Objective To investigate the effects of synovial fluid from patients with rheumatoid arthritis(RA)on the polarization of normal neutrophils and the MEK/ERK signaling pathway,thereby providing experimental evidence to elucidate the pathological mechanisms of RA and offering novel insights into its treatment.Methods Synovial fluid samples were collected from 20 RA patients and 20 osteoarthritis(OA)patients.ELISA was used to measure the levels of interferon-gamma(IFN-γ),tumor necrosis factor-alpha(TNF-α),transforming growth factor-beta(TGF-β)and interleukin-8(IL-8)in the synovial fluid.Neutrophils were isolated from the peripheral blood of healthy donors,and their purity was confirmed by flow cytometry.Neutrophils were then treated with synovial fluid and divided into four groups:Control(untreated),RA(treated with 10％RA synovial fluid),OA(treated with 10％OA synovial fluid),and anti-IFN-β(treated with 20 μg anti-IFN-β antibody as an N2-type control).Western blot was used to assess the changes in the expression of IFN-γ,TNF-α,TGF-β,MEK,p-MEK,ERK and p-ERK.Additionally,the MEK inhibitor PD0325901 was used to block the MEK/ERK pathway,and subsequent changes in IFN-γ and TGF-β expression in neutrophils were evaluated.Results The levels of IFN-γ,TNF-α,TGF-β and IL-8 in the synovial fluid of RA patients were significantly higher than those in OA patients.After intervention with RA synovial fluid,the relative expression of IFN-γ and TNF-α in neutrophils were significantly increased compared to those in the untreated control group and the OA group.While,TGF-β expression in the RA group was lower than that in both the control and anti-IFN-β groups.The relative expression of p-MEK and p-ERK in the RA group were significantly higher than those in the control,OA and Anti-IFN-β groups.Upon addition of the MEK inhibitor,the relative expression of p-MEK and p-ERK were reduced.Furthermore,in both the RA and OA groups,the relative expression levels of IFN-γ decreased,while the expression of TGF-β increased.Conclusion Synovial fluid from RA patients contains higher of IFN-γ,TNF-αand TGF-β,which activate neutrophils through phosphorylation of the MEK/ERK signaling pathway,promoting their polarization toward the pro-inflammatory N1 phenotype.

The accumulation of deoxyribonucleic acid (DNA) oxidative damage mediated by reactive oxygen species (ROS) is closely associated with liver diseases. 8-Oxoguanine (8-OxoG), a prevalent DNA oxidation product, plays a significant role in liver disease progression. The base excision repair (BER) pathway, comprising over 30 proteins including 8-OxoG DNA glycosylase1 (OGG1), MutY homolog (MUTYH), and MutT homolog protein 1 (MTH1), is responsible for the clearance and mismatch repair of 8-OxoG. Abnormally high levels of 8-OxoG and dysregulated expression and function of 8-OxoG repair enzymes contribute to the onset and development of liver diseases. Consequently, targeting the 8-OxoG production and repair system with agonists or inhibitors may offer a promising approach to liver disease treatment. This review summarizes the impact of 8-OxoG accumulation and dysregulated repair enzymes on various liver diseases, including viral liver disease, alcoholic liver disease (ALD), metabolic dysfunction-associated steatotic liver disease (MASLD), cholestatic liver disease (CLD), liver fibrosis, cirrhosis, and liver cancer. Additionally, we review natural constituents as potential therapeutic agents that regulate 8-OxoG production, repair enzymes, and repair system-related signal pathways in oxidative damage-induced liver diseases.
Humans ; Liver Diseases/genetics* ; Biological Products/pharmacology* ; DNA Repair/drug effects* ; Guanine/metabolism* ; Animals ; Disease Progression ; DNA Damage ; Oxidative Stress

Objective To investigate the effects of extracellular vesicles(EVs)derived from synovial fluid of rheumatoid arthritis(RA)patients on angiogenesis of human umbilical vein endothelial cells,and to preliminarily explore the underlying mechanisms.Methods Synovial fluid samples of knee joint were collected from 20 patients with RA and 20 patients with osteoarthritis(OA)in this study.EVs were purified using ultracentrifugation.The morphology and size of EVs were observed by transmission electron microscopy and nanoparticle tracking analysis.CD9,CD63,cytochrome c(Cyt-c),vascular endothelial growth factor(VEGF),lysyl oxidase(LOX),matrix metalloproteinase 2(MMP2),tumour necrosis factor alpha(TNF-α),transforming growth factor beta1(TGF-β1)in EVs were detected using Western blot.Human umbilical vein endothelial cells(HUVEC)were treated with the EVs.The growth,migration and angiogenesis of HUVEC were observed by CCK8 assay,TranswellTM chamber assay,scratch test and matrigel angiogenesis assay,respectively.The effect of EVs on the PI3K/AKT pathway in HUVEC was assessed using Western blot.Results Both EVs from RA synovial fluid(RA-EVs)and OA synovial fluid(OA-EVs)were cup-shaped,mainly between 30-400 nm in diameter,expressing CD63 and CD9,but not Cyt-c.RA-EVs carried more VEGF,LOX,MMP2,TNF-α and TGF-β1 than OA-EVs.Compared with the OA-EVs intervention,RA-EVs significantly promoted the proliferation,migration,and angiogenesis of HUVECs,as well as upregulated PI3K/AKT phosphorylation.The inhibitor of PI3K suppressed angiogenesis induced by EVs.Conclusion EVs in synovial fluid of RA carried more cytokines and enzymes that related angiogenesis and inflammation.These EVs exert their pro-angiogenic effects by activating the PI3K/AKT pathway,then contributing to the pathological progression of RA.

Objective To investigate the effects of synovial fluid from patients with rheumatoid arthritis(RA)on the polarization of normal neutrophils and the MEK/ERK signaling pathway,thereby providing experimental evidence to elucidate the pathological mechanisms of RA and offering novel insights into its treatment.Methods Synovial fluid samples were collected from 20 RA patients and 20 osteoarthritis(OA)patients.ELISA was used to measure the levels of interferon-gamma(IFN-γ),tumor necrosis factor-alpha(TNF-α),transforming growth factor-beta(TGF-β)and interleukin-8(IL-8)in the synovial fluid.Neutrophils were isolated from the peripheral blood of healthy donors,and their purity was confirmed by flow cytometry.Neutrophils were then treated with synovial fluid and divided into four groups:Control(untreated),RA(treated with 10％RA synovial fluid),OA(treated with 10％OA synovial fluid),and anti-IFN-β(treated with 20 μg anti-IFN-β antibody as an N2-type control).Western blot was used to assess the changes in the expression of IFN-γ,TNF-α,TGF-β,MEK,p-MEK,ERK and p-ERK.Additionally,the MEK inhibitor PD0325901 was used to block the MEK/ERK pathway,and subsequent changes in IFN-γ and TGF-β expression in neutrophils were evaluated.Results The levels of IFN-γ,TNF-α,TGF-β and IL-8 in the synovial fluid of RA patients were significantly higher than those in OA patients.After intervention with RA synovial fluid,the relative expression of IFN-γ and TNF-α in neutrophils were significantly increased compared to those in the untreated control group and the OA group.While,TGF-β expression in the RA group was lower than that in both the control and anti-IFN-β groups.The relative expression of p-MEK and p-ERK in the RA group were significantly higher than those in the control,OA and Anti-IFN-β groups.Upon addition of the MEK inhibitor,the relative expression of p-MEK and p-ERK were reduced.Furthermore,in both the RA and OA groups,the relative expression levels of IFN-γ decreased,while the expression of TGF-β increased.Conclusion Synovial fluid from RA patients contains higher of IFN-γ,TNF-αand TGF-β,which activate neutrophils through phosphorylation of the MEK/ERK signaling pathway,promoting their polarization toward the pro-inflammatory N1 phenotype.

Objective To investigate the effects of extracellular vesicles(EVs)derived from synovial fluid of rheumatoid arthritis(RA)patients on angiogenesis of human umbilical vein endothelial cells,and to preliminarily explore the underlying mechanisms.Methods Synovial fluid samples of knee joint were collected from 20 patients with RA and 20 patients with osteoarthritis(OA)in this study.EVs were purified using ultracentrifugation.The morphology and size of EVs were observed by transmission electron microscopy and nanoparticle tracking analysis.CD9,CD63,cytochrome c(Cyt-c),vascular endothelial growth factor(VEGF),lysyl oxidase(LOX),matrix metalloproteinase 2(MMP2),tumour necrosis factor alpha(TNF-α),transforming growth factor beta1(TGF-β1)in EVs were detected using Western blot.Human umbilical vein endothelial cells(HUVEC)were treated with the EVs.The growth,migration and angiogenesis of HUVEC were observed by CCK8 assay,TranswellTM chamber assay,scratch test and matrigel angiogenesis assay,respectively.The effect of EVs on the PI3K/AKT pathway in HUVEC was assessed using Western blot.Results Both EVs from RA synovial fluid(RA-EVs)and OA synovial fluid(OA-EVs)were cup-shaped,mainly between 30-400 nm in diameter,expressing CD63 and CD9,but not Cyt-c.RA-EVs carried more VEGF,LOX,MMP2,TNF-α and TGF-β1 than OA-EVs.Compared with the OA-EVs intervention,RA-EVs significantly promoted the proliferation,migration,and angiogenesis of HUVECs,as well as upregulated PI3K/AKT phosphorylation.The inhibitor of PI3K suppressed angiogenesis induced by EVs.Conclusion EVs in synovial fluid of RA carried more cytokines and enzymes that related angiogenesis and inflammation.These EVs exert their pro-angiogenic effects by activating the PI3K/AKT pathway,then contributing to the pathological progression of RA.

Objective:To explore the assessment value of liver enzyme changes before and after hepatectomy on the prognosis of patients with primary hepatocellular carcinoma(HCC). Methods:A retrospective analysis was conducted on the clinical data of 421 patients with primary HCC who received at the First and Second Affiliated Hospitals of Xi'an Jiaotong University from January 2016 to December 2020.The univariate and multivariate COX regression analyses were used to screen the risk factors affecting the prognosis of patients with primary HCC,and the optimal cut-off value was selected to group the most valuable indicators among them,further analyzing the different factors among subgroups.The Kaplan-Meier method was used to assess the prognosis of the subgroups,and the survival curves were plotted and statistically tested. Results:Multivariate COX regression analysis revealed that Barcelona Clinic Liver Cancer(BCLC)stage,tumor number,alpha-fetoprotein(AFP),the quantitative change of γ-glutamyl transpeptidase(GGT)and total bilirubin(TBIL)before and after surgery were independent risk factors affecting the prognosis of patients with primary HCC(P＜0.01),and there were differences in hepatitis B virus surface antigen(HBsAg),TNM stage,BCLC stage,tumor size,tumor number,tumor differentiation degree,AFP,surgery type,intraoperative bleeding volume,postoperative albumin-bilirubin(ALBI)and white blood cell count between the high and low GGT variation groups(P＜0.05).Kaplan-Meier survival analysis showed that the prognosis of patients with primary HCC between the high and low GGT variation groups was significantly different,with a hazard ratio of 2.603 for mortality and 1.449 for recurrence(both P＜0.01). Conclusion:The quantitative change of GGT before and after hepatectomy has an evaluation value for the prognosis of patients with primary HCC,and the quantitative change of GGT greater than 29.5 is an independent risk factor for the prognosis of patients with primary HCC.

Objective To explore the expression and clinical significance of lysine oxidase (Lox) in primary lesion of esophageal carcinoma (ESCA) and bone metastasis lesion based on bioinformatics. Methods The Cancer Genome Atlas (TCGA) and Gene Expression Profiling Interactive Analysis (GEPIA) databases were used to screen for differentially expressed genes between ESCA and normal esophageal tissues. Follow-up information of patients with surgery for esophageal cancer in the Fourth Hospital of Hebei Medical University from January 2016 to December 2020 were screened, and the clinical materials of patients diagnosed as bone metastasis during the follow-up period were collected. Western blot was used to verify the expression of Lox in ESCA and normal esophageal tissues; immunohistochemical staining was used to detect the expression of Lox in human ESCA tissue and normal tissue; the impact of Lox expression on survival was explored by Kaplan-Meier curve and Cox regression. Results Through the analysis of ESCA data in GEPIA and TCGA databases, it was found that the expression of Lox in ESCA lesions was significantly higher than that in normal tissues (P<0.05); the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses found that Lox may be involved in the information conduction of various signaling pathways; the Western blot result showed that the expression of Lox in cancer tissue was higher than that in adjacent normal tissue (P<0.05); the analysis of follow-up data found that patients with high expression of Lox were more likely to have multiple bone metastases; survival analysis revealed that patients with high Lox expression had significantly shorter bone metastasis free survival and overall survival compared to patients with low Lox expression (P<0.05); Cox regression found that Lox was an independent risk factor for prognosis of patients with esophageal cancer bone metastasis. Conclusion Lox is highly expressed in ESCA and significantly related to clinical prognosis, which can be used as an effective target for the diagnosis and treatment of ESCA.

Objective:To analyze the epidemiology, and infant outcomes of congenital diaphragmatic hernia (CDH) in Beijing.Methods:This retrospective study involved cases diagnosed with CDH among 877 230 fetuses/infants monitored by the Beijing Birth Defects Monitoring System from January 2018 to December 2022. Data on the occurrence of CDH, prenatal diagnosis, and infant mortality were analyzed. The statistical method was the Chi-square test (or Fisher's exact test). Results:(1) A total of 254 CDH cases were included in this study. During the study period, the incidence of CDH throughout the pregnancy and the perinatal period was 0.29 per thousand (254/877 230) and 0.17 per thousand (153/877 230), respectively. (2) Among the 254 cases, 79 (31.1%) underwent prenatal genetic diagnosis, 6.3%(5/79) copy number varations were detected, including one case of like pathogenic and four cases with types of variation unreported. The proportions of prenatal genetic diagnosis from 2018 to 2022 showed no statistically significant difference ( χ2trend=2.86, P=0.091). The gestational age at prenatal diagnosis for 241 cases was (23.0±6.4) weeks, ranging from 13 to 38 weeks; 130 cases (53.9%) were diagnosed before 25 weeks of gestation. The proportion of genetic diagnosis was higher in the group diagnosed by ultrasound before 25 weeks (43.1%, 56/130) compared to the group diagnosed at or after 25 weeks (20.7%, 23/111) ( χ2=12.59, P<0.001). (3) A total of 146 live births were recorded, with the proportion of continuing pregnancy to live birth after prenatal diagnosis being 55.2% (133/241), with no significant difference across the years. The proportion of continuing pregnancy to live birth after prenatal diagnosis in the isolated CDH group was 64.5% (100/155), higher than the 38.3% (33/86) in the non-isolated group ( χ2=15.57, P<0.001). The live birth rate for those diagnosed prenatally at<25 weeks of gestation (26.2%, 34/130) was lower than for those diagnosed at≥25 weeks of gestation (89.2%, 99/111) ( χ2=93.67, P<0.001). (4) Among the 146 live-born infants, 28 (19.2%) died during infancy. All 28 cases were prenatally diagnosed with CDH. The mortality rate of CDH infants born in tertiary obstetric institutions (17.9%, 24/134) showed a trend towards being lower than that in secondary institutions (4/12), but the difference was not statistically significant (Fisher's exact test, P=0.245). Among the 118 surviving infants, seven were abandoned from treatment, and of the remaining 111, 104 (93.7%) were treated at the Children's Hospital, Capital Institute of Pediatrics. The infant mortality rate at this institution (15.4%, 16/104) showed a trend towards being lower than that at other institutions (3/7), but the difference was not statistically significant (Fisher's exact test, P=0.096). The infant mortality rate for those diagnosed prenatally at<25 weeks of gestation (38.2%, 13/34) was higher than for those diagnosed at≥25 weeks of gestation (15.2%, 15/99) ( χ2=6.78, P=0.009). Conclusions:Beijing achieved good secondary and tertiary prevention outcomes in prenatal diagnosis and reducing infant mortality. The concentration of neonatal surgical resources is beneficial for accumulating surgical experience and reducing infant mortality.

Objective:To analyze the epidemiology, and infant outcomes of congenital diaphragmatic hernia (CDH) in Beijing.Methods:This retrospective study involved cases diagnosed with CDH among 877 230 fetuses/infants monitored by the Beijing Birth Defects Monitoring System from January 2018 to December 2022. Data on the occurrence of CDH, prenatal diagnosis, and infant mortality were analyzed. The statistical method was the Chi-square test (or Fisher's exact test). Results:(1) A total of 254 CDH cases were included in this study. During the study period, the incidence of CDH throughout the pregnancy and the perinatal period was 0.29 per thousand (254/877 230) and 0.17 per thousand (153/877 230), respectively. (2) Among the 254 cases, 79 (31.1%) underwent prenatal genetic diagnosis, 6.3%(5/79) copy number varations were detected, including one case of like pathogenic and four cases with types of variation unreported. The proportions of prenatal genetic diagnosis from 2018 to 2022 showed no statistically significant difference ( χ2trend=2.86, P=0.091). The gestational age at prenatal diagnosis for 241 cases was (23.0±6.4) weeks, ranging from 13 to 38 weeks; 130 cases (53.9%) were diagnosed before 25 weeks of gestation. The proportion of genetic diagnosis was higher in the group diagnosed by ultrasound before 25 weeks (43.1%, 56/130) compared to the group diagnosed at or after 25 weeks (20.7%, 23/111) ( χ2=12.59, P<0.001). (3) A total of 146 live births were recorded, with the proportion of continuing pregnancy to live birth after prenatal diagnosis being 55.2% (133/241), with no significant difference across the years. The proportion of continuing pregnancy to live birth after prenatal diagnosis in the isolated CDH group was 64.5% (100/155), higher than the 38.3% (33/86) in the non-isolated group ( χ2=15.57, P<0.001). The live birth rate for those diagnosed prenatally at<25 weeks of gestation (26.2%, 34/130) was lower than for those diagnosed at≥25 weeks of gestation (89.2%, 99/111) ( χ2=93.67, P<0.001). (4) Among the 146 live-born infants, 28 (19.2%) died during infancy. All 28 cases were prenatally diagnosed with CDH. The mortality rate of CDH infants born in tertiary obstetric institutions (17.9%, 24/134) showed a trend towards being lower than that in secondary institutions (4/12), but the difference was not statistically significant (Fisher's exact test, P=0.245). Among the 118 surviving infants, seven were abandoned from treatment, and of the remaining 111, 104 (93.7%) were treated at the Children's Hospital, Capital Institute of Pediatrics. The infant mortality rate at this institution (15.4%, 16/104) showed a trend towards being lower than that at other institutions (3/7), but the difference was not statistically significant (Fisher's exact test, P=0.096). The infant mortality rate for those diagnosed prenatally at<25 weeks of gestation (38.2%, 13/34) was higher than for those diagnosed at≥25 weeks of gestation (15.2%, 15/99) ( χ2=6.78, P=0.009). Conclusions:Beijing achieved good secondary and tertiary prevention outcomes in prenatal diagnosis and reducing infant mortality. The concentration of neonatal surgical resources is beneficial for accumulating surgical experience and reducing infant mortality.

OBJECTIVE: Digital therapy is important in treating motor system disease. The outcome of digital therapy in post-operative rehabilitation of knee anterior cruciate ligament (ACL) reconstruction is assessed. METHODS: 142 patients are treated with digital rehabilitation therapy after ACL reconstruction. Patients' pain score, joint motion, lower limb function score, anxiety score are statistically analyzed. Patients' satisfaction, device usage and adverse events are documented. RESULTS: At post-operative 1st day, 8th weeks, 12th weeks, pain score are 4, 2, 1, knee joint range of motion are 55°, 110°, 143°, lower limb function score are 18, 56, 76, anxiety score are 32.5, 26, 23.5 respectively. Patients' satisfaction are 9.4. Mean duration of device usage is (177.6±38.0) minutes per week. Rehabilitation-related and device-related adverse event does not happen. CONCLUSIONS Digital therapy promotes post-operative rehabilitation after ACL reconstruction.
Humans ; Knee Joint ; Lower Extremity ; Medicine ; Anterior Cruciate Ligament Reconstruction ; Pain