1.Effect of RUNX3 on the activation, proliferation, and migration capabilities of hepatic stellate cells
Hui LING ; Xianchen WANG ; Junbo YOU ; Jiahao FAN ; Xiao CUI ; Jiming SHA ; Liquan YU
Acta Universitatis Medicinalis Anhui 2026;61(2):277-284
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.
2.RUNX3 regulates FAP to influence the proliferation of mouse lung primary fibroblasts
Junbo YOU ; Xianchen WANG ; Hui LING ; Jiahao FAN ; Qi CHEN ; Hui TAO ; Jiming SHA
Acta Universitatis Medicinalis Anhui 2026;61(4):606-611
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.
3.Assessment of perioperative pulmonary fluid volume using remote dielectric sensing (ReDSTM) non-invasive lung fluid measurement technology in transcatheter tricuspid valve-in-valve implantation: The first case report
Yuliang LONG ; Yuan ZHANG ; Xiaochun ZHANG ; Peng WANG ; Xiaotong CUI ; Wenzhi PAN ; Daxin ZHOU ; Junbo GE
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(04):571-574
One of its primary surgical treatments of tricuspid regurgitation is tricuspid valve biological valve replacement. Catheter tricuspid valve-in-valve implantation is a novel interventional alternative for biological valve failure. The non-invasive lung fluid measuring device remote dielectric sensing (ReDSTM) has been increasingly incorporated into clinical practice as a means of monitoring chronic heart failure in recent years. This report describes the process and outcomes of the first instance of perioperative lung fluid volume evaluation following transcatheter tricuspid valve implantation utilizing ReDSTM technology. The patient has a short-term, substantial increase in postoperative lung fluid volume as compared to baseline.
4.Free inferior gluteal perforator flap for immediate breast reconstruction: a case report and literature review
Lan MU ; Junbo PAN ; Guisheng HE ; Xiuxiu CHEN ; Tao SONG ; Haohao JIAN ; Zuolei YANG ; Sisi WANG ; Huangfu WU ; Yazhen ZHANG ; Kun XIE ; Chuanwei SUN ; Wentian XU ; Guanghua FU ; Junzhang CHEN ; Bo LI ; Hengyu CHEN ; Yilian XU ; Mingmei HE ; Jinhui HUANG ; Peng LI
Chinese Journal of Microsurgery 2025;48(2):161-166
Objective:To explore the possibility of using a inferior gluteal artery perforator flap (IGAPF) for breast reconstruction in the patient who did not have suitable donor site in back and abdomen.Methods:In November 2024, a 25-year-old unmarried and childless woman with right breast cancer received immediate right breast reconstruction by a right free IGAPF after modified right mastectomy in the Department of Breast and Thyroid Surgery, Second Affiliated Hospital of Hainan Medical University. The locations of perforators were confirmed by both Multi-detector computed tomography angiography (MDCTA) and portable Doppler blood flow detector before surgery. The IGAPF was designed to take the inferior gluteal wrinkle as the lower edge, the axis of the flap was parallel to the inferior gluteal wrinkle, and the width of the flap was estimated where the incision could be directly closed. The size of right IGAPF was 6.0 cm×19.0 cm. Sharp dissection was performed between the sarcolemma and muscle fibres of gluteus, then the perforators were dissected along the direction of muscle fibres of gluteus. The vascular pedicle was kept at about 8.0 cm in length. The diameter of artery was about 2.0 mm and that for the veins was about 1.5 mm. End-to-end anastomoses with the right thoracodorsal artery and vein were successfully carried out. The donor site was directly closed, and it was hidden in the inferior gluteal wrinkle. Postoperative outpatient clinical review was made.Results:Pathological examination reported: an invasive carcinoma of right breast, axillary lymph node metastasis (2/10). The patient recovered well and the flap survived without any complication, i.e. ischemic necrosis, infection and haematoma. The patient was off-bed at 3 days and discharged at 13 days after surgery. At the 40 days of postoperative follow-up, the patient achieved a good recovery and the lower limb activity was not affected by the surgery. The patient was satisfied with the reconstructed breast and donor site recovery. The patient followed with scheduled chemotherapy and subsequent radiotherapy. The volume of reconstructed breast was smaller than the other breast, of which the patient was fully informed before the surgery.Conclusion:A free IGAPF provides an alternative donor sites for achieving a breast reconstruction due to the reliable pedicle vessels and invisible donor scars.
5.Identification of endothelial cell key genes associated with pathogenesis and invasion of human venous malformations using single-nucleus RNA sequencing-based co-expression network analysis
Wenbo LIU ; Junjie LIN ; Meijuan ZHANG ; Chunjie YUAN ; Xiaojuan FENG ; Wenting JIAO ; Junbo QIAO ; Wenqiu WANG ; Bin FANG ; Changkuan CHEN
Chinese Journal of Preventive Medicine 2025;59(4):458-467
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.
6.Association between platelet function and in-hospital mortality in patients with acute aortic dissection undergoing emergency surgical procedures
Haixiu XIE ; Feng YANG ; Xiaomeng WANG ; Xing HAO ; Feilong HEI ; Junbo FENG ; Fuhua HUANG ; Xiaotong HOU
Chinese Journal of Thoracic and Cardiovascular Surgery 2025;41(5):276-283
Objective:To evaluate the correlation between preoperative platelet function and in-hospital mortality in patients with acute Stanford A aortic dissection(ATAAD) undergoing emergency surgery.Methods:ATAAD emergency surgical patients who underwent preoperative thromboelastography(TEG) examination at three cardiovascular disease hospitals from January 2018 to December 2023 were consecutively selected in this study. The patients were divided into two groups according to whether the patient survived to discharge.Results:867 patients were included in this study. The in-hospital mortality was 11.2%(97 cases). Compared with the DG group(97 cases), the age, pericardial effusion volume, lactate concentration, and MA value of patients in the SG group(770 cases) were higher( P<0.001). Factors independently associated with in-hospital mortality were age(>60 years old), coronary hypoperfusion, pericardial effusion volume(>200.00 ml), and MA value(<60.6 mm). There was a correlation between MA value and in-hospital mortality( P=0.012), and the mediating effect between MA value and platelet count was not significant. Conclusion:There is a correlation between preoperative platelet function and in-hospital in ATAAD patients, and improving platelet function may be one of the important ways to improve the clinical prognosis of those patients.
7.A clinical study of deep learning-based artificial intelligence model for precise identification of early gastric cancer boundaries in narrow-band and near focus narrow-band endoscopic images
Xiaozhe MAO ; Kaicheng HONG ; Yunbo GUO ; Bilin WANG ; Junbo LI ; Rui LI
Chinese Journal of Digestive Endoscopy 2025;42(9):707-714
Objective:To develop and validate artificial intelligence (AI) models based on deep learning for precise boundary identification of early gastric cancer (EGC) in narrow-band imaging (NBI) and near focus narrow-band imaging (NF-NBI) endoscopic images.Methods:Endoscopic submucosal dissection (ESD) images from 282 patients diagnosed as having EGC by postoperative pathology at the Department of Gastroenterology, the First Affiliated Hospital of Soochow University were retrospectively collected from February 2016 to June 2024. The images were randomly divided into the training set and the validation set at an approximate 8∶2 ratio. In the NBI modality, 980 images from 171 patients were used for training, 235 images from 61 patients were used for validation. In the NF-NBI modality, 1 273 images from 128 patients were used for training, and 373 images from 35 patients were used for validation. This study trained a total of six convolutional neural network (CNN) models: two independent CNN1 models, two independent CNN2 models, and two fused CNN3 models. Using expert-delineated EGC boundaries based on post-ESD pathological findings as the gold standard, the intersection over union (IOU) value of the CNN3 models was compared against junior (<5 years experience), mid-level (5-10 years), and senior (>10 years) endoscopists.Results:In NBI validation set, the IOU value of CNN3 model was significantly higher than that of junior (0.732 VS 0.489, Z=11.528, P<0.001) and mid-level endoscopists (0.732 VS 0.521, Z=11.184, P<0.001). However, no significant difference was observed between CNN3 model and senior endoscopists (0.732 VS 0.739, Z=0.593, P=0.554). Similarly, in NF-NBI validation set, CNN3 model outperformed junior (0.757 VS 0.537, Z=15.944, P<0.001) and mid-level endoscopists (0.757 VS 0.597, Z=9.722, P<0.001), while matching senior endoscopists (0.757 VS 0.769, Z=0.854, P=0.394). Conclusion:The fused CNN3 model achieves senior expert-level accuracy in delineating EGC boundaries in both NBI and NF-NBI images, demonstrating potential to assist less-experienced endoscopists in precise identification of EGC boundaries.
8.Determination of 10 drugs including morphine in hair using triple quadrupole tandem mass spectrometry
Meiting LIN ; Xueyan ZHU ; Jian LI ; Quanlu DOU ; Xin WANG ; Ping XIANG ; Junbo ZHAO
Chinese Journal of Forensic Medicine 2025;40(3):330-337
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.
9.Analysis of the clinical efficacy of percutaneous transluminal angioplasty in treating dysfunction of autogenous arteriovenous fistula
Jingcun SU ; Haiyang WANG ; Junbo LI ; Huawen XIA
Journal of Practical Radiology 2025;41(4):656-659
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.
10.Identification of endothelial cell key genes associated with pathogenesis and invasion of human venous malformations using single-nucleus RNA sequencing-based co-expression network analysis
Wenbo LIU ; Junjie LIN ; Meijuan ZHANG ; Chunjie YUAN ; Xiaojuan FENG ; Wenting JIAO ; Junbo QIAO ; Wenqiu WANG ; Bin FANG ; Changkuan CHEN
Chinese Journal of Preventive Medicine 2025;59(4):458-467
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.

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