Objective To investigate whether Piceatannol(PIC)improves diabetic retinopathy(DR)mediated by microglial polarization and to elucidate the underlying molecular mechanisms.Methods Network pharmacology and bioinformatics were used to analyze the common targets of DR,PIC,and microglia.Human retinal vascular endothelial cells(HRVECs)were cultured in vitro and randomly divided into the NG-HRVECs group,HG-HRVECs group,and HG+PIC-HRVECs group.Cell viability was assessed by the CCK-8 assay,apoptosis was detected by the TUNEL assay,and the concentrations of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were measured using ELISA kits.BV-2 cells were cultured in vitro and randomly divided into the NG-BV-2 group,HG-BV-2 group,HG+PIC-BV-2 group,HG+Si-NC-BV-2 group,HG+Si-CXCR4-BV-2 group,and HG+Ibrutinib-BV-2 group.The levels of arginase-1(Arg-1)and inducible ni-tric oxide synthase(iNOS)were measured using ELISA kits.Furthermore,conditioned medium(CM)from BV-2 cells of each group was collected to treat HRVECs,after which the viability,apoptosis rate,and TNF-α and IL-6 concentrations of the HRVECs were measured.A DR rat model was established and intervened with PIC to investigate the ameliorative effects of PIC on retinal pathology.Results Bioinformatics analysis identified CXCR4 as the key target of this study.Compared with the NG-HRVECs group,the apoptosis rate and the concentrations of TNF-α and IL-6 were increased in the HG-HRVECs group.Compared with the HG-HRVECs group,the HG+PIC-HRVECs group showed a decreased apoptosis rate and reduced concentrations of TNF-α and IL-6(all P＜0.05).Compared with the NG-BV-2 group,the HG-BV-2 group ex-hibited decreased Arg-1 levels and increased iNOS levels.Compared with the HG-BV-2 group,the HG+PIC-BV-2,HG+Si-CXCR4-BV-2,and HG+Ibrutinib-BV-2 groups all showed increased Arg-1 levels and decreased iNOS levels(all P＜0.05).Compared with the NG-BV-2-CM group,the HG-BV-2-CM group led to decreased viability,increased apoptosis rate,and in-creased concentrations of TNF-α and IL-6 in HRVECs.In contrast,the HG+PIC-BV-2-CM,HG+Si-CXCR4-BV-2-CM,and HG+Ibrutinib-BV-2-CM groups reversed the effects induced by HG-BV-2-CM on HRVECs(all P＜0.05).Animal experiment results showed that compared with DR model rats,rats treated with different doses of PIC exhibited significantly ameliora-ted retinal histopathological damage,and the protein expressions of Arg-1,iNOS,CXCR4,and p-BTK were reversed.Con-clusion PIC ameliorates DR progression mediated by microglial polarization by inhibiting the CXCR4/BTK pathway.

Objective To search,evaluate and summarise the best evidence on pre-rehabilitation before surgery for elderly patients with hip fracture so as to provide an evidence-based basis for further standardisation in clinical practice.Methods Based on the"6S Pyramid"evidence model,desktop searches were conducted across databases and websites including Medlive,BMJ Best Practice,UpToDate,the World Health Organization,the Guidelines International Network(GIN),British National Institute for Health and Care Excellence(NICE),American National Guideline Clearinghouse(NGC),Canadian Medical Association:Clinical Practice Guidelines Infobase(CMA),Scottish Intercollegiate Guidelines Network(SIGN),the Registered Nurses'Association of Ontario(RNAO),American College of Sports Medicine(ACSM),American Academy of Orthopaedic Surgeons(AAOS),European Society for Clinical Nutrition and Metabolism(ESPEN),American Society for Parenteral and Enteral Nutrition(ASPEN),the Enhanced Recovery After Surgery Society(ERASS),Cochrane Library,Joanna Briggs Institute,PubMed,CINAHL,Web of Science,Embase,Scopus,Science Direct,CNKI,Wanfang Data,Vip,and SinoMed.The research targeted evidence on pre-rehabilitation in elderly patients with hip-fracture before surgery,encompassing clinical decisions,guidelines,evidence summaries,best practices,expert consensus,systematic reviews and randomized controlled trials(RCTs).The search items spanned from the inception of database to 31st December,2023.Two researchers trained with evidence-based knowledge evaluated the quality of the included literature and extracted and summarised the evidence.Results A total of 14 documents were included,consisting of 2 clinical decisions,4 guidelines,1 best practice,4 expert consensus and 3 systematic reviews.Evidence was summarised into 7 domains from 25 pieces of evidence:multidisciplinary management,preoperative assessment,preoperative health education,multi-modal analgesia,nutritional management,exercise intervention and assessment and prevention of complication risks.Conclusion The best evidence summarised in this study provides a basis for pre-rehabilitation in elderly patients with hip fracture before surgery.Nurses should adapt the findings to specific contexts,standardise the preoperative rehabilitation procedures,and improve clinical outcomes for the patients.

Objective To search,evaluate and summarise the best evidence on pre-rehabilitation before surgery for elderly patients with hip fracture so as to provide an evidence-based basis for further standardisation in clinical practice.Methods Based on the"6S Pyramid"evidence model,desktop searches were conducted across databases and websites including Medlive,BMJ Best Practice,UpToDate,the World Health Organization,the Guidelines International Network(GIN),British National Institute for Health and Care Excellence(NICE),American National Guideline Clearinghouse(NGC),Canadian Medical Association:Clinical Practice Guidelines Infobase(CMA),Scottish Intercollegiate Guidelines Network(SIGN),the Registered Nurses'Association of Ontario(RNAO),American College of Sports Medicine(ACSM),American Academy of Orthopaedic Surgeons(AAOS),European Society for Clinical Nutrition and Metabolism(ESPEN),American Society for Parenteral and Enteral Nutrition(ASPEN),the Enhanced Recovery After Surgery Society(ERASS),Cochrane Library,Joanna Briggs Institute,PubMed,CINAHL,Web of Science,Embase,Scopus,Science Direct,CNKI,Wanfang Data,Vip,and SinoMed.The research targeted evidence on pre-rehabilitation in elderly patients with hip-fracture before surgery,encompassing clinical decisions,guidelines,evidence summaries,best practices,expert consensus,systematic reviews and randomized controlled trials(RCTs).The search items spanned from the inception of database to 31st December,2023.Two researchers trained with evidence-based knowledge evaluated the quality of the included literature and extracted and summarised the evidence.Results A total of 14 documents were included,consisting of 2 clinical decisions,4 guidelines,1 best practice,4 expert consensus and 3 systematic reviews.Evidence was summarised into 7 domains from 25 pieces of evidence:multidisciplinary management,preoperative assessment,preoperative health education,multi-modal analgesia,nutritional management,exercise intervention and assessment and prevention of complication risks.Conclusion The best evidence summarised in this study provides a basis for pre-rehabilitation in elderly patients with hip fracture before surgery.Nurses should adapt the findings to specific contexts,standardise the preoperative rehabilitation procedures,and improve clinical outcomes for the patients.

Objective To investigate the potential role of plasma adiponectin concentration in predicting the response of patients with depression to electroconvulsive therapy(ECT).Methods This study enrolled depressive patients scheduled for ECT at the First Hospital of Shanxi Medical University between January 2022 and March 2024.Patients were categorized as either ECT responders(>50%reduction)or non-responders(≤50%reduction)based on post-ECT reduction rate of the 24-item Hamilton depression scale(HAMD)scores from baseline.Plasma adiponectin levels were measured in all patients before their first ECT treatment.To identify predictors of ECT response in patients with depression,the predictive value of plasma adiponectin was evaluated using receiver operating characteristic(ROC)curve analysis.Results A total of 80 patients with depression were enrolled including 44 in the ECT-responsive group and 36 in the non-responsive group.The plasma adiponectin level was significantly higher in the responsive group than in the non-responsive group[5.67(3.64,10.55)μg/mL vs.4.01(2.59,5.04)μg/mL,P=0.002].Pearson correlation analysis revealed a significant positive correlation between plasma adiponectin levels and the HAMD reduction rate(r=0.300,P=0.007).Multivariate logistic regression analysis showed that plasma adiponectin was independently associated with the response to ECT in patients with depression(OR=1.218,95%CI:1.009-1.470,P=0.040).Furthermore,ROC curve analysis demonstrated that the area under the curve(AUC)for adiponectin in predicting ECT response was 0.748(95%CI:0.583-0.813).Conclusion Pretreatment plasma adiponectin may serve as a potential biomarker for predicting response to ECT in patients with depression.

Purpose This study aims to analyze genetic mutations in patients with BCR ∷ABL negative myelopro-liferative neoplasms(MPN)and to explore their relationship with clinical features.Methods We retrospectively ana-lyzed the clinical data of 208 patients diagnosed with BCR ∷ABL negative MPN,which included 34 patients with poly-cythemia vera(PV),33 with essential thrombocytopenia(ET),and 141 with primary myelofibrosis(PMF).Mutations in driver genes were assessed in all patients.A total of 72 patients underwent next-generation sequencing(NGS)with 69-gene panel,and the relationship between gene mutations and clinical features were analyzed.Results Among the 208 MPN patients,at least one driver gene mutation(JAK2,CALR,MPL)was detected in 96.15％(200/208)of the patients.Only 0.48％(1/208)of the patients exhibited both JAK2 and CALR driver mutations.We analyzed the clinical data of 136 patients with only driver gene mutations to compare the relationship between the most common JAK2 mutations(identified in 110 patients)and clinical outcomes.The JAK2 mutation group demonstrated higher white blood cell(WBC)counts and lower platelet(PLT)counts compared to the group without JAK2 mutations.173 muta-tions in 40 genes were detected in 72 patients,per capita carried(2.40±1.40)mutations.TET2,ASXL1,and TP53 are the most prevalent non-driver gene mutations,with 44.4％(32/72)of patients exhibiting at least one mutation in these three genes.In comparison to patients without detected mutations in TET2,ASXL1,and TP53,those with muta-tions in these genes demonstrated lower hemoglobin(HGB)levels,a higher incidence of splenomegaly,and more se-vere bone marrow fibrosis.High-molecular risk category(HMR)mutations were detected in 22.22％(16/72)of the patients,and patients with HMR exhibited lower hemoglobin(HGB)levels,lower PLT counts,a higher likelihood of peripheral blood primitive cell percentage ≥ 1％,a greater incidence of splenomegaly,and more severe myelofibrosis.Mutations in the ASXL1 gene were exclusively observed in patients with PMF.Among the PMF patients with ASXL1 mutations(12 patients),there was a higher likelihood of having a peripheral blood primitive cell percentage of ≥1％,as well as a more severe degree of myelofibrosis.Conclusion Approximately 97％of patients with myeloproliferative neoplasms(MPN)exhibit positivity for driver genes,with a notably high mutation rate of the JAK2 gene.Each sub-group of MPN is characterized by distinct gene mutation patterns.Notably,ASXL1 mutations are exclusive to patients with primary myelofibrosis(PMF).Furthermore,PMF patients harboring ASXL1 mutations tend to demonstrate more pronounced bone marrow fibrosis and a greater proportion of blast cells in peripheral blood.

Background and purpose:In recent years,chimeric antigen receptor T(CAR-T)cell therapy has achieved breakthrough progress in cancer treatment.γδ T cells,with their non-major histocompatibility complex(MHC)-restricted antigen recognition,broad antitumor activity,and low risk of graft-versus-host disease(GVHD),have garnered significant interest in CAR-γδ T cell therapy.However,critical challenges including suboptimal in vitro expansion and low viral transduction efficiency severely hinder the research and clinical application of CAR-γδ T cells.This study aimed to establish an efficient platform for preparing CAR-γδ T cells by optimizing the in vitro expansion conditions of γδ T cells and refining lentiviral transduction strategies.Methods:We first optimized the expansion protocol for γδ T cells by screening various cytokine combinations and the concentrations of individual cytokines within combination,and evaluating cell purity,viability,fold expansion,and expressions of cytotoxicity and exhaustion markers to identify the optimal culture conditions.Subsequently,the transduction conditions for CAR-γδ T cells were improved by determining the optimal activation duration of γδ T cells prior to gene transfer,as well as the optimal multiplicity of infection(MOI)for lentiviral transduction.Finally,CAR-γδ T cells were successfully generated using the optimized protocol,and their cytotoxic activity against target cells was validated via calcein-release assay and flow cytometry,with a preliminary assessment of the potential risk of GVHD induction.Results:Experimental data demonstrated that,compared with the interleukin(IL)-2-only culture,the IL-2+IL-7+IL-15 combination significantly enhanced the expansion capacity of γδ T cells(876.50±238.35-fold vs 1 627.50±472.15-fold),cell purity(73.67%±1.53%vs 90.69%±2.00%),and cell viability(63.01%±7.05%vs 89.00%±3.61%).It also increased the expression of the cytotoxicity marker CD16(4.20%±1.73%vs 14.66%±0.58%)and reduced the expression of the exhaustion marker programmed death-1(PD-1)(35.67%±6.26%vs 21.10%±6.49%).A cytokine concentration gradient orthogonal assay further identified 10 ng/mL IL-7 and 10 ng/mL IL-15 as the optimal concentrations within the IL-2+IL-7+IL-15 combination.Gene transduction performed 96-120 h after activation using a multiplicity of infection(MOI)of 5-10 resulted in the highest transduction efficiency for CAR-γδ T cells(96 h:12.87%±4.35%;120 h:11.37%±2.35%).CAR-γδ T cells generated using the optimized system exhibited specific cytotoxic effects against tumor cells expressing the target antigen,and no evidence of GVHD induction was observed.Conclusion:CAR-γδ T cells produced using the IL-2+IL-7(10 ng/mL)+IL-15(10 ng/mL)regimen combined with a 96-120 h activation period prior to transduction using a multiplicity of infection(MOI)of 5-10 significantly outperformed conventional methods in terms of expansion efficiency,cell purity,and transduction efficiency.The synergistic antitumor effects mediated by both natural immune receptors and CAR-specific recognition,along with the initial absence of GVHD risk,provide critical technical support for the clinical translation of CAR-γδ T cell therapy,establishing a solid theoretical and practical foundation.

Objective·To investigate an optimal severe periodontitis mouse model by comparing two induction methods:simple ligature and ligature combined with injection of Porphyromonas gingivalis lipopolysaccharide(P.g.LPS).Methods·Fifteen C57BL/6 mice were divided into three groups:a healthy control group,a simple ligature-induced periodontitis group,and a ligature combined with P.g.LPS injection-induced periodontitis group.After 14 d,the following evaluations were conducted:tooth mobility and probing depth under a stereomicroscope;alveolar bone resorption[bone volume fraction,bone mineral density,the distance from the cemento-enamel junction(CEJ)to the alveolar bone crest(ABC),and the area between CEJ and ABC]analyzed via micro computed tomography(Micro-CT)and stereomicroscopic examination.The serum levels of inflammatory cytokines interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were assessed by enzyme-linked immunosorbent assay(ELISA).Results·Compared with the simple ligature group,mice in the ligature with P.g.LPS injection group exhibited significantly increased tooth mobility[(2.20±0.45)vs(1.40±0.55)]and probing depth[(1.05±0.21)mm vs(0.58±0.39)mm],with statistically significant differences(P＜0.05).The ligature with P.g.LPS injection group also demonstrated significantly reduced bone volume fraction[(16.44%±3.35%)vs(28.97%±7.90%)]and bone mineral density[(0.42±0.04)g/cm3 vs(0.55±0.08)g/cm3],as well as increased distance from CEJ to ABC[(0.88±0.03)mm vs(0.74±0.12)mm]and area between CEJ and ABC[(0.34±0.01)mm2 vs(0.30±0.02)mm2],all with statistically significant differences(all P＜0.05).Additionally,serum levels of TNF-α and IL-1β were significantly elevated in the ligature with P.g.LPS injection group compared to the simple ligature group(both P＜0.05).Conclusion·The method of ligature combined with continuous P.g.LPS injection is more effective for constructing a severe periodontitis mouse model,making it suitable for studying the progression and treatment of severe periodontitis.

Background and purpose:In recent years,chimeric antigen receptor T(CAR-T)cell therapy has achieved breakthrough progress in cancer treatment.γδ T cells,with their non-major histocompatibility complex(MHC)-restricted antigen recognition,broad antitumor activity,and low risk of graft-versus-host disease(GVHD),have garnered significant interest in CAR-γδ T cell therapy.However,critical challenges including suboptimal in vitro expansion and low viral transduction efficiency severely hinder the research and clinical application of CAR-γδ T cells.This study aimed to establish an efficient platform for preparing CAR-γδ T cells by optimizing the in vitro expansion conditions of γδ T cells and refining lentiviral transduction strategies.Methods:We first optimized the expansion protocol for γδ T cells by screening various cytokine combinations and the concentrations of individual cytokines within combination,and evaluating cell purity,viability,fold expansion,and expressions of cytotoxicity and exhaustion markers to identify the optimal culture conditions.Subsequently,the transduction conditions for CAR-γδ T cells were improved by determining the optimal activation duration of γδ T cells prior to gene transfer,as well as the optimal multiplicity of infection(MOI)for lentiviral transduction.Finally,CAR-γδ T cells were successfully generated using the optimized protocol,and their cytotoxic activity against target cells was validated via calcein-release assay and flow cytometry,with a preliminary assessment of the potential risk of GVHD induction.Results:Experimental data demonstrated that,compared with the interleukin(IL)-2-only culture,the IL-2+IL-7+IL-15 combination significantly enhanced the expansion capacity of γδ T cells(876.50±238.35-fold vs 1 627.50±472.15-fold),cell purity(73.67%±1.53%vs 90.69%±2.00%),and cell viability(63.01%±7.05%vs 89.00%±3.61%).It also increased the expression of the cytotoxicity marker CD16(4.20%±1.73%vs 14.66%±0.58%)and reduced the expression of the exhaustion marker programmed death-1(PD-1)(35.67%±6.26%vs 21.10%±6.49%).A cytokine concentration gradient orthogonal assay further identified 10 ng/mL IL-7 and 10 ng/mL IL-15 as the optimal concentrations within the IL-2+IL-7+IL-15 combination.Gene transduction performed 96-120 h after activation using a multiplicity of infection(MOI)of 5-10 resulted in the highest transduction efficiency for CAR-γδ T cells(96 h:12.87%±4.35%;120 h:11.37%±2.35%).CAR-γδ T cells generated using the optimized system exhibited specific cytotoxic effects against tumor cells expressing the target antigen,and no evidence of GVHD induction was observed.Conclusion:CAR-γδ T cells produced using the IL-2+IL-7(10 ng/mL)+IL-15(10 ng/mL)regimen combined with a 96-120 h activation period prior to transduction using a multiplicity of infection(MOI)of 5-10 significantly outperformed conventional methods in terms of expansion efficiency,cell purity,and transduction efficiency.The synergistic antitumor effects mediated by both natural immune receptors and CAR-specific recognition,along with the initial absence of GVHD risk,provide critical technical support for the clinical translation of CAR-γδ T cell therapy,establishing a solid theoretical and practical foundation.

In recent years, idiopathic macular hole (IMH) surgery has a very high success rate due to the development of small-gauge vitrectomy, while the recovery of visual acuity is not satisfactory.Current studies have found that continuity of retinal blood vessels in the macular area and changes in retinal and choroidal perfusion are closely related to the formation and healing of MH, and have a significant impact on the visual prognosis in IMH patients.This article reviews the research progress on variations of parameters for retinal and choroidal vessels in IMH patients, as well as their relationship with visual prognosis.For retinal parameters, the changes in the foveal avascular zone, superficial and deep capillary plexus density, paracentral intraretinal cystoid spaces, retinal vessels position are discussed.Relevant research related to choroidal blood flow perfusion changes and the effects of different surgical methods on macular perfusion in patients with IMH are summarized to provide basis for further understanding the etiology and prognosis of IMH.

Objective To investigate whether Piceatannol(PIC)improves diabetic retinopathy(DR)mediated by microglial polarization and to elucidate the underlying molecular mechanisms.Methods Network pharmacology and bioinformatics were used to analyze the common targets of DR,PIC,and microglia.Human retinal vascular endothelial cells(HRVECs)were cultured in vitro and randomly divided into the NG-HRVECs group,HG-HRVECs group,and HG+PIC-HRVECs group.Cell viability was assessed by the CCK-8 assay,apoptosis was detected by the TUNEL assay,and the concentrations of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were measured using ELISA kits.BV-2 cells were cultured in vitro and randomly divided into the NG-BV-2 group,HG-BV-2 group,HG+PIC-BV-2 group,HG+Si-NC-BV-2 group,HG+Si-CXCR4-BV-2 group,and HG+Ibrutinib-BV-2 group.The levels of arginase-1(Arg-1)and inducible ni-tric oxide synthase(iNOS)were measured using ELISA kits.Furthermore,conditioned medium(CM)from BV-2 cells of each group was collected to treat HRVECs,after which the viability,apoptosis rate,and TNF-α and IL-6 concentrations of the HRVECs were measured.A DR rat model was established and intervened with PIC to investigate the ameliorative effects of PIC on retinal pathology.Results Bioinformatics analysis identified CXCR4 as the key target of this study.Compared with the NG-HRVECs group,the apoptosis rate and the concentrations of TNF-α and IL-6 were increased in the HG-HRVECs group.Compared with the HG-HRVECs group,the HG+PIC-HRVECs group showed a decreased apoptosis rate and reduced concentrations of TNF-α and IL-6(all P＜0.05).Compared with the NG-BV-2 group,the HG-BV-2 group ex-hibited decreased Arg-1 levels and increased iNOS levels.Compared with the HG-BV-2 group,the HG+PIC-BV-2,HG+Si-CXCR4-BV-2,and HG+Ibrutinib-BV-2 groups all showed increased Arg-1 levels and decreased iNOS levels(all P＜0.05).Compared with the NG-BV-2-CM group,the HG-BV-2-CM group led to decreased viability,increased apoptosis rate,and in-creased concentrations of TNF-α and IL-6 in HRVECs.In contrast,the HG+PIC-BV-2-CM,HG+Si-CXCR4-BV-2-CM,and HG+Ibrutinib-BV-2-CM groups reversed the effects induced by HG-BV-2-CM on HRVECs(all P＜0.05).Animal experiment results showed that compared with DR model rats,rats treated with different doses of PIC exhibited significantly ameliora-ted retinal histopathological damage,and the protein expressions of Arg-1,iNOS,CXCR4,and p-BTK were reversed.Con-clusion PIC ameliorates DR progression mediated by microglial polarization by inhibiting the CXCR4/BTK pathway.