, IL-6, IL1β in the liver, increased ABCG5 and ABCG8 expressions in the intestines, and decreased ICAM-1 and VCAM-1 expressions in the aortic plaques. CONCLUSIONS JZHBD improves atherosclerotic vascular damage and plaque formation possibly by regulating hepatic reverse cholesterol transport and inflammation via modulating the hepatic PPARγ/LXRα/NF-κB signaling pathway.
Animals ; Drugs, Chinese Herbal/therapeutic use* ; Mice, Inbred C57BL ; Plaque, Atherosclerotic/metabolism* ; Liver/metabolism* ; Mice ; Atherosclerosis/metabolism* ; Cholesterol/metabolism* ; PPAR gamma/metabolism* ; Male ; Diet, High-Fat ; Biological Transport

BACKGROUND:Mitochond rial biogenesis in skeletal muscle and its regulatory mechanisms during exercise have become focal points of research.Pathways such as AMP-activated protein kinase,peroxisome proliferator-activated receptory coactivator 1α,mitogen-activated protein kinase,calcium-regulated signaling play profound roles in exercise-induced mitochondrial biogenesis,impacting muscle metabolic optimization,enhanced athletic performance,and the prevention of metabolic diseases.However,the interactions among these pathways,their regulatory mechanisms,and their comprehensive effects on exercise-induced mitochondrial biogenesis in skeletal muscle remain unclear.OBJECTIVE:To explore the signaling pathways related to mitochondrial biogenesis in skeletal muscle,precisely analyze the induction and regulatory details of exercise within these pathways,and clearly elucidate the principles by which exercise-activated signaling pathways promote mitochondrial generation and functional enhancement.This will establish a theoretical foundation for improving muscle metabolism,enhancing exercise efficiency,and preventing metabolic diseases.METHODS:An extensive literature search was conducted using China National Knowledge Infrastructure(CNKI),WanFang,VIP,PubMed,and Web of Science.The latest publications related to mitochondrial biogenesis in skeletal muscle and its regulatory mechanisms were collected from inception to August 2024.By integrating findings from multiple signaling pathways,the regulatory mechanisms of exercise on mitochondrial biogenesis were systematically reviewed,with a focus on the interactions and synergistic mechanisms of AMP-activated protein kinase,peroxisome proliferator-activated receptor y coactivator 1α,protein kinase A,mitogen-activated protein kinase,calcium-regulated signaling pathways.RESULTS AND CONCLUSION:(1)Mitochondrial biogenesis in skeletal muscle is a complex biological process involving the coordinated regulation of multiple signaling pathways.This process aims to optimize the metabolic capacity,fatigue resistance,and overall athletic performance of skeletal muscle in response to changes in energy demand and external stress.The core mechanisms include the interactions and regulation of key factors such as AMP-activated protein kinase,peroxisome proliferator-activated receptor gamma coactivator 1α,and mitogen-activated protein kinase.(2)AMP-activated protein kinase senses the cellular energy status and activates peroxisome proliferator-activated receptor gamma coactivator 1α,thereby promoting mitochondrial biogenesis.Peroxisome proliferator-activated receptor gamma coactivator 1α,as the main regulator of mitochondrial biogenesis in skeletal muscle,can modulate the synthesis of mitochondrial proteins and DNA,enhance the antioxidant stress response,and improve mitochondrial function.(3)The mitogen-activated protein kinase signaling pathway,particularly p38 mitogen-activated protein kinase,further promotes mitochondrial generation by activating peroxisome proliferator-activated receptor gamma coactivator 1α during stress responses.(4)Additionally,calcium signaling and protein kinase A pathways play significant roles in the metabolic regulation of skeletal muscle.(5)Exercise can significantly enhance mitochondrial biogenesis capacity in skeletal muscle by activating these multiple signaling pathways,optimizing cellular metabolic efficiency,increasing muscle endurance,and improving athletic performance.(6)Future research should focus on in-depth exploration of the interaction mechanisms among AMP-activated protein kinase,peroxisome proliferator-activated receptor gamma coactivator1α,mitogen-activated protein kinases,and calcium signaling under different exercise intensities and modalities;strengthen studies across diverse age groups,genders,and health conditions;validate the universality and population-specificity of research findings;investigate the intricate mechanisms of emerging regulatory factors such as FNIP1 and PERM1 and their potential in exercise interventions;and promote the translation of exercise health research outcomes into clinical applications.

Objective: To explore the independent and interactive effects of school green space exposure and outdoor activity duration on screening myopia among primary and secondary school students, so as to provide theoretical support for the prevention and control of screening myopia in children and adolescents. Methods: From September to November 2023, 117 487 primary and secondary school students from 497 schools were selected using a cluster random sampling method, covering 98 counties (cities, districts) in Jiangsu Province. Data on the students screening myopia status and associated health influencing factors were collected and analyzed. School green space exposure was quantified using the normalized difference vegetation index (NDVI), which was extracted with ArcGIS Pro software; meanwhile, information on students outdoor activity duration was gathered through self reported questionnaires. Multivariate Logistic regression was applied to assess the independent and interactive effects of green space exposure and outdoor activity duration on screening myopia among primary and secondary school students. Results: Univariate analysis showed that there were statistically significant differences in screening myopia detection rates among primary and secondary school students of different genders, NDVI groups, every outdoor activity duration, monitoring points, school stages, parents educational level, and whether they lived on campus or had parents with screening myopia ( χ 2=88.91-1 950.08, all P <0.05); as the school age and sedentary time increased, the detection rate of screening myopia in primary and secondary school students also increased ( χ 2 trend =8 410.15, 2 028.91, both P <0.05). Independent effects showed that compared to the low NDVI group, the medium and high NDVI groups had lower risks of screening myopia ( OR =0.93, 0.95, both P <0.05). Compared to those with outdoor activity duration<2 h/d, students with outdoor activity duration≥2 h/d had a lower risk of screening myopia ( OR =0.96, P <0.05). When stratified by school level, compared to the low NDVI group, the medium NDVI group had lower risks of screening myopia in primary and junior high schools (primary school: OR =0.91; junior high school: OR =0.88, both P <0.05). Compared to those with outdoor activity duration<2 h/d, junior high school students with outdoor activity duration≥2 h/d had a lower risk of screening myopia ( OR = 0.90, P <0.05). When stratified by monitoring site, urban primary and secondary school students in the medium and high NDVI groups and those with outdoor activity duration≥2 h/d had lower risks of screening myopia ( OR =0.92, 0.92, 0.93, all P <0.05). Interactive effects showed that when medium or high NDVI was combined with outdoor activity duration≥2 h/d, the risks of screening myopia among primary and secondary school students were lower (medium NDVI×≥2 h/d: OR =0.89; high NDVI×≥ 2 h/d : OR =0.89, both P <0.05), and the combined effect was superior to that of a single factor. Conclusion Green space exposure and outdoor activity duration have negative correlations with screening myopia among primary and secondary students, and the combined effect is better than that of a single factor.

Objective: To investigate the association between outdoor light at night (LAN) exposure and sleep quality among primary and secondary school students, and to examine the pathways of negative affects including depressive, stress and anxiety symptoms, so as to provide a theoretical basis for optimizing the school environment and enhancing the physical and mental well being of students. Methods: In December 2024, a total of 36 885 students from 154 primary and secondary schools in Suzhou, Nantong, and Changzhou were included via a stratified cluster sampling method. Sleep quality and negative affect were assessed by using the Pittsburgh Sleep Quality Index (PSQI), Center for Epidemiologic Studies Depression Scale (CES-D), Generalized Anxiety Disorder-7 (GAD-7), and Depression, Anxiety and Stress Scale-21 (DASS-21), respectively. Outdoor LAN exposure data were obtained from the national polar orbiting partnership visible infrared imaging radiometer suite nighttime light(NPP-VIIRS NTL) dataset provided by the National Earth System Science Data Center. Multivariate Logistic regression model was employed to analyze the association between LAN exposure and sleep quality across different regions, with stratification by monitoring site. Spearman correlation analysis was used to examine the relationships between LAN, negative affect, and sleep quality. The mediating role of negative affect was tested by using Model 4 of the PROCESS macro. Results: The detection rates among students were 13.95%( n =5 147) for depressive symptom, 16.72%( n =6 166) for stress symptom, and 17.49%( n =6 451) for anxiety symptom. The median outdoor LAN exposure was 28.85(19.10, 41.44)nW/(cm 2 · ( sr). After adjusting for confounders, multivariate Logistic regression analysis showed that high LAN exposure ( Q 4) was positively associated with sleep problems (urban areas: OR =1.28, 95% CI = 1.17- 1.41; rural areas: OR =1.21, 95% CI =1.07-1.36; both P <0.05). Spearman correlation analysis revealed significant positive correlations between LAN exposure, depressive symptoms, stress symptoms, anxiety symptoms, and sleep quality ( r =0.03-0.75, all P < 0.01). The mediation analysis indicated that all dimensions of negative affect significantly mediated the relationship between high LAN exposure and poor sleep quality (all P <0.01). Specifically, the indirect effects were 0.03 (95% CI =0.02-0.05) for depressive symptom, 0.05(95% CI =0.03-0.08) for stress symptom, and 0.07(95% CI =0.05-0.09) for anxiety symptom. Overall, 31.9% of the total effect was mediated by negative affect, with anxiety (14.89%) being the strongest mediator, followed by stress (10.64%) and depression(6.38%). Conclusion Reducing high levels of outdoor LAN exposure and implementing interventions targeting negative affect may contribute to improved sleep quality among primary and secondary school students.