1.Baicalein inhibits proliferation,migration and invasion of human pancre-atic cancer Aspc-1 and Bxpc-3 cells via AKT/β-catenin signaling pathway
Xiaoliang CHEN ; Qiong WANG ; Lin TANG ; Lili YIN ; Jinsheng YU ; Yun-feng BAI ; Yi QIN ; Feng FENG
Chinese Journal of Pathophysiology 2025;41(9):1738-1747
AIM:To investigate the antitumor activity and targets of baicalein(Bai)in pancreatic cancer using network pharmacology combined with in vitro and in vivo experiments.METHODS:The targets of Bai and pancreatic can-cer were analyzed via multi-data screening.A protein-protein interaction network was constructed using STRING,and core targets were identified via Cytoscape.Functional enrichment was analyzed by Gene Ontology(GO)and Kyoto Ency-clopedia of Genes and Genomes(KEGG).Antitumor effects of Bai were assessed in pancreatic cancer cells Aspc-1 and Bxpc-3 using MTT and colony formation assays for proliferation,flow cytometry for apoptosis and cell cycle analysis,and Transwell assays for migration and invasion.A xenograft tumor model was established to evaluate tumor proliferation,im-munohistochemistry was performed to detect the protein expression of AKT in tumor tissues,and Western blot was used to analyze the expression levels of AKT,β-catenin,N-cadherin and Slug.RESULTS:A total of 108 overlapping targets were identified between Bai and pancreatic cancer.Among these,7 core targets were recognized,including proto-onco-gene tyrosine-protein kinase Src,heat shock protein 90 alpha family class A member 1(HSP90AA1),estrogen receptor 1(ESR1),tumor protein p53(TP53),epidermal growth factor receptor(EGFR),AKT1,and mitogen-activated protein ki-nase 3(MAPK3).The GO analysis revealed significant enrichment in oxidative stress response,protein phosphorylation,and serine/threonine kinase activity.The KEGG analysis primarily enriched the PI3K/AKT,MAPK and Ras signaling pathways.The MTT and colony formation assays showed that Bai inhibited the viability of Aspc-1 and Bxpc-3 cells in a dose-dependent manner(72 h IC50 values were 73.6 μmol/L and 83.4 μmol/L,respectively)and reduced cell colony for-mation(P<0.05 or P<0.01).Flow cytometry confirmed that Bai induced apoptosis of Aspc-1 and Bxpc-3 cells(P<0.01)and blocked the cell cycle at the G0/G1 phase(P<0.05 or P<0.01).Transwell experiments indicated that Bai inhibited the migration and invasion of Aspc-1 and Bxpc-3 cells(P<0.05 or P<0.01).In vivo,Bai significantly inhibited the growth of Aspc-1 cell xenograft tumors(P<0.01).Immunohistochemistry revealed a significant reduction in AKT expression in tu-mor tissues(P<0.01),and Western blot showed decreased expression of AKT,β-catenin,N-cadherin and Slug in both Aspc-1 and Bxpc-3 cells(P<0.01).CONCLUSION:Baicalein inhibits pancreatic cancer cell proliferation,migration,and invasion,potentially through down-regulation of AKT,β-catenin,N-cadherin,and Slug expression.
2.Efficacy of neuromuscular electrical stimulation combined with limb rehabilitation therapy in the treatment of hemiplegia due to stroke
Weixi WANG ; Lintao GAO ; Xiaoliang YANG ; Wenbin HUANG
Chinese Journal of Primary Medicine and Pharmacy 2025;32(7):1065-1071
Objective:To investigate the efficacy of neuromuscular electrical stimulation (NMES) combined with limb rehabilitation therapy in improving limb function of patients with hemiplegia due to stroke.Methods:A randomized controlled study was conducted on 110 patients with hemiplegia due to stroke admitted to the 987 Hospital of the PLA Joint Logistic Support Force from January 2021 to January 2023. These patients were divided into a control group and an observation group, with 55 cases each, using a random number table method. The control group received conventional rehabilitation therapy, while the observation group received NMES combined with limb rehabilitation therapy. All patients were treated for 3 months. The Trunk Impairment Scale, Brunnstrom Motor Function Scale, Surface Electromyography, Brunel Balance Scale, Simple Gait Test Scale, and Modified Ashworth Scale were used to evaluate trunk control function, limb control function, neuromuscular disorders, balance control function (forward-backward inclination angle, left-right inclination angle), gait temporal-spatial parameters, symmetry parameters, and muscle tone recovery in both groups before and after treatment.Results:After treatment, the Total Impairment Scale and Brunnstrom scores for upper and lower limbs in the observation group were higher than those in the control group [(18.31 ± 1.61) vs. (14.38 ± 1.42), (3.78 ± 0.41) vs. (3.01 ± 0.63), (3.75 ± 0.42) vs. (3.28 ± 0.63), t = 13.58, 7.60, 4.60, all P < 0.05]. Additionally, the scores for tibialis anterior, fibularis brevis, fibularis longus, medial gastrocnemius, lateral gastrocnemius muscle control were higher in the observation group compared with the control group. The scores for Berg Balance Scale, forward-backward inclination angle, left-right inclination angle, step frequency, step length, step speed, and step width in the observation group were also superior to those in the control group. All differences were statistically significant ( t = 17.88, 20.15, 24.62, 19.53, 22.80, 3.31, 8.66, 7.88, 6.08, 4.21, 7.43, 3.35, all P < 0.05). Furthermore, the deviations in step length, the ratio of support on the affected side to the unaffected side, and the ratio of swing on the affected side to the unaffected side in the observation group were lower than those in the control group [(3.53 ± 2.55) vs. (5.35 ± 3.21), (1.11 ± 0.04) vs. (1.14 ± 0.03), (1.10 ± 0.06) vs. (1.24 ± 0.09), t = 3.29, 4.45, 9.60, all P < 0.05]. Additionally, the level of muscle tone recovery in the observation group was higher than that in the control group ( Z = 2.53, P = 0.011). Conclusions:In the treatment of hemiplegia due to stroke, NMES combined with limb rehabilitation therapy can effectively improve patients' trunk and limb control, enhance balance function, and greatly promote muscle strength recovery.
3.The association between clinical drug utilization and the risk of nosocomial infections among inpatients:a comprehensive dose-response analysis
Xiaoliang ZHANG ; Fangbin LI ; Xiaolong YUAN ; Yujuan FENG ; Haimo WANG ; Xiaoyong LIN ; Bingpeng WEI ; Lei WANG ; Haojun ZHANG
Chinese Journal of Infection and Chemotherapy 2025;25(2):121-126
Objective To analyze the relationship between clinical drug utilization and the risk of nosocomial infections among hospitalized patients,and provide evidence for the prevention and control of nosocomial infections.Methods This study adopted a retrospective case-control design.The case group included 209 patients with nosocomial infection reported from January 2023 to December 2023 in a tertiary hospital.The control group included 209 patients without nosocomial infection during the same period.The patients in the control group were selected by stratified sampling based on Charlson Comorbidity Index(CCI).Results Univariate analysis showed that proton pump inhibitors,antacids,immunosuppressants and prior antimicrobial combination therapy increased the risk of nosocomial infection(P<0.05).Multivariate log-binomial regression analysis showed that proton pump inhibitors,immunosuppressive drugs,and prior antimicrobial combination therapy were correlated with nosocomial infection.The corresponding relative risk(RR)was 1.31(95%CI:1.07-1.60),1.40(95%CI:1.02-1.91),and 1.66(95%CI:1.01-2.74),respectively.Further analysis indicated that the patients with nosocomial infection had longer time in use of proton pump inhibitors and prior antimicrobial combination therapy than the patients in the control group(Z=-6.331,P<0.001;Z=-2.667,P=0.008).The trend Chi-square test showed that there was a dose-response relationship for proton pump inhibitors(x2=73.869,P<0.001),immunosuppressive drugs(x2=16.530,P<0.001),and prior antimicrobial combination therapy(x2=35.107,P<0.001).Conclusions The use of immunosuppressants,proton pump inhibitors and antimicrobial combination therapy increases the risk of nosocomial infections in hospitalized patients.The prolonged use of these drugs will further increase the risk of nosocomial infection.
4.Baicalein inhibits proliferation,migration and invasion of human pancre-atic cancer Aspc-1 and Bxpc-3 cells via AKT/β-catenin signaling pathway
Xiaoliang CHEN ; Qiong WANG ; Lin TANG ; Lili YIN ; Jinsheng YU ; Yun-feng BAI ; Yi QIN ; Feng FENG
Chinese Journal of Pathophysiology 2025;41(9):1738-1747
AIM:To investigate the antitumor activity and targets of baicalein(Bai)in pancreatic cancer using network pharmacology combined with in vitro and in vivo experiments.METHODS:The targets of Bai and pancreatic can-cer were analyzed via multi-data screening.A protein-protein interaction network was constructed using STRING,and core targets were identified via Cytoscape.Functional enrichment was analyzed by Gene Ontology(GO)and Kyoto Ency-clopedia of Genes and Genomes(KEGG).Antitumor effects of Bai were assessed in pancreatic cancer cells Aspc-1 and Bxpc-3 using MTT and colony formation assays for proliferation,flow cytometry for apoptosis and cell cycle analysis,and Transwell assays for migration and invasion.A xenograft tumor model was established to evaluate tumor proliferation,im-munohistochemistry was performed to detect the protein expression of AKT in tumor tissues,and Western blot was used to analyze the expression levels of AKT,β-catenin,N-cadherin and Slug.RESULTS:A total of 108 overlapping targets were identified between Bai and pancreatic cancer.Among these,7 core targets were recognized,including proto-onco-gene tyrosine-protein kinase Src,heat shock protein 90 alpha family class A member 1(HSP90AA1),estrogen receptor 1(ESR1),tumor protein p53(TP53),epidermal growth factor receptor(EGFR),AKT1,and mitogen-activated protein ki-nase 3(MAPK3).The GO analysis revealed significant enrichment in oxidative stress response,protein phosphorylation,and serine/threonine kinase activity.The KEGG analysis primarily enriched the PI3K/AKT,MAPK and Ras signaling pathways.The MTT and colony formation assays showed that Bai inhibited the viability of Aspc-1 and Bxpc-3 cells in a dose-dependent manner(72 h IC50 values were 73.6 μmol/L and 83.4 μmol/L,respectively)and reduced cell colony for-mation(P<0.05 or P<0.01).Flow cytometry confirmed that Bai induced apoptosis of Aspc-1 and Bxpc-3 cells(P<0.01)and blocked the cell cycle at the G0/G1 phase(P<0.05 or P<0.01).Transwell experiments indicated that Bai inhibited the migration and invasion of Aspc-1 and Bxpc-3 cells(P<0.05 or P<0.01).In vivo,Bai significantly inhibited the growth of Aspc-1 cell xenograft tumors(P<0.01).Immunohistochemistry revealed a significant reduction in AKT expression in tu-mor tissues(P<0.01),and Western blot showed decreased expression of AKT,β-catenin,N-cadherin and Slug in both Aspc-1 and Bxpc-3 cells(P<0.01).CONCLUSION:Baicalein inhibits pancreatic cancer cell proliferation,migration,and invasion,potentially through down-regulation of AKT,β-catenin,N-cadherin,and Slug expression.
5.Efficacy of neuromuscular electrical stimulation combined with limb rehabilitation therapy in the treatment of hemiplegia due to stroke
Weixi WANG ; Lintao GAO ; Xiaoliang YANG ; Wenbin HUANG
Chinese Journal of Primary Medicine and Pharmacy 2025;32(7):1065-1071
Objective:To investigate the efficacy of neuromuscular electrical stimulation (NMES) combined with limb rehabilitation therapy in improving limb function of patients with hemiplegia due to stroke.Methods:A randomized controlled study was conducted on 110 patients with hemiplegia due to stroke admitted to the 987 Hospital of the PLA Joint Logistic Support Force from January 2021 to January 2023. These patients were divided into a control group and an observation group, with 55 cases each, using a random number table method. The control group received conventional rehabilitation therapy, while the observation group received NMES combined with limb rehabilitation therapy. All patients were treated for 3 months. The Trunk Impairment Scale, Brunnstrom Motor Function Scale, Surface Electromyography, Brunel Balance Scale, Simple Gait Test Scale, and Modified Ashworth Scale were used to evaluate trunk control function, limb control function, neuromuscular disorders, balance control function (forward-backward inclination angle, left-right inclination angle), gait temporal-spatial parameters, symmetry parameters, and muscle tone recovery in both groups before and after treatment.Results:After treatment, the Total Impairment Scale and Brunnstrom scores for upper and lower limbs in the observation group were higher than those in the control group [(18.31 ± 1.61) vs. (14.38 ± 1.42), (3.78 ± 0.41) vs. (3.01 ± 0.63), (3.75 ± 0.42) vs. (3.28 ± 0.63), t = 13.58, 7.60, 4.60, all P < 0.05]. Additionally, the scores for tibialis anterior, fibularis brevis, fibularis longus, medial gastrocnemius, lateral gastrocnemius muscle control were higher in the observation group compared with the control group. The scores for Berg Balance Scale, forward-backward inclination angle, left-right inclination angle, step frequency, step length, step speed, and step width in the observation group were also superior to those in the control group. All differences were statistically significant ( t = 17.88, 20.15, 24.62, 19.53, 22.80, 3.31, 8.66, 7.88, 6.08, 4.21, 7.43, 3.35, all P < 0.05). Furthermore, the deviations in step length, the ratio of support on the affected side to the unaffected side, and the ratio of swing on the affected side to the unaffected side in the observation group were lower than those in the control group [(3.53 ± 2.55) vs. (5.35 ± 3.21), (1.11 ± 0.04) vs. (1.14 ± 0.03), (1.10 ± 0.06) vs. (1.24 ± 0.09), t = 3.29, 4.45, 9.60, all P < 0.05]. Additionally, the level of muscle tone recovery in the observation group was higher than that in the control group ( Z = 2.53, P = 0.011). Conclusions:In the treatment of hemiplegia due to stroke, NMES combined with limb rehabilitation therapy can effectively improve patients' trunk and limb control, enhance balance function, and greatly promote muscle strength recovery.
6.Evaluation of the effect of vitamin D on improving autism symptoms based on the microbiota-gut-brain axis
Haoyu HUANG ; Xiaoliang DU ; Jing WANG ; Jinting WU ; Zhuo ZOU ; Yingjuan CHEN ; Yun LIU
Chinese Journal of Nervous and Mental Diseases 2025;51(3):149-155
Objective To analyze the symptomatic improvement effects of vitamin D in children with autism spectrum disorder(ASD)based on the microbiota-gut-brain axis.Methods Seventy-two children with ASD were randomly divided into an observation group and a control group,with 36 cases in each group.Three cases dropped out in the control group.The observation group received 1200 IU/day of vitamin D supplementation in addition to conventional rehabilitation training,while the control group received only conventional rehabilitation training.The intervention lasted for 12 weeks.Assessments were conducted before and after the intervention using the childhood autism rating scale(CARS),autism behavior checklist(ABC),and repetitive behavior scale-revised(RBS-R).Resting-state functional connectivity of the brain was measured using near-infrared functional imaging,and serum levels of 25(OH)D3,inflammatory cytokines,and gut microbiota were analyzed.The differences in these indicators before and after the intervention were compared between the two groups to evaluate clinical efficacy.Results The between-group differences in pre-and post-intervention changes showed that the observation group had significantly greater improvements than the control group in the following measures:CARS scores(-5.92±1.40 vs.-2.55±1.43),RBS-R scores(-5.99±1.01 vs.-3.10±1.47),resting-state brain functional connectivity(0.19±0.15 vs.0.10±0.18),serum 25(OH)D3 levels[(34.89±8.18)ng/mL vs.(0.68±6.73)ng/mL],serum interleukin-6(IL-6)levels[(-6.60±6.07)pg/mL vs.(-0.74±9.45)pg/mL],IL-1β levels[(-2.56±1.33)pg/mL vs.(-0.04±2.13)pg/mL],and tumor necrosis factor-α(TNF-α)levels[(-4.09±3.85)pg/mL vs.(0.21±4.05)pg/mL](P<0.05).Post-intervention,significant differences in gut microbial β-diversity were observed between the two groups(R2=0.030,P=0.040,Adonis).LEfSe analysis revealed that the observation group exhibited enrichment in Clostridia(LDA=4.747,P=0.003),Clostridiales(LDA=4.747,P=0.003),Clostridiaceae(LDA=3.476,P=0.001),Lachnospiraceae(LDA=4.709,P=0.004),Odoribacteraceae(LDA=3.458,P=0.027),Odoribacter(LDA=3.458,P=0.027),Burkholderiales(LDA=3.339,P=0.038),Firmicutes(LDA=4.764,P=0.003),and Betaproteobacteria(LDA=3.338,P=0.037).Conclusion Vitamin D supplementation can modulate gut microbial diversity in children with ASD,significantly influence the abundance of specific gut microbiota,reduce systemic inflammatory cytokines,enhance brain functional connectivity,and alleviate clinical symptoms of ASD.
7.Characteristics of cardiometabolic risk in patients with different subtypes of non-alcoholic fatty liver disease
Lili GAO ; Yong WANG ; Huafang YAN ; Xiaoliang WANG ; Yunfeng WANG
Journal of Clinical Hepatology 2025;41(1):63-68
ObjectiveTo investigate the difference in the risk of cardiovascular diseases between patients with different subtypes of non-alcoholic fatty liver disease (NAFLD) from the perspective of metabolism, since cardiovascular events induced by metabolic disorders are the leading cause of death in NAFLD. MethodsThe cluster sampling method was used to conduct a multicenter cross-sectional study among three representative hospitals in Pudong New Area of Shanghai, China. A total of 37 122 sets of physical examination data from July 2022 to June 2023 were collected and stratified according to body mass index (BMI). The chi-square test was used for comparison of continuous data between groups, and a multivariable Logistic regression analysis was used to investigate the association between NAFLD subtypes and cardiometabolic risk factors. ResultsA total of 9 372 cases of NAFLD were detected, with a detection rate of 25.25%, and more than 97% of these patients were diagnosed with metabolic associated fatty liver disease (MAFLD). The subgroup analysis showed that the detection rates of lean, overweight, and obese NAFLD were 7.72%, 33.99%, and 63.56%, respectively. Compared with the patients with lean or overweight NAFLD, the patients with obese NAFLD showed a significantly higher proportion of patients with abnormalities in blood pressure, blood glucose, triglyceride (TG), high-density lipoprotein (HDL) or uric acid (all P<0.001). Among related risk factors, lean NAFLD was associated with the increase in total cholesterol (TC)(P<0.05), while overweight NAFLD and obese NAFLD were not associated with TC abnormalities (P>0.05); obese NAFLD was not associated with TG abnormalities, while lean NAFLD and overweight NAFLD were associated with TG abnormalities (both P<0.05); all types of NAFLD were associated with the abnormalities of waist-hip ratio, blood pressure, blood glucose, low-density lipoprotein, HDL, and uric acid (all P<0.05). ConclusionThe detection rates of different subtypes of NAFLD in Shanghai Pudong are close to those reported in China and globally, and the epidemiologic data of NAFLD can be used analogously for MAFLD. There are certain differences in the distribution and association of cardiometabolic risk factors between different subtypes of NAFLD, and targeted interventions should be formulated based on the metabolic characteristics of each type of NAFLD.
8.Measurement and analysis of activity concentrations of varying forms of 131I in nuclear medicine workplaces
Shuo WANG ; Fei TUO ; Jianfeng ZHANG ; Xiaoliang LI ; Baolu YANG ; Qiang ZHOU
Chinese Journal of Radiological Medicine and Protection 2025;45(5):465-471
Objective:To understand the activity concentrations of varying chemical forms of 131I in nuclear medicine workplaces and assess the internal irradiation doses of 131I to workers. Methods:A high-volume air sampler was used for air sampling of 131I. Glass fiber filters, activated carbon filters, and iodine cartridges, which were connected in series, were employed to collect aerosol iodine, gaseous inorganic iodine, and gaseous organic iodine, respectively. A method for analyzing the activity of 131I unevenly distributed in the iodine cartridge was developed, and an HPGe γ spectrometer was used to determine the activity of 131I in samples collected from the nuclear medicine workplaces of 15 hospitals. Results:The concentrations of aerosol iodine, inorganic iodine, and organic iodine in 15 hospitals were determined at 0.19-206.67, 0.27-138.45, and 2.35-3 821.11 Bq/m 3, respectively, with arithmetic means of 22.04, 12.79 and 365.08 Bq/m 3, respectively. The maximum annual committed effective doses of varied forms of 131I inhaled by workers were determined at 0.19, 0.19, and 3.81 mSv, respectively, with a maximum total committed effective dose of 4.13 mSv. Conclusions:Gaseous organic iodine is identified as the primary form of 131I in the air within nuclear medicine workplaces. Therefore, it is necessary to highlight the monitoring and protection of gaseous organic iodine.
9.Association Between Selvester QRS Score and Myocardial Fibrosis in Patients With Hypertrophic Cardiomyopathy
Baojing SUN ; Juan WANG ; Xiaoliang LUO ; Haobo XU ; Dong LIU ; Hanyang LIANG ; Jiansong YUAN ; Shubin QIAO
Chinese Circulation Journal 2025;40(11):1103-1110
Objectives:The aim of this study was to investigate the value of assessing Selvester QRS score in evaluating myocardial fibrosis in hypertrophic cardiomyopathy(HCM)patients.Methods:A total of consecutive 149 patients with HCM,who were admitted in Fuwai Hospital from October 2008 to September 2013,were enrolled in this study.Qualitative and quantitative analyses of myocardial fibrosis were conducted based on late gadolinium enhancement(LGE)derived from enhanced cardiac magnetic resonance imaging(CMR).According to the qualitative analysis of myocardial fibrosis by CMR-LGE,the patients were divided into the LGE-negative group(n=46)and the LGE-positive group(n=103).Standard 12-lead electrocardiogram examination was performed and the Selvester QRS score was calculated.Multivariate binary Logistic regression analysis was used to analyze the predictive factors of positive LGE,and the correlation analysis between the Selvester QRS score and the LGE score was also performed.Results:Compared with the LGE-negative group,the proportion of patients with a family history of HCM(12.8%vs.28.2%),the proportion of patients with a positive Selvester QRS score(65.2%vs.83.5%)and the Selvester QRS score([1.65±1.81]points vs.[5.71±3.80]points)were higher,the maximum thickness of the left ventricular wall([20.5±3.2]mm vs.[24.5±5.0]mm)and the percentage of left ventricular myocardial scar area([5.0±5.5]%vs.[17.1±11.4]%)were larger in the LGE-positive group(all P<0.05).Multivariate binary logistic regression analysis showed that the Selvester QRS score(OR=1.429,95%CI:1.128-1.802,P=0.003)and the maximum left ventricular wall thickness(OR=1.257,95%CI:1.029-1.537,P=0.025)were independent predictors of positive LGE.Pearson correlation analysis showed that in all HCM patients,the Selvester QRS score was positively correlated with the LGE score(r=0.682,P<0.001),and the percentage of left ventricular myocardial scar area was positively correlated with the percentage of enhanced area to the total mass of left ventricular myocardium(LGE%)(r=0.682,P<0.001);In the LGE positive group,the Selvester QRS score was positively correlated with the LGE score(r=0.569,P<0.001).Receiver operating characteristic(ROC)curve analysis showed that the optimal cut-off values of Selvester QRS score and maximum left ventricular wall thickness for predicting positive LGE were≥3.5 points and≥21 mm,respectively.Conclusions:In HCM patients,the Selvester QRS score is significantly associated with myocardial fibrosis,and the total score≥3.5 serves as a good predictive cutoffvalue for identifying the presence of LGE.
10.Evaluation of the effect of vitamin D on improving autism symptoms based on the microbiota-gut-brain axis
Haoyu HUANG ; Xiaoliang DU ; Jing WANG ; Jinting WU ; Zhuo ZOU ; Yingjuan CHEN ; Yun LIU
Chinese Journal of Nervous and Mental Diseases 2025;51(3):149-155
Objective To analyze the symptomatic improvement effects of vitamin D in children with autism spectrum disorder(ASD)based on the microbiota-gut-brain axis.Methods Seventy-two children with ASD were randomly divided into an observation group and a control group,with 36 cases in each group.Three cases dropped out in the control group.The observation group received 1200 IU/day of vitamin D supplementation in addition to conventional rehabilitation training,while the control group received only conventional rehabilitation training.The intervention lasted for 12 weeks.Assessments were conducted before and after the intervention using the childhood autism rating scale(CARS),autism behavior checklist(ABC),and repetitive behavior scale-revised(RBS-R).Resting-state functional connectivity of the brain was measured using near-infrared functional imaging,and serum levels of 25(OH)D3,inflammatory cytokines,and gut microbiota were analyzed.The differences in these indicators before and after the intervention were compared between the two groups to evaluate clinical efficacy.Results The between-group differences in pre-and post-intervention changes showed that the observation group had significantly greater improvements than the control group in the following measures:CARS scores(-5.92±1.40 vs.-2.55±1.43),RBS-R scores(-5.99±1.01 vs.-3.10±1.47),resting-state brain functional connectivity(0.19±0.15 vs.0.10±0.18),serum 25(OH)D3 levels[(34.89±8.18)ng/mL vs.(0.68±6.73)ng/mL],serum interleukin-6(IL-6)levels[(-6.60±6.07)pg/mL vs.(-0.74±9.45)pg/mL],IL-1β levels[(-2.56±1.33)pg/mL vs.(-0.04±2.13)pg/mL],and tumor necrosis factor-α(TNF-α)levels[(-4.09±3.85)pg/mL vs.(0.21±4.05)pg/mL](P<0.05).Post-intervention,significant differences in gut microbial β-diversity were observed between the two groups(R2=0.030,P=0.040,Adonis).LEfSe analysis revealed that the observation group exhibited enrichment in Clostridia(LDA=4.747,P=0.003),Clostridiales(LDA=4.747,P=0.003),Clostridiaceae(LDA=3.476,P=0.001),Lachnospiraceae(LDA=4.709,P=0.004),Odoribacteraceae(LDA=3.458,P=0.027),Odoribacter(LDA=3.458,P=0.027),Burkholderiales(LDA=3.339,P=0.038),Firmicutes(LDA=4.764,P=0.003),and Betaproteobacteria(LDA=3.338,P=0.037).Conclusion Vitamin D supplementation can modulate gut microbial diversity in children with ASD,significantly influence the abundance of specific gut microbiota,reduce systemic inflammatory cytokines,enhance brain functional connectivity,and alleviate clinical symptoms of ASD.

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