Objective To investigate the effect of lysosome biogenesis complex 1 subunit 1(GCN5L1)on myocardial inflammatory injury and regulatory T cell(Treg)infiltration in mice with myocardial infarction(MI).Methods The differentially expressed gene GCN5L1 was screened from the GSE66360 dataset using whole blood samples from healthy controls and MI patients.The correlation between GCN5L1 and IL-10,IL-35 or Treg marker FOXP3 was analyzed.A model of MI was established in C57BL/6 mice by left anterior descending coronary artery ligation.Thirty mice were divided into five groups:sham,MI model,AAV9-NC(control adeno-associated virus),AAV9-GCN5L1 and AAV9-GCN5L1+FOXP3 inhibitor(epirubicin)groups,with 6 mice in each group.MI area was assessed by TTC staining,and histopathological changes were evaluated via HE staining.Immunofluorescence was used to quantify GCN5L1 and FOXP3 positivity;ELISA was used to measure IL-10 and IL-35 levels;Western blot was used to analyze the levels of GCN5L1,Bax,Bcl-2 and PI3K/AKT pathway proteins;transcriptomic sequencing was performed on AAV9-NC and AAV9-GCN5L1 groups,while differential genes subjected to GO and KEGG enrichment analysis.Results GSE66360 dataset showed that GCN5L1 levels were significantly lower in MI patients than in healthy controls,and positively correlated with IL-10,IL-35 and FOXP3 levels(P＜0.01).Compared to the sham group,the MI group exhibited increased infarct area,severe histopathology,reduced GCN5L1/IL-10/IL-35 levels,elevated Bax,and decreased Bcl-2,which could be reversed by AAV9-GCN5L1(P＜0.01).Transcriptomics revealed the enrichment of differential genes in Treg infiltration and PI3K/AKT pathways.The AAV9-GCN5L1 group showed higher FOXP3 positivity and PI3K/AKT expression than the MI group.Co-treatment with AAV9-GCN5L1+epirubicin reduced FOXP3 positivity compared to AAV9-GCN5L1 alone(P＜0.01).Conclusion GCN5L1 overexpression alleviates myocardial inflammatory injury post-MI,activates the PI3K/AKT pathway,and enhances Treg infiltration.

Objective To investigate the expressions of IL-18,IL-18 binding protein isoform a(IL-18BPa)and IL-18Rα in blood CD4+Th1 cells of patients with allergic asthma(AA),and with allergic rhinitis and asthma syndrome(ARA),and the effects of allergens on their expressions.Methods Blood samples were collected from patients with AA,ARA and healthy control(HC)subjects.Flow cytometry was used to evaluate the actions of allergens on the expressions of IL-18,IL-18BPa and IL-18Rα in CD4+Th1 cells.Ovalbumin(OVA)-induced AA mouse models were established,and the expression level of IL-18Rα along with the influence of IL-18 on its expression in blood and lung Th1 cells was also explored by flow cytometry.Results Compared with HC,we observed increased IL-18 while decreased IL-18BPa expression in blood Th1 cells of AA and ARA patients.Moreover,allergens upregulated the expression levels of IL-18,IL-18BPa and IL-18Rα in Th1 cells of patients with AA and ARA.Additionally,intratracheal challenge with OVA plus IL-18 increased the proportions of blood and lung Th1 cells of HC and AA mice,and upregulated IL-18Rα expression on blood and lung Th1 cells of HC mice.Conclusion Allergens may be involved in the pathogenesis of airway allergic diseases by inducing the expressions of IL-18 and IL-18Rα in CD4+Th1 cells.

Objective To investigate the expression of serum homeobox C6(HOXC6),C-C motif chemokine ligand 18(CCL18),and soluble CD86(sCD86)in multiple myeloma patients and their correlation with prognosis.Methods Totally 110 patients with multiple myeloma accepted by our hospital from January 2020 to September 2023 were regarded as research group and followed up for one year.They were assigned into a good prognosis group(n=69)and a poor prognosis group(n=41),according to prognositic outcome.Additionally,135 healthy people who underwent physical checkups were recruited as control group.qRT-PCR was used to determine the relative expression of serum HOXC6 mRNA;ELISA was used to measure serum CCL18 and sCD86;K-M method was used to analyze the relationship of serum HOXC6,CCL18,sCD86 with overall survival(OS)and progression-free survival(PFS)in multiple myeloma patients.Receiver operating characteristic(ROC)curves were plotted to analyze the prognostic value of serum HOXC6,CCL18 and sCD86 in patients with multiple myeloma;COX regression model was used to analyze the factors influencing the prognosis of patients with multiple myeloma.Results Compared with the control group,the research group had higher levels of serum HOXC6,CCL18 and sCD86(P＜0.05).The median OS time and median PFS of patients with high and low expression of HOXC6,CCL18 and sCD86 were significantly different(P＜0.05).The poor prognosis group had a higher proportion of ISS stage Ⅲ,and higher levels of serum HOXC6,CCL18,sCD86 than those of good prognosis group(P＜0.05).The AUC value of combination of serum HOXC6,CCL18 and sCD86 for predicting poor prognosis in multiple myeloma patients was greater than that of serum HOXC6,CCL18 or sCD86 alone(P＜0.05).Conclusion The combination of serum HOXC6,CCL18 and sCD86 has a high predictive power for poor prognosis in patients with multiple myeloma,and all three are risk factors for poor prognosis.

Objective To investigate the relationship between the expression levels of mir-155/Th17 and mir-23b-3p/Tfh in peripheral blood and the prognostic value of secondary atopic dermatitis(AD)in AIDS patients.Methods In this study,80 AIDS patients were treated in our hospital from February 2021 to December 2023.The subjects were divided into AD group and non-AD group according to whether they had secondary AD.The levels of mir-155,Th17,mir-23b-3p and Tfh in peripheral blood of AD group and non-AD group were compared.Spearman method was used to analyze the relationship between levels of mir-155,Th17,mir-23b-3p and Tfh in peripheral blood and secondary AD in AIDS patients.At the same time,according to the prognosis of patients,the subjects were divided into good prognosis group and poor prognosis group,and the risk factors affecting the prognosis of patients were screened by Logistic regression analysis.Receiver operating characteristic(ROC)curve was drawn to analyze the predictive value of peripheral blood mir-155,Th17,mir-23b-3p and Tfh levels on the prognosis of AIDS patients.Results Compared with non-AD group,the levels of miR-155,Th17 and Tfh in peripheral blood of AD group were higher(P＜0.05),and the levels of miR-23b-3p were lower(P＜0.05).The level of miR-155,Th17 and Tfh in peripheral blood were positively correlated with secondary AD of AIDS(r=0.487,0.492,0.495,P＜0.05),and the level of miR-23b-3p in peripheral blood was negatively correlated with secondary AD of AIDS(r=-0.503,P＜0.05).The levels of HIV load(high dose),lymphoma,PLT,NLR,CRP,miR-155,Th17 and Tfh in the poor prognosis group were higher than those in the good prognosis group(P＜0.05),while the levels of albumin and miR-23b-3p in the poor prognosis group were lower than those in the good prognosis group(P＜0.05).Logistic regression analysis showed that lymphoma,PLT,NLR,low albumin,CRP,miR-155,Th17,Tfh and low miR-23b-3p were all risk factors for poor prognosis of AIDS patients(P＜0.05).ROC curve results showed that the AUC(95％CI)and sensitivity of combined detection of peripheral blood miR-155,Th17,Tfh and miR-23b-3p in predicting poor prognosis of AIDS patients were 0.883(0.805～0.992)and 96.70％,respectively.All of them were higher than the above peripheral blood indexes alone(P＜0.05).Conclusion The abnormal expression of miR-155,Th17,Tfh,and miR-23b-3p in peripheral blood is closely related to secondary AD in AIDS patients,and is a risk factor affecting the poor prognosis of AIDS patients.The combined detection of these indicators has a high predictive value for the poor prognosis of AIDS patients.

Objective To explore the positive distribution of antinuclear antibody(ANA)and antinuclear antibody spectrum(ANAs)specific antibody detection in patients with chronic kidney disease(CKD)and its clinical application.Methods Indirect immunofluorescence(IIF)and line immunoassay(LIA)were used to detect ANA and ANAs in the sera of 3 968 CKD patients.Results The positive rate of IIF-ANA was 14.34％,that of LIA-ANAs was 26.84％,and the coincidence rate of the two detection results was 74.65％.In IIF-ANA,the distribution of nuclear patterns showed that the highest proportion was the nuclear speckled type(38.87％),followed by the cytoplasmic speckled type(20.06％).In LIA-ANAs,anti-SSB antibody showed the highest positive rate(13.40％),followed by anti-SSA antibody(13.33％).Additionally,the positive rates of IIF-ANA and LIA-ANAs were 20.44％and 30.35％respectively in females,significantly higher than those of males at 9.98％and 24.33％(P＜0.05),and also showed a relatively high positive rate in patients under 20 years old at 46.15％and 38.46％(P＜0.05).Among patients with stage 5 CKD(CKD5),the positive detection rate of ANA was the highest at 15.30％,and the specific antibody indicators with relatively high detection rates in stages 1 to 5 mainly included anti-SSB antibody(average 5.12％)and anti-SSA antibody(average 5.09％).Conclusion The positive detection rates and consistent coincidence rates of autoantibodies in CKD patients,detected by IIF and LIA,are relatively high,especially in young women,middle-aged and elderly women,and patients who have progressed to stage 5.The positive rate of IIF-ANA increases with the progression of the disease.Therefore,routine detection of autoantibodies in patients is of great value and significance for the early identification of autoimmune diseases and can provide assistance for the treatment and prevention of CKD.

Objective A random forest model was used to analyze the risk of immune reconstitution dysfunction(INR)after antiviral therapy(HAART)in HIV/AIDS patients.Methods A total of 67 HIV/AIDS patients treated in our hospital from January 2019 to December 2022 were selected and all of them received HAART treatment.After 18 months of follow-up,patients were divided into INR group and non-INR group according to prognosis.The clinical data of the two groups were compared,and the importance of the indicators affecting INR in patients was ranked by random forest model,and the related factors affecting INR in HIV/AIDS patients after HAART were analyzed by Logistic regression model.Results During the follow-up period of 18 months,a total of 13(19.40％)of 67 HIV/AIDS patients developed INR.There were statistically significant differences in the time interval from diagnosis to treatment and baseline CD4+T lymphocyte count between INR group and non-INR group(P＜0.05).Random-forest model results showed that variables were ranked in descending order of importance as baseline CD4+T lymphocyte count and time interval between diagnosis and treatment.Logistic multivariate regression analysis showed that more than 1 year between diagnosis and treatment and baseline CD4+T lymphocyte count≤ 200/μl are all risk factors for INR in HIV/AIDS patients after HAART treatment(P＜0.05).Conclusion More than 1 year between diagnosis and treatment and baseline CD4+T lymphocyte count≤200/μl are all risk factors for INR in HIV/AIDS patients after HAART.In addition,the random forest model is of great significance for health care workers to identify high-risk groups of INR after HAART and formulate intervention plans.

Objective To explore the mechanism of TubA in the treatment of ulcerative colitis in mice,and to lay a foundation for the treatment strategy of ulcerative colitis.Methods Twenty C57BL/6 mice aged 6-8 weeks were randomly divided into 4 groups(n=5):the control group drank pure water every day,the model group and the treatment groups drank 3％dextran sulfate sodium(DSS)every day,and the treatment groups were injected with 10 mg/kg TubA and 20 mg/kg TubA every day from the third day,respectively.The weight changes of mice in all groups were recorded.Nine days later,the serum of mice was collected,and the expression levels of inflammatory factors IL-1β,IL-6 and IL-10 in serum were detected by ELISA.HE staining was used to observe the pathological changes of the mouse colon.The expression of myeloperoxidase(MPO)was detected by immunohistochemistry,the mRNA levels of inflammatory factors IL-1β,IL-6 and IL-10 were detected by RT-qPCR,and the expressions of GPX4 and FTH were detected by immunohistochemistry.The mRNA and protein expression levels of GPX4,GCLM,FTH,Nrf2,Keap1 and HO-1 in colon tissues were detected by RT-qPCR and Western blotting.Results Compared with the control group,the body weight and colon length of the model group decreased significantly.HE staining showed that inflammatory cells infiltrated the mucosa and submucosa of colon tissue,goblet cells were lost and crypt structure disordered and disappeared.Immunohistochemistry showed that the expression of MPO and FTH proteins were significantly increased,while the expression of GPX4 protein was significantly decreased(P＜0.05).The mRNA and protein expression levels of IL-1β and IL-6 were significantly increased,while the mRNA and protein expression levels of IL-10 were significantly decreased(P＜0.05).The mRNA and protein expression levels of FTH,Nrf2 and HO-1 were significantly increased,while the mRNA and protein expression levels of GPX4,GCLM and Keap1 were significantly decreased(P＜0.05).After TubA treatment,compared with the model group,all these changes mentioned above suppressed(P＜0.05).Conclusion TubA may reduce ulcerative colitis symptoms by inhibiting ferroptosis,providing new ideas for the treatment of ulcerative colitis.

Objective To investigate the effect of resveratrol on airway remodeling induced by ovalbumin(OVA)in asthmatic young mice based on the sphingosine-1-phosphate(S1P)/transforming growth factor-β1(TGF-β1)pathway.Methods Young mice were randomly assigned into normal group,model group,S1P group,resveratrol group,and resveratrol+S1P group,with 12 mice in each group.A mouse model of asthma induced by OVA was established.The Wright Giemsa staining was applied to detect the number of inflammatory cells in bronchoalveolar lavage fluid(BALF).ELISA was applied to detect the levels of interleukin-4(IL),IL-5,and IL-13 in BALF.The colorimetric method was applied to detect the activities of SOD and CAT and the content of MDA in lung tissue.HE staining was applied to observe pathological changes in lung tissue,and the wall area of bronchial tube(Wat)and perimeter of basement membrane(Pbm)were analyzed.Masson staining was applied to observe the area of collagen fibers in lung tissue.Immunohistochemistry was applied to detect the expression of α-SMA in lung tissue.Western blot was applied to detect the expression of E-cadherin,N-cadherin,Snail,SP1,and TGF-β1 proteins in lung tissue.Results Compared with the normal group,the number of eosinophils,lymphocytes,neutrophils,levels of IL-4,IL-5,and IL-13 in BALF,lung tissue MDA content,Wat/Pm ratio,collagen fiber area,α-SMA positive expression,the protein levels of N-cadherin,Snail,SP1,TGF-β1 increased in the model group,while the activities of SOD,CAT,and the protein level of E-cadherin in lung tissue decreased(P＜0.05).Compared to the model group,the number of eosinophils,lymphocytes,neutrophils,levels of IL-4,IL-5,and IL-13 in BALF,lung tissue MDA content,Wat/Pm ratio,collagen fiber area,α-SMA positive expression,the protein levels of N-cadherin,Snail,SP1,TGF-β1 decreased in the resveratrol group,while the activities of SOD,CAT,and the protein level of E-cadherin in lung tissue increased(P＜0.05).The S1P recombinant protein could weaken the inhibitory effect of resveratrol on airway remodeling in asthmatic young mice.Conclusion Resveratrol can inhibit oxidative stress,airway inflammation,and epithelial mesenchymal transition,thereby suppressing airway remodeling in asthmatic young mice.Its effect is related to the inhibition of the S1P/TGF-β1 signaling pathway.

Objective To investigate the effects of asperuloside(ASP)on airway inflammation and the interleukin-6/Janus kinase 2/signal transducer and activator of transcription 3(IL-6/JAK2/STAT3)signaling pathway in juvenile rats with bronchial asthma.Methods A juvenile rat model of bronchial asthma was constructed and randomly separated into a Model group,low,medium,and high dose asperuloside groups(ASP-L,ASP-M,ASP-H groups),and high-dose asperuloside+pathway activator group(ASP-H+CA1 group),with 12 rats in each group.An additional 12 healthy rats were included as Control group.All rats were evaluated for lung function.The number of inflammatory cells in bronchoalveolar lavage fluid(BALF),and the levels of inflammatory factors in BALF and serum were detected.HE staining was applied to observe the pathological morphology of lung tissue.Western blot was applied to detect the expression of proteins related to the IL-6/JAK2/STAT3 signaling pathway.Results Compared with Control group,the lung tissue of rats in the Model group showed severe damage and higher levels of inflammatory cell infiltration,the PEF,Cdyn,and FEV/FVC indexes were greatly decreased,the numbers of inflammatory cells(WBC,EOS,NEU,LYM),levels of inflammatory factors(IL-1β,TNF-α,and IL-6)in BALF and serum,and the expression levels of signaling pathway proteins(IL-6,p-JAK2/JAK2,p-STAT3/STAT3)were obviously increased(P＜0.05).Compared with the Model group,with the increase of ASP dosage,the degree of lung tissue damage and inflammatory cell infiltration of rats in the ASP-L,ASP-M,and ASP-H groups were significantly reduced,the PEF,Cdyn,and FEV/FVC indexes were gradually increased,the numbers of inflammatory cells(WBC,EOS,NEU,LYM),levels of inflammatory factors(IL-1β,TNF-α,and IL-6)in BALF and serum,and the expression levels of signaling pathway proteins(IL-6,p-JAK2/JAK2,p-STAT3/STAT3)were gradually decreased(P＜0.05).Compared with the ASP-H group,the lung tissue of rats in the ASP-H+CA1 group showed severe damage,the infiltration of inflammatory cells increased,the PEF,Cdyn,and FEV/FVC indexes were decreased,the numbers of inflammatory cells(WBC,EOS,NEU,LYM),levels of inflammatory factors(IL-1β,TNF-α,and IL-6)in BALF and serum,and the expression levels of signaling pathway proteins(IL-6,p-JAK2/JAK2,p-STAT3/STAT3)were greatly increased(P＜0.05).Conclusion Asperuloside can improve lung tissue damage,reduce inflammation levels,and improve lung ventilation function in rats with bronchial asthma.Its effect may be related to the regulation of the IL-6/JAK2/STAT3 signaling pathway.

Objective To investigate the effects of salidroside(Sal)on inflammatory response,lung injury,and high mobility group protein B1/Toll-like receptor 4/nuclear factor kappa B(HMGB1/TLR4/NF-κB)signaling pathway in mice with mycoplasmal pneumonia(MP).Methods An MP mouse model was constructed and randomly assigned into a Model group,a positive control-Azithromycin group(Azithromycin group),low and high dose salidroside groups(Sal-L,Sal-H groups),and high dose salidroside+pathway activator group(Sal-H+rHMGB1 group),with 12 mice in each group.Another 12 healthy mice were included as the control group.The total number of inflammatory cells and levels of inflammatory factors(IL-1β,TNF-α,and IL-6)in mice bronchoalveolar lavage fluid(BALF),levels of inflammatory factors in serum,levels of oxidative stress factors in lung tissue(ELISA),degree of lung tissue injury(HE staining),apoptosis of lung tissue cells(TUNEL staining),and expression levels of HMGB1/TLR4/NF-κB signaling pathway related proteins(Western blot)were measured.Results The lung tissue of mice in the Model group showed obvious pathological injury and infiltration of inflammatory cells,the total inflammatory cell count,inflammatory cytokine levels in BALF,the serum inflammatory cytokine levels,the MDA and NO levels in lung tissue,apoptosis rate,HMGB1 and Bax protein expression levels and p-TLR4/TLR4,p-NF-κB p65/NF-κB p65 values were obviously higher,while the SOD and GSH-Px activities,and the Bel-2 protein expression level in lung tissue were obviously lower(P＜0.05).Compared with the Model group,with the increase of Sal dose,the degree of lung tissue injury and the infiltration of inflammatory cells were reduced in the Sal-L,Sal-H,and Azithromycin groups,the total cell count,inflammatory cytokine levels in BALF,the MDA and NO levels in lung tissue,apoptosis rate,HMGB1 and Bax protein expression levels and p-TLR4/TLR4,p-NF-κB p65/NF-κB p65 values were obviously lower,while the SOD and GSH-Px activities,and the Bcl-2 protein expression level in lung tissue were obviously higher(P＜0.05).Compared with the Sal-H group,the mice in the Sal-H+rHMGB1 group showed severe lung tissue injury,the total inflammatory cell count,inflammatory cytokine levels in BALF,the serum inflammatory cytokine levels,the MDA and NO levels in lung tissue,apoptosis rate,HMGB1 and Bax protein expression levels and p-TLR4/TLR4,p-NF-κB p65/NF-κB p65 values were obviously higher,while the SOD and GSH-Px activities,and the Bcl-2 protein expression level in lung tissue were obviously lower(P＜0.05).Conclusion Sal can inhibit inflammation and oxidative stress responses,improve lung tissue injury in MP mice,and its effect may be related to the inhibition of HMGB1/TLR4/NF-κB signaling pathway activation.