Objective To explore the effect of modified Buzhong Yiqi decoction(MBZYQ)on thyroid function in rats with Hashimoto's thyroiditis(HT)by regulating the Toll-like receptor 2(TLR2)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway.Methods The HT rat model was constructed.The successfully modeled rats were divided into the HT group,L-MBZYQ,M-MBZYQ,H-MBZYQ(intragastric administration of 11.35,22.70,and 45.40 g/kg MBZYQ granules,respectively),and the MBZYQ+Pam3CSK4 group(intragastric administration of 45.40 g/kg MBZYQ granules+intraperitoneal injection of 20 μg TLR2 agonist Pam3CSK4),with 10 rats in each group.Another 10 normally raised rats were selected as the control group.The pathological changes of thyroid tissues and the apoptosis of thyroid tissue cells in each group were observed by HE staining and TUNEL staining respectively.The expressions of serum thyroglobulin antibody(TGAb),thyroid peroxidase antibody(TPOAb),free triiodothyronine(FT3),free thyroxine(FT4),thyroid stimulating hormone(TSH),interleukin-17(IL-17),and interleukin-10(IL-10)were detected by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the protein expression of TLR2,MyD88 and NF-κB in thyroid tissues.Results The pathological damage of thyroid tissues in the L-MBZYQ,M-MBZYQ and H-MBZYQ groups was alleviated.The apoptosis rate of cells,the expressions of TGAb,TPOAb,TSH,IL-17,TLR2,MyD88 and NF-κB were lower than those in the HT group,while the expressions of FT3,FT4 and IL-10 were higher than those in the HT group.Moreover,the degree of decrease or increase in the M-MBZYQ and H-MBZYQ groups was greater than that in the L-MBZYQ group,and the changes in the H-MBZYQ group were greater than those in the M-MBZYQ group(P<0.05).The pathological damage of thyroid tissue in the MBZYQ+Pam3CSK4 group was further aggravated.The apoptosis rate of cells,the expressions of TGAb,TPOAb,TSH,IL-17,TLR2,MyD88 and NF-κB were higher than those in the H-MBZYQ group,while the expressions of FT3,FT4 and IL-10 were lower than those in the H-MBZYQ group(P<0.05).Conclusion MBZYQ can protect the thyroid function of HT rats,and its mechanism of action may be achieved by inhibiting the TLR2/MyD88/NF-κB signaling pathway.

Objective To analyze the effect of Midazolam(MID)on visceral pain in diarrheal irritable bowel syndrome(IBS-D)rats based on the forkhead box protein O1/forkhead box transcription factor O3a(FoxO1/FoxO3a)pathway.Methods An IBS-D rat model was constructed,and successfully modeled rats were randomly assigned into IBS-D group,low MID(MID-L)group,high MID(MID-H)group,and MID-H+AS1842856 group,with 12 rats in each group.Additionally,12 normal healthy rats were included as the Control group.The Control group and the IBS-D group were intraperitoneally injected with an equal volume of 0.9%sodium chloride.The MID-L and MID-H groups were intraperitoneally injected with 30 and 90 mg/kg MID respectively,while the MID-H+AS1842856 group was intraperitoneally injected with 90 mg/kg MID and given 50 mg/kg AS1842856 by gavage.Rats in each group were evaluated for abdominal wall withdrawal response(AWR)score.Enzyme-linked immunosorbent assay(ELISA)was applied to detect the levels of oxidative stress and pain factors in serum,and HE staining was applied to detect pathological changes in colonic tissue.Immunohistochemistry was applied to detect the expression of glial fbrillary acidic protein(GFAP)and nerve growth factor(NGF)in colon tissue,and Western Blot method was applied to detect the expression of proteins related to the FoxO1/FoxO3a pathway.Results The pathological damage in the colonic tissue in the IBS-D group was more severe than that in the Control group,and the AWR score,malondialdehyde(MDA),substance P(SP),5-hydroxytryptamine(5-HT),and GFAP and NGF expression elevated,while the superoxide dismutase(SOD)level and the expression of FoxO1 and FoxO3a decreased(P<0.05).The colonic tissue damage in the MID-L and MID-H groups was less severe compared with the IBS-D group,and the AWR score,MDA,SP,5-HT,and GFAP and NGF expression decreased,while SOD level and the expression of FoxO1,FoxO3a increased;the changes were more significant in the MID-H group(P<0.05).The colonic tissue damage in the MID-H+AS1842856 group was more severe than that in the MID-H group,and the AWR score,MDA,SP,5-HT,and GFAP and NGF expression increased,while SOD level level and the expression of FoxO1,FoxO3a decreased(P<0.05).Conclusion MID can improve visceral pain in IBS-D rats,and its mechanism of action is related to the activation of the FoxO1/FoxO3a pathway.

Objective To analyze the therapeutic effect and mechanism of Neferine(Nef)on psoriasis mice based on the nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.Methods A mouse model of psoriasis was constructed.The 50 mice with successful modeling were divided into the model group,the L-Nef group,the M-Nef group,the H-Nef group,and the H-Nef+ML385 group,with 10 mice in each group according to random number table method.Additionally,10 normal mice were selected as the control group.The L-Nef group,M-Nef group and H-Nef group were respectively administered 5,10 and 20 mg/kg Nef by gavage.The H-Nef+ML385 group was administered 20 mg/kg Nef by gavage and intraperitoneally injected with 30 mg/kg Nrf2/HO-1 signaling pathway inhibitor ML385.The control group and the model group were given the same amount of normal saline.The changes of skin lesions in each group of mice were observed and the psoriasis area and severity index(PASI)scoring was conducted.HE staining and toluidine blue staining were used to observe the pathological changes of the skin and the number of mast cells in each group,and immunohistochemical staining was used to observe the expression of Ki-67 in the lesion sites of mice in each group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammation and oxidative stress factors in the skin lesion tissues of mice in each group,and Western blot was used to detect the expression of Nrf2/HO-1 signaling pathway-related proteins in the skin lesion tissues of mice in each group.Results Compared with the control group,the mice in the model group presented symptoms such as skin erythema and scales,and the pathological changes of the skin lesion tissue were severe.The PASI score,the number of mast cells,Ki-67,tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),interleukin-6(IL-6),interleukin-17A(IL-17A),and malondialdehyde expressions were increased,while the expressions of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),Nrf2 and HO-1 were decreased(P<0.05).Compared with the model group,the symptoms such as skin erythema and scales and the pathological changes of skin lesion tissues in mice of the L-Nef group,M-Nef group and H-Nef group were alleviated,and the PASI score,the number of mast cells,the expressions of Ki-67,TNF-α,IFN-γ,IL-6,IL-17A and malondialdehyde were significantly reduced;however,the expressions of SOD,GSH-Px,Nrf2 and HO-1 were significantly increased(P<0.05).Compared with the H-Nef group,the symptoms such as skin erythema and scales and the pathological changes of skin lesion tissues in the H-Nef+ML385 group of mice were more severe.The PASI score,the number of mast cells,and the expressions of Ki-67,TNF-α,IFN-γ,IL-6,IL-17A,and malondialdehyde were significantly increased.while the expressions of SOD,GSH-Px,Nrf2 and HO-1 were significantly decreased(P<0.05).Conclusion Nef may have therapeutic effects by activating the Nrf2/HO-1 signaling pathway to improve the inflammation and oxidative stress responses in the lesion tissues of mice with psoriasis.

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.

Objective To explore the mechanism by which the early growth response protein 1(EGR1)/AMP-activated protein kinase(AMPK)/nuclear factor erythroid 2-related factor 2(Nrf2)pathway improves Neuro-2a cell injury via Mendelian randomization(MR),bioinformatics and in vitro models of ischemic stroke(IS).Methods The blood proteins associated with genetic susceptibility to IS were identified based on MR analysis.Then,the GSE22255 dataset was comprehensively analyzed,including GSEA,immunoinfiltration and differential expression analysis.The transcription factors closely related to the occurrence and development of IS were identified after joint analysis with MR results,and the biological functions of key genes were further verified by in vitro experiments.Results A total of 712 proteins related to IS susceptibility were identified by MR analysis.A total of 2357 differentially expressed genes were identified from the GSE22255 dataset,and 75 intersection genes and 34 transcription factors were identified after combined analysis with MR results.In vitro experiments showed that knockdown of EGR1 expression could significantly inhibit oxygen-glucose deprivation/reperfusion(OGD/R)-induced Neuro-2a cell damage,and the mechanism might be related to the up-regulation of p-AMPK and Nrf2 protein expression.Conclusion This study integrates MR analysis,transcriptomics and in vitro experiments to reveal the potential role of 75 IS susceptibility genes and transcription factor EGR1 in the pathogenesis of IS,which provides theoretical basis for the development of therapeutic drugs for IS.

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.

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.

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.

Knee osteoarthritis(KOA)is a chronic joint disease characterized by degeneration of articular cartilage and joint pain.Its pathogenesis is complex,involving cell death,inflammatory responses and the interaction of various signaling pathways.Apoptosis is activated by Caspase family proteins and DNA endonucleases,leading to cellular dysfunction and promoting the degeneration of articular cartilage.Pyroptosis is a form of programmed cell death triggered by inflammasome activation,involving the activation of nucleotide-binding oligomeric domain-like receptor protein 3 inflammasome,which leads to the release of pro-inflammatory cytokines such as interleukin(IL)-6,IL-1 β and tumor necrosis factor-α.This release intensifies local inf/lammation and accelerates the progression of the disease.Ferroptosis is driven by iron-catalyzed lipid peroxidation,which induces the rupture of chondrocyte cell membranes and the degradation of matrix,thereby promoting joint degeneration.In addition,autophagy plays a dual role in KOA:moderate autophagy supports chondrocyte repair,while excessive autophagy may promote joint destruction,cartilage matrix degradation and subchondral bone sclerosis.The PI3K/AKT,NF-κB,Wnt/β-catenin and Hedgehog signaling pathways can regulate the growth,differentiation,proliferation,migration and apoptosis of articular chondrocytes.Single-cell RNA sequencing technology has provided new insights into the cellular heterogeneity and molecular mechanisms in the pathogenesis of KOA.This review explores the mechanisms of cell death in KOA,with a focus on the roles of pro-inflammatory cytokines,signaling pathway,and single-cell RNA sequencing techniques in the occurrence and progression of KOA.

The incidence of thyroid cancer has significantly increased in the past decade or so.The tumor tissues,stromal cells and immune cells that exist in thyroid cancer itself constitute the dynamic and changing tumor microenvironment of thyroid cancer.The tumor microenvironment plays a decisive role in the occurrence,development and treatment of thyroid cancer,among which B lymphocytes have received increasing attention.This review systematically summarizes the latest research progress of B lymphocytes in the tumor microenvironment of thyroid cancer,to better explore the characteristics of the tumor microenvironment of thyroid cancer.