Aim To investigate the effect of ginkgolide B on TLR4 expression in glucose-treated endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs)were stimulated by high concentra-tion of glucose.TLR4,inflammatory protein expression and Akt phosphorylation were analyzed by Western blot.Transcription factor NF-κB nuclear translocation was analyzed by immunofluorescence.Results The expression of TLR4 and PAF receptor was increased in high glucose-treated HUVECs. In contrast, both ginkgolide B and CV3988 dose-dependently decreased TLR4 and PAF receptor expression in high glucose-treated cells,respectively.Ginkgolide B decreased in-flammatory protein ICAM-1 ,VCAM-1 expression.Mo-reover,ginkgolide B potently abolished Akt phospho-rylation and NF-κB p65 nuclear translocation.Conclu-sion Ginkgolide B can reduce TLR4,PAF receptor, ICAM-1 and VCAM-1 expression in high dose of glu-cose-treated HUVECs,the mechanism might be linked to inhibition of Akt phosphorylation and NF-κB activa-tion.

Aim To investigate the effect of resveratrol on ROS level and PECAM-1 expression in ox-LDL-stimulated platelets. Methods The expression of PE-CAM-1 , Sirt1 and p38 MAPK phosphorylation in ox-LDL-stimulated platelets was determined by Western blot. The level of ROS was measured by immunofluo-rescence kit. Results ox-LDL induced platelet aggre-gation by 14%, whereas resveratrol inhibited platelet aggregation by 50%. Resveratrol decreased ROS level by 3 . 2 fold and completely suppressed PECAM-1 expression in ox-LDL-treated platelets. Resveratrol re-covered Sirt1 expression in ox-LDL-treated platelets. EX527 ( a Sirt1 inhibitor ) increased ROS level and PECAM-1 expression in ox-LDL-stimulated platelets. Meanwhile, resveratrol also suppressed p38MAPK phosphorylation induced by ox-LDL. Conclusion Resveratrol can inhibit platelet aggregation, decrease ROS production and PECAM-1 expression in ox-LDL-stimulated platelets. The mechanism maybe associated with recovery of Sirt1 expression. Moreover, resveratrol can decrease PECAM-1 expression, which may be linked to abolishing p38MAPK phosphorylation.

Objective To investigate the molecular mechanisms of protective effects of thioredoxin (Trx) on human vascular endothelial cells in atherosclerosis.Methods The cell models of Trx-overexpressing cells (Ad Trx) and the control cells (Ad-con) were established by adenovirus vector gene transfer technology in human umbilical vein endothelial cells (HUVECs).The oxidized low density lipoprotein,a risk factor of atherosclerosis,was used as a stimulator.Western blot and indirect immunofluorescence were used to detect the protein expression levels and the cellular localization of Trx,adhesion molecules (ICAM-1,VCAM-1) and the upstream signal pathways.Trx activity was detected by insulin disulfide reduction assay,and cellular reactive oxygen species (ROS)production was detected by fluorescent probe DCFH-DA.Results As compared with control group,Trx protein expression level was enhanced in Ad-trx group and the Trx activity in Ad-Trx group was upregulated by (26.2 ±3.3)％.The result of ROS detection showed that overexpression of Trx significantly inhibited the cellular ROS generation.As compared with control group,overexpression of Trx obviously inhibited the adhesion molecules expression but markedly promoted the phosphorylation of Smad3 in endothelial cells with or without oxLDL stimulation (P＜0.05).Pretreatment of cells with SIS3,a specific inhibitor of Smad3 phosphorylation,reversed Trx-induced inhibition of adhesion molecules expression.Further studies showed that pretreatment of cells with SIS3 enhanced oxLDL-induced AP-1 subunit c-fos nuclear expression.Conclusions The enhancement of Smad3 phosphorylation and c-Fos nuclear expression are mainly responsible for the Trx-induced downregulation of adhesion molecules.

Aim Toinvestigatetheeffectofginkgolide B on apoptosis in high glucose-treated endothelial cells.Methods Humanumbilicalveinendothelial cells(HUVECs)were used in the present study.The level of transmigration of HUVECs was analyzed by Tr-answell experiment.Apoptosis was detected by flow cy-tometry.Reactive Oxygen Species (ROS ) was meas-ured by immunofluorescence kit.The protein expres-sionwasanalyzedbyWesternblot.Result Highglu-cose treatment resulted in a reduction in transmigration of HUVECs and ginkgolide B recovered the phenome-non in glucose-treated endothelial cells.The level of ROS generation was increased in high glucose-treated group,whereas ginkgolide B inhibited ROS genera-tion.Immunofluorescence data showed high glucose in-creased apoptosis,whereas ginkgolide B inhibited ap-optosis in high glucose-treated HUVECs.Moreover, the expressions of Bax and caspase-3 were increased and Bcl-2 was reduced in high glucose-treated group. In contrast,ginkgolide B abolished the expressions of Bax and caspase-3 and increased Bcl-2 expression. Moreover,high glucose enhanced the expression and phosphorylation of p53,while ginkgolide B suppressed the expression and phosphorylation of p53 induced by highglucose.Conclusions GinkgolideBcaninhibit apoptosis and improve transmigration function in high glucose-treated HUVECs.Ginkgolide B has protection against high glucose-induced endothelial cell injury.

Objective:To investigate the feasibility and clinical effects of using free perforator propeller myocutaneous flap from buttock in repairing complex wounds in the buttock with deep dead cavity.Methods:A retrospective observational study was conducted. From June 2020 to June 2022, 9 patients with complex wounds in the buttock with deep dead cavity who met the inclusion criteria were admitted to Lanzhou University Second Hospital, including 6 males and 3 females, aged 26 to 62 years, with original wound area ranging from 4.0 cm×3.0 cm to 8.0 cm×7.0 cm and dead cavity depth of 7 to 11 cm. All the wounds were repaired with free perforator propeller myocutaneous flap from buttock, with flap area of 6.0 cm×2.5 cm to 13.0 cm×7.0 cm and muscle flap length of 6 to 11 cm. All the wounds in the donor area were closed and sutured directly. Postoperative myocutaneous flap survival, complications, as well as donor and recipient wound healing were observed, and the shape of donor and recipient areas were followed up.Results:Congestion occurred under the myocutaneous flap of one patient due to poor drainage on post surgery day 2, which was healed after 15 days of drainage and dressing change. The myocutaneous flaps of other patients survived successfully after surgery. The wounds in the donor and recipient areas were all well healed. During the follow-up of 3 to 10 months, the donor and recipient areas were full in shape, with little difference from the healthy side, and were able to bear pressure.Conclusions:The free perforator propeller myocutaneous flap from buttock can repair the deep dead cavity and surface wounds at the same time. The use of this myocutaneous flap in repairing complex wounds in the buttock with deep dead cavity results in minimal damage to the donor area, allows pressure-bearing of the donor and recipient areas after surgery, and ensures a full buttock shape.

Objective To investigate the relationship between systemic immune-inflammation index (SII)and lower extremity vascular disease in patients with type 2 diabetes mellitus (T2DM).Methods A cross-sectional study was conducted on 390 T2DM patients admitted in our department from January 2013 to January 2024.According to the diagnostic criteria for lower extremity vascular disease in T2DM patients,they were divided into a lower extremity vascular disease group (n=158)and a control group (n=232).General data and results of laboratory tests were compared between the 2 groups.Spearman correlation analysis was used to identify the related factors for lower extremity vascular diseases in T2DM patients.The correlation between SII and lower extremity vascular diseases in T2DM patients was analyzed using the Row Mean Scores and Cochran-Armitage Trend analysis.Multivariate logistic regression analysis was applied to identify the risk factors for lower limb vascular lesions in T2DM patients.Receiver operating characteristic (ROC)curve was plotted to evaluate the diagnostic efficacy of SII for lower extremity vascular disease in the patients.Results Compared with T2DMpatients without lower extremity vascular disease,those with lower extremity vascular disease were older,had higher levels of total cholesterol (TC),low-density lipoprotein cholesterol (LDL-C),SII,larger proportion of carotid vascular lesions,and increased proportion of no-taking statins.The lower extremity vascular disease in T2DM patients was positively correlated with SII/100 (r=0.429,P<0.001),age (r=0.517,P<0.001),TC (r=0.161,P=0.001),LDL-C (r=0.117,P=0.021),carotid artery lesions (r=0.101,P=0.047),no-taking statins (r=0.266,P<0.001).Logistic regression analysis showed that SII,age,LDL-C,and no-taking statins were the risk factors for lower extremity vascular lesions in T2DM patients (P<0.01).The area under the curve (AUC)value of SII combined with age,LDL-C,and no-taking statins in predicting lower extremity vascular disease in T2DM patients was 0.896.Conclusion SII is not only a risk factor,but also a simple marker for lower extremity vascular disease in T2DM patients,suggesting that inflammatory response plays an important role in the occurrence and development of lower extremity vascular disease in T2DM.

Objective:To investigate the regulatory effect of WNT1-inducible signaling pathway protein 2(WISP2)on macrophage polarization in palmitic acid(PA)and lipopolysaccharide(LPS)-induced inflammation.Methods:The macrophage cell line RAW264.7 was treated with different concentrations of WISP2 protein, and cell viability was determined by means of luminescence assay using Cell-Titer Glo to determine the concentration of WISP2.The cells were divided into control group, palmitic acid group, palmitic acid combined with different concentrations of WISP2 group(10 μg/L and 100 μg/L)and lipopolysaccharide group, lipopolysaccharide combined with different concentrations of WISP2 group(10 μg/L and 100 μg/L). mRNA expression of M1 and M2 macrophages phenotype of each group were detected by real-time quantitative polymerase chain reaction.The protein expression of important inflammatory factors, TNF-α and IL-6, were evaluated by ELISA.Results:Compared with the control group, both 10 μg/L and 100 μg/L WISP2 groups had no effect on the activity of RAW264.7 cells, but significantly up-regulated the expression of various inflammatory factors, including Tnfα(1.877±0.039, 2.202±0.034, F=309.7, P<0.001), Il6(1.418±0.056, 1.506±0.059, F=81.39, P<0.001), Mcp1(1.620±0.014, 1.982±0.125, F=71.45, P<0.001), Ccl3(1.892±0.118, 1.942±0.132, F=32.93, P<0.001), and iNos(1.691±0.201, 1.548±0.090, F=13.60, P<0.05). mRNA in macrophages, and significantly down-regulated the expression of anti-inflammatory factors, including Tgfβ(1.376±0.025, 2.152±0.107, F=1.846, P<0.05), CD206(2.123±0.031, 3.139±1.663, F=8.037, P<0.05), Il4(2.098±0.464, 2.494±0.141, F=48.68, P<0.01), and Il10(1.303±0.216, 1.574±0.274, F=5.774, P<0.05)mRNA, causing M1 type macrophage polarization.Compared with the control group, 100 μmol/L palmitic acid could mildly but significantly increase the expression of inflammatory factors such as TNF-α and IL-6 at the transcriptional and protein levels.Compared with palmitic acid stimulation alone, the combination of palmitic acid and WISP2 further promoted the protein expression of macrophage inflammatory factors TNF-α[(589.4±17.0)ng/L, (692.6±83.4)ng/L, F=56.38, P<0.05], IL-6[(15.13±1.14)ng/L, (13.33±1.22)ng/L, F=23.32, P<0.001]and the mRNA expression of chemokines Mcp1(160±9.796, 140±18.91, F=141.1, P<0.0001)and C cl3(17.76±1.92, 14.41±1.27, F=125.2, P<0.0001). Compared with the control group, 100 μg/L lipopolysaccharide strongly stimulated the expression of inflammatory factors such as TNF-α[(3444±423)ng/L, F=71.20, P<0.0001]and IL-6[(497.0±41.2)ng/L, F=63.50, P<0.0001]in macrophages at the protein level.Compared with lipopolysaccharide stimulation alone, the combination of lipopolysaccharide and WISP2 further significantly up-regulated the mRNA expression of chemokines Mcp1(106.8±8.7, 118.7±4.6, F=251.5, P<0.0001)and Ccl3(35.3±12.5, 116.4±4.5, F=160.1, P<0.0001). Conclusions:The adipokine WISP2 can promote M1 macrophage polarization in palmitic acid and lipopolysaccharide-induced inflammation, and it had distinct regulation in macrophage polarization under different inflammatory response conditions.

Objective: To evaluate the accuracy and feasibility of coronary artery calcium score ( CACS) on virtual non-contrast scan ( VNC) images obtained from coronary artery CT angiography ( CCTA) scan with dual －layer spectral detector CT (SDCT) ． Methods : The data of 197 patients who underwent CCTA scan in hospital were analyzed retrospectively，and 88 patients with CACS ＞0 were further analyzed． Linear regression analysis of CACS and coronary artery calcium volume ( CACV) of true non-contrast (TNC) images and VNC images ( CACS-TNC， CACS-VNC，CACV-TNC，CACV-VNC) was performed to obtain linear regression equation and correction coefficients λ 1AVG and λ2AVG ．CACS-VNC and CACV-VNC were corrected by the corresponding regression equation and recorded as CCACS-VNC and CCACV-VNC，respectively．Spearman correlation coefficient was used for correlation analysis and Bland-Altman plot was used for consistency test．Mann-Whitney U test was used to compare the difference between the two groups. Results : For the total coronary artery，there was a strong correlation between CACS- TNC and CACS-VNC (rs = 0. 952，P ＜0. 001 ，λ 1AVG = 2. 19 ) ，CACV-TNC and CACV-VNC ( rs = 0. 954，P < 0. 001，λ2AVG = 1. 93) ．The results of Mann-Whitney U test showed that there was no significant difference between CACS-TNC and CCACS-VNC or between CACV-TNC and CCACV-VNC，and the Bland-Altman plot showed good consistency between CACS-TNC and CCACS-VNC ，CACV-TNC and CCACV-VNC． Conclusion VNC images based on SDCT can accurately measure CACS and be used for cardiovascular risk classification，which is expected to replace TNC scan and reduce the radiation dose of patients.