BACKGROUND:During bone remodeling,bone formation and bone resorption are spatially and temporally coordinated,involving intricate interactions between osteoclasts and osteoblasts.Isoginkgetin,a flavonoid found in Ginkgo biloba,has a wide range of anticancer activity and anti-reactive oxygen species activity;however,the effect of isoginkgetin on osteoclast differentiation is unknown.OBJECTIVE:To study the effect and mechanism of action of isoginkgetin on osteoclastogenesis.METHODS:In vitro studies were performed on mouse bone marrow-derived macrophages,and cell counting kit-8 cytotoxicity assay was used to detect the effect of isoginkgetin on cell viability of bone marrow-derived macrophages.Macrophage colony-stimulating factor and receptor activator of nuclear factor kappa-B ligand were used to induce the differentiation of bone marrow-derived macrophages to osteoclasts.Network pharmacology and molecular docking and molecular dynamics simulations were used to predict the processes and targets of the effects of isoginkgetin on the differentiation of osteoclasts.Tartrate-resistant acid phosphatase staining and F-actin staining were used to detect the effects of isoginkgetin on the differentiation and function of osteoclasts.Western blot and RT-PCR were used to detect the effects of isoginkgetin on the expression of genes and proteins related to osteoclast differentiation,reactive oxygen species,and PI3K/AKT pathways.Fluorescent probes were used to detect cellular and mitochondrial reactive oxygen species levels.Flow cytometry technology was used to detect reactive oxygen species levels in cells.RESULTS AND CONCLUSION:(1)Network pharmacology results showed that isoginkgetin affected osteoporosis mainly through the PI3K-AKT pathway and cellular response to drugs and hypoxia,and GSK3β,ESR1,MCL1 and CCNA2 were the key targets.(2)Cell counting kit-8 and tartrate-resistant acid phosphatase staining results showed that isoginkgetin at 8 μmol/L had the most significant inhibitory effect on osteoclastogenesis in vitro,and F-actin results showed that isoginkgetin inhibited osteoclast cytoskeletal actin ring formation in a concentration-dependent manner.(3)Molecular dynamics simulations showed that isoginkgetin bound well to osteoclastogenesis marker proteins(NFATc1,c-Fos,CTSK,and MMP9).Western blot and RT-PCR results indicated that isoginkgetin inhibited the expression of osteoclastogenesis marker proteins and genes(NFATc1,c-Fos,CTSK,and MMP9).(4)Western blot results showed that isoginkgetin inhibited the phosphorylation level of PI3K/AKT/GSK3β and suppressed osteoclastogenesis by activating the PI3K-AKT-GSK3β pathway.(5)The results of reactive oxygen species assay showed that isoginkgetin significantly reduced receptor activator of nuclear factor kappa-B ligand-induced cellular and mitochondrial reactive oxygen species production,and inhibited the differentiation of bone marrow-derived macrophages to osteoclasts.

BACKGROUND:During bone remodeling,bone formation and bone resorption are spatially and temporally coordinated,involving intricate interactions between osteoclasts and osteoblasts.Isoginkgetin,a flavonoid found in Ginkgo biloba,has a wide range of anticancer activity and anti-reactive oxygen species activity;however,the effect of isoginkgetin on osteoclast differentiation is unknown.OBJECTIVE:To study the effect and mechanism of action of isoginkgetin on osteoclastogenesis.METHODS:In vitro studies were performed on mouse bone marrow-derived macrophages,and cell counting kit-8 cytotoxicity assay was used to detect the effect of isoginkgetin on cell viability of bone marrow-derived macrophages.Macrophage colony-stimulating factor and receptor activator of nuclear factor kappa-B ligand were used to induce the differentiation of bone marrow-derived macrophages to osteoclasts.Network pharmacology and molecular docking and molecular dynamics simulations were used to predict the processes and targets of the effects of isoginkgetin on the differentiation of osteoclasts.Tartrate-resistant acid phosphatase staining and F-actin staining were used to detect the effects of isoginkgetin on the differentiation and function of osteoclasts.Western blot and RT-PCR were used to detect the effects of isoginkgetin on the expression of genes and proteins related to osteoclast differentiation,reactive oxygen species,and PI3K/AKT pathways.Fluorescent probes were used to detect cellular and mitochondrial reactive oxygen species levels.Flow cytometry technology was used to detect reactive oxygen species levels in cells.RESULTS AND CONCLUSION:(1)Network pharmacology results showed that isoginkgetin affected osteoporosis mainly through the PI3K-AKT pathway and cellular response to drugs and hypoxia,and GSK3β,ESR1,MCL1 and CCNA2 were the key targets.(2)Cell counting kit-8 and tartrate-resistant acid phosphatase staining results showed that isoginkgetin at 8 μmol/L had the most significant inhibitory effect on osteoclastogenesis in vitro,and F-actin results showed that isoginkgetin inhibited osteoclast cytoskeletal actin ring formation in a concentration-dependent manner.(3)Molecular dynamics simulations showed that isoginkgetin bound well to osteoclastogenesis marker proteins(NFATc1,c-Fos,CTSK,and MMP9).Western blot and RT-PCR results indicated that isoginkgetin inhibited the expression of osteoclastogenesis marker proteins and genes(NFATc1,c-Fos,CTSK,and MMP9).(4)Western blot results showed that isoginkgetin inhibited the phosphorylation level of PI3K/AKT/GSK3β and suppressed osteoclastogenesis by activating the PI3K-AKT-GSK3β pathway.(5)The results of reactive oxygen species assay showed that isoginkgetin significantly reduced receptor activator of nuclear factor kappa-B ligand-induced cellular and mitochondrial reactive oxygen species production,and inhibited the differentiation of bone marrow-derived macrophages to osteoclasts.

Iron is one of the essential trace elements for the human body,which is crucial for the growth and development of newborns,especially premature infants.It participates in the generation of hemoglobin,affects the activity of various enzymes,and subsequently affects neurometabolism,neurochemistry,neuroanatomy,and gene/protein composition,thereby having a lasting impact on the development of the central nervous system.This article reviews the research progress on the relationship between iron metabolism and neurodevelopment in premature infants in recent years,aiming to provide scientific basis for clinical management and preventive intervention of premature infants.

Objective To discuss the causes and the therapeutic strategy of acute cardiac tamponade(ACT)occurring as a complication of transcatheter aortic valve replacement(TAVR)so as to improve the success rate of the surgery and to make a further understanding of this complication.Methods The general clinical data,surgical procedures,and postoperative follow-up results of five patients,who received TAVR at the Affiliated Northern Jiangsu People's Hospital of Yangzhou University of China and developed ACT from March 2018 to September 2024,were retrospectively analyzed.Results After developing ACT,all the 5 patients received pericardiocentesis together with other adjuvant therapies including blood volume expansion with infusion,vasopressors,heparin neutralization,and blood transfusion.However,due to no obvious reduction in drainage volume and unstable hemodynamics all the 5 patients had eventually to receive open-chest surgery to identify the source of bleeding and to make hemostasis.Surgical exploration revealed that the perforation or rupture of cardiac structures caused by the temporary pacemaker lead or a super-stiff guide wire during the procedure was the main cause of ACT.Finally,after active treatment four patients recovered and discharged,and one patient died.The discharged patients were followed up for 3-12 months,and no procedure-related complications such as acute coronary artery occlusion,severe arrhythmia,exacerbation of heart failure symptoms,valve displacement,or stroke occurred.Conclusion As a severe complication occurring during the TAVR procedure,ACT requires to get a rapid diagnosis and management.Improvement of surgical techniques and operative methods,comprehensive preoperative assessment,and close intraoperative monitoring are crucial points for the prevention of ACT.

AIM:To investigate the role and mechanism of the glucagon-like peptide-1 receptor agonist lira-glutide(Lira)in regulating ferroptosis of mouse insulinoma MIN6 cells induced by high glucose and high fat.METHODS:The mouse insulinoma MIN6 cells were exposed to 30 mmol/L glucose and 500 μmol/L palmitic acid to establish an islet β cell injury model.On this basis,a ferroptosis inducer erastin,a ferroptosis inhibitor ferrostatin-1(Fer-1),and low and high concentrations of Lira were administered.Cell viability of different treatment groups were detected by CCK-8 assay.The malondialdehyde(MDA)kit was used to determine the changes in intracellular MDA content.The reactive oxygen species(ROS)kit was used to detect the changes in the ROS level of cells.The Fe2+fluorescence probe FerroOrange and mitochondrial membrane potential(JC-1)were used to detect the intracellular Fe2+levels and mitochondrial functions in different treatment groups.The mouse insulin ELISA kit was used to detect the insulin secretion of cells.RT-qPCR was used to detect the changes in the expression levels of key ferroptosis genes and insulin secretion genes in different treat-ment groups.Western blot was used to detect the expression levels of key ferroptosis proteins,glutathione peroxidase 4(GPX4)and solute carrier family 7 member 11(SLC7A11)in different treatment groups.RESULTS:Compared with the cells treated with high glucose and high fat,after treatment with Fer-1 and high-dose Lira,the cell viability,insulin secre-tion of the cells,and mitochondrial membrane potential all increased significantly,the levels of ROS,MDA and Fe2+were decreased(P<0.05).The results of RT-qPCR showed that Fer-1 and high-dose Lira significantly upregulated the expres-sion of genes promoting insulin secretion(P<0.05).The results of Western blot showed that Fer-1 and high-dose Lira sig-nificantly upregulated the expression of ferroptosis-inhibiting proteins GPX4 and SLC7A11(P<0.05).CONCLUSION:Liraglutide inhibits ferroptosis of mouse insulinoma MIN6 cells by regulating the SLC7A11/GPX4 signaling pathway,there-by improving the damage and dysfunction of MIN6 cells induced by high glucose and high fat.

Objective:To investigate the surgical techniques and clinical efficacy of arthroscopic-assisted posterior malleolus reduction for the management of ankle fracture-dislocation.Methods:A retrospective analysis was performed on the clinical data of 27 patients who underwent arthroscopy-assisted posterior malleolar reduction via the posterior approach for the surgical management of ankle fracture-dislocation at the Third Affiliated Hospital of Nanjing Medical University (Changzhou No.2 People's Hospital) between January 2022 and June 2023. The cohort comprised 17 males and 10 females, with a mean age of 43.67±9.56 years (range, 25-63 years). Based on the Bartonícek and Rammelt classification, there were 15 type II cases, 9 type III cases, and 3 type IV cases. The operation time, posterior ankle arthroscopy duration, and postoperative complications, such as neurovascular injury, wound infection, or skin necrosis, were recorded. X-ray and 3D CT imaging were utilized to assess joint surface reduction quality, tibiofibular matching and fracture healing status. The ankle plantarflexion, dorsiflexion and hallux flexion contractures were recorded at the last follow-up. Functional outcomes were measured using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and the Olerud-Molander ankle score (OMAS), while pain was evaluated using the visual analogue scale (VAS).Results:All 27 patients were followed up postoperatively, with a mean follow-up duration of 14.30±1.38 months (range, 12-18 months). Postoperatively, one patient experienced wound exudation, while another developed intermuscular venous thrombosis in the calf. No cases of neurovascular injury, wound infection, skin necrosis, or hallux flexion contracture occurred, and no reduction loss was observed. The mean operation time was 96.11±11.55 min (range, 80-120 min), and the posterior ankle arthroscopy duration was 35.74±5.67 min (range, 30-45 min). Postoperative X-ray evaluations demonstrated no loss of fracture reduction, and all fractures achieved bony union. The mean fracture healing time was 3.78±0.75 months (range, 3-5 months). Postoperative CT evaluations showed no joint surface malalignment, and distal tibiofibular matching was satisfactory. At the final follow-up, mean ankle plantarflexion was 46.74°±4.73° (range, 33°-50°), and dorsiflexion was 20.96°±3.29° (range, 14°-26°). There was no hallux flexion contracture occurred. The mean AOFAS ankle-hindfoot score was 92.11±7.19 (range, 74-100), with 20 excellent, 5 good, and 2 fair, yielding an excellent-good rate of 93%. The mean OMAS score was 94.44±7.25 (range, 75-100), comprising 20 excellent and 7 good results, yielding an excellent-good rate of 100%. The mean VAS score was 0.70±0.95 (range, 0-3).Conclusion:Arthroscopy-assisted posterior malleolar reduction in ankle fracture-dislocation surgery provides optimal soft tissue protection and ensures precise fracture reduction and fixation.

Objective:To investigate the predictive value of endothelial activation and stress index(EASIX)for the prognosis of patients with mantle cell lymphoma(MCL).Methods:A retrospective analysis was conducted to assess prognosis and compare the clinical features of patients diagnosed with MCL who were admitted to the Affiliated Hospital of Xuzhou Medical University from January 2010 to June 2023,had therapeutic indications and received standard treatment.Results:A total of 66 patients were included and divided into high EASIX group and low EASIX group,according to a cutoff value of 0.97 determined by the receiver operating characteristic(ROC)curve.Multivariate Cox regression analysis showed that prealbumin＜0.2 g/L,high EASIX,and ECOG PS score ≥2 were independent risk factors influencing overall survival(OS)in MCL patients.The median OS of patients in the high and low EASIX group was 13.0 and 37.5 months,and the median progression-free survival was 8.8 and 26.0 months,respectively.The proportions of patients with ECOG PS score ≥2 and prealbumin＜0.2 g/L at onset significantly increased in the high EASIX group compared to those in the low EASIX group.Conclusion:At the time of initial diagnosis,EASIX can serve as an independent prognostic indicator impacting OS in patients with MCL.Furthermore,patients in the high EASIX group experience a poorer prognosis and shorter survival duration compared with those in the low EASIX group.

Objective:To investigate the significance of serum β2-microglobulin(β2-MG)for survival and relapse of patients with diffuse large B-cell lymphoma(DLBCL)in the rituximab era.Methods:Clinical data of 92 patients with DLBCL admitted from December 2003 to July 2015 were retrospectively analyzed.The optimal cutoff value of β2-MG levels for predicting prognosis of the DLBCL patients was determined using receiver operating characteristic(ROC)curve.Kaplan-Meier analysis was used to estimate progression-free survival(PFS)and overall survival(OS).Cox logistic regression analysis was used to explore potential prognostic factors associated with survival.Binary logistic regression analysis was used to analyze the relationship between various factors and relapse.Results:The most discriminative cutoff value for β2-MG level was determined to be 2.25 mg/L by the ROC curve.Subgroup analysis showed that patients in the elevated β2-MG(＞2.25 mg/L)group had significantly worse PFS(P=0.006)and a trend toward worse OS compared with those in the low β2-MG(≤2.25 mg/L)group(P=0.053).Univariate analysis showed that elevated β2-MG,age＞60 years,Ann Arbor stage Ⅲ-Ⅳ,as well as IPI score ≥ 3 were associated with worse PFS.Binary logistic regression analysis showed that age＞60 years and β2-M G＞2.25 mg/L were potential influencing factors for relapse of DLBCL patients.Conclusion:Serum β2-MG might be an important predictor for the survival and relapse of DLBCL patients in the rituximab era.

Objective:To investigate the surgical techniques and clinical efficacy of arthroscopic-assisted posterior malleolus reduction for the management of ankle fracture-dislocation.Methods:A retrospective analysis was performed on the clinical data of 27 patients who underwent arthroscopy-assisted posterior malleolar reduction via the posterior approach for the surgical management of ankle fracture-dislocation at the Third Affiliated Hospital of Nanjing Medical University (Changzhou No.2 People's Hospital) between January 2022 and June 2023. The cohort comprised 17 males and 10 females, with a mean age of 43.67±9.56 years (range, 25-63 years). Based on the Bartonícek and Rammelt classification, there were 15 type II cases, 9 type III cases, and 3 type IV cases. The operation time, posterior ankle arthroscopy duration, and postoperative complications, such as neurovascular injury, wound infection, or skin necrosis, were recorded. X-ray and 3D CT imaging were utilized to assess joint surface reduction quality, tibiofibular matching and fracture healing status. The ankle plantarflexion, dorsiflexion and hallux flexion contractures were recorded at the last follow-up. Functional outcomes were measured using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and the Olerud-Molander ankle score (OMAS), while pain was evaluated using the visual analogue scale (VAS).Results:All 27 patients were followed up postoperatively, with a mean follow-up duration of 14.30±1.38 months (range, 12-18 months). Postoperatively, one patient experienced wound exudation, while another developed intermuscular venous thrombosis in the calf. No cases of neurovascular injury, wound infection, skin necrosis, or hallux flexion contracture occurred, and no reduction loss was observed. The mean operation time was 96.11±11.55 min (range, 80-120 min), and the posterior ankle arthroscopy duration was 35.74±5.67 min (range, 30-45 min). Postoperative X-ray evaluations demonstrated no loss of fracture reduction, and all fractures achieved bony union. The mean fracture healing time was 3.78±0.75 months (range, 3-5 months). Postoperative CT evaluations showed no joint surface malalignment, and distal tibiofibular matching was satisfactory. At the final follow-up, mean ankle plantarflexion was 46.74°±4.73° (range, 33°-50°), and dorsiflexion was 20.96°±3.29° (range, 14°-26°). There was no hallux flexion contracture occurred. The mean AOFAS ankle-hindfoot score was 92.11±7.19 (range, 74-100), with 20 excellent, 5 good, and 2 fair, yielding an excellent-good rate of 93%. The mean OMAS score was 94.44±7.25 (range, 75-100), comprising 20 excellent and 7 good results, yielding an excellent-good rate of 100%. The mean VAS score was 0.70±0.95 (range, 0-3).Conclusion:Arthroscopy-assisted posterior malleolar reduction in ankle fracture-dislocation surgery provides optimal soft tissue protection and ensures precise fracture reduction and fixation.