OBJECTIVES: To elucidate the underlying mechanisms of macrophage-mediated inflammation and tissue injury in diabetic foot ulcer (DFU). METHODS: Skin tissue samples were collected from patients with DFU and with non-DFU. A total of 79 272 high-quality cell transcriptomes were obtained using single-cell RNA sequencing. An unbiased clustering approach was employed to identify cell subpopulations. Seurat functions were used to identify differentially expressed genes between DFU and non-DFU groups, and gene ontology (GO) enrichment analysis was used to reveal gene function. Furthermore, cell-cell communication network construction and ligand-receptor interaction analysis were performed to reveal the mechanisms underlying cellular interactions and signaling regulation in the DFU microenvironment from multiple perspectives. RESULTS: The results revealed a significant expansion of myeloid cells in DFU tissues, alongside a marked reduction in structural cells such as endothelial cells, epithelial cells, and smooth muscle cells. Major cell types underwent functional reprogramming, characterized by immune activation and impaired tissue remodeling. Specifically, macrophages in DFU skin tissues exhibited a shift toward a pro-inflammatory M1 phenotype, with upregulation of genes associated with inflammation and oxidative stress. Cell communication analysis further demonstrated that M1 macrophages served as both primary signal receivers and influencers in the COMPLEMENT pathway mediated communication network, and as key signal senders and mediators in the secreted phosphoprotein 1 (SPP1) pathway mediated communication network, actively shaping the inflammatory microenvironment. Key ligand-receptor interactions driving macrophage signaling were identified, including C3-(ITGAM+ITGB2) and SPP1-CD44. CONCLUSIONS This study establishes a comprehensive single-cell atlas of DFU, revealing the role of macrophage-driven cellular networks in chronic inflammation and impaired healing. These findings may offer potential novel therapeutic targets for DFU treatment.
Humans ; Macrophages/immunology* ; Diabetic Foot/pathology* ; Single-Cell Analysis ; Transcriptome ; Gene Expression Profiling ; Inflammation ; Skin ; Cell Communication ; Signal Transduction ; Cellular Reprogramming

Objective:To study the feasibility of design and harvest of free medial sural artery perforator flap with the "two-point and two-line method".Methods:From September 2022 to June 2023, Suzhou Ruihua Orthopaedic Hospital implemented the "two-point and two-line method" to guide preoperative perforator positioning and flap design. Thirty medial sural artery perforator flaps were successfully harvested with the method, and 21 wounds of hand and 9 of foot and ankle were reconstructed with the flaps. The size of soft tissue defects were 2.5 cm×2.5 cm-7.0 cm×14.5 cm, and the flaps size were 3.0 cm×3.0 cm-7.5 cm×15.0 cm. All donor sites were directly closed or by skin grafting. All patients were entered in 6-15 months of postoperative outpatient follow-up, and the recovery of donor and recipient sites was assessed by the comprehensive evaluation scales. The sensory function of the flaps was evaluated using the sensory function evaluation standard of British Medical Research Council (BMRC).Results:All perforators were successfully located with 47 perforators in total, and all of them were musculocutaneous perforator. It was found that there was 1 perforator in 14 flaps, 2 perforators in 15 flaps and 3 perforators in 1 flap. All 30 flaps survived after surgery, beside 2 flaps that had arterial insufficiency but survived successfully after surgical exploration. All donor sites healed in one stage. Comprehensive evaluation scale of flap was employed to evaluate the flaps and the scores were 84 points to 96 points with an average score of 92.5 points. The excellent and good grades were achieved in 27 flaps and 3 flaps, respectively, with a combined excellent and good rate at 100%. Sensation of the flaps was evaluated by BMRC with 1 flap of S 1, 17 of S 2 and 12 of S 3. Conclusion:The "two-point and two-line method" has been used in design of the perforator flap of medial sural artery. This method is simple and accurate, and is feasible and ideal in design of flaps before surgery.

Objective To investigate the relationship between non-high-density lipoprotein cho-lesterol(non-HDL-C)and cardiovascular disease(CVD)in maintenance hemodialysis(MHD)pa-tients.Methods A total of 124 MHD patients were enrolled and divided into CVD group(53 pa-tients)and non-CVD group(71 patients)based on the presence or absence of CVD.Clinical data be-tween the two groups were compared.Additionally,patients were divided into severe calcification group[coronary artery calcification score(C ACS)≥ 400,40 patients]and non-severe calcification group(CACS＜400,84 patients)based on CACS,and clinical data between these two groups were also compared.Multivariate Logistic regression analysis was used to explore the independent risk fac-tors for CVD in MHD patients,and the predictive performance of related indicators for CVD in MHD patients was assessed using receiver operating characteristic(ROC)curves.Results The levels of serum total cholesterol,low-density lipoprotein cholesterol(LDL-C),and non-HDL-C were higher,while the level of high-density lipoprotein cholesterol(HDL-C)was lower in the CVD group compared with the non-CVD group(P＜0.05).The levels of serum total cholesterol,LDL-C,and non-HDL-C were higher,and the level of HDL-C was lower in the severe calcification group compared with the non-severe calcification group(P＜0.05).Multivariate Logistic regression analysis showed that high levels of LDL-C and non-HDL-C were both independent risk factors for CVD in MHD patients(P＜0.05).ROC curve analysis showed that the areas under the curve for predicting CVD in MHD patients were 0.858 and 0.723 for non-HDL-C and LDL-C,respectively,and non-HDL-C had high-er specificity and Youden index than LDL-C.Conclusion Elevated non-HDL-C level is an inde-pendent risk factor and has high predictive performance for CVD in MHD patients.

BACKGROUND:The occurrence of neuronal axonal injury can result in neurological dysfunction,and the facilitation of axonal elongation is anticipated to play a pivotal role in the treatment of diseases affecting the nervous system.OBJECTIVE:To investigate whether ectomesenchymal stem cells-derived extracellular vesicles can promote neuronal axonal elongation.METHODS:(1)Ectomesenchymal stem cells were obtained from nasal mucosa using the tissue adherence method,and the specific markers of were identified through immunofluorescence.Ectomesenchymal stem cells-derived extracellular vesicles were acquired via ultracentrifugation and identified.(2)Ectomesenchymal stem cells-derived extracellular vesicles(0,0.5,1.0,1.5 mg/mL)were incubated with PC12 cells for 72 hours.The cytotoxicity and proliferation of ectomesenchymal stem cells-derived extracellular vesicles on PC12 cells were assessed using the CCK-8 assay.(3)Ectomesenchymal stem cells-derived extracellular vesicles(1.0 mg/mL)were incubated with PC12 cells or neurons for 72 hours.The changes in axon length were observed using microscopic analysis.The expression levels of axon-related markers β3-tubulin(early stage),growth associated protein 43(middle stage),and neurofilament 200(mature stage)were analyzed through real-time fluorescence quantitative PCR and Western blotting.These investigations aimed to explore the potential of ectomesenchymal stem cells-derived extracellular vesicles in promoting neurite elongation within PC12 cells or neurons.RESULTS AND CONCLUSION:(1)The majority of the acquired ectomesenchymal stem cells exhibited a spindle-shaped morphology,while a minority displayed irregular shapes,and demonstrated high expression levels of mesenchymal stem cell-specific markers Nestin,CD44,and Vimentin.The obtained ectomesenchymal stem cells-derived extracellular vesicles fulfilled the biological criteria for extracellular vesicles.(2)Within the detected protein concentration range of 0.5 to 1.5 mg/mL,the proliferation of PC12 cells was promoted by ectomesenchymal stem cells-derived extracellular vesicles,and this effect was further enhanced with increasing concentrations.(3)Ectomesenchymal stem cells-derived extracellular vesicles increased the length of axons in PC12 cells and neurons and the expression of axon-related markers β3-tubulin,growth associated protein 43,and neurofilament 200.Above findings suggest that ectomesenchymal stem cells-derived extracellular vesicles have the potential to enhance neuronal axonal elongation.

BACKGROUND:The occurrence of neuronal axonal injury can result in neurological dysfunction,and the facilitation of axonal elongation is anticipated to play a pivotal role in the treatment of diseases affecting the nervous system.OBJECTIVE:To investigate whether ectomesenchymal stem cells-derived extracellular vesicles can promote neuronal axonal elongation.METHODS:(1)Ectomesenchymal stem cells were obtained from nasal mucosa using the tissue adherence method,and the specific markers of were identified through immunofluorescence.Ectomesenchymal stem cells-derived extracellular vesicles were acquired via ultracentrifugation and identified.(2)Ectomesenchymal stem cells-derived extracellular vesicles(0,0.5,1.0,1.5 mg/mL)were incubated with PC12 cells for 72 hours.The cytotoxicity and proliferation of ectomesenchymal stem cells-derived extracellular vesicles on PC12 cells were assessed using the CCK-8 assay.(3)Ectomesenchymal stem cells-derived extracellular vesicles(1.0 mg/mL)were incubated with PC12 cells or neurons for 72 hours.The changes in axon length were observed using microscopic analysis.The expression levels of axon-related markers β3-tubulin(early stage),growth associated protein 43(middle stage),and neurofilament 200(mature stage)were analyzed through real-time fluorescence quantitative PCR and Western blotting.These investigations aimed to explore the potential of ectomesenchymal stem cells-derived extracellular vesicles in promoting neurite elongation within PC12 cells or neurons.RESULTS AND CONCLUSION:(1)The majority of the acquired ectomesenchymal stem cells exhibited a spindle-shaped morphology,while a minority displayed irregular shapes,and demonstrated high expression levels of mesenchymal stem cell-specific markers Nestin,CD44,and Vimentin.The obtained ectomesenchymal stem cells-derived extracellular vesicles fulfilled the biological criteria for extracellular vesicles.(2)Within the detected protein concentration range of 0.5 to 1.5 mg/mL,the proliferation of PC12 cells was promoted by ectomesenchymal stem cells-derived extracellular vesicles,and this effect was further enhanced with increasing concentrations.(3)Ectomesenchymal stem cells-derived extracellular vesicles increased the length of axons in PC12 cells and neurons and the expression of axon-related markers β3-tubulin,growth associated protein 43,and neurofilament 200.Above findings suggest that ectomesenchymal stem cells-derived extracellular vesicles have the potential to enhance neuronal axonal elongation.

Objective:To study the feasibility of design and harvest of free medial sural artery perforator flap with the "two-point and two-line method".Methods:From September 2022 to June 2023, Suzhou Ruihua Orthopaedic Hospital implemented the "two-point and two-line method" to guide preoperative perforator positioning and flap design. Thirty medial sural artery perforator flaps were successfully harvested with the method, and 21 wounds of hand and 9 of foot and ankle were reconstructed with the flaps. The size of soft tissue defects were 2.5 cm×2.5 cm-7.0 cm×14.5 cm, and the flaps size were 3.0 cm×3.0 cm-7.5 cm×15.0 cm. All donor sites were directly closed or by skin grafting. All patients were entered in 6-15 months of postoperative outpatient follow-up, and the recovery of donor and recipient sites was assessed by the comprehensive evaluation scales. The sensory function of the flaps was evaluated using the sensory function evaluation standard of British Medical Research Council (BMRC).Results:All perforators were successfully located with 47 perforators in total, and all of them were musculocutaneous perforator. It was found that there was 1 perforator in 14 flaps, 2 perforators in 15 flaps and 3 perforators in 1 flap. All 30 flaps survived after surgery, beside 2 flaps that had arterial insufficiency but survived successfully after surgical exploration. All donor sites healed in one stage. Comprehensive evaluation scale of flap was employed to evaluate the flaps and the scores were 84 points to 96 points with an average score of 92.5 points. The excellent and good grades were achieved in 27 flaps and 3 flaps, respectively, with a combined excellent and good rate at 100%. Sensation of the flaps was evaluated by BMRC with 1 flap of S 1, 17 of S 2 and 12 of S 3. Conclusion:The "two-point and two-line method" has been used in design of the perforator flap of medial sural artery. This method is simple and accurate, and is feasible and ideal in design of flaps before surgery.

Objective:To introduce the application of highly selective arterial indocyanine green angiography (hereinafter referred to as highly selective arterial angiography) in the design of anterolateral thigh free flap.Methods:This study was a retrospective observational study. From November 2023 to April 2024, 29 patients with wounds in extremities which were repaired by anterolateral thigh free flaps designed under the assistance of highly selective arterial angiography and met the inclusion criteria were admitted to the Department of Hand Surgery and Department of Wound Repair Surgery of Suzhou Ruihua Orthopedic Hospital, including 26 males and 3 females, aged 16 to 71 years. The wound area after debridement ranged from 8.0 cm×4.5 cm to 27.0 cm×16.0 cm. During the surgery, highly selective arterial angiography was used to assist in flap design. The fluorescence development range of the source arteries or perforators of flaps was observed. The blood supply range of the source arteries or perforators of flaps was determined based on the fluorescence development of the skin, and the excision position of the flap was adjusted. The flap incision area ranged from 9.0 cm×6.0 cm to 29.0 cm×16.0 cm. During the surgery, the number of highly selective arterial angiography, the type of source artery of perforators for puncture, and changes in the excision position of flaps were observed and recorded. After surgery, the blood supply and survival of flaps, the healing of wounds and the survival of skin grafts in the flap donor sites, and the angiography-related complications were observed.Results:All the 32 flaps of 29 patients were successfully excised. The highly selective arterial angiography was performed 37 times, including 13 cases of puncture of the oblique branch of the lateral circumflex femoral artery, 6 cases of puncture of the descending branch, 8 cases of double puncture of the oblique and descending branches, and 2 cases of puncture of arteries from other branches. During the surgery, the excision position of 28 flaps did not change, the excision position of 3 flaps moved towards proximal extremity of the thigh, and the excision position of 1 flap moved towards distal extremity of the thigh. All the flaps survived successfully after the surgery, and there was no partial necrosis of the flaps at the proximal or distal ends. The wounds in the flap donor sites healed, and all skin grafts survived. No angiography-related complications occurred.Conclusions:Highly selective arterial angiography can be used to determine the blood supply range of the source artery and perforators of the anterolateral thigh free flaps during the surgery. It can evaluate the blood supply of flaps more intuitively and objectively. Its application in assisting flap design can avoid partial flap necrosis caused by unreasonable preoperative design to a certain extent, and it is safe and reliable.

Objective:To construct a scientific and systematic indicator system for the ideological and political construction of medical professional courses in military medical universities, and to provide a reference for the teaching management department of military medical universities to guide and manage the ideological and political construction of courses, as well as for medical professional course instructors to carry out the ideological and political teaching of courses.Methods:On the basis of literature research and case studies, a framework of the indicator system was established for the ideological and political construction of medical professional courses in military medical universities, and six experts in the field of military medical education were invited for pre-consultation. A preliminary draft of the indicator system was formed after discussion within the research group. A total of 25 experts, who had intermediate or above professional titles and long-term experience in military medical education, teaching and management, military academy education management, and military ideological and political education in military academies, were invited to conduct two rounds of Delphi expert consultation, and a statistical analysis was performed for the consultation results to determine the indicator system for the ideological and political construction of medical professional courses in military medical universities. The percentage weight method and the product method were used to calculate the weight coefficient of each indicator and combined weight coefficient.Results:In the two rounds of expert consultation, the expert positivity coefficient was 100%, the expert authority coefficient was 0.883, and the Kendall harmony coefficients were 0.254 and 0.268, respectively ( P<0.05). Finally, an indicator system was established for the ideological and political construction of medical professional courses in military medical universities, which consisted of 5 primary indicators, 12 secondary indicators, and 31 tertiary indicators. The 5 primary indicators were the teaching objectives, teaching team, teaching resources, teaching implementation, and teaching effect of curriculum ideology and politics, with a weight coefficient of 0.210, 0.202, 0.197, 0.202, and 0.189, respectively. Conclusions:The indicator system constructed in this study is scientific, reliable, systematic, and comprehensive and has a certain guiding significance for the ideological and political construction of medical professional courses in military medical universities.

Objective:To explore the application of a scenario-based onsite first-aid skills station in objective structured clinical examination (OSCE).Methods:Based on common scenarios and cases in medical practice, an evaluation framework of the OSCE onsite first-aid skills station—containing assessment indicators, exam room setting, examiner training, and assessment process—was designed to evaluate the onsite first-aid competencies of medical graduates of the five-year program for three consecutive years. SPSS 24.0 was used to perform the Kruskal-Wallis test and Pearson correlation analysis to calculate the correlation between course examination scores and OSCE onsite first-aid skills station assessment scores. Excel was used to calculate the difficulty index and discrimination index of test items.Results:The graduates' OSCE onsite first-aid skills station assessment scores were improved year by year, with a mean score of about 80 points. The station assessment items showed a moderate difficulty level (0.7-0.8), a good discrimination level (>0.4), and good internal consistency (Cronbach's α>0.7). The examiners and examinees had a high recognition of the design and effectiveness of this station assessment method. There was a positive correlation between the OSCE scores and corresponding course scores (2016, r=0.245, P=0.001; 2017, r=0.108, P=0.026; 2018, r=0.198, P=0.006). Conclusions:Through scientific scoring and strict examination management, the OSCE scenario-based onsite first-aid skills station can effectively evaluate examinees' injury treatment competencies in different situations, which can provide a reference for course teaching.

Objective:To investigate the correlation between serum hypoxia inducible factor 1 alpha (HIF-1α), vascular endothelial growth factor (VEGF), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), and inflammatory factors and incision infection after anal fistula surgery.Methods:A retrospective analysis was conducted on the clinical data of 120 patients who underwent anal fistula thread hanging surgery at the First People′s Hospital of Shuangliu District, Chengdu from June 2022 to April 2023. The patients were divided into an infected group (36 cases) and a non infected group (84 cases) based on their postoperative incision infection status. The differences in serum HIF-1 α, VEGF, sTREM-1 and inflammatory factors [C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor α (TNF-α)] between the infected and non infected groups were compared, and the correlation between CRP, IL-6, TNF-α, HIF-1 α, VEGF, sTREM-1 and postoperative infection of anal fistula was analyzed by Pearson correlation coefficient; The correlation between CRP, IL-6, TNF-α and HIF-1 α, VEGF, sTREM-1. And the efficacy of serum HIF-1 α, VEGF, sTREM-1, inflammatory factors and their individual and combined diagnosis of incision infection after anal fistula surgery was analyzed by receiver operating characteristic (ROC) curve analysis.Results:The levels of serum HIF-1 α, sTREM-1, CRP, IL-6, and TNF-α in the infected group were higher than those before surgery and higher than those in the uninfected group 3 days after surgery; VEGF levels were lower than preoperative levels and lower than those in the non infected group (all P<0.05). Pearson correlation analysis showed that HIF-1α, sTREM-1, CRP, IL-6, TNF-α were positively correlated with postoperative infection in anal fistula ( r=0.456, 0.494, 0.455, 0.510, 0.363, all P<0.05), while VEGF was negatively correlated with postoperative infection in anal fistula ( r=-0.462, P<0.05). CRP, IL-6, TNF-α were positively correlated with HIF-1 α and sTREM-1 ( r=0.574/0.611/0.653, 0.661/0.608/0.610, all P<0.05), while CRP, IL-6, TNF-α were negatively correlated with VEGF ( r=-0.200, -0.207, -0.254, all P<0.05). The area under the curve (AUC) of HIF-1 α, VEGF, sTREM-1, CRP, IL-6, and TNF-α for diagnosing incision infection after anal fistula surgery were 0.716, 0.787, 0.741, 0.678, 0.792, and 0.688, respectively. The AUC of the combined diagnosis of inflammatory factors and 6 data points for postoperative incision infection in anal fistula surgery were 0.836 and 0.921, respectively. Conclusions:Serum levels of HIF-1 α, sTREM-1, CRP, IL-6, and TNF-α are abnormally high in patients with incision infection after anal fistula surgery, while VEGF is abnormally low in expression. HIF-1 α, VEGF, sTREM-1, and inflammatory factors can be used as effective indicators for clinical diagnosis of incision infection after anal fistula surgery, and their combined diagnostic value is better. HIF-1 α and sTREM-1 are positively correlated with inflammatory factors, while VEGF is negatively correlated with inflammatory factors.