OBJECTIVE To investigate the effects and potential mechanism of curcumin on neurological injury in neonatal rats with bacterial meningitis based on the signal transducer and activator of transcription 1（ STAT1）/ nucleotide-binding domain leucine- rich repeat and pyrin domain-containing receptor 3 （NLRP3） signaling pathway. METHODS Neonatal rats， with an equal number of males and females， were randomly divided into control group， model group， curcumin low-dose （Cur-L）， medium-dose （Cur- M） and high-dose （Cur-H） groups， and Cur-H+STAT1 transcription enhancer [2-（1，8-naphthyridin-2-yl）phenol] group （Cur-H+2- NP group）， with 15 rats in each group. Except for the control group， rats in other groups were injected with a suspension of group B Streptococcus （1×104 cfu/mL， 10 μL） into the cerebellomedullary cistern to establish a bacterial meningitis model. After successful model establishment， rats in Cur-L， Cur-M and Cur-H groups were intraperitoneally injected with 1.25， 2.5 and 5 mg/kg curcumin， respectively， and those in the Cur-H+2-NP group were intraperitoneally injected with 5 mg/kg curcumin and 0.5 mg/kg 2- NP， once a day， for 3 consecutive weeks. After the last administration， modified Loeffler score was conducted， white blood cells （WBC） count in cerebrospinal fluid as well as the contents of inflammatory factors [tumor necrosis factor-α， interleukin-6 （IL-6）， IL-1β and IL-18]， brain water content and blood-brain barrier permeability were detected； the histopathological changes of hippocampus and cortex tissues were observed. The percentage of apoptosis in hippocampal/cortical tissue cells， the positive expression of ionized calcium-binding adapter molecule-1 （Iba-1）， the co-localization of Iba-1 and NLRP3， as well as the expressions of proteins related to the STAT1/NLRP3 signaling pathway （phosphorylated STAT1， NLRP3， apoptosis-associated speck-like protein containing a CARD， gasdermin D， caspase-1， IL-1β and IL-18） were examined. RESULTS Compared with the control group， the neurons in the hippocampal/cortical tissues of rats in the model group exhibited significant morphological abnormalities， accompanied by neuronal edema and necrosis， as well as infiltration of inflammatory cells. The modified Loeffler score and the number of Nissl bodies were significantly decreased/reduced in the model group， while the WBC count， levels of inflammatory factors， brain water content， blood-brain barrier permeability， HE staining score， number of degenerated neurons， percentage of apoptotic cells， positive expression of Iba-1， percentage of Iba-1 and NLRP3 co-localization- positive cells， and expressions of pathway-related proteins were all significantly rose/increased/upregulated （P＜0.05）. Compared with the model group， the histopathological changes in the hippocampal/cortical tissues of rats in all curcumin dosage groups were alleviated to varying degrees， with significant improvements in all quantitative indicators （P＜0.05）； conversely， 2-NP significantly reversed the ameliorative effects of curcumin on these quantitative indicators （P＜0.05）. CONCLUSIONS Curcumin can alleviate cerebral edema and blood-brain barrier damage， reduce neuroinflammation， inhibit cell apoptosis and pyroptosis， and thereby alleviate neuronal injury in neonatal rats with bacterial meningitis. The underlying mechanism may be related to the inhibition of the STAT1/NLRP3 signaling pathway.

Objective:To investigate the current status of insomnia symptoms and executive function (EF) impairments among adolescents from regions with different economic development levels, and to analyze their relationship with depressive symptoms, so as to provide clues for improved depressive symptoms screening practices.Methods:This population-based cross-sectional study employed a multistage, stratified cluster random sampling method. During November 2017 to January 2018 and December 2018 to January 2019, a total of 2 495 adolescents aged 11 to 18 years were selected from Shanghai, representing a highly developed economic region, and 2 704 adolescents aged 11 to 18 years were selected from Shangrao city, Jiangxi province, representing a less developed economic region. The depressive symptoms were assessed using the short version of the 21-item depression, anxiety, and stress scale, based on which participants were categorized into groups with or without depressive symptoms. Insomnia symptoms and EF impairments were measured using a self-designed insomnia scale and the behavior rating inventory of executive function, respectively. Participants were further classified into 4 subgroups: neither insomnia nor EF impairment, EF impairment only, insomnia only, and comorbid insomnia and EF impairment. Chi-square test was used to compare the differences in basic information of adolescents from different regions. Multivariate Logistic regression models were applied to examine the associations between insomnia, EF impairment, and their combination with depressive symptoms as well as the differences in gender and school-stage among each subgroup.Results:A total of 2 305 adolescents were recruited from Shanghai (1 192 boys and 1 113 girls, 1 266 junior high school students and 1 039 senior high school students) and 2 250 adolescents from Shangrao (1 126 boys and 1 124 girls, 1 146 junior high school students and 1 104 senior high school students). The numbers of adolescents with depressive symptoms, insomnia symptoms and EF impairment in Shanghai were 460 adolescents (20.0%), 907 adolescents (39.3%), and 411 adolescents (17.8%), respectively, all of which were fewer than those in Shangrao, which were 616 adolescents (27.4%), 1 251 adolescents (55.6%), and 524 adolescents (23.3%), respectively (all P<0.001). In Shanghai, the numbers of adolescents with EF impairment only, insomnia only, and comorbid insomnia and EF impairment were 219 adolescents (9.5%), 670 adolescents (29.1%), and 237 adolescents (10.3%), respectively. And in Shangrao, the corresponding numbers were 193 adolescents (8.6%), 865 adolescents (38.4%), and 386 adolescents (17.2%), respectively. Compared to adolescents in Shanghai with neither EF impairment nor insomnia, the risk of depressive symptoms was all higher in adolescents with EF impairment only, insomnia only, and comorbid EF impairment-insomnia ( OR=2.86, 6.48, 20.10; 95% CI 1.57-5.22, 5.09-8.26, 13.66-29.58; all P<0.01). Similar results were observed in adolescents in Shangrao ( OR=3.22, 4.82, 10.91; 95% CI 1.66-6.28, 3.09-7.51, 7.26-16.40; all P<0.01). The analysis of gender and educational stage differences showed that, compared to the group neither EF impairment nor insomnia, the risk of depressive symptoms all higher in the groups with EF impairment only, insomnia only (all P<0.05), and comorbid EF impairment-insomnia, and the risk in comorbid EF impairment-insomnia group was the highest (all P<0.05). Conclusions:Compared with adolescents in regions with underdeveloped economies, those in economically developed regions had lower rates of insomnia, EF impairment, and depression. Both insomnia and EF impairment significantly increase the risk of depressive symptoms. Their coexistence confers the highest risk and therefore warrants particular attention for prevention and intervention efforts.

Melatonin is widely used as an over-the-counter medication to treat insomnia in children with autism spectrum disorder (ASD) and neurogenetic disorders (NGD). However, there is still a lack of research on its efficacy and safety, and clinical practice standards are to be established. In response, the International Pediatric Sleep Association (IPSA) convened an expert panel and developed a consensus statement:"Melatonin Use in Managing Insomnia in Children with Autism and Other Neurogenetic Disorders-an Assessment by the International Pediatric Sleep Association (IPSA)", which was published in Sleep Medicine, April 2024. The consensus focused on the efficacy and adverse effects of melatonin treatment for insomnia in children with ASD and NGD-including Smith-Magenis syndrome, Rett syndrome, Angelman syndrome, and tuberous sclerosis complex. It systematically reviews randomized controlled trials (RCTs) conducted between 2012 and 2022, and integrates current best clinical practices to formulate 10 consensus recommendations. Despite these contributions, the consensus has limitations: a small number of included RCTs, a lack of grading for evidence quality, and recommendation strength. Furthermore, the study population is primarily composed of children from Western countries. This article seeks to interpret the consensus to improve standardized use of melatonin for insomnia in Chinese children with ASD and NGD, and to provide a reference for the future development of localized evidence-based guidelines.

Objective To explore the regulation of the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)-mediated inflammatory response in chronic obstructive pulmonary disease(COPD)by modified Shengxian Decoction through the lung-gut axis.Methods Thirty rats were divided into three groups:Control group,COPD group,and COPD+modified Shengxian Decoction(SXT)group,with 10 rats in each.The COPD model was established using passive smoking combined with intratracheal instillation of lipopolysaccharide(LPS).General symptoms and signs of the rats were monitored during the modeling and intervention periods.Hematoxylin and eosin(HE)staining and immunohistochemistry(IHC)were used to observe lung tissue structure.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory cytokines interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in lung tissues.Flow cytometry was used to detect the number of type Ⅱ innate lymphoid cells(nILC2)and type 2 innate lymphoid cells(iILC2)in lung and intestinal tissues.Illumina MiSeq sequencing technology was used to perform 16S rRNA gene sequencing on rat feces to analyze the gut microbiota structure.Gas chromatography-mass spectrometry(GC-MS)was used to determine the content of short-chain fatty acids(SCFAs)in rat feces.Western blotting was used to detect the expressions of related proteins in the PI3K/AKT pathway.Results Compared with the Control group,the COPD group showed significantly reduced lung function indicators,increased heart rate and decreased body mass,while the SXT group showed significant improvement in lung function and general signs(P＜0.05).HE staining showed that the COPD group had lung tissue damage filled with inflammatory cells,while the SXT group had significantly fewer inflammatory cells.IHC results showed that the SXT group had significantly reduced expression of caspase-3 protein(P＜0.05).ELISA results showed that the levels of IL-6 and TNF-α were significantly increased in the COPD group,while the SXT group showed significant improvement in inflammatory damage.The ratio of nILC2 to iILC2 in lung and intestinal tissues was significantly reduced in the COPD group,indicating a significant inflammatory response,while the SXT group showed significant improvement(P＜0.05).The levels of ILC2 cytokines IL-13 and IL-4 were significantly increased in the COPD group,while the SXT group had significantly reduced IL-13 and IL-4 levels.The relative abundance of lung and gut microbiota in the SXT group was significantly higher than that in the Control and COPD groups(P＜0.05).Beta diversity index analysis showed significant differences in species diversity among the three groups(P＜0.05).GC-MS detected six types of SCFAs in rat feces:acetic acid,propionic acid,isobutyric acid,butyric acid,isovaleric acid,and valeric acid.Their levels were lower in the COPD group than in the Control group,but the levels in the SXT group were higher than those in the COPD group.Western blotting results showed that the expressions of p-PI3K,PI3K,p-AKT,AKT,p-NF-κB,and NF-κB proteins were significantly reduced in the SXT group compared to the COPD group(P＜0.05).ELISA results showed that the SXT group had significantly downregulated expression levels of IL-1β and IL-10 compared to the COPD group(P＜0.05).Conclusion Modified Shengxian Decoction can alleviate COPD inflammation.It may mediate the inflammatory response in COPD by inhibiting iILC2 cell activity and expressions of related proteins in the PI3K/AKT signaling pathway through gut microbiota metabolism.

Objective:To investigate the impact of the number of examined lymph nodes (ELN) on survival outcomes in gastric cancer patients with postoperative pathological stage pN3b.Methods:This retrospective cohort study included 279 pN3b gastric cancer patients who underwent D2 gastrectomy at Nanfang Hospital, Southern Medical University (September 2008 to April 2023), with 35 patients receiving combination chemotherapy and anti-PD-1 therapy (immunotherapy group) and 244 receiving adjuvant chemotherapy alone (nonimmunotherapy group). Additionally, 422 patients with pN3b from the SEER database (2005 to 2020) were collected as an external validation cohort to determine the optimal cutoff value for the number of lymph nodes examined in the nonimmunotherapy group. The primary endpoints were overall survival (OS) and recurrence-free survival (RFS) in the nonimmunotherapy group of the Nanfang Hospital cohort, stratified by whether the number of examined lymph nodes was above or below the ELN optimal cutoff value. These findings were subsequently validated in the SEER cohort.Results:The optimal ELN cutoff value (34 nodes) was determined using X-tile software and by constructing an ELN-HR fitting model with inflection point identification. In the nonimmunotherapy group, patients with ELN >34 exhibited significantly prolonged survival compared to ELN ≤34 (median OS: 25.0 (95%CI:20.5-29.5) to 17.0 (95%CI:12.7-21.3) months, P=0.004; median RFS: 19.0 (95%CI:15.6-22.4) to 13.0 (95%CI:9.5-16.5) months, P=0.048). Multivariate Cox analysis also showed ELN >34 to be an independent protective factor for both OS (HR=0.576, 95%CI: 0.397-0.836) and RFS (HR=0.701, 95%CI: 0.492-0.998). In the SEER cohort, ELN >34 was associated with a 5-month OS extension (19 to 14 months, P=0.065), with multivariate analysis supporting its independent prognostic significance (HR=0.729, 95%CI: 0.580-0.915, P=0.006). Notably, in the immunotherapy group, patients with ELN >34 ( n=30) achieved a median OS of 41 months, but the median OS had not been reached in the ELN ≤34 group ( n=5) (1 death at 48 months). Conclusion:Higher ELN (>34) correlates with improved survival in nonimmunotherapy-treated pN3b gastric cancer patients. However, in pN3b gastric cancer patients treated with immunotherapy, the optimal ELN threshold requires further exploration to determine.

Objective To explore the regulation of the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)-mediated inflammatory response in chronic obstructive pulmonary disease(COPD)by modified Shengxian Decoction through the lung-gut axis.Methods Thirty rats were divided into three groups:Control group,COPD group,and COPD+modified Shengxian Decoction(SXT)group,with 10 rats in each.The COPD model was established using passive smoking combined with intratracheal instillation of lipopolysaccharide(LPS).General symptoms and signs of the rats were monitored during the modeling and intervention periods.Hematoxylin and eosin(HE)staining and immunohistochemistry(IHC)were used to observe lung tissue structure.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory cytokines interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in lung tissues.Flow cytometry was used to detect the number of type Ⅱ innate lymphoid cells(nILC2)and type 2 innate lymphoid cells(iILC2)in lung and intestinal tissues.Illumina MiSeq sequencing technology was used to perform 16S rRNA gene sequencing on rat feces to analyze the gut microbiota structure.Gas chromatography-mass spectrometry(GC-MS)was used to determine the content of short-chain fatty acids(SCFAs)in rat feces.Western blotting was used to detect the expressions of related proteins in the PI3K/AKT pathway.Results Compared with the Control group,the COPD group showed significantly reduced lung function indicators,increased heart rate and decreased body mass,while the SXT group showed significant improvement in lung function and general signs(P＜0.05).HE staining showed that the COPD group had lung tissue damage filled with inflammatory cells,while the SXT group had significantly fewer inflammatory cells.IHC results showed that the SXT group had significantly reduced expression of caspase-3 protein(P＜0.05).ELISA results showed that the levels of IL-6 and TNF-α were significantly increased in the COPD group,while the SXT group showed significant improvement in inflammatory damage.The ratio of nILC2 to iILC2 in lung and intestinal tissues was significantly reduced in the COPD group,indicating a significant inflammatory response,while the SXT group showed significant improvement(P＜0.05).The levels of ILC2 cytokines IL-13 and IL-4 were significantly increased in the COPD group,while the SXT group had significantly reduced IL-13 and IL-4 levels.The relative abundance of lung and gut microbiota in the SXT group was significantly higher than that in the Control and COPD groups(P＜0.05).Beta diversity index analysis showed significant differences in species diversity among the three groups(P＜0.05).GC-MS detected six types of SCFAs in rat feces:acetic acid,propionic acid,isobutyric acid,butyric acid,isovaleric acid,and valeric acid.Their levels were lower in the COPD group than in the Control group,but the levels in the SXT group were higher than those in the COPD group.Western blotting results showed that the expressions of p-PI3K,PI3K,p-AKT,AKT,p-NF-κB,and NF-κB proteins were significantly reduced in the SXT group compared to the COPD group(P＜0.05).ELISA results showed that the SXT group had significantly downregulated expression levels of IL-1β and IL-10 compared to the COPD group(P＜0.05).Conclusion Modified Shengxian Decoction can alleviate COPD inflammation.It may mediate the inflammatory response in COPD by inhibiting iILC2 cell activity and expressions of related proteins in the PI3K/AKT signaling pathway through gut microbiota metabolism.

Objective:To investigate the impact of the number of examined lymph nodes (ELN) on survival outcomes in gastric cancer patients with postoperative pathological stage pN3b.Methods:This retrospective cohort study included 279 pN3b gastric cancer patients who underwent D2 gastrectomy at Nanfang Hospital, Southern Medical University (September 2008 to April 2023), with 35 patients receiving combination chemotherapy and anti-PD-1 therapy (immunotherapy group) and 244 receiving adjuvant chemotherapy alone (nonimmunotherapy group). Additionally, 422 patients with pN3b from the SEER database (2005 to 2020) were collected as an external validation cohort to determine the optimal cutoff value for the number of lymph nodes examined in the nonimmunotherapy group. The primary endpoints were overall survival (OS) and recurrence-free survival (RFS) in the nonimmunotherapy group of the Nanfang Hospital cohort, stratified by whether the number of examined lymph nodes was above or below the ELN optimal cutoff value. These findings were subsequently validated in the SEER cohort.Results:The optimal ELN cutoff value (34 nodes) was determined using X-tile software and by constructing an ELN-HR fitting model with inflection point identification. In the nonimmunotherapy group, patients with ELN >34 exhibited significantly prolonged survival compared to ELN ≤34 (median OS: 25.0 (95%CI:20.5-29.5) to 17.0 (95%CI:12.7-21.3) months, P=0.004; median RFS: 19.0 (95%CI:15.6-22.4) to 13.0 (95%CI:9.5-16.5) months, P=0.048). Multivariate Cox analysis also showed ELN >34 to be an independent protective factor for both OS (HR=0.576, 95%CI: 0.397-0.836) and RFS (HR=0.701, 95%CI: 0.492-0.998). In the SEER cohort, ELN >34 was associated with a 5-month OS extension (19 to 14 months, P=0.065), with multivariate analysis supporting its independent prognostic significance (HR=0.729, 95%CI: 0.580-0.915, P=0.006). Notably, in the immunotherapy group, patients with ELN >34 ( n=30) achieved a median OS of 41 months, but the median OS had not been reached in the ELN ≤34 group ( n=5) (1 death at 48 months). Conclusion:Higher ELN (>34) correlates with improved survival in nonimmunotherapy-treated pN3b gastric cancer patients. However, in pN3b gastric cancer patients treated with immunotherapy, the optimal ELN threshold requires further exploration to determine.

Objective:To investigate the current status of insomnia symptoms and executive function (EF) impairments among adolescents from regions with different economic development levels, and to analyze their relationship with depressive symptoms, so as to provide clues for improved depressive symptoms screening practices.Methods:This population-based cross-sectional study employed a multistage, stratified cluster random sampling method. During November 2017 to January 2018 and December 2018 to January 2019, a total of 2 495 adolescents aged 11 to 18 years were selected from Shanghai, representing a highly developed economic region, and 2 704 adolescents aged 11 to 18 years were selected from Shangrao city, Jiangxi province, representing a less developed economic region. The depressive symptoms were assessed using the short version of the 21-item depression, anxiety, and stress scale, based on which participants were categorized into groups with or without depressive symptoms. Insomnia symptoms and EF impairments were measured using a self-designed insomnia scale and the behavior rating inventory of executive function, respectively. Participants were further classified into 4 subgroups: neither insomnia nor EF impairment, EF impairment only, insomnia only, and comorbid insomnia and EF impairment. Chi-square test was used to compare the differences in basic information of adolescents from different regions. Multivariate Logistic regression models were applied to examine the associations between insomnia, EF impairment, and their combination with depressive symptoms as well as the differences in gender and school-stage among each subgroup.Results:A total of 2 305 adolescents were recruited from Shanghai (1 192 boys and 1 113 girls, 1 266 junior high school students and 1 039 senior high school students) and 2 250 adolescents from Shangrao (1 126 boys and 1 124 girls, 1 146 junior high school students and 1 104 senior high school students). The numbers of adolescents with depressive symptoms, insomnia symptoms and EF impairment in Shanghai were 460 adolescents (20.0%), 907 adolescents (39.3%), and 411 adolescents (17.8%), respectively, all of which were fewer than those in Shangrao, which were 616 adolescents (27.4%), 1 251 adolescents (55.6%), and 524 adolescents (23.3%), respectively (all P<0.001). In Shanghai, the numbers of adolescents with EF impairment only, insomnia only, and comorbid insomnia and EF impairment were 219 adolescents (9.5%), 670 adolescents (29.1%), and 237 adolescents (10.3%), respectively. And in Shangrao, the corresponding numbers were 193 adolescents (8.6%), 865 adolescents (38.4%), and 386 adolescents (17.2%), respectively. Compared to adolescents in Shanghai with neither EF impairment nor insomnia, the risk of depressive symptoms was all higher in adolescents with EF impairment only, insomnia only, and comorbid EF impairment-insomnia ( OR=2.86, 6.48, 20.10; 95% CI 1.57-5.22, 5.09-8.26, 13.66-29.58; all P<0.01). Similar results were observed in adolescents in Shangrao ( OR=3.22, 4.82, 10.91; 95% CI 1.66-6.28, 3.09-7.51, 7.26-16.40; all P<0.01). The analysis of gender and educational stage differences showed that, compared to the group neither EF impairment nor insomnia, the risk of depressive symptoms all higher in the groups with EF impairment only, insomnia only (all P<0.05), and comorbid EF impairment-insomnia, and the risk in comorbid EF impairment-insomnia group was the highest (all P<0.05). Conclusions:Compared with adolescents in regions with underdeveloped economies, those in economically developed regions had lower rates of insomnia, EF impairment, and depression. Both insomnia and EF impairment significantly increase the risk of depressive symptoms. Their coexistence confers the highest risk and therefore warrants particular attention for prevention and intervention efforts.

Melatonin is widely used as an over-the-counter medication to treat insomnia in children with autism spectrum disorder (ASD) and neurogenetic disorders (NGD). However, there is still a lack of research on its efficacy and safety, and clinical practice standards are to be established. In response, the International Pediatric Sleep Association (IPSA) convened an expert panel and developed a consensus statement:"Melatonin Use in Managing Insomnia in Children with Autism and Other Neurogenetic Disorders-an Assessment by the International Pediatric Sleep Association (IPSA)", which was published in Sleep Medicine, April 2024. The consensus focused on the efficacy and adverse effects of melatonin treatment for insomnia in children with ASD and NGD-including Smith-Magenis syndrome, Rett syndrome, Angelman syndrome, and tuberous sclerosis complex. It systematically reviews randomized controlled trials (RCTs) conducted between 2012 and 2022, and integrates current best clinical practices to formulate 10 consensus recommendations. Despite these contributions, the consensus has limitations: a small number of included RCTs, a lack of grading for evidence quality, and recommendation strength. Furthermore, the study population is primarily composed of children from Western countries. This article seeks to interpret the consensus to improve standardized use of melatonin for insomnia in Chinese children with ASD and NGD, and to provide a reference for the future development of localized evidence-based guidelines.

Objective Based on TLR4/NF-κB/MLCK pathway,the therapeutic effect and mechanism of Baihe Dihuang Decoction on insomnia with intestinal flora disturbance in mice induced by p-chlorophenlalanine(PCPA)and multi-factor stimulation were studied.The aim is to provide theoretical basis for clinical use.Methods Eighty-four KM mice were randomly divided into normal group,model group,positive group(Diazepam,1.38 mg·kg-1),Baihe group(2.25 g·kg-1),Dihuang group(2.25 g·kg-1)and Baihe Dihuang Decoction group(4.5 g·kg-1).Insomnia mouse model was established by intraperitoneal injection of PCPA for 2 days combined with 4 weeks of multi-factor stimulation,including stimulating the tail with forceps clip for 2 minutes,reversing day and night for 24 hours,wetting the padding for 24 hours,tilting the cage at 45° for 24 hours,alternating the cages for 24 hours,fasting food for 24 hours,and getting cold bath for 3 minutes,etc.After successfully modeling,corresponding drug treatment was given.The anxiety-like behavior of mice was observed by elevated cross maze system.The latency and duration of sleep were observed by righting reflex experiment.16sRNA sequencing was used to analyze the composition and structure of intestinal flora in mice.The concentration changes of γ-aminobutyric acid(GABA),glutamic acid(Glu),tryptophan(Trp),5-hydroxytryptophan(5-HTP)and 5-hydroxytryptamine(5-HT)in brain and colon were detected by liquid chromatography-mass spectrometry(LC-MS).Immunohistochemical method was used to detect the expressions of zona atresia protein 1(ZO-1)and Occludin in colon.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the genes including interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),ZO-1,Occludin,Toll-like receptor 4(TLR4),nuclear factor κB(NF-κB)and myosin light chain kinase(MLCK)in colonic tissue.Western Blot was used to detect the expressions of TLR4,NF-κB,phosphorylated nuclear factor κB(p-NF-κB),MLCK,myosin light chain(MLC)and phosphorylated myosin light chain(p-MLC)in colonic epithelial tissue.Results Compared with the normal group,the distance of entering the open arm and the duration of stay in the open arm of model group in the elevated cross maze were significantly shortened(P＜0.05).The sleep latency was significantly prolonged,and the sleep duration was significantly shortened(P＜0.05).The richness and uniformity of intestinal flora were decreased(P＜0.05,P＜0.01).The concentration of neurotransmitters GABA,Trp,5-HTP and 5-HT in the brain decreased significantly(P＜0.01),while the concentration of Glu increased significantly(P＜0.01).The concentration of GABA and Glu in colon decreased significantly(P＜0.01),while the concentration of Trp,5-HTP and 5-HT increased significantly(P＜0.01).The expression levels of inflammatory factors IL-1β,IL-6 and TNF-α in colonic tissue were significantly increased(P＜0.01),and the expression levels of ZO-1 and Occludin genes and proteins were significantly decreased(P＜0.01).The gene expression levels of TLR4,NF-κB and MLCK were significantly increased(P＜0.01),and the protein expression levels of TLR4,p-NF-κB,MLCK and p-MLC were significantly increased(P＜0.01).Compared with the model group,Baihe Dihuang Decoction could significantly prolong the distance of entering the open arm and the duration of stay in the open arm of insomnia mice in the elevated cross maze(P＜0.05).The sleep latency was significantly shortened,and the sleep duration was significantly increased(P＜0.05).The richness and uniformity of intestinal flora were increased(P＜0.05,P＜0.01).The concentration of neurotransmitters GABA,Trp,5-HTP and 5-HT in brain was increased(P＜0.05,P＜0.01),and the concentration of Glu was decreased(P＜0.01).The concentration of GABA and Glu in colon was increased(P＜0.01),while the concentration of Trp,5-HTP and 5-HT was decreased(P＜0.05,P＜0.01).The expression levels of IL-1β,IL-6 and TNF-α genes were down-regulated(P＜0.01),the expression levels of ZO-1 and Occludin genes and proteins were up-regulated(P＜0.01),TLR4,NF-κB,MLCK gene expression levels and TLR4,p-NF-κB,MLCK,p-MLC protein expression levels were down-regulated in the pathway(P＜0.01).Conclusion Baihe Dihuang Decoction can effectively treat insomnia with intestinal flora disorders.Its mechanism of action may be related to the regulation of brain and intestinal neurotransmitter disorders,down-regulating TLR4/NF-κB/MLCK signaling pathway,and up-regulating tight junction proteins expression,reducing inflammatory responses,and then repairing the mechanical barrier of intestinal mucosa.