ObjectiveTo investigate the effect of mitochondrial calcium uniporter （MCU） on the cytoskeleton of pancreatic ductal epithelial cells in a mouse model of acute pancreatitis （AP） induced by caerulein （CAE）， to analyze the role of MCU in the development of AP， and to provide a theoretical basis for clinical treatment. MethodsIn the in vivo experiment， wild-type male C57BL6/J mice， aged 4 weeks， were randomly divided into control group and AP group， with 6 mice in each group. The mice in the AP group were given intraperitoneal injection of CAE to establish a model of AP， and those in the control group were given intraperitoneal injection of an equal volume of normal saline. Serum and pancreatic tissue samples were collected after 24 hours of modeling. HE staining was used to observe pancreatic histopathological changes； Western Blot was used to measure the expression levels of MCU， glutathione peroxidase 4 （GPX4）， and acyl-CoA synthetase long chain family member 4 （ASCL4）； kits were used to measure the serum level of amylase. In the in vitro experiment， the human pancreatic ductal epithelial cell line HPDE6-C7 was co-cultured with CAE for 24 hours to establish an in vitro AP model， and the cells were divided into control group， CAE group， RR （an MCU activity inhibitor） group， CAE+RR group， Fer-1 （an ferroptosis inhibitor） group， CAE+Fer-1 group， Erastin （an ferroptosis inducer） group， and CAE+Erastin group. CCK-8 assay was used to observe the influence of different agents on cell viability； Western Blot was used to measure the expression levels of MCU， GPX4， and ASCL4； immunofluorescence assay was used to measure reactive oxygen species （ROS）， actin cytoskeleton， and monolayer permeability； kits were used to measure the concentrations of malondialdehyde （MDA）， glutathione （GSH）， Fe²⁺， and total iron. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for comparison between two groups. ResultsIn the in vivo experiment， compared with the control group， the AP group had significant increases in pancreatic histopathological score， the serum level of amylase， and the expression levels of MCU and ASCL4， as well as a significant reduction in the expression of GPX4 （all P<0.05）. In the in vitro experiment， compared with the control group， the CAE group had significant increases in the expression levels of MCU and ASCL4， a significant reduction in the expression of GPX4， and significant increases in the concentrations of Fe²⁺， total iron， and MDA， the green fluorescence intensity of ROS， and monolayer permeability， as well as a significant reduction in the concentration of GSH （all P<0.05）， with the presence of actin cytoskeleton disruption. Compared with the CAE group， the CAE+RR group had a significant increase in the expression level of GPX4， a significant reduction in the expression level of ASCL4， and significant reductions in the concentrations of Fe²⁺， total iron， and MDA， the green fluorescence intensity of ROS， and monolayer permeability and a significant increase in the concentration of GSH （all P<0.05）， with alleviation of actin cytoskeleton disruption. Compared with the CAE group， the CAE+Fer-1 group had significant reductions in the concentrations of Fe²⁺， total iron， and MDA， the green fluorescence intensity of ROS， and monolayer permeability and a significant increase in the concentration of GSH （all P<0.05）， with alleviation of actin cytoskeleton disruption. Compared with the CAE group， the CAE+Erastin group had significant increases in the concentrations of Fe²⁺， total iron， and MDA， the green fluorescence intensity of ROS， and monolayer permeability and a significant reduction in the concentration of GSH （all P<0.05）， with aggravation of actin cytoskeleton disruption. ConclusionDuring the onset of AP， MCU mediates oxidative stress-induced ferroptosis and leads to the disruption of the pancreatic ductal epithelial barrier， which may be one of the possible pathogeneses of AP.

Objective To explore the role and related mechanism of neutrophil membrane-coated poly(lactic-co-glycolic acid) (PLGA) nanoparticles (Neu-NP) in cardiac repair after acute myocardial ischemia-reperfusion (MI/R) injury in mice. Methods The male C57 mouse model of acute MI/R injury was established and randomly divided into three groups: PBS control group (injection of 200 μL PBS), NP treatment group (injection of 0.5 mg/mL NP 200 μL), and Neu-NP treatment group (injection of 0.5 mg/mL Neu-NP 200 μL). Neutrophil membranes were extracted and fused with PLGA nanoparticles to construct biomimetic Neu-NP. The in vivo homing ability of Neu-NP was assessed using ex vivo imaging technology in the MI/R injury model, and the expression levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the myocardium were measured using enzyme linked immunosorbent assay one day and three days after administration. Echocardiography was used to determine cardiac function indicators of MI/R injured mice 28 days post-administration. Immunofluorescence staining was used to observe angiogenesis repair and inflammatory cell infiltration in mouse heart tissue. Results Neu-NP, engineered by integrating neutrophil membranes with nanoparticles, inherited surface receptors (TNF-αR and IL-6R) and functioned as decoys for inflammatory targeting. Compared with the PBS control group and NP treatment group, the secretion levels of TNF-α and IL-6 in the damaged myocardium of the Neu-NP treatment group were significantly decreased one and three days after administration (P＜0.05); 28 days after administration, the cardiac ejection fraction in the Neu-NP treatment group was significantly higher than that in the other two groups (P＜0.05). Immunofluorescence staining indicated a significant increase in the proportion of angiogenesis in the myocardial infarction area and a significant reduction in inflammation cell infiltration (P＜0.05). Conclusions Neu-NP plays an important role in cardiac tissue repair after MI/R injury by alleviating inflammatory factors in the damaged area and promoting angiogenesis.

Objective To explore the safety and effects of alpha-lipoic acid (ALA) in patients with ischemic heart failure (IHF). Methods A randomized, double-blind, placebo-controlled trial was designed (ClinicalTrial.gov registration number NCT03491969). From January 2019 to January 2023, 300 patients with IHF were enrolled in four medical centers in China, and were randomly assigned at a 1∶1 ratio to receive ALA (600 mg daily) or placebo on top of standard care for 24 months. The primary outcome was the composite outcome of hospitalization for heart failure (HF) or all-cause mortality events. The second outcome included non-fatal myocardial infarction (MI), non-fatal stroke, changes of left ventricular ejection fraction (LVEF) and 6-minute walking distance (6MWD) from baseline to 24 months after randomization. Results Finally, 138 patients of the ALA group and 139 patients of the placebo group attained the primary outcome. Hospitalization for HF or all-cause mortality events occurred in 32 patients (23.2%) of the ALA group and in 40 patients (28.8%) of the placebo group (HR＝0.753, 95%CI 0.473-1.198, P＝0.231; Figure 1A-1C). The absolute risk reduction (ARR) was 5.6%, the relative risk reduction (RRR) associated with ALA therapy was approximately 19.4% compared to placebo, corresponding to a number needed to treat (NNT) of 18 patients to prevent one event. In the secondary outcome analysis, the composite outcome of the major adverse cardiovascular events (MACE) including the hospitalization for HF, all-cause mortality events, non-fatal MI or non-fatal stroke occurred in 35 patients (25.4%) in the ALA group and 47 patients (33.8%) in the placebo group (HR＝0.685, 95%CI 0.442-1.062, P＝0.091; Figure 1D). Moreover, greater improvement in LVEF (β＝3.20, 95%CI 1.14-5.23, P＝0.002) and 6MWD (β＝31.7, 95%CI 8.3-54.7, P＝0.008) from baseline to 24 months after randomization were observed in the ALA group as compared to the placebo group. There were no differences in adverse events between the study groups. Conclusions These results show potential long-term beneficial effects of adding ALA to IHF patients. ALA could significantly improve LVEF and 6MWD compared to the placebo group in IHF patients.

Objective:To summarize and analyze the clinical efficacy and experience of ultrasound-guided radiofrequency ablation (RFA) in the treatment of Kasabach-Merritt syndrome (KMS).Methods:A retrospective analysis was conducted on the data of pediatric patients with KMS who underwent ultrasound-guided RFA in Department of Hemangioma Surgery, the Third Affiliated Hospital of Zhengzhou University, between March 2018 and March 2024. Preoperative laboratory tests and imageological examination were performed. Under general anesthesia, the working tip of the RFA electrode needle was precisely reached the bottom of the lesion under ultrasound guidance. The electrode needle was then gradually withdrawn until the entire lesion area was covered by hyperechoic signals, indicating complete ablation. Postoperative symptomatic and supportive treatments, such as ice pack application and dressing changes, were administered to the surgical area. Platelet detection was performed immediately after the operation. Complications were closely monitored and regular follow-ups were carried out.Results:A total of 30 pediatric patients were included, comprising 14 males and 16 females, from 10 min to 5 months and 29 d after birth, with a median time of 6 d. Lesions were located in the limbs and trunk in 27 cases, and head and neck region in 3 cases, with lesion volumes ranged from 2.4 cm×2.3 cm×1.2 cm to 14.4 cm×9.3 cm×3.3 cm. The mean preoperative platelet count was 43×10 9/L, among them, the platelet values of 11 cases were (10-30) ×10 9/L, and those of 6 cases were lower than 10×10 9/L, other 13 cases with progressive thrombocytopenia. All patients successfully underwent RFA, achieving complete lesion ablation and normalization of platelet counts postoperatively. Platelet counts recovered to above 300×10 9/L in 15 patients, with no severe complications observed. The RFA area became slightly hardened within 7 d postoperatively but gradually returned to normal after consistent dressing changes for 2 weeks. During the follow-up period of 6 months to 2 years, complete lesion ablation was confirmed, with disappearance of the mass, no recurrence, good local function, mild local scar formation, and satisfactory cosmetic appearance. Conclusion:Ultrasound-guided RFA for KMS has advantages of favorable therapeutic outcomes, minimal tissue damage, no significant complications, and satisfactory cosmetic result.

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.

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 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.

Objective:To investigate the effects of oxycodone on perioperative pain and cognitive function in elderly patients with colorectal cancer.Methods:A prospective randomized-controlled trial was conducted. A total of 88 elderly patients with colorectal cancer who underwent elective laparoscopic surgery at Benxi Central Hospital from June 2022 to December 2023 were selected. According to random number table method, all patients were randomly divided to the experimental group and the control group, with 44 cases in each group. Anesthesia in the experimental group was induced with oxycodone 0.2-0.3 mg/kg, intraoperative oxycodone was maintained at 0.1-0.15 mg·kg -1·h -1; oxycodone was intravenously injected with analgesia pump after surgery. In the control group, anesthesia was induced with sufentanil 0.2-0.3 μg/kg, remifentanil was intraoperatively maintained at 0.1-0.3 μg·kg -1·min -1, sufentanil was intravenously injected with analgesia pump after surgery. Blood pressure and heart rate before anesthesia induction (T 0), at tracheal intubation (T 1), at skin incision (T 2), and at extubation (T 3) of both groups were compared; numerical rating scale (NRS) at 30 minutes, 6 h, 24 h, and 48 h after surgery were scored, the confusion assessment method (CAM) and the mini-mental state examination (MMSE) score at day 1, 3, 7 after surgery were used to assess the postoperative early cognitive decline and other adverse reactions. Results:Finally, 81 patients were included in the study. There were 41 cases in the experimental group including 20 males and 21 females with the age of (76±3) years, and 40 cases in the control group including 19 males and 21 females with the age of (75±2) years. There were no statistically significant differences in gender composition, age, body mass index, operative time, intraoperative blood loss (all P > 0.05). There were statistically differences in blood pressure [(91±8) mmHg (1 mmHg = 0.133 kPa) vs. (89±10) mmHg at T 0, (92±9) mmHg vs. (90±8) mmHg at T 2, (93±9) mmHg vs. (92±9) mmHg at T 3] and heart rate [(70± 15) times/min vs.(69±16) times/min at T 0, (68±12) times/min vs. (67±12) times /min at T 2, (70± 15) times/min vs. (69±14) times/min at T 3] between the experimental group and the control group (all P > 0.05). Blood pressure and heart rate [(101±9) mmHg, (83±15) times /min] at T 1 in the experimental group were higher than those in the control group [(93±11) mmHg, (70±17) times /min], and the differences were statistically significant ( t values were 3.73, 3.77; all P < 0.001). There were no statistically significant differences in NRS scores [(2.6±1.2) scores vs. (2.8±1.1) scores at 30 min, (2.8±1.6) scores vs. (2.9±1.3) scores at 6 h, (1.8±1.2) scores vs. (2.1±1.3) scores at 24 h, and (1.5±0.7) scores vs. (1.5±0.7) scores at 48 h after surgery] between the experimental group and the control group (all P > 0.05). The incidence of postoperative early cognitive decline [4.9% (2/41) vs. 22.5% (9/40)], nausea and vomiting after surgery [7.3% (3/41) vs. 25.0% (10/40)] in the experimental group was lower than that in the control group, and the difference was statistically significant ( χ2 values were 5.36, 4.70; P values were 0.021, 0.030, respectively). Conclusions:The application of oxycodone during laparoscopic surgery can meet the needs of perioperative analgesia and improve the postoperative early cognitive function of elderly patients with colorectal cancer.

Objective:To utilize single-nucleus RNA sequencing(snRNA-seq) technology to investigate the primary cell subpopulations in human limb venous malformations (VMs) tissue and the role of the key gene RhoJ.Methods:Surgical resection specimens of VMs tissues and surrounding normal vein tissues were collected from 100 clinically diagnosed and screened patients with limb VMs at the Department of Hemangioma Surgery of the Third Affiliated Hospital of Zhengzhou University from January 2021 to December 2023. (1) Transcriptome analysis: Three patient samples were randomly selected for snRNA-seq studies, with the surgically removed VMs tissue serving as the experimental group and the surrounding normal vein tissue as the control group. A gene expression matrix for cell nuclei was established, followed by data quality control, dimensionality reduction, clustering, and cell type annotation. Cell-to-cell communication analysis was performed using the R language CellChat package to identify dominant cell subpopulations. The FindMarkers function was utilized to screen for differentially expressed genes (DEGs) between the dominant cell subpopulations of the experimental and control groups, and functional enrichment analysis was conducted. (2) Tissue experiments: An additional 35 patient samples from both the experimental and control groups were randomly selected. The mRNA and protein expression levels of the RhoJ gene were measured using real-time quantitative PCR (RT-qPCR) and Western blotting, respectively. (3) Validation experiments with human umbilical vein endothelial cells (HUVECs): HUVECs were transfected with pcDNA3.1-NC (blank control) and pcDNA3.1-RhoJ (plasmid expression vector carrying the RhoJ gene), respectively. The biological behavior differences between the two groups of cells were examined using the CCK-8 cell proliferation assay, Transwell invasion assay, and Matrigel angiogenesis assay. Measurement data conforming to a normal distribution were expressed as Mean±SD, and comparisons between the two groups were performed using an independent samples t-test. Results:Through CellChat intercellular communication analysis, it was discovered that endothelial cells were the predominant cell subpopulation in both the experimental and control groups, exhibiting strong communication links with other cell subpopulations. In the analysis of DEGs, it was found that the RhoJ gene in endothelial cells was significantly involved in the biological processes of angiogenesis and regulation. In tissue experiments, RT-qPCR and Western bloting results indicated that the relative expression levels of RhoJ mRNA (4.48±1.29 vs. 1.01±0.17) and protein (1.22±0.03 vs. 0.51±0.20) in the experimental group were significantly higher than those in the control group, with statistically significant differences ( P<0.01 for both). The results of the HUVECs validation experiment showed that the cell proliferation, invasion, and angiogenesis abilities of the pcDNA3.1-RhoJ group were significantly enhanced compared to the pcDNA3.1-NC group. Conclusion:Endothelial cells represent the dominant cell subpopulation during the occurrence and locally invasive progression of VMs, playing a crucial role in this process. The RhoJ gene is significant in regulating the biological behavior of VMs endothelial cells.

Objective To explore the diagnostic performance of prostate imaging reporting and data system(PI-RADS)V2.1 score combined with prostate specific antigen(PSA)-related parameters in transition zone prostate cancer(PCa)in patients with PSA levels of 4-20 ng/mL.Methods The clinical data of 129 patients who underwent prostate biopsy were retrospectively analyzed,including age,total prostate specific antigen(tPSA),free prostate specific antigen(fPSA),f/t PSA,prostate volume(PV),and prostate specific antigen density(PSAD).Among 129 patients,52 were diagnosed with transition zone PCa and 77 with benign prostate hyperplasia.The MRI images of all patients were scored according to the PI-RADS V2.1 criteria.The clinical and imaging parameters between transition zone PCa and benign prostate hyperplasia were compared using univariate logistic regression analysis.Independent predic-tors were identified via multivariate logistic regression,and a combined predictive model was constructed.Diagnostic performance of the model was evaluated using the receiver operating characteristic(ROC)curves.Results Univariate logistic regression analysis showed that the transition zone PCa exhibited significantly higher PI-RADS V2.1 score,tPSA,and PSAD than the benign prostate hyperplasia(P＜0.05).Multivariate logistic regression analysis identified PI-RADS V2.1 score and PSAD were independent predictors for predicting transition zone PCa(P＜0.05).Optimal diagnostic thresholds were PI-RADS V2.1 score＞3 points and PSAD＞0.23 ng/mL2.The combined prediction model achieved an area under the curve(AUC)of 0.863,outperforming PI-RADS V2.1 score(AUC=0.821)and PSAD(AUC=0.779)alone(P＜0.05).Conclusion The combination of PI-RADS V2.1 score and PSAD has a high predictive value for transition zone PCa in patients with PSA levels of 4-20 ng/mL.Thresholds of PI-RADS V2.1 score＞3 points and PSAD＞0.23 ng/mLL2 significantly improve diagnostic accuracy.