ObjectiveTo investigate the role of runt-related transcription factor 3 （RUNX3） in transforming growth factor-β₁ （TGF-β₁）-induced activation of mouse primary pulmonary fibroblasts （PFs）， and its effects on fibroblast activation protein （FAP） expression， cell proliferation， and collagen synthesis. MethodsPFs were isolated from C57BL/6 mice and cultured. A RUNX3 knockdown model was established using small interfering RNA （siRNA）. Cells were assigned to the control group （Control）， TGF-β₁-treated group （TGF-β₁）， negative control group （TGF-β₁+siRNA-NC）， and RUNX3-silenced group （TGF-β₁+si-RUNX3）. In addition， a RUNX3 overexpression rescue experiment was performed based on TGF-β₁ stimulation. Protein and mRNA levels of RUNX3， FAP， and typeⅠcollagen （COL1A1） were measured by Western blot and reverse transcription quantitative real-time PCR （RT-qPCR）. Cell proliferation was assessed using CCK-8 and EdU assays. Co-expression of COL1A1 and FAP was examined by double immunofluorescence staining. ResultsCompared with the Control group， RUNX3， FAP， and COL1A1 expression levels were upregulated in PFs in the TGF-β₁ group （P<0.01）. The CCK-8 assay showed that the absorbance value was reduced in the RUNX3 knockdown group compared with the negative control group （P<0.01）. Consistently， the EdU assay demonstrated a lower proportion of EdU-positive cells in the RUNX3 knockdown group than in the negative control group （P<0.01）. Immunofluorescence double staining revealed decreased fluorescence intensities of COL1A1 and FAP in the RUNX3 knockdown group relative to the negative control. Under RUNX3 overexpression conditions， these fluorescence signals exhibited a partial rebound （P<0.01）. ConclusionRUNX3 in TGF-β1-induced PFs may promote cell proliferation and collagen synthesis by positively regulating FAP expression. Targeting the RUNX3/FAP axis may represent a potential therapeutic strategy for pulmonary fibrosis.

ObjectiveTo investigate the effects of targeted silencing of Runt-related Transcription Factor 3 （RUNX3） on the proliferation and migration of Mouse Hepatic Stellate Cells （HSCs）， as well as subsequent collagen deposition. MethodsMouse hepatic stellate cell line （JS-1） was selected and then morphologically observed and identified under a microscope. After the cells had fully adhered， they were treated with 5 ng/mL of transforming growth factor beta 1 （TGF-β₁） for 24 hours to induce hepatic stellate cell activation. Furthermore， a RUNX3 silencing model was established using RUNX3 lentiviral infection. The experiment was divided into four groups： Control group， TGF-β₁ group， TGF-β₁+siRNA-NC group， and TGF-β₁+siRNA-RUNX3 group. Protein expression changes of RUNX3， alpha-smooth muscle actin （α-SMA）， and Alpha 1 type I collagen （Collagen I） were detected using Western blot method. Cellular immunofluorescence assays were employed to investigate the deposition changes of α-SMA and RUNX3 in hepatic stellate cells. RT-qPCR was utilized to examine the mRNA expression changes of RUNX3， α-SMA， and Collagen I. The proliferative capacity of hepatic stellate cells was assessed using Edu staining. The migratory ability of hepatic stellate cells was evaluated through wound healing assays and Transwell migration experiments. ResultsCompared with Control group， a significant elevation in RUNX3 was observed in the TGF-β₁-induced activated HSCs （P<0.01）. Meanwhile， the protein and mRNA levels of fibrosis-related markers and α-SMA and Collagen I were significantly upregulated （P<0.001）. Additionally， the proliferation and migration capabilities of HSCs were significantly enhanced （P<0.001）. In contrast， when compared to TGF-β₁+siRNA-NC group， TGF-β₁+siRNA-RUNX3 group exhibited a notable decrease in RUNX3 and other related indicators， such as the protein and mRNA levels of α-SMA and Collagen I （P<0.05）. Concurrently， the proliferation and migration capabilities of HSCs were significantly inhibited in TGF-β₁+siRNA-RUNX3 group （P<0.01）. ConclusionSilencing RUNX3 can inhibit the deposition of collagen and the proliferation and migration of hepatic stellate cells. Conversely， RUNX3 promotes the proliferation and migration capabilities of HSCs， thereby facilitating the activation of HSC.

ObjectiveTo observe the effectiveness and safety of Jiuzihuichun decoction combined with spleen- strengthening moxibustion in patients with asthenospermia infertility with spleen-kidney deficiency pattern. MethodsA total of 82 patients with asthenospermia of spleen-kidney deficiency pattern in Shanghai Seventh People's Hospital were included. The patients were randomly divided into an observation group and a control group， with 41 patients in each group. The control group received oral administration of WuziYanzong pills combined with spleen-strengthening moxibustion. The dosage of Wuzi Yanzong pills was 1 bag each time， and it was taken twice a day. The spleen-strengthening moxibustion was carried out once a week. The observation group， on the other hand， took Jiuzi Huichun decoction orally combined with spleen-strengthening moxibustion. The Jiuzi Huichun Decoction was taken 200 mL each time， twice a day， with one dose in the morning and one in the evening. The spleen-strengthening moxibustion for the observation group was also performed once a week. The treatment course for both groups was 12 weeks， and they were followed up for an additional 12 weeks. During the treatment process，12 cases were either lost to follow-up or excluded. Eventually， 70 cases were available for evaluation，with 35 cases in the control group and 35 cases in the observation group. The pregnancy status of the patients' spouse within 6 months was recorded. The traditional Chinese medicine （TCM） syndrome scores of spleen-kidney deficiency pattern before and after treatment were evaluated. The semen volume，semen routine parameters，normal sperm morphology，sperm DNA fragmentation index，seminal plasma fructose，seminal plasma acid phosphatase， and seminal plasma α-glucosidase levels of the two groups were detected before and after treatment. In addition， the safety indicators related to liver and kidney functions of the two groups were detected before and after treatment. ResultsDuring the 6-month observation period， when compared with the situation before treatment in their respective groups，the semen volume of the observation group and the control group increased. In contrast， the sperm concentration，sperm motility，proportion of a+b-grade sperm，normal sperm morphology，seminal plasma fructose，seminal plasma acid phosphatase，seminal plasma α-Glucosidase，the proportion of a-grade sperm，linear sperm motility，linear sperm concentration， and linear sperm count all increased significantly（P<0.05）. At the same time， the sperm DNA fragmentation index and the TCM syndrome scores of the spleen-kidney deficiency pattern decreased significantly（P<0.05）. When the observation group was compared with the control group after treatment， the clinical efficacy of the observation group was better（Z=-2.276，P<0.05）. The pregnancy rate of the observation group's spouses was 14.3%，which was higher than the 2.9% of the control group. The sperm motility， the proportion of a+b-grade sperm，seminal plasma fructose，the proportion of a-grade sperm，normal sperm morphology，α-glucosidase， and linear sperm motility in the observation group were higher than those in the control group （P<0.05）. Moreover， the sperm DNA fragmentation index and the TCM syndrome scores of spleen-kidney deficiency pattern in the observation group were lower than those in the control group （P<0.01）. No serious adverse reactions occurred in the two groups，and no abnormalities were found in the safety indicators after treatment. ConclusionJiuzi Huichun decoction combined with spleen-strengthening moxibustion can enhance sperm viability and sperm concentration. It can also improve the TCM-related symptoms of asthenospermia of spleen-kidney deficiency pattern and sperm morphology. Additionally， it can reduce the sperm DNA fragmentation index and regulate the level of seminal plasma bioenzyme in patients with male asthenospermia infertility of spleen-kidney deficiency pattern. Therefore， it is worthy of further promotion and application in clinical practice.

Objective: To investigate the effect of targeted silencing of DNA methyltransferase 3A(DNMT3A) on collagen deposition, proliferation and migration activity of mouse lung fibroblasts(PFs). Methods: In order to ensure the proliferation and migration activity of primary fibroblasts, the lung tissues of neonatal C57 suckling mice were taken, PFs were extracted after being sheared, and the morphology was observed and identified under the microscope. PFs cells were activated by 5 ng/ml TGF-β1for 24 h after cell attachment, and DNMT3A silencing model was constructed by small interfering RNA; The experiment was divided into control group, TGF-β1group, TGF-β1+ siRNA-NC group and TGF-β1+ siRNA-DNMT3A group. The protein expressions of DNMT3A, α-smooth muscle actin(α-SMA) and Collagen Ⅰ were detected by Western blot; Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression changes ofDNMT3A,α-SMAandCollagenⅠ. The proliferation ability of PFs was detected by CCK-8 and EdU staining; the migration ability of PFs was detected by scratch test and Transwell migration test. Results: Compared with the control group, TGF-β1induced the increase of DNMT3A in the activated PFs cell group(P<0.01), the protein and mRNA levels of fibrosis and proliferation related indicators α-SMA and Collagen Ⅰ also increased(allP<0.05), and the proliferation and migration ability of PFs increased(allP<0.000 1). Compared with the siRNA-NC group, the protein expression levels of DNMT3A(P<0.000 1) and related indicators α-SMA(P<0.01) and Collagen Ⅰ(P<0.01) significantly decreased in the DNMT3A silencing group by Western blot, and the mRNA levels ofDNMT3A,α-SMAandCollagenⅠby RT-qPCR also decreased(allP<0.001), and the proliferation(P<0.01) and migration ability(P<0.05) of PFs cells decreased compared with the control group. Conclusion Silencing DNMT3A can inhibit the deposition of collagen and the proliferation of PFs. DNMT3A can promote the proliferation and migration of PFs, and then promote the activation of PFs and the development of pulmonary fibrosis. This process may be regulated by DNA methylation modification.

Objectives:This study aimed to investigate the impact of quantitative flow ratio(QFR)-guided revascularization on outcome of elderly patients with coronary artery disease(CAD)undergoing valve surgery.Methods:We retrospectively analyzed 750 consecutive patients with angiographically confirmed CAD(≥50%stenosis)who underwent valve surgery at Zhongshan Hospital,Fudan University,between January 2016 and December 2021.According to the patients'ages,they were divided into the younger group(age<70 years old,n=532)and the elderly group(age≥70 years old,n=218).Revascularization strategies were evaluated using anatomical(angiography-based)and functional(QFR-based)criteria.Anatomical complete revascularization(CR)was defined as bypass grafting for all lesions with≥70%diameter stenosis in major coronary arteries or≥50%stenosis in the left main coronary artery.Functional CR referred bypass grafting for all lesions with QFR≤0.80.Incomplete revascularization(ICR)was defined as failure to meet CR criteria.According to the anatomical and functional definitions,the younger group and the elderly group were further divided into the incomplete revascularization subgroup and the complete revascularization subgroup respectively.Major adverse cardiovascular events(MACE),including death,myocardial infarction,repeat revascularization,and stroke,were assessed as the composite endpoint.Results:Over a follow-up of(3.7±1.8)years,the overall MACE rate was 13.3%.The younger group exhibited significantly lower MACE rates than the elderly group(10.7%vs.19.7%,P=0.001).In the younger group,anatomical ICR did not increase MACE risk(HR=1.46,95%CI:0.81-2.62,P=0.164),whereas functional ICR significantly increased MACE risk(HR=2.27,95%CI:1.24-4.15,P=0.001).In the elderly group,neither anatomical ICR(HR=1.22,95%CI:0.62-2.41,P=0.540)nor functional ICR(HR=1.52,95%CI:0.78-2.96,P=0.172)was associated with increased MACE risk.Conclusions:In patients undergoing valve surgery with CAD,functional ICR correlated with adverse outcomes in the younger group,whereas neither anatomical nor functional ICR significantly affected prognosis in elderly patients.These findings suggest that a moderately conservative revascularization strategy may be more appropriate for elderly populations.

Objective:To construct life context by modifying instructional settings and explore its application effectiveness in teaching the "dietary survey" practice course.Methods:The investigation was carried out on the senior students majoring in preventive medicine at medical colleges. The study was conducted using a pilot-control methodology. The experimental group ( n=76) engaged in a hands-on activity where students independently selected and weighed food in the campus cafeteria, followed by a paired 24-hour dietary recall survey conducted in class. The control group ( n=84) received traditional teaching method involving the 24-hour dietary recall survey only within the classroom setting. The scores, questionnaires, and qualitative interviews were used to compare the learning outcomes and professional competence of the two groups. SPSS 22.0 was used for chi-square test and t test. Results:Compared with the median scores of the experimental reports and thinking questions of the students in the control group (90 and 70), that median scores of the students in the experimental group were significantly improved (95 and 80) ( P<0.001). The score ratios of the dietary survey in the test paper in the two groups were 85.53% (65/76) (experimental group) and 65.48% (55/84) (control group), showing a statistically significant difference ( P=0.003). Moreover, the results showed that the scores of knowledge, experimental skills, and professional competence in the experimental group were significantly higher than those in the control group ( P<0.01). Over 90% of the students expressed satisfaction with the teaching. Conclusions:The life context-based teaching method applied in dietary survey practice course can improve the teaching effectiveness and professional competence.

Objective:This study aimed to identify key genes in endothelial cell (EC) associated with the pathogenesis and progression of human venous malformations (VMs) through bioinformatics analysis, providing potential biomarkers for early screening and targeted therapy of VMs.Methods:A case-control study was conducted using surgically resected tissue specimens from VMs patients at the Third Affiliated Hospital of Zhengzhou University (from September 2021 to September 2023), with malformed venous tissues as the experimental group and distal normal venous tissues as controls. Single-nucleus RNA sequencing (snRNA-seq) was performed on paired experimental and control samples from four VM patients. High-dimensional weighted gene co-expression network analysis (hdWGCNA), combined with gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), and protein-protein interaction (PPI) network analysis, identified critical genes. Validation experiments included 15 additional VM cases and controls using reverse transcription quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry (IHC), and Western blot.Results:A total of 55 430 nuclei were captured using snRNA-seq, with 30 391 nuclei from the experimental group and 25 039 nuclei from the control group. Cluster analysis identified 22 distinct cell populations, which were annotated into 8 cell types. hdWGCNA revealed four modules associated with invasion, which were enriched in angiogenesis, integrin signaling, and cell adhesion according to GO analysis. KEGG pathway analysis indicated that the PI3K-AKT signaling pathway and focal adhesion are key regulatory mechanisms. PPI network analysis combined with cytoscape identified EGFL7, TEK, and FLT1 as key genes. RT-qPCR results demonstrated that the relative mRNA expression levels of these three genes in the experimental group (6.66±2.31, 1.86±0.62, 3.49±0.58) were significantly higher than those in the control group (1.05±0.14, 1.00±0.14, 1.06±0.25), with statistically significant differences ( t=9.37, 4.27, 11.20, P<0.05). Immunohistochemical analysis showed that the relative protein expression levels of these three genes in the cytoplasm of the experimental group (0.84±0.15, 0.68±0.14, 0.85±0.12) were also significantly higher than those in the control group (0.19±0.05, 0.23±0.06, 0.30±0.05), with statistically significant differences ( t=16.62, 5.93, 11.68, P<0.05). Western blot analysis confirmed that the relative protein expression levels of these three genes in the experimental group (0.35±0.04, 0.36±0.09, 0.31±0.04) were significantly higher than those in the control group (0.19±0.01, 0.13±0.02, 0.14±0.04), with statistically significant differences ( t=7.05, 4.61, 5.93, P<0.05). Conclusion:EGFL7, FLT1, and TEK in EC may play crucial roles in the occurrence and invasion of VMs.

Objective To investigate the efficacy of percutaneous transluminal angioplasty(PTA)in the treatment of dysfunction of autogenous arteriovenous fistula(AVF),and to analyze the factors influencing 12-month patency.Methods The data from maintenance hemodialysis patients who underwent PTA for AVF stenosis were collected.The technical success rate,clinical success rate,complications,and short-to medium-term AVF patency were evaluated.Univariate analysis and binary logistic regression were used to identify predictors of 12-month post-procedural patency.Results A total of 70 patients were included in this study.The technical and clinical success rates were 94.29%and 95.71%,respectively.The 3-,6-,and 12-month post-procedural patency rates of AVF were 97.1%,75.7%,and 61.4%,respectively.Independent protective factors for 12-month AVF patency included age＜60 years[odds ratio(OR)=0.277,95%confidence interval(CI)0.097-0.792]and statin use(OR=0.299,95%CI 0.101-0.887).Independent risk factors included diabetes mellitus(OR=5.167,95%CI 1.824-14.64),AVF usage≥1 year(OR=2.885,95%CI 1.061-7.840),and non-use of aspirin(OR=2.782,95%CI 1.016-7.615).Conclusion PTA is a safe and effective treatment for AVF stenosis,though long-term patency rates require further improvement.

Objective To establish a sensitive,accurate and rapid detection method for 10 drugs and metabolites in hair with triple quadrupole tandem mass spectrometry,addressing the identification of drugs in real hair samples.Methods After cryogenic grinding and ultrasonic extraction,hair was separated by Restek Allure PFPP column(100 mm × 2.1 mm,5 μm).The mobile phase A was 20 mmol/L ammonium acetate,0.1％formic acid and 5％acetonitrile aqueous solution.The mobile phase B was acetonitrile.An electrospray ionization source was used for data acquisition in scheduled MRM mode.Results The method showed good linearity for all analytes within the validated range(R2＞0.995),with the limits of detection ranging from 0.5 to 6 pg/mg,the limits of quantification ranging from 2.5 to 10 pg/mg.Accuracy ranged from 89.1％to 114.6％,with intra-day precision ranging from 0.2％to 11.7％,inter-day precision ranging from 4.0％to 15.8％.the matrix effects ranging from 89.4％to 118.4％,the recoveries ranging from 63.6％～112.1％.Totally 20 cases were detected positive in 196 actual hair samples.Conclusion The time-scheduled scanning method exhibits high stability and sensitivity,enabling high-throughput detection,and is suitable for forensic toxicology laboratories to identify drugs in hair.

Objective To explore the correlation between serum circularRNA-HOMER1(circHOMER1),microRNA(miR)-23a-3p levels with clinical stages and oxidative stress in patients with diabetic retinopathy(DR).Methods From January 2023 to July 2024,75 DR patients treated in Handan Central Hospital were included as the DR group.According to the clinical staging of DR,they were divided into non proliferative DR(NPDR group,n=43)and proliferative DR(PDR group,n=32).In addition,75 patients with simple type 2 diabetes who came to Handan Central Hospital were included as non DR group.The levels of serum circHOMER1,miR-23a-3p,malondialdehyde(MDA),superoxide dismutase(SOD),and reduced glutathione(GSH)were detect-ed.Clinical data of the subjects were collected.The TargetScan website was used to predict the targeting relationship between circHOMER1 and miR-23a-3p.Pearson method was used to analyze the correlation between serum circHOMER1,miR-23a-3p and MDA,SOD,GSH.Univariate and multivariate Logistic regression were used to analyze the influencing factors of progression of DR in type 2 diabetes patients.Receiver operating characteristic(ROC)carve was used to analyze the predictive value of serum circHOMER1 and miR-23a-3p in the progression of DR in patients with type 2 diabetes.Results There was a targeted relationship between circHOMER1 and miR-23a-3p.The serum MDA(28.66±4.52ng/ml)and circHOMER1(1.24±0.16)levels in the DR group were higher than those in the non DR group(16.95±3.27ng/ml,1.02±0.11),while SOD(45.39±7.84U/L),GSH(135.82±21.23μg/mL)and miR-23a-3p(0.88±0.07)levels were lower than those in the non DR group(81.65±11.47U/L,207.44±25.95μg/mL,1.01±0.09),and differences were statistically significant(t=9.813～22.602,all P<0.001).The serum MDA(33.28±4.96ng/ml)and circHOMER1(1.36±0.20)levels in the PDR group were higher than those in the NPDR group(25.23±3.58ng/ml,1.15±0.17),while SOD(34.39±7.15U/L),GSH(113.50±20.17μg/ml)and miR-23a-3p(0.79±0.07)levels were lower than those in the NPDR group(53.27±8.44U/L,152.43±23.99μg/ml,0.94±0.08),and the differences were statistically significant(t=4.906～10.376,all P<0.001).Spearman analysis showed that serum MDA and circHOMER1 were positively correlated with the severity of DR(r=0.533,0.473,all P<0.001),while SOD,GSH,miR-23a-3p were negatively correlated with the severity of DR(r=-0.552,-0.515,-0.529,all P<0.001).Pearson analysis showed that serum circHOMER1 was negatively correlated with miR-23a-3p,SOD,GSH,and positively correlated with MDA(r=-0.475,-0.460,-0.455,0.462,all P<0.001).Serum miR-23a-3p was positively correlated with SOD and GSH,and negatively correlated with MDA(r=0.428,0.437,-0.439,all P<0.001).Logistic regression analysis showed that high MDA,low SOD,low GSH,high circHOMER1,low miR-23a-3p,high FPG and high HbA1c were the risk factors of progression of DR in type 2 diabetes patients(OR=0.214～3.556,all P<0.05).The area under curve(AUC)of serum circHOMER1 and miR-23a-3p alone and jointhy predicting the progression of DR in type 2 diabetes patients were 0.751,0.797 and 0.903 respectively.The combined prediction was higher than that of serum circHOMER1 and miR-23a-3p alone(Z=3.179,2.335,P=0.002,0.020).Conclusion Serum MDA and circHOMER1 levels are higher in DR patients,while serum SOD,GSH and miR-23a-3p levels are lower.Abnormal expression of circHOMER1 and miR-23a-3p in serum is associated with progression of DR and oxidative stress.Combined detection of circHOMER1 and miR-23a-3p in serum can predict the progression of DR in patients with type 2 diabetes.