Abstract:Objective To observe the changes of invariant natural killer T(iNK T)cells in visceral adipose tissue and blood lipids of high-fat diet-fed mice,and to analyze the correlation between iNK T cells and lipid metabolism.Methods Fifty-two C57BL/6 mice were selected as the high-fat diet group,and 51 C57BL/6 mice as the normal control group.The high-fat diet intervention lasted for 12 weeks.At weeks 1,4,8,and 12,the epididymal and perirenal fats of mice in both groups were collected and weighed to record the visceral fat mass(VFM),and the changes in body fat content(BFC)were calculated.Flow cytometry and laser scanning confocal microscopy were used to detect the changes of invariant natural killer T(iNK T)cells in visceral adipose tissue.An automatic biochemical analyzer was used to measure the lipid levels in mice,and the correlations of iNKT cells in visceral adipose tissue with VFM,BFC,and serum lipid levels were analyzed.Results At 12 weeks after high-fat diet feeding,the body weight,VFM,BFC,serum total cholesterol(TC),triglyceride(TG)and high-density lipoprotein cholesterol(HDL-C)increased significantly,while the content of invariant natural killer T(iNK T)cells in visceral adipose tissue decreased obviously in the high-fat diet group,as compared with the control group(P＜0.01).The iNKT cell number in visceral adipose tissue of mice was negatively correlated with VFM,BFC,serum HDL-C and serum TG(r=-0.293,-0.289,-0.337,-0.199,P＜0.05),and was not correlated with serum TC and LDL-C(r=-0.122,-0.082,P＞0.05).Conclution VFM is increased and iNK T cell number is decreased in in adipose tissue of high-fat diet-fed mice.The number of iNK T cells is negatively correlated with VFM,BFC,serum HDL-C and TG.

Abstract:Objective To investigate the key targets and pathways of Juhongtai formula granules in improving acute lung injury(ALI).Methods Juhongtai formula granules and their drug-containing serum components were analyzed by UPLC-QTOF-MS/MS,and the active ingredients were screened based on the"Lipinski Rule of Five".The action targets of the above active ingredients were obtained through the TCMSP,Swiss Target Prediction and SuperPred databases,and the ALI-related targets were obtained from the GeneCards,OMIM,PharmGKB,CTD databases and GEO chip.The target protein-protein interaction network was constructed using the String database and Cytoscape to analyze the key targets.The DAVID database was used for GO functional annotation and KEGG pathway enrichment analysis,and the CB-dock2 platform was used for molecular docking verification.After one week of adaptive feeding,the experimental SD rats were randomly divided into the blank group,the model group,the positive control group,the negative control group and the Juhongtai formula granules group,with 6 rats in each group.The rat model of ALI was replicated by tracheal infusion of lipopolysaccharide.The pathological morphology of lung tissues in each group of rats was observed by HE staining,and the mRNA expressions of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1 β(IL-1β)and nitric oxide were detected by reverse transcription quantitative polymerase chain reaction(RT-qPCR).Results In total,44 components of the Juhongtai formula granules were identified,including 26 drug-containing serum components,and 21 key compounds were screened.A total of 22 intersection targets were obtained.The pathway enrichment results indicated the action of these targets on the advanced glycation end product receptor(AGE-RAGE)and TNF signaling pathways.The docking results showed that limonin,kaempferol and naringenin could be matched with prostaglandin peroxidase 2,nitric oxide synthase 2 and TNF.Animal experiments confirmed that the Juhongtai formula granules can alleviate inflammatory infiltration in lung tissue and reduce the expressions of TNF-α,IL6,IL-1β mRNA and nitric oxide.Conclusion Juhongtai formula granules can inhibit ALI-related inflammatory targets through the synergistic effects of multiple components such as limonin and kaempferol,regulate signaling pathways such as AGE-RAGE and TNF,reduce the production of inflammatory factors and nitric oxide,and improve ALI.

Abstract:Objective To investigate the impact of Epstein-Barr virus(EBV)infection on immune responses among systemic lupus erythematosus(SLE)patients.Methods AA total of 103 SLE patients(SLE group),50 rheumatoid arthritis(RA)patients(RA group),and 120 healthy physical examinees(control group)were enrolled from May 2022 to April 2024.Anti-EBV CA IgG,IgA,IgM,anti-EBV EA IgM,and anti-EBV NA IgG,IgA antibody levels were measured using direct chemiluminescent magnetic particle-based indirect immunoassay,and positive rates were compared among groups.SLE patients were further divided into reactivated infection and past infection groups based on anti-EBV antibody profiles.Non-specific and specific immune response markers were assessed.Peripheral blood T lymphocytes from two groups were isolated via fluorescence-activated cell sorting(FACS)for transcriptomic sequencing.Results The positive rate of anti-EBV CA IgA antibody in the SLE group[22.30％(23/103)]was higher than that in the RA group[6.00％(3/50)]and the control group[5.00％(6/120)],showing significant difference(P＜0.05).In SLE patients,anti-EBV EA IgM antibody was positively correlated with interferon-α(IFN-α)(r=0.1984,P＜0.05);anti-EBV CA IgM antibody was positively correlated with interleukin(IL)-2,IL-5,and IL-1β(r=0.1980,0.2553,0.1797,P＜0.05);anti-EBV CA IgG antibody was positively correlated with IL-5(r=0.2769,P＜0.05),IL-2(r=0.1820,P＜0.05),IL-8(r=0.1920,P＜0.05),and interferon-γ(IFN-γ)(r=0.1807,P＜0.05).Anti-EBV NA IgG antibody was positively correlated with IL-4(r=0.2015,P＜0.05),IL-8(r=0.2395,P＜0.05),and IL-17(r=0.1795,P＜0.05),and negatively correlated with IL-5(r=-0.2212,P＜0.05).Anti-EBV NA IgG antibody in SLE patients was positively correlated with IgG(r-0.2731,P＜0.05),and anti-EBV CA IgM antibody was positively correlated with IgM(r=0.2614,P＜0.05);anti-EBV EA IgM antibody was negatively correlated with white blood count(r=-0.2742,P＜0.05),neutrophil count(r=-0.2249,P＜0.05),lymphocyte count(r=-0.2723,P＜0.05),and monocyte count(r=-0.2275,P＜0.05),and anti-EBV NA IgA antibody was positively correlated with IgA results(r=0.3231,P＜0.05).The anti-EBV CA IgA antibody-positive SLE group showed elevated levels of the nonspecific immune response marker IgA and decreased levels of C3 and C4,as compared with the anti-EBV CA IgA antibody-negative SLE group,RA group,and control group(P＜0.05).Additionally,when compared with the RA group and control group,the anti-EBV CA IgA antibody-positive SLE group had reduced specific immune response parameters,including CD16+56+,CD 19,and the CD4/CD8 ratio(P＜0.05).Results from transcriptomic sequencing of peripheral blood T lymphocytes revealed upregulation of IL-2,IL-5,IL-8 and IFN-γ in reactivated infection group,as compared with the past infection group(P＜0.05).Results of the differential gene enrichment analysis between the two groups revealed that the differentially expressed genes were primarily associated with inflammatory signaling pathways,such as"inflammatory response activation,""inflammasome formation,"and"inflammatory cytokine release."Conclusion SLE patients demonstrate higher EBV activity.EBV infection may exacerbate SLE's inflammatory microenvironment by activating Th1/Th2 immune responses and promote humoral immune activation.

Abstract:Objective To investigate the correlation between urinary β2-microglobulin(β2-MG)and the clinicopathological characteristics of IgA nephropathy(IgAN).Methods A retrospective analysis was conducted on 77 patients diagnosed with IgAN by renal biopsy from October 2019 to October 2024.The research group was defined as those with a urinary β2-MG value higher than or equal to the median,and the control group was defined as those with a value lower than the median.The clinicopathological indicators of the two groups were compared.The Spearman method was used to analyze the correlation of the estimated glomerular filtration rate(eGFR)and pathological indicators of IgAN patients with urinary β2-MG.Multivariate logistic regression was used to analyze the clinicopathological factors related to urinary β2-MG.The receiver operating characteristic(ROC)curve and the area under the curve(AUC)were used to evaluate the ability of urinary β2-MG to predict related pathological damage.Result Compared with the control group,the research group had older age,elevated levels of serum creatinine and blood urea,increased 24-hour urine protein quantification,elevated levels of urine α 1-microglobulin and urinary β2-MG,elevated urine microalbumin,elevated levels of total cholesterol and D-dimer,and decreased levels of eGFR,serum albumin,hemoglobin,and complement C3(P＜0.05).Compared with the control group,the research group had more severe pathological damage in terms of renal tubular atrophy and renal interstitial fibrosis(T),as well as cellular and fibrocellular crescents(C)(P＜0.05).There was a negative correlation between the urinary β2-MG level and eGFR in patients with IgAN(P＜0.01),and a significant positive correlation with T and C(P＜0.05).After controlling for confounding factors,eGFR(OR=0.983),serum albumin(OR=0.889),triglycerides(OR=0.099),total cholesterol(OR=2.677),and pathological damage T(OR=11.358)and C(OR=12.018)were independent influencing factors associated with high urinary β2-MG levels(P＜0.05).The AUC of urinary β2-MG level for predicting the pathological indicator T in patients with IgAN was 0.784(95％CI:0.660,0.907),with the corresponding sensitivity of 0.717,and the specificity of 0.882.Conclusion The level of urinary β2-MG is closely related to multiple clinicopathological characteristics of patients with IgAN.It can be used as a potential biomarker for evaluating renal structural damage in patients,especially tubulointerstitial and renal vascular lesions,and has a certain predictive ability for tubulointerstitial injury.

Abstract:Objective To investigate the effect of vitamin D deficiency on the balance of peripheral blood T helper cell 17(Th 17)and regulatory T cell(Treg)in patients with psoriasis and its relationship with the severity of the disease.Methods A total of 169 patients with psoriasis were selected from June 2020 to June 2023.According to the results of vitamin D measurement,the patients were divided into normal vitamin D group(n=61),vitamin D insufficiency group(n=57),and vitamin D deficiency group(n=51).The percentages of Th17 and Treg cells in peripheral blood were compared among the three groups,and the correlation between vitamin D and Th17/Treg balance was analyzed.COX regression was used to analyze the relationship between vitamin D level and the severity of the disease,followed by a trend test.The restricted cubic spline method was used to explore the dose-response relationship between vitamin D,Th17/Treg and disease severity.The interaction between vitamin D and Th17/Treg on disease severity was analyzed.Results The level of Treg in the serum of patients with psoriasis decreased with the increase of vitamin D level,while the level of Th17 increased with the increase of vitamin D level(P＜0.01).Vitamin D deficiency could lead to the imbalance of Th17/Treg.With the decrease of vitamin D level,the Treg level decreased,while the level of Th17 increased.COX regression analysis showed that after adjusting for confounding factors,vitamin D levels were closely related to disease severity(P＜0.001).Restricted cubic spline model analysis showed that there was a significant nonlinear dose-response relationship of vitamin D and Th17/Treg levels with the severity of disease.With the decrease of vitamin D level,the decrease of Treg level and the increase of Th 17 level,the severity of the disease showed a corresponding increasing trend.Patients with severe psoriasis had the highest proportion given vitamin D deficiency and Th17/Treg imbalance(P＜0.05).At different vitamin D levels,the higher the Th17/Treg ratio,the more significant the increase in the risk of psoriasis,showing a significant interaction[OR(95％CI):3.54(2.45,3.83)](P＜0.05).Conclusion Vitamin D deficiency can lead to the imbalance of Th17/Treg in peripheral blood of patients with psoriasis,thereby aggravating the severity of the disease.

Abstract:Objective To investigate the therapeutic effect of Formononetin on mice with Mycoplasma pneumoniae pneumonia(MPP)by regulating the mitogen-activated protein kinase(MAPK)/nuclear factor κB(NF-κB)signaling pathway.Methods Six-week-old SPF-grade male BALB/c mice were selected.The MPP mouse model was established as the model group by instillating Mycoplasma pneumoniae bacterial solution through the nose.On the second day after successful modeling,mice were intraperitoneally injected with 15,30,and 60 mg/kg of Formononetin and 60 mg/kg of Formononetin+20 mg/kg of Anisomycin respectively as the low-dose,medium-dose,and high-dose Formononetin groups and the high-dose Formononetin+Anisomycin group,with 12 mice in each group.Another 12 mice were intraperitoneally injected with the same amount of 0.9％sodium chloride injection as the control group.The cough frequency of mice in each group was detected through the cough induction test.The partial pressure of carbon dioxide(PCO2)and partial pressure of oxygen(PO2)in each group of mice were detected by a blood gas analyzer,and the oxygenation index(OI)was calculated.The levels of inflammatory factors in each group were detected by ELISA,and the apoptosis of lung tissue cells in each group of mice was detected by TUNEL.HE staining was performed to observe the pathological changes of lung tissues in each group.The expression of MAPK/NF-κB pathway-related proteins in the lung tissues of mice in each group was detected by Western blot method.Results The lung tissue morphology of the mice in the control group was normal.The alveolar ducts and alveolar structures of mice in the model group were damaged,the alveolar septa thickened,and there was a large amount of inflammatory cell infiltration.Compared with the model group,the lung tissue morphology was improved in the low-dose,medium-dose and high-dose Formononetin groups.The lung tissue injury in the high-dose Formononetin+Anisamycin group was more severe compared with the high-dose Formononetin group.Compared with the control group,the cough latency period in the model group was shortened,and PO2,OI,and interleukin-10(IL-10)were decreased,while the frequency of coughing,PCO2,interleukin-18(IL-18),tumor necrosis factor-α(TNF-α),apoptosis rate,and the ratios of p-P38 MAPK/P38 MAPK,p-NF-κB p65/NF-κB p65,p-extracellular signal-regulated kinase 1/2(ERK1/2)/ERK1/2,and p-C-jun N-terminal kinase(p-JNK)/JNK increased(P＜0.05).Compared with the model group,the low-,medium-and high-dose Formononetin groups had improved lung tissue morphology,prolonged cough latency,increased PO2,OI and IL-10,and reduced cough frequency.The PCO2,IL-18,TNF-α,apoptosis rate,and the ratios of p-p38 MAPK/P38 MAPK,p-NF-κB p65/NF-κB p65,ERK1/2/ERK1/2,and p-JNK/JNK decreased(P＜0.05).In the low-,medium-,and high-dose Formononetin groups,with the increase of Formononetin dose,the cough latency gradually prolonged,the cough frequency gradually decreased,and the PO2,oxygenation index,and IL-10 gradually increased.The PCO2,IL-18,TNF-α,apoptosis rate and the ratios of p-p38 MAPK/P38 MAPK,p-NF-κB p65/NF-κB p65,p-ERK1/2/ERK1/2,and p-JNK/JNK gradually decreased(P＜0.05).Compared with the high-dose Formononetin group,the high-dose Formononetin+Anisamycin group had shorter cough latency,lower PO2,OI and IL-10.The frequency of coughing,PCO2,IL-18,TNF-α,apoptosis rate,and the ratios of p-p38 MAPK/P38 MAPK,p-NF-κB p65/NF-κB p65,p-ERK1/2/ERK1/2,and p-JNK/JNK increased(P＜0.05).Conclusion Formononetin may improve lung injury in MPP mice by inhibiting the MAPK/NF-κB signaling pathway.

Abstract:Breast cancer is the leading cause of cancer-related mortality among women worldwide and has drawn extensive research attention.Owing to its molecular heterogeneity,drug resistance,and low therapeutic response,single-modality treatments often fail to achieve satisfactory efficacy or broad applicability.Combination therapy,designed based on the pathophysiological characteristics,related signaling pathways,and biomarkers of breast cancer,has emerged as a promising approach for improving therapeutic outcomes.With the advancement of research on combination strategies,the understanding of their molecular mechanisms—particularly key signaling pathways and biomarkers—has become increasingly important.However,comprehensive reviews addressing these molecular mechanisms and synergistic strategies remain scarce.This article summarizes recent advances in combination therapy for breast cancer,providing a comprehensive review of recent combination therapies for breast cancer and their underlying molecular mechanisms,and focusing on key signaling pathways involved in combination therapy and synergistic strategies,thereby providing theoretical insights and reference for researchers,graduate students,and clinicians engaged in the development of novel combination therapeutic strategies for breast cancer and related malignancies.

Abstract:Objective To investigate the impact of neoadjuvant chemotherapy combined with transurethral resection of bladder tumor(TURBT)on the phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt)signaling pathway in patients with bladder cancer.Methods A total of 118 patients with bladder cancer who were admitted from February 2022 to February 2025 were selected as the research subjects.According to different treatment methods,they were divided into the control group(n=76)for TURBT treatment and the observation group(n=42)for neoadjuvant chemotherapy combined with TURBT treatment.The perioperative indicators,clinical efficacy and adverse reactions of the two groups were compared,and the mRNA expression levels of tumor markers,inflammatory factors and PI3K/Akt signaling pathway indicators before and after treatment were also compared,to analyze the correlation between inflammatory factors and indicators of the PI3K/Akt signaling pathway.Results Both total effective rate and disease control rate of the observation group were higher than those of the control group,and the incidence of adverse reactions was lower than that of the control group(P<0.05).The duration of operation,length of hospital stay and duration of gross hematuria in the observation group were all lower than those in the control group(P<0.05).After treatment in the observation group,carcinoembryonic antigen,carbohydrate antigen 125,carbohydrate antigen 19-9,cytokeratin protein 19 fragment antigen 21-1,interleukin-6(IL-6),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),C reactive protein(CRP),prostaglandin E2(PGE2),and the expression levels of B-cell lymphoma-2(Bcl-2),PI3K and Akt mRNA were all lower than those in the control group,while the expression levels of Caspase-3 and Bcl-2 associated X protein(Bax)mRNA were significantly higher than those in the control group(P<0.05).The levels of IL-6,IL-10,TNF-α,CRP and PGE2 in patients with bladder cancer after treatment had stable positive correlations with the expression levels of Bcl-2,PI3K and Akt mRNA,respectively(P<0.05),and a stable negative correlation with the expression levels of Caspase-3 and Bax mRNA(P<0.05).Conclusion Neoadjuvant chemotherapy can attenuate the proliferation of bladder cancer cells by inhibiting the abnormal activation of the PI3K/Akt signaling pathway,while TURBT can eliminate residual tumor tissue and reduce the risk of recurrence.The combined application of the two can achieve effective control at the molecular level,improving the survival rate and quality of life of patients.

Abstract:Objective To explore the clinical effect of stereotactic body radiotherapy(SBRT)combined with dendritic cell-cytokine-induced killer cell(DC-CIK)immunotherapy in the treatment of centrally-located locally advanced non-small cell lung cancer(LA-NSCLC).Methods A total of 113 patients with centrally-located LA-NSCLC admitted from January 2022 to March 2024 were divided into the observation group(n=56)and the control group(n=57)by using the random number table method.The observation group was treated with SBRT combined with DC-CIK immunotherapy,while the control group was treated with SBRT.The clinical efficacy of the two groups after treatment,as well as the levels of tumor markers,immune function indicators and inflammatory indicators before and after treatment,was compared.The adverse reactions during the treatment period and the short-term survival during the 12-month follow-up of the two groups were recorded.Results Compared with the control group,both objective response rate and disease control rate of the observation group were higher(P<0.05).After treatment,the levels of serum carcinoembryonic antigen,cytokeratin 19 fragment and cancer antigen 125 in both groups were lower than those before treatment,and lower in the observation group than in the control group(P<0.05,P<0.01).After treatment,the levels of CD3+,CD4+,and CD4+/CD8+in the control group were lower than those before treatment,while the level of CD8+was higher than that before treatment(P<0.05).After treatment,the levels of CD3+,CD4+and CD4+/CD8+in the observation group were higher than those before treatment and also higher than those in the control group,while the level of CD8+was lower than that before treatment and also lower than that in the control group(P<0.05,P<0.01).After treatment,the interferon-γ level in the observation group was higher than that before treatment and also higher than that in the control group,while the levels of tumor necrosis factor-α and interleukin-6 were lower than those before treatment and also lower than those in the control group(P<0.05,P<0.01).There was no significant difference in the incidence of adverse reactions between the two groups during the treatment period(P>0.05).The overall survival rate of the observation group at 12-month follow-up was higher,and the median survival time was longer,as compared with the control group(P<0.05).Conclusion SBRT combined with DC-CIK immunotherapy has a definite effect in the treatment of patients with centrally-located LA-NSCLC.It can significantly improve the immune function of patients,reduce the levels of tumor markers,alleviate inflammatory responses,increase short-term survival rates,prolong survival time,and does not significantly increase adverse reactions,offering good safety and reliability.

Abstract:Objective To investigate the protective effect of valeric acid on Doxorubicin-induced myocardial injury in mice.Methods C57BL/6J mice and AC16 cells were randomly divided into control group,injury group,and low-,medium-,and high-dose valeric acid groups.Doxorubicin was used to treat mice and myocardial cells to establish myocardial injury models.Hematoxylin-eosin(HE)staining was used to analyze the pathological changes of myocardial tissue in mice,and enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of myocardial injury markers and inflammatory factors in mice from each group.Cell counting kit was used to detect the viability of myocardial cells in each group,and spectrophotometry was used to detect the levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in the serum and myocardial cells in mice from each group.Reactive oxygen species(ROS)fluorescence probe was used to detect the levels of reactive oxygen species in each group,and flow cytometry was used to detect the apoptosis rate of each group.Western blot was used to detect the expression levels of nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase 1(HO-1)proteins in myocardial tissue and myocardial cells of mice from each group.Results Compared with the injury group,the myocardial injury in the low-,medium-,and high-dose valeric acid groups was ameliorated,the levels of myocardial injury markers gradually decreased,the levels of SOD in the body,and the expression levels of Nrf2 and HO-1 proteins gradually increased,and the levels of inflammatory factors,MDA,and apoptosis rate gradually decreased(P<0.05).Compared with myocardial cells in the injury group,the viability of myocardial cells,the levels of SOD,and the expression levels of Nrf2 and HO-1 proteins in the low-,medium-,and high-dose valeric acid groups gradually increased,while the levels of inflammatory factors,MDA,and apoptosis rate gradually decreased(P<0.05).Conclusion Valeric acid can inhibit inflammation and oxidative stress to improve Doxorubicin-induced myocardial injury,which may be related to the activation of Nrf2/HO-1 signaling pathway by valeric acid.

Abstract:Chronic pain impairs functions across multiple dimensions,including physical,psychological,and social well-being.The neuro-immune mechanisms involve complex interactions between the immune system and nerve fibers at one or multiple levels of the peripheral or central nervous system.In recent years,research on the neuro-immune circuit mechanisms underlying the generation,transmission,modulation,and integration of chronic pain has advanced rapidly.However,the overall interactive dynamics of neuro-immune mechanisms remain unclear.This article systematically reviews recent advances in the neuro-immune mechanisms of chronic pain,elucidating the cascading transformation from peripheral to central sensitization.These findings provide a theoretical basis for a deeper understanding of the overall pathophysiological basis of pain chronicity and the exploration of effective clinical strategies.

Abstract:Objective To explore the correlations between interleukin-6(IL-6),interleukin-8(IL-8),antimicrobial peptide LL-37 and the microbial abundance and the degree of mucosal injury in patients with trichomonal vaginitis(TV).Methods A total of 120 TV patients admitted from January 2022 to December 2024 were selected and divided into the occurrence group(n=63)and the non-occurrence group(n=57)according to presence and absence of mucosal injury.The general data,IL-6,IL-8,LL-37 and microbial abundance were compared between the two groups.According to the degree of mucosal injury,the occurrence group was divided into the mild subgroup(n=34),the moderate subgroup(n=20),and the severe subgroup(n=9).Pearson correlation analysis was applied to investigate the correlations between IL-6,IL-10,antimicrobial peptide LL-37,microbial abundance and the degree of mucosal injury.Multivariate logistic regression was used to analyze the influencing factors of mucosal injury in TV patients.The receiver operating characteristic curve(ROC)was used to evaluate the predictive value of relevant indicators for mucosal injury in TV patients.Result The levels of IL-6,IL-8,antimicrobial peptide LL-37,and the abundance of Actinobacteria and Gardnerella in the occurrence group were higher than those in the non-occurrence group,while the abundance of Firmicutes and Lactobacillus was lower than that in the non-occurrence group(P<0.05,P<0.01).Pearson correlation analysis showed that IL-6,IL-8,antimicrobial peptide LL-37,and abundance of Actinobacteria and Gardnerella were positively correlated with the degree of mucosal injury(r=0.543,0.713,0.352,0.409,0.659,P<0.01),while the abundance of Firmicutes and Lactobacillus was negatively correlated with the degree of mucosal injury(r=-0.540,-0.504,P<0.01).Multivariate logistic regression analysis showed that IL-6,IL-8,antimicrobial peptide LL-37,and abundance of Actinobacteria were risk factors for mucosal injury in TV patients,while the abundance of Firmicutes and the abundance of Lactobacillus were protective factors(P<0.05).The ROC curve showed that the area under the ROC curve(AUC)of the combined detection of IL-6,IL-8,antibacterial peptide 37,and the abundance of Actinobacteria,Firmicutes,and Lactobacillus in evaluating mucosal injury in TV patients was higher than that of the single detection(P<0.01).Conclusion IL-6,IL-8,antimicrobial peptide LL-37 and microbial abundance are closely related to the degree of mucosal injury in TV patients.The combined detection has a high value in evaluating the occurrence of mucosal injury in TV patients.

Abstract:Objective To investigate the efficacy of Ustekinumab combined with fecal microbiota transplantation in the treatment of inflammatory bowel disease(IBD)and its impact on intestinal mucosal barrier and immune function.Methods A total of 106 IBD patients treated from January 2023 to March 2024 were selected and divided into the Usteinumab group and the Usteinumab combined with fecal microbiota transplantation group(combination group)by random number table method,with 53 patients in each group.The Ustekinumab group was treated with Ustekinumab,while the combination group was treated with Ustekinumab combined with fecal microbiota transplantation.The clinical efficacy,intestinal mucosal barrier indicators,immune function indicators,scores of the Inflammatory Bowel Disease Quality of Life Questionnaire(IBDQ)and the total incidence of adverse reactions were compared between the two groups.Results The total effective rate of the combination group was 94.34%(50/53),which was higher than that of the Ustekinumab group[73.58%(39/53)](P<0.05).After treatment,the levels of endotoxin,D-lactic acid and diamine oxidase in the combination group were all lower than those in the Ustekinumab group,while the levels of CD3+,CD4+,CD4+/CD8+and the scores of IBDQ were all higher than those in the Ustekinumab group(P<0.05).There was no significant difference in the total incidence of adverse reactions between the two groups(P>0.05).Conclusion The clinical efficacy of Ustekinumab combined with fecal microbiota transplantation is remarkable in the treatment of IBD.It can effectively alleviate intestinal mucosal injury,promote the recovery of intestinal mucosal barrier function,improve the intestinal microbial environment,enhance the immune function of patients,effectively control the progression of the disease,and has high safety.

Abstract:Objective To investigate the relationship of C-reactive protein-albumin-lymphocyte(CALLY)index,serum autotaxin and serine protease 2(PRSS2)with postoperative recurrence and metastasis in gastric cancer patients.Methods A total of 188 patients who underwent radical gastrectomy for gastric cancer from December 2019 to December 2021 were selected as the research subjects.According to the postoperative situation,they were divided into the recurrence and metastasis group(n=72)and the non-recurrence and metastasis group(n=116).C reactive protein(CRP)was detected by immunoturbidimetry,albumin was detected by bromocresol green method,lymphocyte count was detected by blood routine analyzer,and CALLY index was calculated.Enzyme-linked immunosorbent assay was used to detect serum levels of autotaxin and PRSS2,and Spearman/Pearson correlation analysis was used to analyze the correlation between CALLY index,serum autotaxin,PRSS2 levels and clinical data.Multivariate logistic regression was used to analyze the influencing factors of recurrence and metastasis in patients with gastric cancer after radical resection,and receiver operating characteristic(ROC)curve was used to analyze the predictive value of CALLY index,serum autotaxin and PRSS2 levels for recurrence and metastasis in patients with gastric cancer after radical resection.Kaplan-Meier method was used to analyze the relationship between CALLY index,serum autotaxin,PRSS2 levels and postoperative recurrence and metastasis.Results Compared with the non-recurrence and metastasis group,the recurrence and metastasis group had a higher proportion of patients with TNM stage Ⅲ,Borrmann type Ⅲ-Ⅳ,lymphovascular invasion,neoadjuvant therapy and postoperative chemotherapy,higher serum levels of autotaxin and PRSS2,and lower CALLY index(P<0.01).TNM stage,Borrmann type,lymphovascular invasion,neoadjuvant therapy and postoperative chemotherapy were negatively correlated with CALLY index,and positively correlated with serum autotaxin and PRSS2 levels,while serum autotaxin and PRSS2 levels were negatively correlated with CALLY index(P<0.01).TNM stage,Borrmann type,lymphovascular invasion,CALLY index,autotaxin and PRSS2 were the influencing factors of recurrence and metastasis in patients with gastric cancer after radical resection(P<0.05,P<0.01).The area under the curve(AUC)of CALLY index,autotaxin and PRSS2 for predicting recurrence and metastasis was 0.962,which was significantly larger than that of CALLY index,autotaxin and PRSS alone(P<0.01).The 3-year recurrence and metastasis rate of patients with low CALLY index expression[56.52%(52/92)]was higher than that of patients with high CALLY index expression[20.83%(20/96)](P<0.01).The 3-year recurrence and metastasis rate of patients with high expression of autotaxin[49.47%(47/95)]was higher than that of patients with low expression[26.88%(25/93)](P<0.01).The 3-year recurrence and metastasis rate of patients with high PRSS2 expression[47.87%(45/94)]was higher than that of patients with low PRSS2 expression[28.72%(27/94)](P<0.01).Conclusion The CALLY index decreases,and serum autotaxin and PRSS2 increase in patients with postoperative recurrence and metastasis of gastric cancer.The combined prediction of the three factors demonstrates higher predictive performance for postoperative recurrence and metastasis.

Abstract:Objective To explore the molecular mechanism by which curcumin enhances the anti-tuberculosis function of macrophages through immune metabolic regulation mediated by liver X receptor α(LXRα)/ABCA1.Methods A model was established by infecting THP-1-derived macrophages with attenuated strain of Mycobacterium bovis(M.bovis).The control group,curcumin group,M.pavis group,M.pavis+LXRα agonist(T0901317)group,M.pavis+LXRα inhibitor(GSK2033)group,M.pavis+curcumin group,M.pavis+curcumin+GSK2033 group and M.pavis+curcumin+T0901317 group were set up.The protein and gene expressions of LXRα/ABCA1 were detected by Western blotting and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).The accumulation of lipid droplets was analyzed by Oil Red O staining and micro-assay.The lipid content of the supernatant was determined by a biochemical analyzer,and cell proliferation was assessed by the MTT method.Bacterial clearance capacity was evaluated by measuring intracellular bacterial load.Results Curcumin significantly upregulated the protein and gene expression of LXRα/ABCA1 in M.Bovis-infected macrophages,reduced intracellular lipid accumulation and promoted lipid efflux,while enhancing cell proliferation and reducing intracellular bacterial load(P<0.05,P<0.01).LXRα inhibitors could reverse the effect of curcumin,while agonists synergistically enhanced its effect.Correlation analysis showed that the expression of LXRα/ABCA1 in cells was negatively correlated with the intracellular bacterial load,while the lipid level was positively correlated with the intracellular bacterial load(P<0.01).Conclusion Curcumin activates the LXRα/ABCA1 pathway,coordinates the metabolic remodeling of macrophages and the enhancement of immune function,and forms a synergistic effect against tuberculosis,providing an experimental basis for the development of a novel host-directed treatment strategy for tuberculosis based on immune-metabolic regulation.

Abstract:Objective To investigate the predictive performance of serum soluble CD40 ligand(sCD40L),interleukin-17(IL-17)and neutrophil extracellular traps(NETs)in combination for intracranial aneurysm rupture.Methods A total of 120 patients with intracranial aneurysms admitted from January 2022 to January 2025 were selected and divided into low-risk group and medium-high-risk group according to different rupture risks.After propensity score matching with 1:1 ratio and caliper matching(caliper=0.02),there were 60 patients in each group.The general data and serum levels of sCD40 L,IL-17 and NETs were compared between the two groups.Multivariate logistic regression analysis was used to analyze the risk factors of intracranial aneurysm rupture.The linear relationship between serum sCD40 L,IL-17,NETs and the risk of intracranial aneurysm rupture was analyzed by smoothing curve fitting.The receiver operating characteristic(ROC)curve,clinical impact curve(CIC)and clinical decision curve analysis(DCA)were used to evaluate the predictive performance of serum sCD40L,IL-17 and NETs for intracranial aneurysm rupture.Results The irregular shape of aneurysms,the proportion of the longest diameter of aneurysms≥7 mm and the levels of serum sCD40L,IL-17 and NETs in the low-risk group were lower than those in the medium-high-risk group(P<0.01).The shape of aneurysm,the longest diameter of aneurysm,sCD40L,IL-17 and NETs were the independent risk factors for intracranial aneurysm rupture(P<0.05).After adjusting for other covariates,serum sCD40L,IL-17 and NETs were positively correlated with the risk of intracranial aneurysm rupture(P<0.05).The ROC curve showed that the area under the curve of serum sCD40L,IL-17 and NETs in combination for predicting the risk of intracranial aneurysm rupture was the largest.The DCA and CIC showed that both the net benefit of the combined prediction model of serum sCD40L,IL-17 and NETs in predicting the risk of intracranial aneurysm rupture and the coincidence rate with the actual situation were relatively high.Conclusion The elevated levels of serum sCD40L,IL-17,and NETs in patients with intracranial aneurysms are associated with their rupture risk.The combination of the three indicators has a high predictive performance for the risk of intracranial aneurysm rupture.

Abstract:Objective To evaluate the predictive value of baseline peripheral blood immune cell profiles for immune-related adverse events(irAEs)following immune checkpoint inhibitors(ICIs)therapy for non-small cell lung cancer(NSCLC)patients.Methods In total,45 NSCLC patients who received ICIs treatment between December 2023 and December 2024 were enrolled.Patients were stratified into irAE(n=19)and non-irAE(n=26)groups based on irAE occurrence during treatment.Pre-treatment peripheral blood samples were collected,and flow cytometry was used to quantify immune subset proportions and activation marker expression.Receiver operating characteristic(ROC)curves were used to evaluate the predictive value of individual cellular markers for irAE development in NSCLC patients after immunotherapy.Results The irAE group exhibited significantly reduced proportion of intermediate monocytes(CD14+CD16+),as compared with the non-irAE group(P<0.05).The expression of CD69,an early activation marker,in CD3+T lymphocytes and the expression of CD154,an early activation marker,in CD8+T lymphocytes were both significantly lower in the irAE group than in the non-irAE group(P<0.05).ROC analysis demonstrated better predictive performance of the three-marker panel(intermediate monocytes/CD69+CD3+T cells and CD154+CD8+T cells;AUC=0.778)compared with individual biomarkers.Conclusion Lower levels of intermediate monocytes,CD69+CD3+T cells,and CD154+CD8+T cells in baseline peripheral blood may serve as potential biomarkers for predicting irAE development in NSCLC patients after immunotherapy.

Abstract:Objective To investigate the effect of Pirfenidone(PFN)on respiratory syncytial virus(RSV)infection-induced damage to bronchial epithelial cells by regulating the high mobility group protein B1(HMGB1)/receptor for advanced glycation end products(RAGE)signaling pathway.Methods Human bronchial epithelial cells(HBE)were divided into Control group(cultured for 24 h under normal conditions),RSV group(inoculated with 4.65×106/mL RSV at 33℃for 2 h);low PFN(L-PFN)group(treated with 0.05 mg/mL PFN for 24 h),moderate PFN(M-PFN)group(treated with 0.10 mg/mL PFN for 24 h),high PFN(H-PFN)group(treated with 0.20 mg/mL PFN for 24 h)and recombinant HMGB1(rHMGB1)group(treated with 1 μg/mL rHMGB1+0.20 mg/mL PFN for 24 h).EdU method was applied to detect the proliferation rate of cells in each group,Hochest33258 staining method was applied to detect apoptosis status of cells in each group,and the migration of cells in each group was evaluated by the scratch experiment.Enzyme linked immunosorbent assay(ELISA)was applied to measure the levels of interferon(IFN)-α,IFN-γ,tumor necrosis factor-α(TNF-α),interleukin(IL)-1α,IL-6 and IL-4 in each group of cells,and Western blot was applied to detect the protein expression of HMGB1,RAGE,B lymphoblastoma-2-associated X protein(Bax),cysteine aspartic protease-3(Caspase-3),and B lymphoblastoma-2(Bcl-2).Results Compared with the RSV group,the cell proliferation rate,scratch closure rate,IL-4 levels,and expression of Bcl-2 in L-,M-,and H-PFN groups increased,while the apoptosis rate,the levels of IFN-α,IFN-γ,TNF-α,IL-1α,IL-6,and the expression of HMGB1,RAGE,Bax,and Caspase-3 reduced(P<0.05);rHMGB1 weakened the effect of H-PFN on the above-mentioned indicators(P<0.05).Conclusion PFN may suppress the apoptosis and inflammatory damage of RSV-infected bronchial epithelial cells by inhibiting the HMGB1/RAGE pathway.Conclusion PFN may suppress the apoptosis and inflammatory damage of RSV-infected bronchial epithelial cells by inhibiting the HMGB1/RAGE pathway.

Abstract:Objective To explore the circRNA expression level in the urine of patients with lupus nephritis(LN),to construct a diagnostic model for LN,and to evaluate and verify its diagnostic efficacy.Methods A total of 120 patients with LN admitted from January 2022 to January 2024 were selected as the observation group and divided into the training group(n=84)and the validation group(n=36)at a ratio of 7:3.Another 100 healthy individuals who underwent physical examination were selected as the control group.The expression levels of circRNA-0001445,circRNA-002059,hsa-circ-0007385 and hsa-circ-0000006 in urine specimens of patients in the training group and the control group were detected by gene sequencing.Multivariate Logistic regression analysis was used to construct a diagnostic model for LN based on the urinary circRNA expression level.The diagnostic value of individual indicators and combined predictive models were evaluated by receiver operating characteristic(ROC)curve analysis,and then the predictive efficacy of the model was verified by the validation group.Results The expression levels of circRNA-0001445,circRNA-002059,hsa-circ-0007385 and hsa-circ-0000006 in the urine of patients in the training group were all higher than those of the control group(P<0.01).Multivariate Logistic regression analysis found that The expression levels of circRNA-0001445(OR=1.094),circRNA-002059(OR=1.040),hsa-circ-0007385(OR=1.033),and hsa-circ-0000006(OR=1.130)in theurine were independent factor for the diagnosis of LN(P<0.01).ROC curve analysis showed that the areas under the curve of combined model constructed based on the circRNA expression level in the urine were 0.985 and 0.911 in the training group and the validation group,respectively,with sensitivities of 95.00%and 91.67%,and specificities of 99.00%and 89.00%,all of which were higher than the diagnostic efficacy of a single urinary circRNA expression level.Conclusion The diagnostic model of LN constructed based on the expression level of circRNA in the urine has a high diagnostic efficacy,which has been confirmed by internal tests and may provide strong support for clinicians in the early diagnosis of LN.

Abstract:Objective To explore the effect of lipoprotein a[Lp(a)]on the inflammatory state and cardiovascular system of patients with essential hypertension.Methods The clinical data of 234 patients with essential hypertension admitted from January 2022 to November 2023 were retrospectively collected.According to the serum Lp(a)concentration,they were divided into the normal Lp(a)group(n=185)and the high Lp(a)group(n=49).The differences in serum inflammatory markers C reactive protein(CRP),neutrophil-to-lymphocyte ratio(NLR),and fibrinogen between the two groups were compared.Pearson correlation analysis was used to analyze the correlation between Lp(a)and inflammatory markers,and the incidence of cardiac target organ damage between the two groups of patients was compared.Results Compared with the normal Lp(a)group,the CRP concentration,NLR and fibrinogen concentration of hypertensive patients in the high Lp(a)group increased(P<0.05,P<0.01),and the concentration of Lp(a)was positively correlated with CRP,NLR and fibrinogen(r=0.168,0.165,0.321,P<0.05).The incidence of myocardial infarction was higher in the high Lp(a)group[22.45%(11/49)]than in the normal Lp(a)group[7.57%(14/185)](P<0.01).Conclusion In patients with essential hypertension,high Lp(a)concentration is associated with a stronger inflammatory response,and elevated Lp(a)concentration is related to an increased incidence of myocardial infarction,suggesting that Lp(a)may affect target organ damage in patients with essential hypertension by promoting inflammation.