Objective:To observe the effects of acupuncture at Houxi(SI3)and Huantiao(GB30)on the expression levels of nuclear factor kappa B(NF-κB),inducible nitric oxide synthase(iNOS),and nitric oxide(NO)of NF-κB inflammatory signaling pathway in L5 spinal nerve root of lumbar disc herniation(LDH)model rats and explore the mechanism of acupuncture in LDH treatment.Methods:Forty specific-pathogen-free healthy male Sprague-Dawley rats were randomly divided into a sham operation group,a model group,acupuncture group 1,and acupuncture group 2,with 10 rats in each group.The non-compression nucleus protrusion model was made by puncturing L4-L5 spinous process space and injecting autologous nucleus suspension.Acupuncture at bilateral Shenshu(BL23),Dachangshu(BL25),and Weizhong(BL40)was carried out in acupuncture group 1,and acupuncture at bilateral Houxi(SI3)and Huantiao(GB30)in acupuncture group 2.All rats were treated with balanced reinforcing and reducing needling manipulations,and the needles were retained for 30 min/time with one episode of needling manipulation every 10 min,once a day,14 times in total.The threshold value of paw withdrawal pain was measured by a thermal stimulation pain instrument;the serum NF-κB,iNOS,and NO levels were measured by enzyme-linked immunosorbent assay.The pathomorphological changes of spinal nerve roots were observed by hematoxylin-eosin(HE)staining;quantitative reverse transcription polymerase chain reaction was used to detect iNOS mRNA expression in spinal nerve roots;the NF-κB and iNOS protein expression in spinal nerve roots was detected by the immunofluorescence method.Results:Compared with the sham operation group,the threshold of paw withdrawal pain in the model group was decreased,and the expression levels of serum NF-κB,iNOS,and NO were increased;HE staining showed many degenerated and dissolved Schwann cells in spinal nerve roots with vacuolar changes;meanwhile,the expression levels of NF-κB and iNOS proteins,and the iNOS mRNA in spinal nerve roots were increased.Compared with the model group,the paw withdrawal pain thresholds in acupuncture group 1 and acupuncture group 2 were increased,and the increase in acupuncture group 2 was greater(P<0.05);the expression levels of serum NF-κB,iNOS,and NO in acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01);the vacuolar changes of spinal nerve roots,and the degeneration and lysis of Schwann cells in acupuncture group 1 and acupuncture group 2 were decreased,which were more obvious in acupuncture group 2;the NF-κB and iNOS protein expression and the iNOS mRNA expression levels in spinal nerve roots of acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01).Conclusion:Acupuncture at Houxi(SI3)and Huantiao(GB30)can improve the morphology of spinal nerve roots,inhibit the NF-κB and iNOS protein expression levels in spinal nerve roots and the serum NO level,and relieve the pain caused by inflammation of spinal nerve roots,which may be one of the mechanisms of acupuncture in LDH treatment.

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.

ObjectiveTo provide a basis for the establishment of an ideal animal model of allergic asthma by statistically analyzing the modeling characteristics and the selection of indicators of the available models. MethodsWe retrieved the relevant articles from China National Knowledge Infrastructure（CNKI）， VIP， Wanfang Data， SinoMed， and PubMed with "allergic asthma" as the keyword and the time interval from January 2019 to January 2024. Through integrating the literature and extracting data， we used Excel 2021 to create a personal database and sorted out the animal strains， genders， allergenic substances， modeling routes， and test indicators and methods. Excel 2021， Cytoscape 3.10.2， and SPSS Modeler 18.0 were then used to analyze the relevant characteristics of the animal models. ResultsA total of 418 articles were included in the database， and the comparative analysis showed that the most frequently used animal strain for modeling was BALB/c mice， and female animals were mostly used. The main modeling method was sensitization by intraperitoneal injection of ovalbumin （OVA）， which was combined with intranasal inhalation. The test indicators mainly included appearance signs， cellular analysis， lung histopathology， lung function indicators， and protein and gene expression in the lung. The test methods mainly involved pathological staining， enzyme-linked immunosorbent assay， immunohistochemistry， immunofluorescence， Western blot， and polymerase chain reaction（PCR） assays. ConclusionThere is no recognized modeling method or evaluation standard for the animal models of allergic asthma. Based on the results of data analysis， the OVA-induced allergic asthma model in BALB/c mice is recommended. The main criteria for evaluating the success of modeling are the general behavioral changes， the morphological changes of the airway and inflammatory cell infiltration in the lung tissue， the changes of pro-inflammatory and anti-inflammatory cytokines in the serum， and the alterations of inflammatory cells in the bronchoalveolar lavage fluid.

Objective:To compare similarities and differences in expression changes of cytotoxic T-lymphocyte antigen-4(CT-LA-4)and programmed death receptor 1(PD-1)in immunosuppressed rabbits under different moxibustion interventions.Methods:Twenty large-eared white rabbits were randomly divided into normal group,model group,moxa stick moxibustion(MSM)group and herbal cake-partitioned moxibustion(HPM)group,with five rabbits in each group.CTX-induced immunosuppressed models were prepared by intraperitoneal injection for 7 consecutive days.After successful modeling,MSM and HPM were performed on alternate days for 10 treatments.Rabbits were anesthetized after treatment,and serum,liver and spleen were collected.Serum PD-1 and PD-L1 contents were detected by ELISA.PD-1 in liver tissue was detected by immunohistochemistry,and CTLA-4 mRNA in liver and spleen tissues were detected by RT-qPCR.Results:Both HPM and MSM could reduce PD-1 and PD-L1 levels due to immunosuppression,and could effectively suppress elevated levels of CTLA-4 in spleen and PD-1 and CTLA-4 in liver,which were statistically different compared with immunosuppression model group(P<0.05).Pearson correlation test showed that CTLA-4 and PD-1 in liver tissue had significant positive correlation(r=0.780 7,P<0.001).Conclusion:HPM can improve body immune function by regulating multiple immunosuppressive sites.

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.