Objective To investigate the role of luteolin(Lut)in regulating reactive oxygen species(ROS)levels through nuclear factor erythroid 2-related factor 2(NFE2L2)/cystine glutamate antitransporter(x-CT)/glutathione peroxidase 4(GPX4)signaling axis to inhibit the viability of glioblastoma and promote apoptosis.Methods U87 MG and U251 cells were cultured in vitro.The CCK-8 assay was used to detect cell survival rates after 48 hours of treatment with different concentrations(0,6.25,12.5,25,50 and 100 μmol/L)of Lut.According to whether cells were treated with Lut,cells were divided into the U87 control group,the U87 Lut group,the U251 control group and the U251 Lut group.The half-maximal inhibitory concentration(IC50)at 48 hours was used as the unified treatment concentration for subsequent experiments.The apoptosis level of cells was detected by flow cytometry double staining method.Changes of reactive oxygen species(ROS)levels in cells were detected by the DCFH-DA method.Molecular docking was conducted using AutoDock software to verify the proteins related to the Lut and oxidative stress pathway.Real-time fluorescence quantitative reverse transcription(RT-qPCR)was used to detect the mRNA levels of NFE2L2 and GPX4.The expression levels of NFE2L2,x-CT and GPX4 proteins were detected by Western blot assay.Results After U87 MG and U251 cells were treated with Lut for 48 hours,the cell viability was significantly inhibited,and with the increase of Lut concentration,the cell viability decreased(P＜0.05).Compared with the U87 control group and the U251 control group respectively,the apoptosis rate of cells increased in the U87 Lut group and the U251 Lut group,the green fluorescence intensity was enhanced,and the intracellular ROS level was upregulated(P＜0.05).Results of molecular docking showed that Lut was tightly bound to NFE2L2,x-CT and GPX4.The results of RT-qPCR and Western blot assay showed that compared with the U87 control group and the U251 control group respectively,the protein and mRNA levels of NFE2L2 and GPX4 in cells of the U87 Lut group and the U251 Lut group,as well as the expression level of x-CT protein,decreased(P＜0.05).Conclusion Lut regulates ROS levels through the NFE2L2/x-CT/GPX4 signaling axis to inhibit the viability of glioblastoma and promote cell apoptosis.

Objective To investigate the role of luteolin(Lut)in regulating reactive oxygen species(ROS)levels through nuclear factor erythroid 2-related factor 2(NFE2L2)/cystine glutamate antitransporter(x-CT)/glutathione peroxidase 4(GPX4)signaling axis to inhibit the viability of glioblastoma and promote apoptosis.Methods U87 MG and U251 cells were cultured in vitro.The CCK-8 assay was used to detect cell survival rates after 48 hours of treatment with different concentrations(0,6.25,12.5,25,50 and 100 μmol/L)of Lut.According to whether cells were treated with Lut,cells were divided into the U87 control group,the U87 Lut group,the U251 control group and the U251 Lut group.The half-maximal inhibitory concentration(IC50)at 48 hours was used as the unified treatment concentration for subsequent experiments.The apoptosis level of cells was detected by flow cytometry double staining method.Changes of reactive oxygen species(ROS)levels in cells were detected by the DCFH-DA method.Molecular docking was conducted using AutoDock software to verify the proteins related to the Lut and oxidative stress pathway.Real-time fluorescence quantitative reverse transcription(RT-qPCR)was used to detect the mRNA levels of NFE2L2 and GPX4.The expression levels of NFE2L2,x-CT and GPX4 proteins were detected by Western blot assay.Results After U87 MG and U251 cells were treated with Lut for 48 hours,the cell viability was significantly inhibited,and with the increase of Lut concentration,the cell viability decreased(P＜0.05).Compared with the U87 control group and the U251 control group respectively,the apoptosis rate of cells increased in the U87 Lut group and the U251 Lut group,the green fluorescence intensity was enhanced,and the intracellular ROS level was upregulated(P＜0.05).Results of molecular docking showed that Lut was tightly bound to NFE2L2,x-CT and GPX4.The results of RT-qPCR and Western blot assay showed that compared with the U87 control group and the U251 control group respectively,the protein and mRNA levels of NFE2L2 and GPX4 in cells of the U87 Lut group and the U251 Lut group,as well as the expression level of x-CT protein,decreased(P＜0.05).Conclusion Lut regulates ROS levels through the NFE2L2/x-CT/GPX4 signaling axis to inhibit the viability of glioblastoma and promote cell apoptosis.

Objective:To analyze the application value of tibial transverse transport (TTT) microvascular reconstruction combined with antibiotic bone cement in the limb salvage of diabetic foot.Methods:Seventy-seven patients with diabetic foot admitted to the Traditional Chinese Medicine Hospital of Zhongshan from January 2018 to January 2021 were enrolled retrospectively, and they were divided into the control group (performed TTT microvascular reconstruction, 47 cases) and the combined group (performed TTT microvascular reconstruction combined with antibiotic bone cement, 30 cases) according to the surgical method. Wound healing rate, limb salvage rate, the ulcer healing time, bone window healing time, wound infection control status, affected side lower limb vascular blood supply situation, and peripheral nerve function recovery were compared between the two groups.Results:The amputation rate between the two groups had no significant difference ( P>0.05), but the healing rate in the combined group was higher than that in the control group: 90.00%(27/30) vs. 68.08%(32/47); and the ulcer healing time, bone window healing time in the combined group were shorter than those in the control group: (12.55 ± 2.14) weeks vs. (15.01 ± 3.17) weeks, (17.47 ± 2.09) weeks vs. (19.00 ± 2.58) weeks, there were statistical differences ( P<0.05). The positive percentage of bacteria culture in wound secretions in the combined group after the surgery for 6 d was lower than that in the control group: 20.00%(6/30) vs. 44.68%(21/47), there was statistical difference ( P<0.05). After the surgery for 3 months, the dorsal skin temperature and ankle brachial index in the combined group were higher than those in the control group: (31.05 ± 1.74) ℃ vs. (29.27 ± 1.66) ℃, 0.93 ± 0.29 vs. 0.80 ± 0.19, there were statistical differences ( P<0.05), but the scores of visual analog scale (VAS) and 10 g nylon line test values between the two groups had no significant differences ( P>0.05). Conclusions:TTT combined with antibiotic bone cement can achieve better wound healing and infection control effect in the limb salvage of diabetic foot, and the blood supply improvement of the affected limb is more significant.

Objective:To investigate the clinical effect of ozone combined with sodium hyaluronate injection in the treatment of knee osteoarthritis and its effect on inflammatory factors and hemorheology in patients.Methods:146 patients with knee osteoarthritis who received treatment in our hospital from October 2016 to February 2019 were selected and divided into observation group and control group according to the random number table method, 73 cases in each group.In the experimental group, intra-articular injection of sodium hyaluronate combined with ozone was used, while in the control group, glucosamine hydrochloride tablets were routinely taken and intra-articular injection of sodium hyaluronate was performed. Patients in both groups were treated once a week for 5 consecutive weeks for a course of treatment. General data of patients in the two groups were analyzed, and the efficacy, visual analogue scale (VAS) score and hospital for special surgery knee score (HSS) before and after treatment, levels of inflammatory factors and changes in hemorheology indexes were compared between the two groups.Results:There was no significant difference in general data between the two groups ( P>0.05). Before treatment, there was no significant difference in VAS score and HSS score between the two groups ( P>0.05). After treatment, VAS score and HSS score in the two groups were significantly decreased and increased respectively ( P<0.05). Compared with the control group, VAS score was significantly decreased and HSS score was significantly increased in the observation group after treatment ( P<0.05). The total effective rate in the observation group was higher than that in the control group, and the difference between the two groups was statistically significant ( P<0.05). Before treatment, there was no significant difference in the levels of inflammatory factors vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF-α), interleukin (IL)-1β, IL-6, hypersensitive C-reactive protein (hs-CRP) and matrix metalloproteinase (MMP)-13 in the articular fluid between the two groups ( P>0.05). After treatment, the above indexs in the observation group were significantly lower than those in the control group ( P<0.05). After treatment, the levels of inflammatory cytokines in the articular fluid of the two groups were lower than before ( P<0.05). There was no significant difference in hemorheology indexes between the two groups before treatment ( P>0.05), and the two groups were significantly improved after treatment, and the degree of improvement in the observation group was significantly better than that in the control group ( P<0.05). Conclusions:Ozone combined with sodium hyaluronate intra-articular injection is more effective in the treatment of knee osteoarthritis, which can effectively reduce the level of inflammatory factors and inflammatory response, improve hemorheology, and then reduce the pain of patients, improve the knee joint function.

Objective: To establish a treatment protocol for severe blepharoptosis. This protocol helps to achieve better accuracy and more stable result. Methods: Evaluate levator muscle function pre-operatively. When levator function ≤ 1 mm, frontalis suspension technique was performed. When levator function more than 1 mm, technique of levator resection, combining excision of tarsus and levator, and tarsus-levator-CFS suspension was performed accordingly until it reaches adequate correction result during the surgery. Results: A total of 275 severe ptosis patients was included from January 2015 to June 2016. 52 cases (388 eyes) received levator resection. 162 cases received combining excision of tarsus and levator. 24 cases received tarsus-levator- CFS suspension. 37 cases received frontalis suspension. 326 eyes achieved adequate correction results. 62 cases were still undercorrected. The asymmetry result showed that 76 cases presented good symmetry. 142 cases presented moderate asymmetry and 57 showed severe asymmetry. Conclusions The new treatment protocol shows a satisfactory result with better accuracy and more stable correction for severe ptosis correction.

Objective To investigate the changes of serum Copeptin and matrix metalloproteinase-9 (MMP-9) in children with chronic heart failure (CHF) and its clinical significance. Methods A total of 186 children with CHF were selected for CHF group, including 78 cases of cardiac function grade Ⅱ, 65 cases of grade Ⅲ, and 43 cases of grade Ⅳ. There were 57 cases of dilated cardiomyopathy, 68 cases of congenital heart disease and 61 cases of other diseases. Another 85 healthy children from health checkup were chosen as controls. The levels of serum Copeptin and MMP-9 were determined by enzyme linked immunosorbent assay (ELISA), and the level of N-terminal pro-brain natriuretic peptide (NT-proBNP) was measured by bidirectional lateral flow immunoassay. The left ventricular end diastolic dimension (LVEDD), left ventricular end systolic dimension (LVESD), left ventricular ejection fraction (LVEF), and left ventricular short fraction shortening (LVFS) were measured by echocardiography. ROC curve was used to analyze the diagnostic value of serum Copeptin and MMP-9 in CHF. The correlation of serum Copeptin and MMP-9 with the cardiac function indices were examined by Pearson correlation analysis. Results The levels of serum copeptin, MMP-9, and NT-proBNP in different cardiac function groups (Ⅱ, Ⅲ, Ⅳ) increased gradually with the aggravation of the cardiac function damage and were higher than those in control group, and the differences were statistically significant (P<0.05). Compared with the control group and cardiac function grade Ⅱ group, the levels of LVESD and LVEDD were increased and the levels of LVEF and LVFS were decreased in the grade Ⅲ and Ⅳ groups. Compared with the grade Ⅲ group, the levels of LVESD and LVEDD were increased and the levels of LVEF and LVFS were decreased in the grade Ⅳ groups. There were significant differences (P<0.05). The ROC curve showed that the area under the curve (AUC) and 95％ CI of serum Copeptin, MMP-9, NT-proBNP and combinations of these three biomarkers in the diagnosis of CHF were 0.845 (0.781～0.914), 0.806 (0.736～0.883), 0.894 (0.828～0.962) and 0.925 (0.846～0.983) respectively, and the optimal thresholds were 12.5 pmol/L, 175.3 μg/L and 2037.0ng/L. The level of serum Copeptin was positively correlated with MMP-9 (r=0.807, P<0.001). Conclusion Serum Copeptin and MMP-9 may be involved in the ventricular remodeling in CHF children and they are expected to be a good indicator for the diagnosis of CHF and cardiac function.

Objective To explore the effects of Bawei Shangke Huoxue Tablets on hypercoagulable state of patients with hip fracture. Methods Seventy-two hip fracture patients at hypercoagulable state were randomly divided into treatment group and control group, 36 cases in each group. Both groups were given conventional orthopedic treatment including raising up the affected limb, external application of Huoxue Zhitong Paste on the affected area, and instructions for affected limb functional exercise. Additionally, the treatment group was given oral use of Bawei Shangke Huoxue Tablets together with subcutaneous injection of low-molecular-weight heparin calcium, and the control group was given subcutaneous injection of low-molecular-weight heparin calcium. Before and after treatment, we observed the plasma D-dimer (D-Di) and blood von Willebrand factor(vWF) levels as well as the incidence of deep venous thrombosis (DVT) of both groups before and after treatment. Results (1) All of the cases of both groups completed the clinical trial, and no severe adverse reaction such as increased bleeding occurred. (2) On the admission day, the D-Di and vWF levels of both groups were higher than the normal levels, and were decreased obviously one day before the operation (P < 0.05). On the first post-operative day, the levels were increased as compared with those before operation (P < 0.05), and then decreased on the twelfth post-operative day (P < 0.05) compared with those on the first post-operative day). The treatment group had better effect on improving D-Di and vWF levels than the control group (P < 0.05). (3) The results of follow-up for 3 months showed that 61 cases got the follow-up, 31 cases from the treatment group and 30 from the control group. The DVT incidence of treatment group was 9.68% and that of the control group was 16.67%, the difference being insignificant(P>0.05). Conclusion Bawei Shangke Huoxue Tablets are effective on improving hypercoagulable state of patients with hip fracture, and can be used at early stage and are beneficial to lowering the risks of anticoagulant drugs. Bawei Shangke Huoxue Tablets plus low-molecular-weight heparin calcium have certain effect on preventing DVT.