Objective Through studying the chemical composition changes before and after the compatibility of Hedysari Radix Paeparata Cum Melle(HRPCM)-vinegar processed Curcumae Rhizoma(VPCR);To discuss the significance of the compatibility of HRPCM and VPCR;To establish the fingerprints before and after their compatibility.Methods ZORBAX Eclipse Plus C18 column was used;acetonitrile-0.01%phosphoric acid water was set as mobile phase,with gradient elution;column temperature was 30℃;detection wavelength was 214 nm;sample injection was 10 μL,which was used to investigate the content difference of major chemical components such as vanillic acid,calycosin-O-β-D-glucopyranoside,ononin,calycosin,onocerin,curdione,cincumol and germacrone,and establish the fingerprint of HRPCM,VPCR and HRPCM-VPCR.Results HPLC chromatographic conditions were established for the determination of 8 components in HRPCM-VPCR.Meanwhile,fingerprints were established before and after the compatibility of HRPCM-VPCR.26 common peaks were identified,among which 11 components such as vanillic acid were derived from HRPCM,14 components such as curcuma zedoariae were derived from VPCR,and 1 component was shared by both.Conclusion The material basis of the compatibility of HRPCM-VPCR differs from that of HRPCM and VPCR.The content of most chemical components decreases while the content of some components increases.The established HPLC method for content determination and fingerprint is simple,stable and reproducible,which can be used to evaluate and control the quality of HRPCM and VPCR.

Objective:To investigate the differences in hippocampal subfield volumes and structural covariance network between participants with subjective cognitive decline (SCD) and healthy individuals, and to analyze the correlations of the volumes of the different subfields and altered covariance brain regions with memory function.Methods:A total of 57 SCD individuals(SCD group) and 44 normal controls(NC group) participants were assessed for memory function using composite scores from the auditory verbal learning test (AVLT) and the Wechsler memory scale visual reproduction (VR) test from June 2022 to October 2023.T1-weighted structural magnetic resonance imaging (MRI) data were collected from all participants, and hippocampal subfields, cortical regions, and subcortical nuclei were segmented using FreeSurfer to measure the gray matter volume of each structure. A structural covariance network was constructed based on the correlation of gray matter volumes across regions. Statistical analysis was performed using R 4.3.1 software. Inter-group differences in hippocampal subfield volumes were compared using multivariate analysis of covariance. Differences in structural covariance connectivity between groups were assessed using Z-test, while network topology differences were compared through permutation testing. Finally, partial correlation analysis was used to examine correlation of the volumes of the differential hippocampal subfields and covariance brain regions with memory function. Results:The SCD group exhibited significantly lower years of education, AVLT-immediate score, AVLT-delayed score, VR-immediate score, VR-delayed score, and memory function Z-score compared to the NC group ( t=2.064, 3.888, 2.622, 3.222, 4.761, 5.184, all P<0.05). The volumes of the right subiculum((387.75±55.20)mm 3, (352.70±70.25)mm 3), left presubiculum((263.12±38.52)mm 3, (239.79±46.02)mm 3), left subiculum((388.12±49.34)mm 3, (351.74±67.30)mm 3) and left CA1((571.01±80.01)mm 3, (526.51±98.80)mm 3) in the SCD group were smaller than the corresponding volumes in NC group ( F=9.139, 8.039, 11.207, 7.266, all P<0.05, FDR correction). Differences in structural covariance connectivity were found between the SCD and NC groups in the following pairs: right CA1-right subiculum, right CA1-left subiculum, right CA3-left parasubiculum and right hippocampus-amygdala transition area-left subiculum ( Z=-3.848, -3.896, -3.597, -3.895, all P<0.05, FDR correction).Partial correlation analysis revealed that in the SCD group, the volume of the left subiculum ( r=0.359, P=0.007), left CA1 ( r=0.430, P=0.001), right entorhinal cortex ( r=0.296, P=0.029), right middle temporal gyrus ( r=0.361, P=0.007), right parahippocampal gyrus ( r=0.313, P=0.021)were positively correlated with the total memory function score. Conclusion:Hippocampal subfields atrophy, as well as alterations in structural covariance network, have been found in SCD individuals. Furthermore, the decline in memory function may be closely associated with atrophy in hippocampal subfields and structurally covariant regions.

Objective:To investigate the differences in hippocampal subfield volumes and structural covariance network between participants with subjective cognitive decline (SCD) and healthy individuals, and to analyze the correlations of the volumes of the different subfields and altered covariance brain regions with memory function.Methods:A total of 57 SCD individuals(SCD group) and 44 normal controls(NC group) participants were assessed for memory function using composite scores from the auditory verbal learning test (AVLT) and the Wechsler memory scale visual reproduction (VR) test from June 2022 to October 2023.T1-weighted structural magnetic resonance imaging (MRI) data were collected from all participants, and hippocampal subfields, cortical regions, and subcortical nuclei were segmented using FreeSurfer to measure the gray matter volume of each structure. A structural covariance network was constructed based on the correlation of gray matter volumes across regions. Statistical analysis was performed using R 4.3.1 software. Inter-group differences in hippocampal subfield volumes were compared using multivariate analysis of covariance. Differences in structural covariance connectivity between groups were assessed using Z-test, while network topology differences were compared through permutation testing. Finally, partial correlation analysis was used to examine correlation of the volumes of the differential hippocampal subfields and covariance brain regions with memory function. Results:The SCD group exhibited significantly lower years of education, AVLT-immediate score, AVLT-delayed score, VR-immediate score, VR-delayed score, and memory function Z-score compared to the NC group ( t=2.064, 3.888, 2.622, 3.222, 4.761, 5.184, all P<0.05). The volumes of the right subiculum((387.75±55.20)mm 3, (352.70±70.25)mm 3), left presubiculum((263.12±38.52)mm 3, (239.79±46.02)mm 3), left subiculum((388.12±49.34)mm 3, (351.74±67.30)mm 3) and left CA1((571.01±80.01)mm 3, (526.51±98.80)mm 3) in the SCD group were smaller than the corresponding volumes in NC group ( F=9.139, 8.039, 11.207, 7.266, all P<0.05, FDR correction). Differences in structural covariance connectivity were found between the SCD and NC groups in the following pairs: right CA1-right subiculum, right CA1-left subiculum, right CA3-left parasubiculum and right hippocampus-amygdala transition area-left subiculum ( Z=-3.848, -3.896, -3.597, -3.895, all P<0.05, FDR correction).Partial correlation analysis revealed that in the SCD group, the volume of the left subiculum ( r=0.359, P=0.007), left CA1 ( r=0.430, P=0.001), right entorhinal cortex ( r=0.296, P=0.029), right middle temporal gyrus ( r=0.361, P=0.007), right parahippocampal gyrus ( r=0.313, P=0.021)were positively correlated with the total memory function score. Conclusion:Hippocampal subfields atrophy, as well as alterations in structural covariance network, have been found in SCD individuals. Furthermore, the decline in memory function may be closely associated with atrophy in hippocampal subfields and structurally covariant regions.

ObjectiveTo explore the mechanism of herbal cake-separated moxibustion using the classical formula Xiaoyaosan in alleviating neuroimmune inflammatory responses in chronic fatigue syndrome （CFS） rats， based on the regulation of the toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear transcription factor-κB （NF-κB） signaling pathway. MethodsFifty SPF-grade SD rats aged 6-8 weeks were randomly divided into five groups： Normal group， model group， sham herbal cake moxibustion group， Chinese medicine intragastric administration group， and herbal cake-separated moxibustion group， with 10 rats in each group. Except for the normal group， all other groups underwent a 21-day modeling process， followed by behavioral testing. The herbal cake-separated moxibustion and sham herbal cake moxibustion groups received interventions at the Shenque （CV8）， Guanyuan （CV4）， Zusanli （ST36）， and Qimen （LR14） acupoints. The Chinese medicine intragastric administration group was treated with a Xiaoyaosan suspension via gavage. Behavioral tests were conducted after 10 days of continuous intervention. Serum levels of interleukin （IL）-1β， IL-6， and tumor necrosis factor-α （TNF-α）， as well as hippocampal levels of IL-1β， IL-6， TNF-α， and NF-κB， were detected by enzyme-linked immunosorbent assay （ELISA）. Morphological changes in the hippocampus were observed using hematoxylin-eosin （HE） staining. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA expression levels of TLR4， MyD88， and NF-κB in the hippocampus. Western blot analysis was performed to detect the relative expression levels of TLR4， MyD88， NF-κB， and p65 proteins in the hippocampus. ResultsCompared with the normal group， the model group showed a significant decrease in upright times during the open field test （P<0.01）， as well as significant reductions in total movement distance， resting time， and center region duration （P<0.01）. In the tail suspension test， immobility time increased （P<0.01）， and struggle times decreased （P<0.01）. Serum and hippocampal levels of IL-1β， IL-6， and TNF-α， as well as hippocampal NF-κB levels and TLR4， MyD88， and NF-κB mRNA expression， were significantly elevated （P<0.01）. After treatment， compared with the model group， the total movement distance and upright times in the open field test were significantly increased in all treatment groups （P<0.01）， while resting time and center region duration were notably prolonged （P<0.05， P<0.01）. Immobility time in the tail suspension test was significantly shortened （P<0.01）， and struggle times significantly increased （P<0.05）. Serum and hippocampal levels of IL-1β， IL-6， TNF-α， hippocampal NF-κB levels， and TLR4 and NF-κB mRNA expression were significantly reduced （P<0.05， P<0.01）. Compared with the sham herbal cake moxibustion group， the herbal cake-separated moxibustion group showed a significant extension in center region duration during the open field test （P<0.05） and a significant increase in upright times （P<0.01）. In the tail suspension test， immobility time was reduced （P<0.01）， and struggle times increased （P<0.01）. Serum TNF-α levels in the Chinese medicine intragastric administration group were significantly reduced （P<0.01）， while serum IL-6 levels， as well as hippocampal levels of IL-1β， TNF-α， NF-κB， and TLR4， MyD88， and NF-κB mRNA expression， were significantly decreased in both the Chinese medicine intragastric administration group and the herbal cake-separated moxibustion group （P<0.05， P<0.01）. Compared with the Chinese medicine intragastric administration group， the herbal cake-separated moxibustion group exhibited significantly increased upright times in the open field test （P<0.01）. In the tail suspension test， immobility time was reduced （P<0.01）， and struggle times increased （P<0.01）. Serum IL-1β， hippocampal TNF-α levels， and TLR4， MyD88， and NF-κB mRNA expression were significantly decreased （P<0.05， P<0.01）. ConclusionHerbal cake-separated moxibustion effectively improves fatigue and memory function in CFS rats， regulates neuroimmune inflammatory responses， and its mechanism may be related to the modulation of the TLR4/MyD88/NF-κB signaling pathway.

BACKGROUND:Immunosuppression leads to impaired body immune function and aggravates the disease.Herb-partitioned moxibustion can effectively regulate immune function and improve immunity in the body,but its regulatory mechanism has not been elucidated. OBJECTIVE:To sequence immunosuppressed model rats treated with herb-partitioned moxibustion using bioinformatics techniques based on transcriptomics and to explore the mechanisms by which it regulates immunity. METHODS:Twenty-four Sprague-Dawley rats were randomly assigned to three groups:control,model,and herb-partitioned moxibustion groups,with eight rats in each group.The model and herb-partitioned moxibustion groups were subjected to establishment of an immune suppression model by intraperitoneal injection of cyclophosphamide at a dose of 35 mg/kg for 3 consecutive days.No interventions were administered to the control and model groups after modeling.In contrast,the herb-partitioned moxibustion group received moxibustion treatment at Zhongwan,Shenque,Guanyuan,and Zusanli acupoints using a combination of moxa and herbal cakes,once a day,for 10 consecutive days,with samples being collected the day after the end of the intervention.Peripheral blood was collected from all groups of rats to measure their white blood cell count.RNA-seq was performed on the Illumina sequencing platform,and differentially expressed genes were selected for bioinformatics analysis using the GO and KEGG databases. RESULTS AND CONCLUSION:Compared with the control group,the model group exhibited a significant decrease in white blood cell count(P<0.001).RNA-seq analysis identified 3 026 differentially expressed genes between the model and control groups,with 1 565 upregulated and 1 461 downregulated.There were 535 differentially expressed genes identified between the herb-partitioned moxibustion group and the model group,with 280 upregulated and 255 downregulated.The Venn diagram analysis revealed that 159 genes were downregulated in the model group compared with the control group.However,after moxibustion with herbal cakes,these genes were upregulated.Protein-protein interaction network analysis identified 10 core targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,Mx1,Syk,Hspa1a,and Ret.According to GO and KEGG analyses,moxibustion with herbal cakes regulated the body through pathways related to immune response,viruses,angiogenesis,and the autoimmune system.To conclude,there is a significant association between herbal cake-separated moxibustion intervention and immune suppression targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,and Mx1.The intervention exhibits regulatory effects in the pathways related to immune responses,viral activities,and angiogenesis.

Objective:To analyze the differentially expressed messenger RNA(mRNA), microRNA(miRNA), and long non-coding RNA(lncRNA)during the browning of mouse subcutaneous adipose tissue, construct a competitive endogenous RNA(ceRNA)network, and provide a theoretical basis for investigating the regulatory mechanisms of white adipose tissue browning.Methods:A cold-stimulated mouse model was established for transcriptome sequencing.Bioinformatics tools were employed to screen for differentially expressed mRNAs, miRNAs, and lncRNAs.An integrated mRNA-miRNA-lncRNA analysis was performed to construct a ceRNA network.Gene Ontology(GO), Kyoto Encyclopedia of Genes and Genomes(KEGG), and Gene Set Enrichment Analysis(GSEA)were conducted on the differentially expressed mRNAs and ceRNA networks to explore transcriptional regulation during the cold-induced browning of subcutaneous adipose tissue.Results:Transcriptomic analysis of the cold-stimulated model identified 4, 256 differentially expressed RNAs, which include 3, 600 mRNAs, 588 lncRNAs, and 68 miRNAs.GO and KEGG analyses revealed that the browning of white adipose tissue involves immune-related processes, such as immune system processes, immune responses, adaptive and innate immune responses, and the positive regulation of T-cell activation.A ceRNA network associated with browning regulation was constructed, comprising 233 nodes(188 mRNAs, 34 miRNAs, and 11 lncRNAs)and 351 edges.Protein-protein interaction(PPI)analysis of the mRNAs within the ceRNA network highlighted pathways including apoptosis, intracellular signaling transduction, hypoxia-inducible factor-1(HIF-1), AMP-activated protein kinase(AMPK), Janus kinase-signal transducer and activators of transcription(JAK-STAT)signaling, carbon metabolism, glycolysis, and thyroid hormone pathways, all of which regulate lipid metabolism, hypoxia, and glycolysis.Cytohubba analysis identified the top 10 hub genes: Bcl2, Src, Cebpb, Creb1, Runx1, Foxo3, Ets1, Socs3, Slc2 a4, and Pkm. Conclusions:The ceRNA network that regulates the browning of white adipose tissue is involved in various pathways, including carbon metabolism, glycolysis, thyroid hormone signaling, growth hormone signaling, prolactin signaling, as well as the HIF-1, AMPK, and JAK-STAT pathways.Key regulatory miRNAs in this context include miR-30e-5p, miR-182-5p, miR-20b-5p, miR-144-3p, miR-363-3p, miR-141-3p, miR-203-3p, and miR-107-3p.These miRNAs may serve as critical targets for inducing browning in response to cold exposure.

Objective:To investigate the differences in hippocampal subfield volumes and structural covariance network properties among patients with mild cognitive impairment (MCI) exhibiting different cognitive outcomes and normal controls (NCs), and to further evaluate the predictive value of these imaging indicators for cognitive deterioration in MCI patients.Methods:A total of 43 NCs, 65 stable MCI (sMCI), and 26 progressive MCI (pMCI) patients enrolled in the Alzheimer′s Disease Neuroimaging Initiative (ADNI) database between December 2012 and May 2016 were included in this study. Baseline demographic information and T 1-weighted magnetic resonance imaging scans were collected. Hippocampal subfield volumes were extracted using freesurfer software, and structural covariance networks of hippocampal subfields were constructed. Multivariate analysis of covariance was used to compare hippocampal subfield volumes among the 3 groups. A general linear model was applied to examine group differences in hippocampal subfield structural covariance network properties. Least absolute shrinkage and selection operator (LASSO)-Logistic regression was employed to identify imaging predictors associated with conversion to Alzheimer′s disease (AD), based on which structural, network-based, and combined predictive models were constructed. Model discrimination was evaluated using the area under the curve (AUC); internal validation was performed using Bootstrap resampling; model calibration was assessed with the Hosmer-Lemeshow test; and clinical utility was evaluated through decision curve analysis. Results:Significant differences in hippocampal subfield volumes (mm3) were observed among the 3 groups (all P<0.05, Bonferroni-corrected). Specifically, left parasubiculum (65.58±13.30, 61.96±17.56, 49.56±11.82, F=9.900), right parasubiculum (65.92±15.21, 59.45±16.65, 47.69±15.48, F=11.612), left presubiculum (277.09±39.85, 258.15±44.86, 224.05±45.05, F=14.513), right presubiculum (262.85±40.43, 247.41±43.27, 209.97±46.11, F=14.500), left subiculum (399.66±32.19, 374.25±55.83, 306.12±51.62, F=32.923), right subiculum (417.93±48.92, 376.59±51.01, 316.82±70.22, F=28.764), left cornu ammonis 1 (CA1) (592.10±83.87, 561.96±94.72, 490.06±86.89, F=13.352), right CA1 (632.15±100.09, 601.24±88.88, 531.05±110.29, F=10.579), left CA3 (191.58±30.08, 180.47±34.66, 155.08±37.82, F=12.182), right CA3 (210.42±28.92, 203.84±34.80, 176.69±41.47, F=9.597), left CA4 (224.61±28.94, 210.49±35.04, 183.98±36.89, F=16.521), right CA4 (238.49±28.14, 227.43±30.65, 200.23±42.74, F=13.702), left granule cell-molecular layer-dentate gyrus (GC-ML-DG) (259.96±36.76, 239.42±41.17, 207.61±41.84, F=19.831), right GC-ML-DG (273.98±35.12, 258.79±36.82, 227.81±49.07, F=14.204), left molecular layer (505.62±66.16, 468.58±75.17, 402.68±75.47, F=22.293), right molecular layer (527.39±72.39, 493.14±70.39, 423.81±88.09, F=19.588), left hippocampal amygdala transition area (HATA) (54.91±9.99, 49.52±9.93, 43.27±9.59, F=13.571), right HATA (58.43±9.83, 54.55±10.80, 47.12±12.54, F=10.037), left fimbria (69.94±25.04, 56.63±23.74, 40.58±19.83, F=14.846), right fimbria (68.61±26.24, 53.95±23.16, 45.25±17.04, F=10.424), left hippocampal tail (488.37±83.44, 463.54±80.33, 393.83±77.73, F=13.570), and right hippocampal tail (519.78±80.22, 498.84±81.68, 419.75±93.29, F=14.339) all showed significant group differences. Significant group differences were also observed in small-worldness metric γ (0.51±0.10, 0.51±0.08, 0.62±0.14, F=9.317), small-worldness metric λ (0.39±0.02, 0.39±0.02, 0.43±0.04, F=9.925), global efficiency (0.19±0.01, 0.20±0.01, 0.18±0.01, F=3.189), local efficiency (0.26±0.02, 0.26±0.01, 0.27±0.01, F=3.068), clustering coefficient (0.23±0.01, 0.23±0.01, 0.24±0.02, F=4.274), and characteristic path length (0.73±0.06, 0.72±0.06, 0.76±0.07, F=4.477) of the hippocampal subfield structural covariance network (all P<0.05). Specifically, the pMCI group exhibited higher γ ( t=3.773, P<0.001), λ ( t=4.060, P<0.001), local efficiency ( t=2.445, P=0.047), and clustering coefficient ( t=2.849, P=0.015) than the NCs group, and higher γ ( t=4.074, P<0.001), λ ( t=4.068, P<0.001), and characteristic path length ( t=2.986, P=0.010) but lower global efficiency ( t=-2.444, P=0.047) than the sMCI group. The AUC of the structural, network, and combined models based on LASSO-Logistic regression was 0.837, 0.861, and 0.899, respectively. After internal validation, the corrected AUC was 0.835, 0.855, and 0.889, respectively. All models demonstrated good calibration ( P>0.05), and decision curve analysis indicated favorable clinical net benefit across models. Conclusions:Both sMCI and pMCI patients exhibit widespread hippocampal subfield atrophy and altered global properties of hippocampal subfield structural covariance networks compared to NCs. The models constructed based on hippocampal subfield volumes and structural covariance networks show strong potential for predicting cognitive decline in MCI patients.

ObjectiveTo explore the mechanism of moxibustion with the classical prescription Xiaoyaosan as the herbal cake in improving cognitive function in the rat model of chronic fatigue syndrome （CFS） by regulating autophagy via phosphatidylinosiol 3-kinase （PI3K）/autophagy key molecule yeast Atg6 homologue 1 （Beclin1） signaling pathway. MethodsFifty SPF-grade SD rats aged 6-8 weeks were randomly grouped as follows： control， model， sham cake-separated moxibustion， intragastric administration with Chinese herbs， and herbal cake-separated moxibustion， each with 10 rats. The other groups except the control group were subjected to modeling by exhaustive swimming and chronic restraint stress for 21 days， and the behavioral test was performed after modeling. Herbal cake-separated moxibustion and sham cake-separated moxibustion were carried out at Shenque （CV8）， Guanyuan （CV4）， Zusanli （ST36）， and Qimen （LR14）. The rats in the intragastric administration with Chinese herbs group were treated with Xiaoyaosan suspension by gavage. After continuous intervention for 10 days， rat behaviors were observed and the levels of interleukin （IL）-4， IL-10， blood urine nitrogen （BUN）， and malondiadehyde （MDA） in the serum and hippocampal tissue of rats in each group were determined by enzyme-linked immunosorbent assay. The morphological changes of the hippocampal tissue were observed by hematoxylin-eosin staining. The relative mRNA and protein levels of PI3K and Beclin1 in the hippocampal tissue were determined by Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultsCompared with the normal group， the model group showed reduced crossings， shortened residence time in the target quadrant， and prolonged average escape latency on days 1， 2， 3， and 4 （P<0.01） in the Morris water maze test. In the new object recognition test， the model group showed decreased recognition index of exploration time and recognition index of exploration times （P<0.01）， lowered levels of IL-4 and IL-10 and elevated levels of BUN and MDA in the serum and hippocampus （P<0.01）， and down-regulated mRNA and protein levels of PI3K and Beclin1 （P<0.01）. After treatment， compared with the model group， each treatment group showed increased crossings， prolonged residence time in the target quadrant， and shortened average escape latency on days 1， 2， 3， and 4 （P<0.05， P<0.01）. In addition， the treatment elevated the levels of IL-4 and IL-10 and lowered the levels of BUN and MDA in the serum and hippocampus （P<0.05， P<0.01）， and up-regulated the mRNA and protein levels of PI3K and Beclin1 （P<0.05， P<0.01）. Compared with the sham cake-separated moxibustion group， the herbal cake-separated moxibustion group and the intragastric administration with Chinese herbs group showed increased crossings， prolonged residence time in the target quadrant （P<0.05）， elevated level of IL-4， lowered level of MDA， and up-regulated relative mRNA and protein levels of PI3K and Beclin1 in the hippocampus （P<0.05， P<0.01）. The serum levels of MDA and BUN in the intragastric administration with Chinese herbs group were significantly decreased. The average escape latency of the herbal cake-separated moxibustion group was significantly shortened on days 2 and 3 （P<0.05， P<0.01）. In addition， this group showed elevated serum level of IL-4， lowered serum level of MDA， and declined BUN level in the hippocampus （P<0.05， P<0.01）. Compared with the intragastric administration with Chinese herbs group， the herbal cake-separated moxibustion group presented risen serum level of IL-4 and up-regulated mRNA level of Beclin1 （P<0.05） and relative protein levels of PI3K and Beclin1 （P<0.05， P<0.01）. ConclusionHerbal cake-separated moxibustion can effectively alleviate fatigue and improve the memory， reduce inflammatory response and oxidative stress， and decrease autophagy of hippocampal neurons in CFS rats by regulating the PI3K/Beclin1 signaling pathway.

ObjectiveTo explore the mechanism of moxibustion with the classical prescription Xiaoyaosan as the herbal cake in improving cognitive function in the rat model of chronic fatigue syndrome （CFS） by regulating autophagy via phosphatidylinosiol 3-kinase （PI3K）/autophagy key molecule yeast Atg6 homologue 1 （Beclin1） signaling pathway. MethodsFifty SPF-grade SD rats aged 6-8 weeks were randomly grouped as follows： control， model， sham cake-separated moxibustion， intragastric administration with Chinese herbs， and herbal cake-separated moxibustion， each with 10 rats. The other groups except the control group were subjected to modeling by exhaustive swimming and chronic restraint stress for 21 days， and the behavioral test was performed after modeling. Herbal cake-separated moxibustion and sham cake-separated moxibustion were carried out at Shenque （CV8）， Guanyuan （CV4）， Zusanli （ST36）， and Qimen （LR14）. The rats in the intragastric administration with Chinese herbs group were treated with Xiaoyaosan suspension by gavage. After continuous intervention for 10 days， rat behaviors were observed and the levels of interleukin （IL）-4， IL-10， blood urine nitrogen （BUN）， and malondiadehyde （MDA） in the serum and hippocampal tissue of rats in each group were determined by enzyme-linked immunosorbent assay. The morphological changes of the hippocampal tissue were observed by hematoxylin-eosin staining. The relative mRNA and protein levels of PI3K and Beclin1 in the hippocampal tissue were determined by Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultsCompared with the normal group， the model group showed reduced crossings， shortened residence time in the target quadrant， and prolonged average escape latency on days 1， 2， 3， and 4 （P<0.01） in the Morris water maze test. In the new object recognition test， the model group showed decreased recognition index of exploration time and recognition index of exploration times （P<0.01）， lowered levels of IL-4 and IL-10 and elevated levels of BUN and MDA in the serum and hippocampus （P<0.01）， and down-regulated mRNA and protein levels of PI3K and Beclin1 （P<0.01）. After treatment， compared with the model group， each treatment group showed increased crossings， prolonged residence time in the target quadrant， and shortened average escape latency on days 1， 2， 3， and 4 （P<0.05， P<0.01）. In addition， the treatment elevated the levels of IL-4 and IL-10 and lowered the levels of BUN and MDA in the serum and hippocampus （P<0.05， P<0.01）， and up-regulated the mRNA and protein levels of PI3K and Beclin1 （P<0.05， P<0.01）. Compared with the sham cake-separated moxibustion group， the herbal cake-separated moxibustion group and the intragastric administration with Chinese herbs group showed increased crossings， prolonged residence time in the target quadrant （P<0.05）， elevated level of IL-4， lowered level of MDA， and up-regulated relative mRNA and protein levels of PI3K and Beclin1 in the hippocampus （P<0.05， P<0.01）. The serum levels of MDA and BUN in the intragastric administration with Chinese herbs group were significantly decreased. The average escape latency of the herbal cake-separated moxibustion group was significantly shortened on days 2 and 3 （P<0.05， P<0.01）. In addition， this group showed elevated serum level of IL-4， lowered serum level of MDA， and declined BUN level in the hippocampus （P<0.05， P<0.01）. Compared with the intragastric administration with Chinese herbs group， the herbal cake-separated moxibustion group presented risen serum level of IL-4 and up-regulated mRNA level of Beclin1 （P<0.05） and relative protein levels of PI3K and Beclin1 （P<0.05， P<0.01）. ConclusionHerbal cake-separated moxibustion can effectively alleviate fatigue and improve the memory， reduce inflammatory response and oxidative stress， and decrease autophagy of hippocampal neurons in CFS rats by regulating the PI3K/Beclin1 signaling pathway.

OBJECTIVE To analyze adverse drug reactions （ADRs） in pediatric patients with Mycoplasma pneumoniae pneumonia （MPP）， and investigate the influential factors associated with the occurrence of ADRs. METHODS Clinical records of pediatric patients with MPP in Xinxiang Central Hospital （hereinafter referred to as “our hospital”） from Oct. 1st， 2023 to Sept. 30th， 2024 collected from the Hospital Information System were retrospectively reviewed using the global trigger tool （GTT）. The positive triggers were further reviewed to confirm the occurrence of ADRs， and the characteristics of those ADRs were analyzed. The children were divided into the ADR group and the non-ADR group. Univariate analysis and multivariate Logistic regression analysis were performed to investigate the potential influential factors for ADRs in MPP children. RESULTS A total of 1 325 MPP children were included. Out of 24 designed triggers， 14 were positive， with a positive trigger rate of 58.3%. There were 1 301 positive trigger cases， involving 846 pediatric patients. After assessment， 399 cases of ADRs were identified in 372 pediatric patients， yielding a positive predictive value of triggers at 30.7%. The main types of ADRs were abnormal blood cells （230 cases， 57.6%）， skin and its appendages damage （78 cases， 19.5%）， and gastrointestinal system damage （42 cases， 10.5%）， which predominantly occurred 4 days or more， within 2-3 days and within 2-3 days after medication， respectively. They were more prevalent in pediatric patients aged＞3-10 years （295 cases， 79.3%）. The major categories of drugs associated with ADRs included intravenous corticosteroids （209 cases）， cephalosporins （180 cases）， macrolides （38 cases）， tetracyclines （28 cases） and traditional Chinese medicine （TCM） injections （28 cases）. These ADR reports were classified into 3 types： 1 serious case， 394 general cases and 4 new cases. In terms of causality assessment， 1 case was assessed as certain， 19 cases as probable， and 379 cases as possible. In terms of ADR outcomes， 151 cases fully recovered， while the outcomes for 248 cases remained unknown. Univariate analysis and multivariate Logistic regression analysis revealed that a history of allergies [OR＝5.231， 95%CI （2.004， 13.656）， P＜0.05]， the number of medications used [OR＝1.064， 95%CI （1.027， 1.103）， P＜0.05]， and intravenous corticosteroid administration [OR＝3.223， 95%CI （2.341， 4.437）， P＜0.05] were independent risk factors of ADRs in pediatric patients with MPP. The use of azithromycin [OR＝0.544， 95%CI （0.376， 0.786）， P＜0.05] was identified as an independent protective factor against the occurrence of ADRs in pediatric patients with MPP. CONCLUSIONS Children with MPP in our hospital exhibit a higher incidence of ADRs， primarily involving abnormal blood cells， skin and its appendages damage， and gastrointestinal system damage. A history of allergies， the number of medications used， intravenous corticosteroid administration， and the use of azithromycin are associated with the occurrence of ADR in MPP pediatric patients.