Objective:To investigate the distribution of pathogenic bacteria in the ear canal secretions of patients with chronic suppurative otitis media（CSOM）, the changes in the levels of interleukin-8（IL-8） and Toll like receptor 4（TLR4） in the ear canal secretions, and their clinical significance. Methods:This study selected 128 CSOM patients who visited our hospital from January 2022 to February 2024 as the study subjects and recorded them as the CSOM group. Additionally, 135 volunteers who underwent physical examinations at our hospital during the same period were regarded as the control group. Video otoscopy was used to collect and cultivate ear canal secretions, and a fully automated microbial identification instrument was used to identify the bacterial species. ELISA was applied to detect levels of IL-8, TLR4. Multivariate logistic regression was employed to examine the factors that affect the occurrence of CSOM. Pearson correlation was applied to analyze the correlation between IL-8, TLR4 levels and various influencing factors. ROC curve was applied to analyze the diagnostic value of IL-8 and TLR4 levels for the occurrence of CSOM. Z-test was applied to compare the differences in AUC. Results:Among 128 patients, the detection rate was 89.06%, and a total of 181 strains of pathogenic bacteria were cultured, among them, Gram positive bacteria accounted for the highest proportion of 54.14%, followed by Gram negative bacteria, accounting for 34.25%, and finally fungi, accounting for 11.60%. The common bacteria were Staphylococcus aureus （20.44%）, Pseudomonas aeruginosa （13.26%）, and Staphylococcus epidermidis （8.29%）. The resistance of Gram-positive bacteria to penicillin, clindamycin, erythromycin, and amoxicillin is high. Gram-negative bacteria are highly resistant to penicillin, ampicillin and erythromycin. Fungi are resistant to ketoconazole and fluconazole. The levels of IL-8 and TLR4 in CSOM group were higher than those in the control group, and gradually increased with the increase of hearing impairment. （P<0.05）. Elevated levels of IL-8, TLR4 were independent risk factors for the occurrence of CSOM（P<0.05）. The AUC of CSOM diagnosed by IL-8 and TLR4 alone was 0.790 and 0.777, respectively, while the AUC of combined diagnosis was 0.898, which was better than their respective individual diagnoses（both P<0.05）. Conclusion:The distribution of pathogenic bacteria in the ear canal secretions of CSOM patients is mainly Gram positive, with common ones being Staphylococcus aureus and Pseudomonas aeruginosa. The levels of IL-8 and TLR4 in CSOM patients are higher than those in the control group. The higher the levels, the higher the degree of hearing loss, which can be used for clinical diagnosis.
Humans ; Toll-Like Receptor 4/metabolism* ; Interleukin-8/metabolism* ; Otitis Media, Suppurative/metabolism* ; Ear Canal/metabolism* ; Chronic Disease ; Male ; Female ; Adult ; Middle Aged ; Clinical Relevance

Objective:To explore the effect, postoperative mucosal pathological changes and molecular biological changes of reboot operation for type 2 inflammation chronic rhinosinusitis with nasal polyps（CRSwNP） patients, and to provide theoretical basis for the clinical application of this kind of operation. Methods:We collected 29 patients who were diagnosed with CRSwNP with type 2 inflammatino response and underwent Reboot surgery from June 2022 to August 2023, and 27 patients who were diagnosed with deviated septum and underwent simple submucosal resection of the septum as the control group. We conducted nasal symptom scoring, endoscopic sinusitis scoring, and CT scanning of the sinuses before and after surgery, as well as HE staining, immunohistochemical staining, and detection of inflammatory factors using Elisa kits at the time of surgery, 1, 3, and 6 months postoperatively. We also observed the ultrastructural changes using transmission electron microscopy and scanning electron microscopy, and performed proteomic analysis of the mucosa in the ethmoid sinus area of the sinusitis patients at the time of surgery and 6 months postoperatively. Results:After 6 months of postoperative follow-up, CT scores of the nasal cavity and sinuses had gradually decreased compared with the preoperative period. The VAS score of main symptoms, SNOT-22 score and Lund-Kennedy endoscopic score were decreased after 12 months follow-up. The histological morphology of the mucosa in the area of the screen was significantly improved compared with the preoperative period, with a reduction in the number of eosinophils. The levels of inflammatory factors such as IL-4 and IL-5 et al. in the mucosa of the area of the screen were gradually reduced compared with the preoperative period. The histological morphology, ultrastructure, and cilia structure of the mucosa in the area of the screen were gradually improved compared with the preoperative period, though not recovered completely. The number of CD4⁺T and CD8⁺T cells not changed significantly before and after the surgery yet. By conducting proteomic analysis of the ethmoidal sinus mucosa before and after surgery, differential proteins were selected, and bioinformatics analysis was conducted on the differentially expressed proteins. By using cytoHubba to identify hub genes and intersecting them with the genes related to chronic sinusitis, we found that MMP9 expression increased in non-type 2 CRS and type 2 CRS in sequence, while ACTC1 expression decreased in non-tpye 2 CRS and type 2 CRS in sequence. Conclusion:Reboot surgery can improve the postoperative symptoms and signs of patients, improve the pathological morphology of the mucosa, and influence the expression of protein after surgery. However, the surgery may not have a significant impact on the distribution of T cell subpopulations and inflammation signal pathway in the nasal mucosa.
Humans ; Sinusitis/metabolism* ; Nasal Polyps/metabolism* ; Nasal Mucosa/ultrastructure* ; Chronic Disease ; Rhinitis/complications* ; Inflammation ; Male ; Female ; Postoperative Period ; Adult ; Interleukin-5/metabolism* ; Interleukin-4/metabolism* ; Middle Aged ; Proteomics ; Rhinosinusitis

Chronic rhinosinusitis（CRS） is a chronic inflammation of the nasal cavity and sinus mucosa, which is mainly characterized by nasal obstruction, runny nose, headache and hyposmia. Due to complex mechanisms, CRS can be divided into different clinical phenotypes and inflammatory endotypes. Approximately 80% of chronic rhinosinusitis with nasal polyps（CRSwNP） patients present with type Ⅱ inflammation. IL-4 and IL-13 are the key factors in the process of type Ⅱ inflammation, and drugs targeting IL-4/IL-13 provide new options for the treatment of CRSwNP. Therefore, this article mainly reviews the application of anti-IL-4 /IL-13 monoclonal antibody in CRSwNP.
Humans ; Nasal Polyps/therapy* ; Sinusitis/therapy* ; Chronic Disease ; Interleukin-13/antagonists & inhibitors* ; Antibodies, Monoclonal/therapeutic use* ; Interleukin-4/antagonists & inhibitors* ; Rhinitis/drug therapy* ; Rhinosinusitis

OBJECTIVES: To investigate the therapeutic effect of electroacupuncture (EA) at Zusanli (ST36) acupoint on hyperlipidemia in mice and explore the underlying mechanisms. METHODS: Thirty C57BL/6J mice were equally randomized into normal diet group, high-fat diet (HFD) group, and EA group. The changes in blood lipids and serum malondialdehyde (MDA) content of the mice were evaluated, and histopathological changes and lipid accumulation in the liver were observed using Oil red O staining (ORO). The expressions of NLRP3, TLR4, and IL-1β proteins in the colon tissues were detected with Western blotting, and gut microbiota changes were analyzed using 16S rDNA sequencing. RESULTS: In mice with HFD feeding, 16 weeks of EA treatment significantly lowered body weight and serum TC, TG, LDL-C and MDA levels, obviously reduced lipid accumulation in the liver, and ameliorated HFD-induced elevations of protein expressions of NLRP3, TLR4, and IL-1β. 16S rRNA sequencing revealed that EA significantly altered gut microbiota composition, and increased the diversity and relative abundance of beneficial bacterial groups such as Muribaculaceae and Lachnospiraceae NK4A136_group. CONCLUSIONS Electroacupuncture at ST36 alleviates blood lipid disorders in hyperlipidemic mice possibly by improving intestinal microbiota structure, promoting degradation of high-caloric carbohydrates, cholesterol lipid metabolism and intestinal mucosa repair, and reducing toxin leakage, lipid peroxides, and liver fat deposition.
Animals ; Electroacupuncture ; Gastrointestinal Microbiome ; Hyperlipidemias/blood* ; Mice, Inbred C57BL ; Mice ; Diet, High-Fat ; Toll-Like Receptor 4/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein ; Acupuncture Points ; Male ; Lipids/blood* ; Interleukin-1beta/metabolism* ; Liver/metabolism*

OBJECTIVES: To explore the therapeutic effect of LuoFuShan Rheumatism Plaster (LFS) on neuropathic pain (NP) and its molecular mechanism. METHODS: Mouse models of sciatic nerve chronic constriction injury (CCI) were treated with low, medium, and high doses (2.2, 4.4, and 8.8 cm², respectively) of LFS by topical application for 14 consecutive days. The therapeutic effects were assessed by evaluating the mechanical withdrawal threshold (MWT), paw withdrawal latency (PWL), plasma IL-6 and TNF-α levels, and histopathology of the sciatic nerve. Network pharmacology and molecular docking were used to identify the key targets and signaling pathways. The key targets were verified by RT-qPCR and immunohistochemistry. The biosafety of LFS was evaluated by measuring the organ indices and damage indicators of the heart, liver, and kidneys. RESULTS: Compared with the CCI group, LFS dose-dependently increased MWT and PWL, reduced plasma IL-6 and TNF-α levels, and alleviated sciatic nerve inflammation in the mouse models. Network pharmacology identified 378 bioactive compounds targeting 279 NP-associated genes enriched in TLR and TNF signaling. Molecular docking showed that quercetin and ursolic acid in LFS could stably bind to TLR4 and TNF‑α. In the mouse models of sciatic nerve CCI, LFS significantly downregulated the mRNA expression levels of Tlr4 and Tnf-α in the spinal cord in a dose-dependent manner and lowered the protein expressions of TLR4 and TNF-α in the sciatic nerve. LFS treatment did not cause significant changes in the organ indices or damage indicators of the heart, liver and kidneys as compared with those in the CCI model group and sham-operated group. CONCLUSIONS LFS alleviates NP in mice by suppression of TLR4/TNF-α-mediated neuroinflammation with a good safety profile.
Animals ; Toll-Like Receptor 4/metabolism* ; Neuralgia/metabolism* ; Mice ; Signal Transduction/drug effects* ; Tumor Necrosis Factor-alpha/metabolism* ; Drugs, Chinese Herbal/pharmacology* ; Sciatic Nerve/injuries* ; Male ; Molecular Docking Simulation ; Disease Models, Animal ; Interleukin-6

Acute lung injury (ALI) is a severe disease caused by viral infection that triggers an uncontrolled inflammatory response. This study investigated the capacity of jasurolignoside (JO), a natural compound, to bind to Toll-like receptor 4 (TLR4) and treat ALI. The anti-inflammatory properties of JO were evaluated in vitro through Western blotting, enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining, and co-immunoprecipitation. The investigation utilized a lipopolysaccharide (LPS)-induced ALI animal model to examine the therapeutic efficacy and mechanism of JO in vivo. JO attenuated inflammatory symptoms in infected cells and tissues by modulating the NOD-like receptor family pyrin domain containing protein 3 (NLRP3) inflammasome and the nuclear factor κB (NF-κB)/mitogen-activated protein kinase (MAPK) pathway. Molecular docking simulations revealed JO binding to TLR4 active sites, confirmed by cellular thermal shift assay. Surface plasmon resonance (SPR) demonstrated direct interaction between JO and TLR4 with a Kd value of 35.1 μmol·L^-1. Moreover, JO inhibited tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and IL-6 secretion and reduced leukocyte, neutrophil, lymphocyte, and macrophage infiltration in ALI-affected mice. JO also enhanced lung function and reduced ALI-related mortality. Immunohistochemical staining demonstrated JO's ability to suppress TLR4 expression in ALI-affected mouse lung tissue. This study establishes that JO can bind to TLR4 and effectively treat ALI, indicating its potential as a therapeutic agent for clinical applications.
Toll-Like Receptor 4/chemistry* ; Animals ; Acute Lung Injury/chemically induced* ; Mice ; Humans ; Ilex/chemistry* ; Molecular Docking Simulation ; Male ; NF-kappa B/immunology* ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein/immunology* ; Tumor Necrosis Factor-alpha/genetics* ; Interleukin-1beta/genetics* ; RAW 264.7 Cells ; Disease Models, Animal

OBJECTIVE: To observe the clinical efficacy of combination therapy of wheat-grained sized cone moxibustion and point-to-point needle insertion with medical ozone penetrating injection for allergic rhinitis (AR) and its effect on inflammation-related indexes. METHODS: Thirty-one patients with persistent AR were enrolled. The patients received medical ozone injection at bilateral Yingxiang (LI20)-to-Shangyingxiang (EX-HN8), and wheat-grained sized cone moxibustion at Dazhui (GV14), twice a week (with a 3-day interval) for 4 consecutive weeks. The total nasal symptoms score (TNSS), total non-nasal symptom score (TNNSS), rhinoconjunctivitis quality of life questionnaire (RQLQ), and rhinitis control assessment test (RCAT) scores were evaluated before treatment, after treatment, and at the 8-week follow-up. Levels of eosinophil (EOS) count, immunoglobulin E (IgE), interleukin (IL)-4, IL-6, and IL-17 were measured before and after treatment. Clinical efficacy was evaluated after treatment, and the recurrence rate was assessed at follow-up. RESULTS: Compared with those before treatment, the TNSS, TNNSS, and RQLQ scores were decreased (P<0.05), while the RCAT score was increased (P<0.05) after treatment and at follow-up. There were no statistically significant differences in above indexes between the post-treatment and follow-up (P>0.05). After treatment, the whole blood EOS count and serum levels of IgE, IL-4, IL-6, and IL-17 were decreased compared with those before treatment (P<0.05). After treatment, 17 cases were markedly effective, 12 cases were effective, and 2 cases were ineffective, resulting in a total effective rate of 93.5%. At follow-up, 2 cases relapsed, and the recurrence rate was 6.9%. CONCLUSION Combination therapy of wheat-grained sized cone moxibustion and point-to-point needle insertion with medical ozone injection can improve AR symptoms, reduce the recurrence rate, and enhance the quality of life. The mechanism may be associated with the regulation of immune-related indexes.
Humans ; Female ; Male ; Moxibustion ; Adult ; Ozone/administration & dosage* ; Middle Aged ; Young Adult ; Acupuncture Points ; Adolescent ; Combined Modality Therapy ; Treatment Outcome ; Immunoglobulin E/blood* ; Rhinitis, Allergic/immunology* ; Interleukin-4/immunology*

BACKGROUND: Electroacupuncture (EA) treatment is efficacious in patients with respiratory disorders, although the mechanisms of its action in lung-function protection are poorly understood. This study aimed to explore the neuroanatomical mechanisms of EA stimulation at the BL13 acupoint (Feishu, EA-BL13) improvement in asthma. METHODS: Allergic asthma was induced by intranasal 2.0% ovalbumin (OVA) instillation combined with intraperitoneal injection of the 10.0% OVA. The levels of interleukin (IL)-4 and IL-5 were detected by enzyme-linked immunosorbent assay. Hematoxylin and eosin and periodic acid-schiff stain were used to evaluate inflammatory cell infiltration and mucus secretion. Cellular oncogene fos induction in neurons after EA stimulation was detected by immunofluorescent staining. The messenger RNA expression levels of adrenergic receptors were quantified with real-time polymerase chain reaction. RESULTS: EA improved airway inflammation and mucus secretion mainly by activating somatosensory-sympathetic pathways ( P <0.001). Briefly, the intermediolateral (IML) nuclei of the spinal cord received signals from somatic EA stimulation and then delivered the information via the sympathetic trunk to the lung. Excited sympathetic nerve endings in lung tissue released large amounts of catecholamines that specifically activated the β2 adrenergic receptor (β2AR) on T cells ( P <0.01) and further decreased the levels of IL-4 and IL-5 ( P <0.001) through the cyclic adenosine monophosphate/protein kinase A signaling pathway. CONCLUSION This study provided a new explanation and clinical basis for the use of EA-BL13 as a treatment for allergic asthma in both the attack and remission stages and other respiratory disorders related to airway inflammation.
Electroacupuncture/methods* ; Animals ; Asthma/immunology* ; Rats ; Rats, Sprague-Dawley ; Male ; Inflammation/therapy* ; Interleukin-4/metabolism* ; Interleukin-5/metabolism*

Objective To explore the effect of Evodiamine (Evo) on the immune function of allergic rhinitis (AR) rats and the regulatory mechanism on C-C motif chemokine ligand 2 (CCL2)/ C-C motif chemokine receptor 2 (CCR2) pathway. Methods The related targets of Evo-AR-immune function were screened by network pharmacology, and the protein interaction network diagram of intersecting targets was constructed. The AR rat model was established by ovalbumin (OVA) combined with aluminium hydroxide, and the rats were divided into six groups: a normal control (NC) group, a model group, a Loratadine (LOR) group, an Evodiamine low dose (Evo-L) group, a Evodiamine high dose (Evo-H) groups, and an Evo-H combined with CCL2 group. After the last administration, the symptoms of rats in each group were scored; ELISA was applied to detect the levels of histamine, immunoglobulin E (IgE), interleukin 4 (IL-4), IL-13 and interferon γ (IFN-γ); Diff-Quick staining solution was applied to detecte the number of cells in the nasal lavage fluid (NALF); hematoxylin eosin (HE) staining was applied to observe the pathological changes of nasal mucosa tissue; real-time quantitative PCR was applied to detect the levels of CCL2 and CCR2 mRNA in tissue; Western blot was applied to detect the expression levels of CCL2, CCR2 and CXC motif chemokine ligand 8 (CXCL8) proteins in nasal mucosa. Results There were eight intersection targets of EVo-AR-immune function, and protein interaction network diagram showed that CXCL8 was the core target. Compared with the NC group, the score of nasal symptoms, the levels of histamine, IgE, IL-4 and IL-13, the numbers of eosinophil, macrophages, neutrophils, lymphocytes and total cells, the mRNA and protein expression levels of CCL2 and CCR2, and the expression of CXCL8 protein in the model group were increased, while the level of IFN-γ was decreased. Compared with the model group, the score of nasal symptoms, the levels of histamine, IgE, IL-4 and IL-13, the numbers of eosinophil, macrophages, neutrophils, lymphocytes and total cells, the mRNA and protein expression levels of CCL2 and CCR2, and the expression of CXCL8 protein in LOR and Evo groups were decreased, while the level of IFN-γ was increased. Further use of CCL2 recombinant protein for compensatory experiments revealed that the improvement effect of Evo on immune function in AR rats was reversed by CCL2. Conclusion Evo can improve the immune function of AR rats, and its mechanism may be related to the inhibition of the CCL2/CCR2 pathway.
Animals ; Receptors, CCR2/immunology* ; Signal Transduction/drug effects* ; Chemokine CCL2/immunology* ; Rats ; Rhinitis, Allergic/metabolism* ; Immunoglobulin E/blood* ; Quinazolines/pharmacology* ; Male ; Interferon-gamma ; Rats, Sprague-Dawley ; Interleukin-13 ; Histamine ; Interleukin-4/immunology* ; Disease Models, Animal

Objective To explore the characteristics of the inhibitory effect of Evodiamine on the proliferation of activated T cells. Methods Mononuclear cells from peripheral blood (PBMCs) were obtained from healthy donors through density gradient centrifugation, and T cells were subsequently purified by using immunomagnetic bead separation. T cell activation was induced by employing anti-human CD3 and anti-human CD28 antibodies. T cells were treated with different concentrations of EVO (0.37, 1.11, 3.33, and 10)μmol/L. Flow cytometry was applied to evaluate the proliferation index, apoptosis rate, viability, CD25 expression levels, and cell cycle distribution of T cells. The expression levels of cytokines IL-2, IL-17A, IL-4, and IL-10 were quantified by using ELISA. Results 1.11, 3.33 and 10 μmol/L EVO effectively inhibited the proliferation of activated T cells, with an IC₅₀ of (1.5±0.3)μmol/L. EVO did not induce apoptosis in activated T cells and affect the survival rate of resting T cells. EVO did not affect the expression of CD25 and the secretion of IL-2 in activated T cells. EVO arrested the T cell cycle at the G2/M phase, resulting in an increase in G2/M phase cells, and exhibited a concentration-dependent effect. EVO did not affect the secretion of IL-4, IL-10 by activated T cells, but significantly inhibited the secretion of IL-17A. Conclusion EVO did not significantly affect the activation process of T cells but inhibited T cell proliferation by arresting the cell cycle at the G2/M phase and significantly suppressed the secretion of the pro-inflammatory cytokine IL-17A, which suggests that EVO has the potential to serve as a lead compound for the development of low-toxicity and high-efficiency immunosuppressants and elucidates the mechanisms underlying the anti-inflammatory and immunomodulatory effects of the traditional Chinese medicine Evodia rutaecarpa.
Humans ; Cell Proliferation/drug effects* ; Quinazolines/pharmacology* ; T-Lymphocytes/metabolism* ; Lymphocyte Activation/drug effects* ; Apoptosis/drug effects* ; Interleukin-4/metabolism* ; Interleukin-10/metabolism* ; Interleukin-2 Receptor alpha Subunit/metabolism* ; Interleukin-17/metabolism* ; Interleukin-2/metabolism* ; Cell Cycle/drug effects* ; Cells, Cultured