Objective·To investigate the effects and molecular mechanisms of phosphatidylethanolamine(PE)on macrophage senescence and its senescence-associated secretory phenotype(SASP),as well as its pathophysiological role in liver injury.Methods·A macrophage senescence model was established using doxorubicin(DOX),followed by PE treatment.A mouse liver injury model was generated via intraperitoneal co-administration of PE and lipopolysaccharide(LPS)to investigate the effects of PE on liver injury.Senescence markers and SASP factors,including senescence-associated β-galactosidase(SA-β-gal),cell cycle inhibitor p21,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6),were evaluated using SA-β-gal staining,quantitative real-time PCR,and Western blotting.RNA sequencing(RNA-seq)was performed,followed by Gene Ontology(GO)cellular component enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,Gene Set Variation Analysis(GSVA),and Gene Set Enrichment Analysis(GSEA),to explore the molecular mechanisms and signaling pathways by which PE promotes macrophage senescence.The expression of endoplasmic reticulum(ER)stress-related proteins,including inositol-requiring enzyme 1 α(IRE1α),spliced X-box binding protein 1(XBP1s),activating transcription factor 6(ATF6),ATF4,and C/EBP homologous protein(CHOP),was analyzed through in vivo and in vitro experiments.Results·PE significantly promoted the expression of senescence markers SA-β-gal,p21,p16 and SASP factors.RNA-seq analysis revealed that ER stress was involved in PE-induced promotion of SASP.Further experiments demonstrated that PE activated the ER stress signaling pathway,promoting macrophage senescence and the expression of SASP factors.In vivo experiments further confirmed that PE exacerbated LPS-induced liver injury in mice through ER stress.Conclusion·PE promotes macrophage senescence and the expression of SASP factors by activating ER stress signaling pathway,thereby aggravating LPS-induced liver injury.

Objective·To explore the mechanism at the atomic level by which the KRASG12D/R68G mutation induces tumor cell resistance to MRTX1133.Methods·The crystal structure data of the KRASG12D-MRTX1133 complex were obtained from the RCSB Protein Data Bank(PDB).PyMOL software was used to mutate arginine at position 68 of KRAS to glycine(R68G),constructing the initial conformations of the KRASG12D-MRTX1133 and KRASG12D/R68G-MRTX1133 complexes.The LEaP module was used to build simulation systems under periodic boundary conditions.The ff19SB force field was applied to standard amino acids in KRAS,GAFF(general AMBER force field)to MRTX1133,and TIP3P(intermolecular potential three point)to water molecules.Energy minimization was performed using the Amber software suite.The systems were then heated to 300 K,followed by NVT(constant volume and temperature)equilibration and NPT(constant pressure and temperature)production.Root mean square deviation(RMSD),root mean square fluctuation(RMSF),principal component analysis(PCA)and solvent-accessible surface area(SASA)of MRTX1133 and GDP were analyzed using cpptraj.The number of hydrogen bonds between regions and the dynamic cross-correlation matrix(DCCM)of amino acid movements were also calculated.Results·RMSD analysis showed greater structural variation in KRAS in the KRASG12D/R68G system compared to the KRASG12D system.RMSF analysis revealed significantly higher fluctuations in the Switch Ⅰ and Switch Ⅱ regions of the KRASG12D/R68G system.PCA indicated that Switch Ⅰ and Switch Ⅱ in the KRASG12D/R68G system were more frequently in an open conformation.The distances between Switch Ⅰ and the P-loop,and between Switch Ⅱ and the P-loop,were larger in the KRASG12D/R68G system,indicating an expanded binding pocket for GDP and MRTX1133 compared to the KRASG12D system.SASA analysis indicated that both GDP and MRTX1133 had increased solvent exposure in the KRASG12D/R68G system.DCCM analysis revealed more decoupled movements among the Switch Ⅰ,Switch Ⅱ and P-loop regions in the KRASG12D/R68G system.Conclusion·The KRASG12D/R68G mutation disrupts the interactions between the Switch Ⅰ and Switch Ⅱregions,leading to their separation and the opening of the MRTX1133 binding pocket.This increases the solvent exposure of MRTX1133,accelerates its dissociation,and ultimately results in KRASG12D/R68G resistance to MRTX1133.

Traditional treatment modalities for thalassemia include regular blood transfusions and allogenic hematopoietic stem cell transplantation(allo-HSCT).In recent years,autologous transplantation of gene-modified hematopoietic stem cells has emerged as a new curative strategy for transfusion-dependent thalassemia(TDT),which has the potential to replace conventional treatments,and provide lifelong benefits for patients.There are two existing technical approaches for gene therapy of β-thalassemia:gene addition,which involves transducing exogenous β-globin genes into hematopoietic stem cells(HSCs),and gene editing,which utilizes CRISPR-Cas9 or other editing systems to re-activate the expression of γ-globin gene.This article summarizes the marketed products and research progress in clinical trials,aiming to analyze the respective advantages and limitations of these two approaches,and discusses the effectiveness and safety of current gene therapies for β-thalassemia,as well as the future directions for associated technologies,including ex vivo HSC expansion with maintenance of stemness and vector-mediated in vivo gene modification.In terms of clinical translational medicine,this article provides in-depth insights into promising solutions for contemporary challenges confronted in clinical trials,including process development challenges,clinical trial conduct,regulatory approval processes,commercialization and payment systems.

Objective·To explore the prognostic value of M2 macrophage-related genes in prostate cancer(PCa),aiming to predict tumor prognosis more accurately and enable personalized treatment.Methods·RNA sequencing(RNA-seq)data of PCa were downloaded from The Cancer Genome Atlas(TCGA)database,and single-cell RNA sequencing(scRNA-seq)data were obtained from the Gene Expression Omnibus(GEO)database.The immune infiltration of TCGA samples was assessed using the CIBERSORTx algorithm.Differential genes in scRNA-seq data were identified using the FindMarkers function,and immune cell subtypes were characterized.M2 macrophage-related pathways and interactions with surrounding cells were explored through Gene Set Enrichment Analysis(GSEA)and the CellChat algorithm.M2 macrophage signature genes were selected to construct a prognostic model for PCa using univariate Cox and LASSO analyses.Based on the risk model,clinical characteristics,immune suppression,drug resistance,and drug sensitivity analyses were conducted.Results·In TCGA samples,patients with high M2 macrophage infiltration exhibited significantly lower progression-free survival(PFS).scRNA-seq analysis identified multiple subpopulations of tumor microenvironment(TME)cells.M2 macrophages interacted with various immune cells in TME,contributing to an immunosuppressive microenvironment and playing a key role in tumor promotion.Based on these findings,a PCa risk model was developed,incorporating TREM2,OTOA,SIGLEC1,and PLXDC1,which showed robust predictive performance in both training and validation cohorts.Patients with higher risk scores demonstrated a more immunosuppressive TME,decreased androgen receptor(AR)signaling activity,and worse clinical characteristics,leading to poorer outcomes.Drug prediction and sensitivity analyses identified six potential therapeutic agents that may offer improved efficacy for patients with higher risk scores.Conclusion·A prognostic model based on M2 macrophage-related genes in the TME has been constructed,providing a theoretical foundation for precision treatment in PCa.

Objective·To study the effect of neodymium-doped yttrium aluminum perovskite(Nd:YAP)laser biostimulation on the proliferation and osteogenic differentiation of human periodontal ligament cells(hPDLCs)and its possible mechanism.Methods·Five premolars removed for orthodontic reasons were collected from Changzhou Stomatological Hospital,and the periodontal ligament tissues from the middle 1/3 of the roots were taken to culture hPDLCs in vitro.The cells were irradiated with the biostimulation function[G(-)mode]of the Nd:YAP laser.According to the irradiation time,the cells were divided into a control group(without laser irradiation),and groups irradiated for 5 s,10 s,15 s,20 s and 30 s.The cell counting kit-8(CCK-8)method was used to detect the proliferation of hPDLCs in each group.After osteogenic differentiation was induced,the alkaline phosphatase(ALP)content and activity level of the cells were detected using an ALP staining kit and an ALP activity detection kit.The calcium salt level of the cells was evaluated by alizarin red S staining and calcium quantitative analysis.The expression of genes and proteins related to the WNT/β-catenin signaling pathway,including dickkopf-related protein 1(DKK-1),β-catenin,and runt-related transcription factor 2(RUNX2),was analyzed by quantitative real-time PCR(qPCR)and Western blotting.Results·The results of CCK-8 showed that the proliferation level of cells in the 10 s,15 s,20 s,and 30 s groups was enhanced from 3 d after irradiation(all P<0.05).After induction of osteogenic differentiation,ALP content,activity,and calcium salt level in the laser irradiation groups increased with the extension of irradiation time(all P<0.05).The results of qPCR and Western blotting analysis showed that the expression levels of the DKK-1 gene and protein in the laser irradiation groups decreased with the extension of irradiation time.However,the expression levels of β-catenin and RUNX2 genes and proteins increased significantly with the extension of irradiation time;there were statistically significant differences between the 15 s,20 s,and 30 s groups and the control group(all P<0.05).Conclusion·Nd:YAP laser biostimulation may promote proliferation and osteogenic differentiation of hPDLCs through WNT/β-catenin signaling pathway.

Objective·To evaluate the effects of normally positioned flap(NPF)and coronally advanced flap(CAF)techniques on clinical results after epulis excision.Methods·A total of 55 patients with epulis who visited the Department of Periodontology,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine from August 2022 to December 2023 were included.The patients were divided into the NPF group and the CAF group.After epulis excision,the surgical area was closed using NPF or CAF technique.The following parameters were recorded:baseline epulis width(EW)and epulis height(EH);papilla width(PW)and papilla height(PH)at 6 months post-surgery;probing depth(PD),attachment loss(AL),and keratinized gingiva width(KGW)at both baseline and 6 months post-surgery.The esthetic outcomes were evaluated using a visual analog scale(VAS)by 2 periodontal specialists.t test was used to compare the differences in periodontal indices between baseline and 6 months post-surgery,as well as between the 2 flap techniques.Results·At 6 months post-surgery,PD of the CAF group was(1.68±0.79)mm,significantly lower than at baseline(P<0.001),but not significantly different from that in the NPF group(P=0.365);the AL in the CAF group was(1.26±1.18)mm,not significantly different from baseline(P=0.746),but significantly lower than in the NPF group(P<0.001).At 6 months post-surgery,PH of the CAF group was(3.74±0.62)mm,significantly higher than that in the NPF group(P<0.001),and the VAS score of the CAF group was significantly higher than that of the NPF group(P<0.001).Conclusion·Compared with NPF,CAF could effectively improve post-surgical KGW and reduce AL,which could prevent periodontal soft tissue defects,and improve esthetic outcome after epulis excision.

Microglia are the resident immune cells in the central nervous system(CNS),and play a dual role in maintaining brain homeostasis and mediating neuroprotection.Under normal conditions,microglia maintain brain homeostasis by monitoring environmental changes.When nerve damage or certain pathological stimuli occur,microglia are rapidly activated and initiate a series of complex immune responses to induce neuroinflammation.This proper activation of microglia can protect the brain by inhibiting or clearing various pathogens,but excessive neuroinflammation can lead to neuronal damage and even death.This imbalance of inflammatory response is one of the core features of pathological development of many CNS inflammatory diseases,such as Alzheimer's disease,Parkinson's disease,sepsis-associated encephalopathy,and ischemic strokes.In recent years,with the rapid development of frontier biotechnology such as single-cell sequencing,proteinomics and gene editing,important progress has been made in understanding the molecular mechanism by which microglia participate in CNS inflammatory diseases,especially in the activation of inflammatory corpuscles,epigenetic modifications,and metabolic reprogramming.However,due to the heterogeneity and duality of microglia under different pathological conditions,therapeutic methods targeting microglia have not yet been widely used in clinical practice.In summary,this article takes microglia as the starting point and introduces the molecular mechanisms of their involvement in the occurrence and development of CNS inflammatory diseases and its targeted regulatory treatment strategy,aiming to provide theoretical reference for the subsequent precise regulation of microglia function and the development of more targeted therapeutic drugs.

Parkinson's disease(PD),the second most prevalent neurodegenerative disease,has shown an increasing incidence in recent years,significantly impacting the quality of life of elderly individuals and their families.The onset of cognitive impairment and Parkinson's disease dementia(PDD)are critical milestones in the progression of PD.Imaging examinations,serving as essential detection and evaluation tools for neurodegenerative diseases,have been increasingly applied to PD-related research.Different imaging techniques demonstrate distinct advantages and features in screening cognitive dysfunction in PD.Choosing a reliable imaging method not only maximizes the advantages of the examination,enhancing the sensitivity and specificity in identifying PD patients at risk of cognitive impairment,but also reduces the frequency of examinations and the radiation dose received by patients.This review summarizes the existing findings on imaging markers of cognitive impairment in PD from three aspects,structural imaging,functional imaging,and multimodal techniques.Furthermore,it explores future research directions,aiming to provide powerful imaging support for the clinical diagnosis and treatment of PD.

The diagnosis of schizophrenia usually relies on the assessment of clinical symptoms,and the search for objective biomarkers is particularly important for the diagnosis and treatment of the disease.Since the retina can reflect the state of the central nervous system,more and more studies are focusing on retina-specific alterations in neuropsychiatric disorders.This review summarizes recent studies on the retinal nerve layer,vascular characteristics,and electrophysiological features in patients with schizophrenia,showing that patients with schizophrenia often have thinner retinal ganglion cell-inner plexiform layer and retinal nerve fiber layer.The changes in the retinal layers vary in different stages of schizophrenia.Studies of the fundus vasculature in schizophrenic patients have also suggested the presence of altered retinal vascular density and microvascular morphology in schizophrenic patients.Studies of electroretinography suggest that patients in the acute phase of schizophrenia tend to exhibit reduced a-wave amplitudes of cone cells,while those at genetic high risk often show a tendency for reduced b-wave amplitudes of rod cells.However,the current retina-related studies in schizophrenia mostly focus on clinical manifestations,with fewer studies on related mechanisms and inconsistent findings.This review attempts to discuss a variety of potential pathophysiological mechanisms,including trans-synaptic retrograde degeneration hypothesis,neurotransmitter disturbance,genetics,brain structural changes,and metabolism,in the context of the retinal nerve layer,microcirculation,and electrophysiology alterations,in order to provide new insights into the pathophysiological mechanisms and objective biomarkers of schizophrenia.

Objective·To optimize iSeroSnFR,a genetically encoded 5-hydroxytryptamine(5-HT)fluorescent sensor based on bacterial periplasmic binding proteins(PBPs),to enhance its performance for both in vivo and in vitro 5-HT detection.Methods·iSeroSnFR1.2 was engineered by replacing the circularly permuted superfolder green fluorescence protein(cpsfGFP)sequence in iSeroSnFR1.0 with that from the acetylcholine sensor iAChSnFR using Gibson assembly.The fluorescence response and kinetic properties of iSeroSnFR1.0 and iSeroSnFR1.2 were compared by overexpressing the sensors in HEK293 cells and puffing with exogenous 5-HT.Additionally,to mimic physiological conditions,cultured mouse cortical neurons infected with Sindbis virus carrying each sensor were electrically stimulated to induce endogenous 5-HT release and further evaluate sensor performance.Results·iSeroSnFR1.2 showed significantly improved performance over iSeroSnFR1.0.In HEK293 cells,it exhibited a 1.5-fold increase in fluorescence response(ΔF/F0)to exogenous 5-HT,along with faster kinetics(rise time:36.3 ms vs 44.9 ms;decay time:1 003.6 ms vs 1 730.4 ms).In cortical neurons,it demonstrated a 2.7-fold increase in response to endogenously released 5-HT,with rise and decay times reduced by 44.0%and 26.7%,respectively.Notably,iSeroSnFR1.2 showed increased basal fluorescence,enabling better imaging in high-background environments.Conclusion·The optimized iSeroSnFR1.2 sensor offers a markedly improved fluorescent response and temporal resolution for 5-HT detection,providing an advanced tool for studying 5-HT dynamics in neuroscience and psychiatric research.