ObjectiveTo explore the potential mechanisms of action of the modified Duhuo Jisheng Mixture （JDJM） in treating synovial lesions in knee osteoarthritis （KOA）. MethodsA total of 43 male New Zealand white rabbits were randomly allocated into a blank group （n=8） and a model group （n=35）. The KOA model was induced by immobilizing the right hind limb with a high-molecular resin plaster bandage， with a modeling period of 6 weeks， resulting in successful modeling in 32 rabbits. These rabbits were then randomly allocated to the model group， celecoxib group， JDJM group and JDJM+740Y-P group， each consisting of 8 rabbits. The celecoxib group received celecoxib via gavage at a single dose of 0.009 3 g·kg^-1， while the JDJM was administered a single dose of 6.8 mL·kg^-1 （4.515 2 g·kg^-1） of the herbal preparation via gavage. The phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） pathway activator + JDJM group received 4.515 2 g·kg^-1 of the herbal preparation via gavage along with an auricular vein injection of 0.15 μmol·kg^-1 740Y-P. For a period of 6 weeks， the remaining groups received an equal volume of physiological saline via gavage daily. After the medication period， the knee joint pain threshold and circumference were measured， and hematoxylin-eosin （HE） staining was performed to assess the pathological changes in the synovial tissues. Enzyme-linked immunosorbent assay （ELISA） measured the levels of interleukin-1β （IL-1β）， interleukin-6 （IL-18） and tumor necrosis factor-α （TNF-α） in the joint fluid. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to assess the mRNA expression of PI3K， Akt， mTOR， NOD-like receptor protein 3 （NLRP3）， cysteine-requiring aspartate protease-1 （Caspase-1） and gasdermin D （GSDMD） in the synovial tissues. Immunohistochemical （IHC） assay was performed to assess the protein expression of NLRP3， Caspase-1 and GSDMD. Western blot was carried out to analyze the protein expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， NLRP3， Caspase-1 and GSDMD. ResultsCompared to the blank group， the model group showed a significant increase in knee joint circumference and decrease in pain threshold， the synovial tissue pathology score was higher （P<0.05）， and the levels of IL-1β， IL-18， and TNF-α in the joint fluid significantly increased （P<0.01）. PI3K， Akt， mTOR phosphorylation as well as mRNA and protein expression increased （P<0.01）， while the mRNA and protein expression levels of NLRP3， Caspase-1 and GSDMD also significantly increased （P<0.01）. Compared to the model group， the celecoxib and JDJM groups exhibited a significant reduction in knee joint circumference and increase in pain threshold， the synovial tissue pathology score was lower （P<0.05）， and the levels of IL-1β， IL-18， and TNF-α in the joint fluid decreased （P<0.01）. The mRNA and protein expression of p-PI3K， p-Akt， p-mTOR， NLRP3， Caspase-1 and GSDMD were reduced （P<0.01）. Compared to the JDJM group， the JDJM+740Y-P group showed a decrease in the improvement of synovial lesions， an increase in knee joint circumference， and a decrease in pain threshold. The synovial tissue pathology score was lower （P<0.05）， and the levels of IL-1β， IL-18， and TNF-α in the joint fluid were higher （P<0.01）. The mRNA and protein expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， NLRP3， Caspase-1 and GSDMD increased （P<0.01）. ConclusionJDJM is effective in treating KOA. Its mechanism may involve modulating the PI3K/Akt/mTOR pathway in synovial tissues， inhibiting pyroptosis， reducing inflammatory factor release， and protecting bony structures.

Objective To observe the effects of Yiqi Huoxue Prescription in regulating METTL3/HAND2/TGF-β1 signaling axis on rats with qi deficiency and blood stasis syndrome of coronary heart disease.Methods Totally 48 male SD rats were randomly divided into sham-operation group,model group,Yiqi Huoxue Prescription group and Western medicine(isosorbide mononitrate)group,with 12 rats in each group.Except for the sham-operation group,the animal models of coronary heart disease with qi deficiency and blood stasis syndrome were prepared by the method of ligation of the left anterior descending branch of coronary artery combined with swimming exhaustion.After modeling,Yiqi Huoxue Prescription group and the Western medicine group were gavaged for 28 d.The general conditions of the rats were observed,and electrocardiograms and echocardiograms were collected to detect the cardiac function of the rats,blood rheology was used to detect the degree of blood stasis,HE staining was used to observe the morphology of myocardial tissue,immunohistochemistry was used to detect the expression of METTL3,TGF-β1,Smad2 and Smad3 in myocardial tissue,RT-PCR and Western blot were used to detect the expression of METTL3,HAND2,TGF-β1,Smad2 and Smad3 mRNA and protein in myocardial tissue.Results Compared with the sham-operation group,the model group rats showed symptoms of qi deficiency such as mental lethargy,slow movement,drowsiness,and decreased body mass,significant elevation of ST segment on electrocardiogram,the left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS)decreased(P＜0.01),the left ventricular end diastolic volume(LVEDV),left ventricular end systolic volume(LVESV),left ventricular end diastolic diameter(LVEDd)and left ventricular end systolic diameter(LVEDs)increased(P＜0.01),the red blood cell aggregation index,Carcasson viscosity and whole blood viscosity increased(P＜0.01);with enlargement and necrosis of myocardial cells,disordered arrangement of myocardial fibers,and infiltration of inflammatory cells into myocardial tissue,the mRNA and protein expressions of METTL3,TGF-β1,Smad2 and Smad3 in myocardial tissue increased(P＜0.05,P＜0.01),while the mRNA and protein expression of HAND2 decreased(P＜0.01).Compared with the model group,the Yiqi Huoxue Prescription group and the Western medicine group showed significant improvement in the mental state of rats,significant ST segment regression on electrocardiogram,LVEF and LVFS increased(P＜0.01),LVEDV,LVESV,LVEDd and LVEDs decreased(P＜0.05,P＜0.01),with red blood cell aggregation index,Carcasson viscosity and whole blood viscosity decreased(P＜0.01);the arrangement of myocardial fibers was neat,the structure and morphology of myocardial cells were improved,the infiltration of inflammatory cells was reduced,the mRNA and protein expressions of METTL3,TGF-β1,Smad2 and Smad3 in myocardial tissue decreased(P＜0.05,P＜0.01),and the mRNA and protein expression of HAND2 increased(P＜0.05,P＜0.01).Conclusion Yiqi Huoxue Prescription can alleviate the pathological damage of myocardial tissue in rats with coronary heart disease of qi deficiency and blood stasis syndrome,improve heart function and inhibit inflammatory response.Its mechanism may be related to the regulation of the METTL3/HAND2/TGF-β1 signaling axis.

Extrachromosomal circular DNA(eccDNA)is a special kind of circular DNA molecules in nucleus of eukaryotes.These molecules are derived from chromosome genomes,and can independently participate in a variety of cell physiological or pathological processes,which are closely associated with the occurrence and development of tumors.The discovery of eccDNA can be traced back to the 1970s,and with the development of molecular biology technology,researchers have gradually revealed its impor-tant function in cancer.The formation of eccDNA is a complex process involving multiple mechanisms,including breakage-fusion-bridge cycle,chromothripsis,translocation-deletion-amplification,episome model and fork stalling and template switching,etc.eccDNA,especially large molecular weight eccDNA(＞1 Mb,also known as ecDNA),plays an important role in promoting the amplification and activation of oncogenes,driving tumor heterogeneity and drug resistance,and has shown broad application potential as a molecular marker for tumor liquid biopsy.This review summarizes the exploration history,biogenesis,functions in tumor biology and detection,specificity in different types of tumors and analysis methods of eccDNA,and also discusses its application value in clinical diagnosis and treatment of tumors,so as to provide reference for further exploration of eccDNA in basic research and clinical practice of tumors.

Objective To describe the clinical features of patients with primary sclerosing cholangitis(PSC)in China based on a nationwide multicenter patient cohort,and to investigate the risk factors for prognosis.Methods A retrospective cohort study was conducted among the patients with a confirmed diagnosis of PSC based on the electronic medical record system of seven grade A tertiary hospitals across the country,and related data were extracted.The Mann-Whitney U test was used for comparison of continuous data between groups,and the chi-square test was used for comparison of categorical data between groups.The Kaplan-Meier method was used to estimate liver transplant-free survival,and the log-rank test was used for comparison of survival rate between PSC patients with different features.The Cox regression model was used to identify independent risk factors for the prognosis of PSC patients and the interactions between key factors.Results A total of 107 patients were enrolled,among whom 55.6%(55/99)had large-duct PSC and 29.0%(31/107)had comorbidity with inflammatory bowel disease(IBD).The positivity rate of anti-neutrophil cytoplasmic antibody(ANCA)was 32.9%(24/73),and 50.0%(40/80)of the patients had an increase in IgG/IgM.The median symptom-to-diagnosis interval was 1 year(<1-4.0),and 38.3%(41/107)of the patients had progressed to decompensated cirrhosis at the time of diagnosis.The median liver transplant-free survival time was 114 months(95%confidence interval[CI]:62-166),with a 5-year survival rate of 65.7%.The multivariate analysis showed that an increase in total bile acid(TBA)(hazard ratio[HR]=1.006,95%CI:1.002-1.010,P=0.001)and a prolonged symptom-to-diagnosis interval(HR=1.252,95%CI:1.059-1.480,P=0.009)were independent risk factors for prognosis.The interaction analysis showed that compared with the female patients with TBA<50 μmol/L,both male and female patients with TBA≥50 μmol/L had a significant increase in the risk of liver transplantation or death(male:HR=16.563,95%CI:2.103-130.449,P<0.001;female:HR=17.009,95%CI:2.113-136.934,P<0.001),and compared with the patients with an age of<45 years and a TBA level of<50 μmol/L,the patients with an age of≥45 years and a TBA level of≥50 μmol/L had a significant increase in the risk of liver transplantation or death(HR=10.729,95%CI:1.325-86.859,P=0.026).Compared with the female patients with an symptom-to-diagnosis interval of≤2 years,the male patients with a symptom-to-diagnosis interval of>2 years had an increased risk of liver transplantation or death(HR=4.825,95%CI:1.725-13.644,P=0.003),and compared with the patients with an age of<45 years and a symptom-to-diagnosis interval of≤2 years,the patients with an age of<45 years and a symptom-to-diagnosis interval of>2 years had an increased risk of liver transplantation or death(HR=4.983,95%CI:1.366-18.173,P=0.015).Conclusion Compared with the reports from Western countries,large-duct PSC is also the main type of PSC in China,but with a relatively low proportion,and there is also a relatively low proportion of patients with IBD or positive ANCA.An increase in TBA and a prolonged symptom-to-diagnosis interval are independent risk factors for prognosis,with significant interactions with age and sex.This suggests that early screening and intervention should be enhanced to improve prognosis.

[Objective]To analyze the clinical characteristics,efficacy of growth hormone(GH)therapy,and follow-up of a child with Rauch-Steindl syndrome(RAUST)caused by NSD2 gene mutation,aiming to enhance pediatricians'understanding of this disorder.[Methods]We summarized the clinical features,gene test results,outcomes of GH therapy,and follow-up data of a child with RAUST syndrome caused by NSD2 mutation admitted to the Pediatric Endocrinology Department of Sun Yat-sen Memorial Hospital in April 2017,and then conducted a comparative analysis with relevant literature.[Results]The 2.9-year-old boy at initial visit was born prematurely at 36 weeks of gestation,with a birth weight of 1.7 kg and a body length of 42.0 cm.Clinical manifestations included intrauterine growth retardation,delayed language and motor development,extreme short stature(82.0 cm,-3.7 SD),emaciation,and distinctive facial features(triangular face,narrow jaw,prominent forehead,arched eyebrows,sparse eyebrows,high anterior hairline,crowded dentition),accompanied by bilateral cryptorchidism.Bone age was delayed by 1.4 years.Karyotyping and chromosomal microarray analysis were normal.GH therapy initiated at 3.8 years old yielded annual growth rates of 4.9-6.6 cm/year.When the treatment was discontinued at the age of 8.0,the boy's height was 113.7 cm(-3.0 SD),with subsequent decline in growth velocity.Whole exome sequencing in July 2024 identified a frameshift variant c.4028del(p.Pro1343Glnfs*49)in NSD2,which was confirmed as de novo pathogenic variation by parental Sanger sequencing.[Conclusions]This study reports the clinical features of RAUST syndrome caused by NSD2 mutation and explores the long-term efficacy of GH therapy.The findings contribute to a better understanding of this rare syndrome and further optimize its diagnosis and management.

AIM To explore the clinical effects of Supplemented Buyang Huanwu Decoction on postoperative patients with lumbar vertebral fracture complicated with spinal cord injury due to Qi Deficiency and Blood Stasis Pattern.METHODS One hundred and twenty patients were randomly assigned into control group(60 cases)for 6-week intervention of conventional treatment,and observation group(60 cases)for 6-week intervention of both Supplemented Buyang Huanwu Decoction and conventional treatment.The changes in clinical effects,TCM syndrome scores,spinal cord conduction signals(SEP amplitude,MEP amplitude),serum neurotrophic factors(NGF,IGF-1,BDNF),coagulation and inflammatory indices(PT,APTT,TNF-α,IL-1 β)and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P＜0.05).After the treatment,the two groups displayed decreased TCM syndrome scores,TNF-α,IL-1β(P＜0.05),increased spinal cord conduction signals,coagulation and inflammatory indices(P＜0.05),and shortened PT,APTT(P＜0.05),especially for the observation group(P＜0.05).No significant difference in incidence of adverse reactions was found between the two groups(P＞0.05).CONCLUSION For the patients with lumbar vertebral fracture complicated with spinal cord injury due to Qi Deficiency and Blood Stasis Pattern,Supplemented Buyang Huanwu Decoction can safely and effectively promote neurological function recovery.

Objective To analyze the imaging features of cerebral small vessel disease in patients with type 2 diabetes mellitus by multimodal MRI.Methods The clinical data of 160 patients with cerebral small vessel disease admitted to our hospital from January to December 2020 were retrospectively analyzed.According to whether they were complicated with type 2 diabetes mellitus,they were divided into the diabetic group and the non-diabetic group,with 80 cases in each group.Both groups underwent multimodal MRI scans.And the severity of lacunar infarction,the severity of subcortical and periventricular white matter lesions,white matter integral and cerebral microbleeds of patients in the two groups were compared.Results The severity of lacunar infarction(χ2=34.076,P=0.001),subcortical white matter lesions(χ2=25.000,P=0.001),periventricular white matter lesions(χ2=22.895,P=0.001)and white matter integral(t=12.370,P=0.001)of patients in the diabetic group were significantly higher than those in the non-diabetic group.No cerebral microbleeds were detected in either group of patients.Conclusion Patients with cerebral small vessel disease and type 2 diabetes mellitus show characteristic multimodal MRI changes.The increase in the number of lacunar infarction lesions and the aggravation of white matter lesions can be used as the characteristic imaging basis for the diagnosis of type 2 diabetes mellitus related cerebral small vessel disease.

[Objective]To investigate and explore the characteristics and influencing factors of glucose metabolism in children with β-thalassemia major(β-TM)after allogeneic hematopoietic stem cell transplantation(allo-HSCT).[Methods]The follow-up data of 41 patients with β-TM who underwent HSCT at Hematopoietic Stem Cell Transplantation Department of Children's Medical Center of Sun Yat-sen Memorial Hospital,Sun Yat-sen University were retrospectively analyzed.Their glucose metabolism characteristics were evaluated through laboratory tests and the related influencing factors were analyzed.[Results]In the study,41.46％(17/41)of patients developed abnormal glucose homeostasis after HSCT.Among them,82.35％(14/17)characterized by insulin resistance,but no cases of diabetes mellitus were found.The results of insulin releasing test and oral glucose tolerance test(OGTT)showed that 45.00％(9/20)of patients had abnormal insulin releasing curve and 40.0％(8/20)had delayed serum glucose peak.The average age of HSCT in abnormal glucose homeostasis group was significantly older than that in the normal glucose homeostasis group[(8.8±3.9)years old vs(6.0±3.1)years old,P=0.015].[Conclusions]Patients with β-TM after HSCT may develop abnormal glucose homeostasis,consists largely of insulin resistance.The elder age of HSCT(≥7 years old)is a risk factor for abnormal glucose homeostasis in β-TM patients after HSCT.It is recommended to regularly monitor glucose metabolism indicators in β-TM children after HSCT,especially in elderly transplant recipients.

Objective:To analyze the predictive role of WT1 expression levels pre-and early post-transplantation on relapse and overall survival(OS)in patients with acute myeloid leukemia(AML)undergoing allogeneic hematopoietic stem cell transplantation(allo-HSCT)during their first complete remission(CR1).Methods:A retrospective analysis was conducted on the clinical data of 107 adult AML patients who underwent allo-HSCT during their CR1 at our center between May 2012 and December 2021.The predictive role of bone marrow WT1 expression levels before transplantation and at 3 and 6 months post-transplantation on relapse and OS was explored in combination with relevant clinical factors.Results:The median follow-up time for the 107 patients was 70(range:11-117)months.Among the patients,15 cases died.Kaplan-Meier survial analysis showed that the 3-year overall survival(OS)rate was 85.0％.20 patients experienced relapse,with a median time to relapse of 8(range:0.5-44)months and a l-year cumulative relapse rate of 13.1％.The overall median value of WT1 before transplantation,3 months after transplantation,and 6 months after transplantation was 0.26％(range:0％-23.64％),with an upper quartile value of 0.74％.No statistically significant differences in WT1 expression levels were observed among the pre-transplantation,3-month post-transplantation,and 6-month post-transplantation time points(P=0.227).Univariate analysis showed that patients with WT1 levels＞0.74％at 3 months post-transplantation had a higher 1-year relapse rate(P=0.029)and lower 3-year OS rate(P＜0.001)compared to patients with WT1 levels ≤0.74％.Other significant factors affecting 1-year relapse included stem cell source(P=0.041)and chronic graft-versus-host disease(cGVHD)(P=0.013).For 3-year OS,additional influencing factors were genetic high risk(P=0.048)and stem cell source(P=0.016).Multivariate analysis revealed that WT1 level＞0.74％at 3 months post-transplantation had a trend to affect 1-year relapse rate(HR=3.309,95％CI:0.958-11.431,P=0.058),while the absence of cGVHD was an independent risk factor for 1-year relapse(HR=3.473,95％CI:0.749-16.100,P=0.037).Only WT1 level＞0.74％at 3 months post-transplantation was an independent risk factor for 3-year OS(HR=6.886,95％CI:2.402-19.738,P＜0.001).Conclusion:High WT1 expression level at 3 months post-transplantation in AML patients undergoing allo-HSCT during CR1 affects the 1-year relapse rate and 3-year OS,and is an independent risk factor affecting 3-year OS.These findings suggest that dynamic monitoring of WT1 expression levels has certain value in prognostic assessment of AML patients who received allo-HSCT during CR1.

Objective To analyze the risk factors and clinical outcomes of cognitive impairment in elderly patients with heart disease after surgery.Methods A total of 156 patients with heart valve diseases undergoing coronary artery bypass surgery admitted to the First Affiliated Hospital of H ainan Medical University from October 2021 to December 2023 were prospectively recruited.At 7 d postoperatively,Montreal cognitive assessment(MoCA)scale was used to assess their cogni-tive function,and based on MoCA score＜26 or not,they patients were divided into a cognitive impairment group(n=61)and a control group(n=95).The clinical features were compared be-tween the two groups,and the risk factors for cognitive impairment were analyzed.All patients were followed up for 1 year to compare the outcomes of the two groups.Results The cognitive impairment group had significantly advanced age,larger proportions of concomitant chronic respiratory diseases and sarcopenia,increased ratio of undergoing open heart surgery,and elevated incidence of intraoperative hypotension than the control group(P＜0.05,P＜0.01).Multivariate logistic regression analysis indicated that age,chronic respiratory diseases,sarcopenia,open heart surgery,and intraoperative hypotension were independent risk factors for postoperative cognitive impairment in elderly patients with heart diseases(OR=1.081,95％CI:1.007-1.161,P=0.030;OR=2.538,95％CI:1.062-6.066,P=0.036;OR=2.650,95％CI:1.174-5.985,P=0.019;OR=3.104,95％CI:1.391-6.929,P=0.006;OR=3.478,95％CI:1.298-9.322,P=0.0013).There was no statistical difference in preoperative MoCA scores between the two groups(27.90±1.40 vs 28.20±1.40,P=0.195).The MoCA score at 7 d and 6 and 12 months after surgery were obviously lower in the cognitive impairment group than the control group(22.90±1.27 vs 27.73±1.08,P=0.000;24.72±1.66 vs 27.73±1.23,P=0.000;25.48±1.73 vs 27.62±1.22,P=0.000).Age was identified as an independent factor affecting the outcome of cognitive function in the patients(OR=1.168,95％CI:1.035-1.318).Conclusion The incidence of postoperative cog-nitive impairment is relatively high in elderly patients with heart disease.So,relevant risk factors should be addressed to strengthen the prevention and management.