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:Rheumatoid arthritis is widely prevalent worldwide,with its high incidence and universality that considerably affects patients' quality of life.Previous studies have focused on a few immune cells or cytokines,whereas this study comprehensively provides a more complete view of the immune mechanisms in rheumatoid arthritis.OBJECTIVE:To explore the causal relationship between 731 immune cell phenotypes and rheumatoid arthritis using the Mendelian randomization method,thereby providing evidence of causality.METHODS:The 731 immune cell phenotypes used in this study were sourced from the GWAScatalog database,jointly developed by the National Human Genome Research Institute(NHGRI)and the European Bioinformatics Institute(EBI).The rheumatoid arthritis data were from the Finngen database,developed by the Finnish Institute for Molecular Medicine(FIMM).The inverse variance weighting method was employed as the primary analytical approach.Additionally,multiple analytical methods,including MR-Egger,weighted mode,simple mode,and weighted median,were concurrently utilized to complement the final results.Sensitivity analyses(Cochran's Q test,MR-Egger regression,and MR-presso analysis)were also conducted to verify the stability and feasibility of the data.RESULTS AND CONCLUSION:(1)After excluding results through heterogeneity testing,the inverse variance weighting analysis indicated that 10 absolute cell counts,15 median fluorescence intensities of surface antigen levels,1 morphological characteristic,and 9 relative cell counts had a causal relationship with the occurrence of rheumatoid arthritis.(2)According to cell classification,this study found that seven types of B cells,seven types of classical dendritic cells,six types of mature T cells,four types of monocytes,three types of myeloid cells,three types of TBNK cells(lymphocyte subset T cells,B cells and natural killer cells),and five types of Tregs had a causal association with the occurrence of rheumatoid arthritis.(3)Through comprehensive bidirectional two-sample MR analysis,we demonstrated the complex causal relationships between multiple immune phenotypes and rheumatoid arthritis,highlighting the intricate interaction patterns between the immune system and rheumatoid arthritis.These results provide new biomarkers for the early screening and diagnosis of rheumatoid arthritis in China,and help to improve the diagnostic accuracy and sensitivity.

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:Rheumatoid arthritis is widely prevalent worldwide,with its high incidence and universality that considerably affects patients' quality of life.Previous studies have focused on a few immune cells or cytokines,whereas this study comprehensively provides a more complete view of the immune mechanisms in rheumatoid arthritis.OBJECTIVE:To explore the causal relationship between 731 immune cell phenotypes and rheumatoid arthritis using the Mendelian randomization method,thereby providing evidence of causality.METHODS:The 731 immune cell phenotypes used in this study were sourced from the GWAScatalog database,jointly developed by the National Human Genome Research Institute(NHGRI)and the European Bioinformatics Institute(EBI).The rheumatoid arthritis data were from the Finngen database,developed by the Finnish Institute for Molecular Medicine(FIMM).The inverse variance weighting method was employed as the primary analytical approach.Additionally,multiple analytical methods,including MR-Egger,weighted mode,simple mode,and weighted median,were concurrently utilized to complement the final results.Sensitivity analyses(Cochran's Q test,MR-Egger regression,and MR-presso analysis)were also conducted to verify the stability and feasibility of the data.RESULTS AND CONCLUSION:(1)After excluding results through heterogeneity testing,the inverse variance weighting analysis indicated that 10 absolute cell counts,15 median fluorescence intensities of surface antigen levels,1 morphological characteristic,and 9 relative cell counts had a causal relationship with the occurrence of rheumatoid arthritis.(2)According to cell classification,this study found that seven types of B cells,seven types of classical dendritic cells,six types of mature T cells,four types of monocytes,three types of myeloid cells,three types of TBNK cells(lymphocyte subset T cells,B cells and natural killer cells),and five types of Tregs had a causal association with the occurrence of rheumatoid arthritis.(3)Through comprehensive bidirectional two-sample MR analysis,we demonstrated the complex causal relationships between multiple immune phenotypes and rheumatoid arthritis,highlighting the intricate interaction patterns between the immune system and rheumatoid arthritis.These results provide new biomarkers for the early screening and diagnosis of rheumatoid arthritis in China,and help to improve the diagnostic accuracy and sensitivity.

Objective To investigate whether exposure pathways influence the distribution pattern and toxicity of polystyrene nanoplastics(PSNPs)in hepatic cells.Methods Male C57BL/6J wild-type healthy mice aged 6 to 8 weeks old and weighed 18 to 22 g were administered with PSNPs via gavage or tail vein injection.Then,we tracked PSNPs distribution in the major organs of mice via an in vivo imaging system(IVIS).After that,we analyzed the cellular accumulation patterns in hepatic cell subpopulations(hepatocytes and Kupffer cells)using immunofluorescence and transmission electron microscopy(TEM).300 nm PSNPs were administered via gastric gavage or tail vein injection,and 70 nm PSNPs were injected via the portal vein.The cellular localization of PSNPs in the liver was analyzed using immunofluorescence.Subsequently,using AML-12 cells,a normal mouse liver cell line,as the parenchymal hepatocyte model,the uptake of PSNPs in AML-12 cells was analyzed by confocal laser scanning microscope(CLSM).Flow cytometry was performed to observe and quantify PSNPs uptake,and to analyze the underlying endocytosis mechanisms.IVIS was used to analyze PSNPs uptake features in vivo.Finally,using mouse macrophage line RAW264.7 as a Kupffer cell model and AML-12 cells as a parenchymal hepatocyte model,the cell-type-specific toxic effects induced by 100 μg/ml PSNPs were examined through transcriptomics and metabolomics analyses.Results IVIS revealed predominant hepatic accumulation of PSNPs regardless of exposure pathways via intragastric gavage or tail vein injection.Immunofluorescence/TEM demonstrated exposure pathway-dependent cellular distribution:intragastric PSNPs were localized mainly in hepatocytes,while intravenous PSNPs were accumulated in Kupffer cells.Changes in particle size(300 nm vs.70 nm)did not alter the cellular distribution pattern,while 70 nm PSNPs injected via the portal vein accumulated in Kupffer cells,which suggested that the cell-type-specific distribution of PSNPs in the liver was independent of PSNPs size and might be related to the transport of PSNPs in the gastrointestinal tract.Flow cytometry showed that PSNPs uptake by AML-12 was time-dependent and that the underlying endocytosis mechanism involved pathways mediated by clathrin(P<0.000 1),macropinocytosis(P=0.002 6),and lipid rafts(P<0.000 1).Findings on PSNPs distribution in blood revealed that the uptake of PSNPs by hepatocytes exhibited a rate saturation phenomenon.Multi-omics analysis identified distinct toxicity patterns:PSNPs disrupted lipid metabolism and neurotransmitter homeostasis in AML-12 cells and induced inflammation and oxidative stress in Kupffer cells.Conclusion Exposure pathways mediate the hepatic cell-type-specific distribution of PSNPs,thereby altering the downstream toxicological consequences induced by exposure to PSNPs.

Objective To explore the role and mechanism of silent information regulator 1(SIRT1)in postoperative cognitive dysfunction(POCD)of aged mice following sevoflurane(SEV)anesthesia.Methods(1)Fifteen-month-old male C57BL/6 mice were randomly divided into control group(n=8)and SEV group(n=24),and SIRT1 expression in hippocampus of mice was assessed using Western blotting on the 1st,3rd and 7th day after 2%SEV exposure.(2)Fifteen-month-old male C57BL/6 mice were randomly divided into AAV-GFP,AAV-SIRT1,SEV+AAV-GFP and SEV+AAV-SIRT1 groups(n=20).AAV-SIRT1 and control AAV-GFP vectors were transfected into the brain of mice respectively.Five days after the transfection,the corresponding groups of mice were exposed to 2%SEV for 5 h.Morris water maze test was used to evaluate the spatial memory of mice before and after SEV exposure,TUNEL staining was applied to assess hippocampal neurons apoptosis,and Western blotting was utilized to measure the expression levels of SIRT1,xCT and glutathione peroxidase 4(GPX4).(3)Hippocampal neurons of mice were divided into control,AAV-SIRT1,Fer-1,SEV,SEV+AAV-SIRT1 and SEV+ferrostatin-1(Fer-1)groups.Neurons in SEV,SEV+AAV-SIRT1 and SEV+Fer-1 groups were exposed to 5%SEV for 4 h.SEV+AAV-SIRT1 and SEV+Fer-1 groups were transfected with AAV-SIRT1 or treated with Fer-1 respectively prior to SEV exposure.Neuronal death was evaluated via propidium iodide(PI)staining.Malondialdehyde(MDA)level and iron content were determined using ELISA,reactive oxygen species(ROS)level was determined using fluorescence probes.Results(1)Western blotting revealed a significant reduction in SIRT1 protein expression levels in the hippocampus tissue of SEV group mice compared to control group(P<0.05).(2)Morris water maze test results showed that,compared with AAV-GFP group,the escape latency of mice in SEV+AAV-GFP and SEV+AAV-SIRT1 groups significantly prolonged(P<0.05),and the frequency of crossing the platform significantly decreased(P<0.05).Compared with SEV+AAV-GFP group,the escape latency of mice in SEV+AAV-SIRT1 group shortened(P<0.05),and the frequency of crossing the platform on the 7th day increased(P<0.05).TUNEL staining,Western blotting and immunohistochemistry indicated that the apoptosis of hippocampal neurons,Bax and cleaved-caspase-3 protein expression levels significantly increased in SEV+AAV-GFP and SEV+AAV-SIRT1 groups compared with those in AAV-GFP group,while the expression of Bcl-2,GPX4,and xCT protein expression levels significantly decreased(P<0.05 or P<0.01 or P<0.001).Compared with SEV+AAV-GFP group,SEV+AAV-SIRT1 group showed that apoptosis of hippocampal neurons,Bax and cleaved-caspase-3 protein expression levels significantly decreased(P<0.05),while Bcl-2,GPX4,and xCT protein expression levels significantly increased(P<0.05).(3)In vitro,PI staining and ELISA demonstrated significantly increased PI positive rate,MDA level and iron content in hippocampus neurons of SEV group compared to control group(P<0.01).Compared with SEV group,the positive rate of PI staining,MDA level,iron content and ROS level in hippocampus neurons of SEV+AAV-SIRT1 and SEV+Fer-1 groups significantly decreased(P<0.05).Conclusions SEV anesthesia leads to a decrease in SIRT1 expression in hippocampus and neurons of aged mice,and the upregulation of SIRT1 could alleviate SEV-induced neuronal death and ferroptosis.

Objective To analyze the acupoint selection law of acupuncture and moxibustion for postherpetic neuralgia(PHN)with R language data mining technology.Methods The clinical research literature on acupuncture and moxibustion treatment of PHN included in CNKI,Wanfang Data,VIP and CBM from January 1,2010 to July 1,2023 was retrieved,and the database was established by Excel 2016.R language was used to statistically analyze the frequency of acupoint usage,meridians,locations,specific acupoints,etc.Through association rule analysis and clustering analysis,the characteristics and law of acupoint selection for acupuncture and moxibustion treatment of PHN were obtained.Results A total of 198 articles were included,including 83 acupoints,with a total frequency of 714 times.The high-frequency acupoints include Ashi acupoint,Jiaji acupoint and Yanglingquan.The commonly used meridians were gallbladder meridian,spleen meridian and large intestine meridiam.The acupoints were mostly in the upper and lower limbs,with the Wushu acupoints,Yuan acupoints and Xiahe acupoints being the most common.The core acupoint was Ashi acupoint,Jiaji acupoint,Hegu,Quchi,and 9 sets of association rules and 5 effective clusters were obtained.Conclusion The most commonly used acupoints for acupuncture and moxibustion treatment of PHN are Ashi acupoint,Jiaji acupoint,Hegu and Quchi,which mainly follow the principle of combining local acupoint selection with distal acupoint selection.

AIM To investigate the effects of Wenyang Jiedu Tongluo Recipe(WYJDTLR)on macrophage recruitment and polarization function in a mouse model of diabetic kidney disease(DKD).METHODS 50 db/db mice were randomly divided into the model group,the valsartan group(10.29 mg/kg)and the high-dose,medium-dose and low-dose WYJDTLR groups(26.52,13.26 and 6.63 g/kg),with 10 mice in each group,in contrast to another 10 db/m mice of the blank group.After 8 weeks of administration,the mice had their levels of fasting blood glucose,24-hour urinary protein quantity(24h-UTP),serum creatinine(Scr)and blood urea nitrogen(BUN)observed;their morphological changes of renal tissues observed by HE staining;their degree of renal glycogen deposition observed by PAS staining;their degree of renal fibrosis observed by Masson staining;their levels of MCP-1 and MCF-1 in serum and TNF-α and IL-1 β in renal tissue detected by ELISA;their renal protein expressions of VCAM-1 and ICAM-1 detected by IHC and Western blot;and their renal expressions of CD86 and CD206 detected by IF.RESULTS Compared with the model group,the WYJDTLR groups displayed decreased levels of fasting blood glucose,24h-UTP,Scr and BUN(P＜0.05,P＜0.01);improved degree of glomerular hypertrophy,mild proliferation of mesangial cells,dilatation of renal tubular,vacuolar degeneration of renal tubular epithelial cells,deposition of glomerular glycogen,and fibrosis of renal tissues(P＜0.01);decreased levels of MCP-1 and MCF-1 in serum and TNF-α and IL-1β in renal tissue(P＜0.05,P＜0.01);decreased renal protein expressions of VCAM-1 and ICAM-1(P＜0.05,P＜0.01),thus reduced the recruitment of macrophages to the kidney;decreased renal CD86 protein expression(P＜0.01);and increased CD206 protein expression(P＜0.01),thus inhibited M1-type polarization of macrophages and promoted M2-type polarization of macrophages.CONCLUSION WYJDTLR can delay the DKD progression in mice by reducing the occurrence of inflammatory reactions through reducing the level of macrophage recruitment factor,inhibiting the M1-type polarization,and promoting the M2-type polarization.

Objective:To investigate the value of tumor and adipose tissue metabolic parameters from baseline 18F-FDG PET/CT in predicting the efficacy and prognosis of immunotherapy in patients with advanced nasopharyngeal carcinoma (NPC). Methods:From February 2019 to February 2022, 112 patients (91 males, 21 females, age 21-73 years) with advanced NPC who were treated with programmed death-1 (PD-1) inhibitors at Sun Yat-Sen University Cancer Center were retrospectively included. All patients underwent baseline PET/CT examination. Tumor and adipose tissue metabolic parameters were measured and calculated. Patients were divided into clinical benefit and non-clinical benefit groups, and Mann-Whitney U test or χ2 test was used to assess the differences between groups. Prognostic analysis of progression-free survival (PFS) was performed using multivariate Cox proportional hazards regression model and a prognostic stratification system was constructed. Results:Of the 112 patients, 85 were in the clinical benefit group and 27 were in the non-clinical benefit group. In non-clinical benefit group and clinical benefit group, the metabolic tumor volume (MTV) of primary tumor (PT-MTV) were 47.7(7.7, 81.2) and 14.0(5.7, 27.1)cm 3, total lesion glycolysis (TLG) of primary tumor (PT-TLG) were 228.9(27.4, 492.8) and 72.7(20.4, 165.5)g, whole-body MTV (WB-MTV) were 94.2(45.9, 215.4) and 61.3(31.6, 104.3)cm 3, whole-body TLG (WB-TLG) were 605.5(214.1, 1 402.5) and 319.2(172.4, 632.8)g, SUV max of visceral adipose tissue (SUV max-VAT) were 0.77(0.55, 0.91) and 0.62(0.48, 0.76), respectively ( Z values: from -2.72 to -1.96, all P<0.05). The proportion of patients with lung metastasis in non-clinical benefit group was higher than that in clinical benefit group (44.4%(12/27) vs 23.5%(20/85); χ2=4.39, P=0.036). PT-MTV (hazard ratio ( HR)=2.807, 95% CI: 1.540-5.118, P=0.001) and the presence of lung metastases ( HR=1.691, 95% CI: 1.012-2.823, P=0.045) were independent predictive factors for PFS in multivariate analysis. The prognostic prediction model based on the two predictive factors was able to significantly differentiate the prognosis in patients. Conclusions:Baseline tumor metabolic parameters and SUV max-VAT are associated with the efficacy of immunotherapy in patients with advanced NPC. PT-MTV and lung metastasis can independently predict PFS. The constructed prediction model can stratify patients′ prognosis.