BACKGROUND:Studies have confirmed that Pterocarya hupehensis skan total flavonoids(PHSTF)can improve the level of collagen-induced arthritis in rats,but there is still a lack of research on the regulation of Wnt/β-catenin signaling pathway in fibroblast-like synoviocytes and its effect on related cell functions.OBJECTIVE:To investigate the effect and mechanism of PHSTF on lipopolysaccharide-induced proliferation,migration and apoptosis of fibroblast-like synoviocytes based on the Wnt/β-catenin signaling pathwayMETHODS:Fibroblast-like synoviocytes were divided into control group,lipopolysaccharide group,lipopolysaccharide+low-,medium-,and high-dose PHSTF groups(10,20,and 40 μg/mL),lipopolysaccharide+Wnt pathway inhibitor DKK1 group,and lipopolysaccharide+Wnt pathway inhibitor DKK1+high-dose PHSTF group(40 μg/mL).The cell counting kit-8 method was used to detect the effect of PHSTF on the viability of fibroblast-like synoviocytes,and the final drug concentration and time were screened.Flow cytometry was used to detect the apoptosis of fibroblast-like synoviocytes.Cell scratch assay,EDU staining and cell cloning assay were used to detect the migration and proliferation of fibroblast-like synoviocytes.Western blot assay was used to detect the protein expression levels of Wnt3a,β-catenin,tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,Bax and Bcl-2 in fibroblast-like synoviocytes.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability decreased significantly when the concentration of PHSTF was＞40 μg/mL(P＜0.01).Therefore,the drug concentration of≤40 μg/mL was selected for subsequent experiments.(2)Compared with the lipopolysaccharide group,the wound healing rate,cell clone formation rate and the number of EDU-positive cells in the low-,medium-and high-dose PHSTF groups were significantly reduced,while the apoptosis rate was significantly increased(P＜0.05-0.01).(3)Western blot results showed that compared with the lipopolysaccharide group,low-,medium-and high-dose PHSTF significantly inhibited cellular Wnt3a,β-catenin,cellular tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,and Bcl-2 protein expression,and promoted the expression of Bax protein(P＜0.01).(4)Compared with the DKK1 group,the combination of DKK1 and high-dose PHSTF significantly inhibited the protein expression of Wnt3a,β-catenin,matrix metalloproteinase 2,matrix metalloproteinase 9 and Bcl-2 protein expression and promoted the protein expression of Bax(P＜0.01).To conclude,PHSTF may inhibit the proliferation and migration of fibroblast-like synoviocytes and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

BACKGROUND:Studies have confirmed that Pterocarya hupehensis skan total flavonoids(PHSTF)can improve the level of collagen-induced arthritis in rats,but there is still a lack of research on the regulation of Wnt/β-catenin signaling pathway in fibroblast-like synoviocytes and its effect on related cell functions.OBJECTIVE:To investigate the effect and mechanism of PHSTF on lipopolysaccharide-induced proliferation,migration and apoptosis of fibroblast-like synoviocytes based on the Wnt/β-catenin signaling pathwayMETHODS:Fibroblast-like synoviocytes were divided into control group,lipopolysaccharide group,lipopolysaccharide+low-,medium-,and high-dose PHSTF groups(10,20,and 40 μg/mL),lipopolysaccharide+Wnt pathway inhibitor DKK1 group,and lipopolysaccharide+Wnt pathway inhibitor DKK1+high-dose PHSTF group(40 μg/mL).The cell counting kit-8 method was used to detect the effect of PHSTF on the viability of fibroblast-like synoviocytes,and the final drug concentration and time were screened.Flow cytometry was used to detect the apoptosis of fibroblast-like synoviocytes.Cell scratch assay,EDU staining and cell cloning assay were used to detect the migration and proliferation of fibroblast-like synoviocytes.Western blot assay was used to detect the protein expression levels of Wnt3a,β-catenin,tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,Bax and Bcl-2 in fibroblast-like synoviocytes.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability decreased significantly when the concentration of PHSTF was＞40 μg/mL(P＜0.01).Therefore,the drug concentration of≤40 μg/mL was selected for subsequent experiments.(2)Compared with the lipopolysaccharide group,the wound healing rate,cell clone formation rate and the number of EDU-positive cells in the low-,medium-and high-dose PHSTF groups were significantly reduced,while the apoptosis rate was significantly increased(P＜0.05-0.01).(3)Western blot results showed that compared with the lipopolysaccharide group,low-,medium-and high-dose PHSTF significantly inhibited cellular Wnt3a,β-catenin,cellular tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,and Bcl-2 protein expression,and promoted the expression of Bax protein(P＜0.01).(4)Compared with the DKK1 group,the combination of DKK1 and high-dose PHSTF significantly inhibited the protein expression of Wnt3a,β-catenin,matrix metalloproteinase 2,matrix metalloproteinase 9 and Bcl-2 protein expression and promoted the protein expression of Bax(P＜0.01).To conclude,PHSTF may inhibit the proliferation and migration of fibroblast-like synoviocytes and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

OBJECTIVES: To investigate the effect of Qianggu Kangshu Formula (QGKSF) for alleviating osteoclast differentiation in rheumatoid arthritis and the underlying mechanism. METHODS: RAW264.7 cells cultured under hypoxic conditions were treated with RANKL to induce osteoclast differentiation and incubated with normal rat serum or sera from rats medicated with methotrexate (MTX) or QGKSF at low and high doses. Cell viability, TRAP-positive multinucleated cells and F-actin ring formation in the treated cells were assessed with CCK-8 assay, TRAP staining and Phalloidin staining, respectively. Autophagy and autophagosomes in the cells were observed with MDC staining and transmission electron microscopy. ELISA was used to measure IL-6 and TNF-α levels in the culture supernatant, and the expressions of HIF-1α, BNIP3, Bcl-2, Beclin1, LC3-I, LC3-II, P62 and TRAP mRNAs and proteins were analyzed using RT-qPCR and Western blotting. RESULTS: In hypoxia- and RANKL-induced RAW264.7 cells treated with normal rat serum, significant increments of TRAP-positive cells and F-actin ring formation were observed with an enhanced autophagic fluorescence intensity and increased autophagosomes. Treatment of the induced cells with rat sera medicated with MTX and low- and high-dose QGKSF obviously reduced the TRAP-positive cells, F-actin rings and autophagosomes as well as the autophagic fluorescence intensity. RANKL treatment significantly increased IL-6 and TNF-α levels in RAW264.7 cells, which were obviously decreased by treatment with MTX- and QGKSF-medicated sera. RANKL also significantly increased the mRNA and protein expression levels of HIF-1α, BNIP3, Bcl-2, Beclin1, LC3 and TRAP and lowered P62 expressions, and these changes were effectively reversed by treatment with MTX- and QGKSF-medicated sera. CONCLUSIONS QGKSF attenuates RANKL-induced osteoclast differentiation in hypoxic RAW264.7 cells by inhibiting the HIF-1α/BNIP3 autophagy signaling pathway, suggesting its potential for treatment of bone destruction in rheumatoid arthritis.
Animals ; Drugs, Chinese Herbal/pharmacology* ; Osteoclasts/drug effects* ; Autophagy/drug effects* ; Mice ; Signal Transduction/drug effects* ; Rats ; Cell Differentiation/drug effects* ; Arthritis, Rheumatoid/pathology* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; RAW 264.7 Cells ; Membrane Proteins/metabolism* ; Mitochondrial Proteins

Objective To compare and analyze the germicidal efficacy of continuous ultraviolet and pulsed ultravio-let(UV)lasers on pathogenic microorganisms.Methods Spore slides of Escherichia coli and Bacillus atrophaeus were irradiated using a 254 nm ultraviolet mercury lamp and UV laser.The carrier quantitative germicidal test was conducted to determine the disinfection efficacy at different irradiation doses.Results When the irradiation dose of the ultraviolet mercury lamp was 245.52 mJ/cm2,the logarithmic killing values of Escherichia coli and Bacillus atro-phaeus spores were 5.00 and 2.92,respectively,and the mean logarithmic killing doses were 49.10 and 84.08 mJ/cm2,respectively;When the UV laser irradiation doses were 208.39 and 206.80 mJ/cm2,the logarithmic killing values for the two microorganisms were 6.29 and 3.32,respectively,and the mean logarithmic killing doses were 33.13 and 62.29 mJ/cm2,respectively.Conclusion Compared with continuous UV radiation,pulsed UV laser has stron-ger penetration ability,better killing efficacy on pathogenic microorganisms at the same radiation dose,and can con-duct targeted disinfection and sterilization on the surface of objects directionally.

Objective To investigate the effect of Qianggu Kangshu Formula(QGKSF)for alleviating osteoclast differentiation in rheumatoid arthritis and the underlying mechanism.Methods RAW264.7 cells cultured under hypoxic conditions were treated with RANKL to induce osteoclast differentiation and incubated with normal rat serum or sera from rats medicated with methotrexate(MTX)or QGKSF at low and high doses.Cell viability,TRAP-positive multinucleated cells and F-actin ring formation in the treated cells were assessed with CCK-8 assay,TRAP staining and Phalloidin staining,respectively.Autophagy and autophagosomes in the cells were observed with MDC staining and transmission electron microscopy.ELISA was used to measure IL-6 and TNF-α levels in the culture supernatant,and the expressions of HIF-1α,BNIP3,Bcl-2,Beclin1,LC3-I,LC3-II,P62 and TRAP mRNAs and proteins were analyzed using RT-qPCR and Western blotting.Results In hypoxia-and RANKL-induced RAW264.7 cells treated with normal rat serum,significant increments of TRAP-positive cells and F-actin ring formation were observed with an enhanced autophagic fluorescence intensity and increased autophagosomes.Treatment of the induced cells with rat sera medicated with MTX and low-and high-dose QGKSF obviously reduced the TRAP-positive cells,F-actin rings and autophagosomes as well as the autophagic fluorescence intensity.RANKL treatment significantly increased IL-6 and TNF-α levels in RAW264.7 cells,which were obviously decreased by treatment with MTX-and QGKSF-medicated sera.RANKL also significantly increased the mRNA and protein expression levels of HIF-1α,BNIP3,Bcl-2,Beclin1,LC3 and TRAP and lowered P62 expressions,and these changes were effectively reversed by treatment with MTX-and QGKSF-medicated sera.Conclusion QGKSF attenuates RANKL-induced osteoclast differentiation in hypoxic RAW264.7 cells by inhibiting the HIF-1α/BNIP3 autophagy signaling pathway,suggesting its potential for treatment of bone destruction in rheumatoid arthritis.

The management of antiretroviral therapy（ART）in pregnant women living with HIV presents considerable challenges，necessitating a comprehensive approach that integrates virological suppression，drug safety，physiological changes during pregnancy，co-infections，and maternal-infant outcomes. To address these issues，the AIDS Committee of the Chinese Society of Tropical Disease and Parasitology，the Perinatal Infection Professional Committee of the Zhejiang Maternal and Child Health Association，and the Committee of AIDS Diagnosis and Treatment Quality Control Management of the Zhejiang Provincial STD/AIDS Prevention Association jointly developed the Expert Consensus on Antiretroviral Therapy for Women Living with HIV During Pregnancy（2025 version）. This consensus synthesizes international guidelines，the latest evidence-based research，and clinical expertise tailored to the management of HIV-infected pregnant women in China. It systematically outlines key aspects of ART management across various clinical scenarios during pregnancy，including treatment initiation or re-initiation，dynamic monitoring，regimen optimization，continuum of perinatal care，and management of co-infections，thereby establishing a clinically applicable decision-making framework. This article interprets the core recommendations and clinical applications of the consensus to assist healthcare providers in optimizing care for this population.

AIM:To investigate the potential effect of Dabuyuan Jian(DBYJ)on peroxisome proliferator-acti-vated receptor γ(PPARγ)/nuclear factor-κB(NF-κB)signaling pathway and related inflammatory proteins in mice with mild cognitive impairment(MCI),and to explore the mechanism of DBYJ in improving the learning and memory ability of MCI mice.METHODS:Forty C57BL/6 male mice were randomly divided into 5 groups,including negative control(NC)group,D-galactose(D-Gal)group,D-Gal+DBYJ group,D-Gal+GW9662(PPARγ inhibitor)group and D-Gal+GW9662+DBYJ group,with 8 mice each.The mice in NC group were subcutaneously injected with 0.9%saline solution on the back of the neck for 8 weeks,while those in the remaining 4 groups were subcutaneously injected with D-Gal(100 mg·kg-1·d-1)on the back of the neck for 8 weeks to establish the MCI model.From week 5 to week 8,the mice in D-Gal+GW9662 and D-Gal+DBYJ+GW9662 groups were intraperitoneally injected with GW9662(1 mg·kg-1·d-1).From week 5,the mice in D-Gal+DBYJ and D-Gal+DBYJ+GW9662 groups were treated with DBYJ(13.2 g/kg)by gavage,while those in the re-maining 3 groups were administered an equal volume of purified water for 4 weeks.The Morris water maze(including posi-tioning navigation experiment and space exploration experiment)was used to assess the learning and memory ability of the mice.The structural integrity of the hippocampus of the mice was assessed by hematoxylin-eosin(HE)staining.Nissl staining was used to evaluate damage to hippocampal neurons in mice,and Western blot was applied to detect the protein levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),PPARγ,P65 and phosphorylated P65(p-P65)in the hippocampus of the mice.RESULTS:Compared with NC group,the escape latency of the mice in D-Gal+D-Gal and D-Gal+GW9662 groups significantly increased(P＜0.01),while the number of platform crossing and the duration of staying in the target quadrant within 60 s significantly decreased(P＜0.01).The number of neurons in the CA3 region remarkably decreased(P＜0.01),and pyramidal cell disarrangement and neuronal shrinkage were observed.The levels of TNF-α,IL-1β and p-P65 were up-regulated,while the expression of PPARγ was down-regulated(P＜0.05).Compared with D-Gal group,the escape latency of the mice in D-Gal+DBYJ group significantly decreased(P＜0.01),while the number of plat-form crossing and the duration of staying in the target quadrant within 60 s remarkably increased(P＜0.01).The number of neurons in the CA3 region increased(P＜0.01),the pyramidal cells were more neatly arranged,and the cytoarchitec-ture was intact.The levels of TNF-α,IL-1β and p-P65 were down-regulated,while the expression of PPARγ was up-regu-lated(P＜0.05).Compared with D-Gal+GW9662 group,significantly decreased escape latency was observed in D-Gal+DBYJ+GW9662 group(P＜0.01),and the number of platform crossing and the duration of staying in the target quadrant within 60 s remarkably increased(P＜0.05).The number of neurons in the CA3 region increased(P＜0.01),the pyrami-dal cells were arranged more neatly,and the cytoarchitecture was relatively intact.The levels of TNF-α,IL-1β and p-P65 were down-regulated,while the expression of PPARγ was up-regulated(P＜0.05).The effects shown in D-Gal+DBYJ+GW9662 group were inferior to those in D-Gal+DBYJ group,indicating that the therapeutic effect of DBYJ was inhibited after the addition of GW9662.CONCLUSION:Dabuyuan Jian improves the learning and memory ability of MCI mice,and the mechanism may be related to the expression of key proteins in the PPARγ/NF-κB signaling pathway.

Objective To compare and analyze the germicidal efficacy of continuous ultraviolet and pulsed ultravio-let(UV)lasers on pathogenic microorganisms.Methods Spore slides of Escherichia coli and Bacillus atrophaeus were irradiated using a 254 nm ultraviolet mercury lamp and UV laser.The carrier quantitative germicidal test was conducted to determine the disinfection efficacy at different irradiation doses.Results When the irradiation dose of the ultraviolet mercury lamp was 245.52 mJ/cm2,the logarithmic killing values of Escherichia coli and Bacillus atro-phaeus spores were 5.00 and 2.92,respectively,and the mean logarithmic killing doses were 49.10 and 84.08 mJ/cm2,respectively;When the UV laser irradiation doses were 208.39 and 206.80 mJ/cm2,the logarithmic killing values for the two microorganisms were 6.29 and 3.32,respectively,and the mean logarithmic killing doses were 33.13 and 62.29 mJ/cm2,respectively.Conclusion Compared with continuous UV radiation,pulsed UV laser has stron-ger penetration ability,better killing efficacy on pathogenic microorganisms at the same radiation dose,and can con-duct targeted disinfection and sterilization on the surface of objects directionally.

Objective To explore the effect of collateral status on prognosis in elderly patients with acute ischemic stroke due to large vessel occlusion(AIS-LVO)after Solitaire stent retriever in combination with the intracranial support catheter aspiration for mechanical thrombectomy(SWIM),and construct a prediction model for prognosis.Methods A retrospective analysis was performed on 240 elderly AIS-LVO patients who underwent SWIM technique in our hospital be-tween February 2019 and February 2024.According to gender,age,occlusion sites and TOAST classifications,they were divided into a modeling group(180 cases)and a verification group(60 cases)in a ratio of 3∶1.Based on the results of modified Rankin scale(mRS)at 3 months after surgery,the patients in the modeling group were further divided into good prognosis subgroup(mRS score:0-2,97 cases)and poor prognosis subgroup(mRS score:3-6,83 cases).Multivari-ate logistic regression analysis was applied to evaluate the relationship between preoperative col-lateral circulation status and prognosis and to identify the influencing factors for prognosis.Then a prediction model for prognosis was constructed,and its performance was evaluated by ROC curve analysis.Results In the modeling group at 3 months of follow-up,the poor prognosis subgroup had significantly larger proportions of posterior circulation occlusion,cardiogenic embolism,ASITN/SIR grades 3-4 and hemorrhage transformation,higher NIHSS score at admission and longer interval from onset to vascular recanalization,while lower ASPECTS score at admission when compared with the good prognosis subgroup(P＜0.01).Multivariate logistic regression analysis showed that occlusion site,TOAST classification,NIHSS score at admission,interval from onset to vascular recanalization and hemorrhage transformation were independent risk fac-tors for poor prognosis,while ASPECTS score at admission and collateral circulation were protec-tive factors of good prognosis in the elderly AIS-LVO patients after SWIM technique(P＜0.01).Hosmer-Lemeshow test showed that the regression equation obtained goodness of fit in the mod-eling group(P=0.435).ROC curve analysis revealed that the AUC,sensitivity and specificity of then constructed prediction model for poor prognosis was 0.855[95％CI(0.797-0.913)],81.93％and 79.38％,respectively.The model was further verified in the data of the verification group(34 cases in good prognosis and 26 cases with poor prognosis),the AUC value,sensitivity and speci-ficity was 0.839[95％CI(0.732-0.947)],84.62％and 79.41％,respectively.Conclusion Our pre-diction model constructed based on screened risk factors for poor prognosis has good validity in patients with AIS-LVO after SWIM technique,which can identify the patients at high risk for poor prognosis.

AIM:To investigate the mechanism of Qianggu-Kangshu formula(QGKSF)in the treatment of rheumatoid arthritis(RA)bone destruction based on network pharmacology,molecular docking,and cell experiments.METHODS:The main active ingredients and potential targets of QGKSF were obtained through TCMSP database and litera-ture search.OMIM database and GeneCards database were used to search the targets related to RA bone destruction,and Venny 2.1.0 was employed to screen the intersected targets of QGKSF and RA bone destruction.The protein-protein inter-action network of potential intersected targets was constructed by STRING database,and topological analysis was carried out by Cytoscape software to screen core targets.The screened targets of QGKSF related to RA bone destruction were evalu-ated through the Metascape system.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology were applied to complete enrichment analysis.AutoDock and PyMOL software was used to carry out molecular docking between the core components and the core proteins.Mouse RAW264.7 macrophages were cultured in vitro,and the cell viability was de-tected by CCK-8 assay.The number of tartrate-resistant acid phosphatase(TRAP)positive multinucleated cells in each group was calculated by TRAP staining.The enzyme activity of the cells was evaluated by determination of TRAP activity.F-actin ring formation was detected by phalloidin staining.Western blot analysis was conducted to detect the protein levels of nuclear factor of activated T cells 1(NFATc1),TRAP,cathepsin K(CTSK),c-Fos,matrix metalloproteinase 9(MMP9),osteoprotegerin(OPG),receptor activator of nuclear factor κB(RANK),RANK ligand(RANKL)and phos-phorylated protein kinase B(p-AKT).RESULTS:A total of 136 active ingredients and 126 targets were selected corre-lated with QGKSF,whil 207 intersected targets of QGKSF and RA bone destruction were screened,175 of which were core targets.There were 199 pathways obtained by GO enrichment,and 20 pathways related to osteoclasts were screened out,including phosphatidylinoinosiol 3-kinase/AKT signaling pathway and osteoclast differentiation,etc.The results of TRAP staining,TRAP enzyme activity determination and phalloidin staining showed less positive cell formation,de-creased enzyme activity and decreased F-actin ring formation in QGKSF and methotrexate(MTX)groups compared with model group.Western blot results showed that compared with model group,the protein levels of NFATc1,TRAP,CTSK,c-Fos,MMP9,p-AKT,RANK and RANKL were decreased(P＜0.05),while the expression of OPG protein was in-creased(P＜0.05)in QGKSF and MTX groups.CONCLUSION:Treatment with QGKSF inhibits RANKL-induced dif-ferentiation of RAW264.7 cells into osteoclasts possibly by inhibiting the over-differentiation of osteoclasts via regulating RANKL/RANK/OPG signaling pathway.