Exosomal miR-320e through wnt2targeted inhibition of the Wnt/β-catenin pathway allevisate cerebral small vessel disease and cognitive impairment

BACKGROUND Exosomal miRNAs play crucial roles in many central nervous system diseases.Cerebral small vessel disease(CVSD)is a small vessel disease that is affected by various factors.This study aimed to investigate the role of exosomal miR-320e in the Wnt/β-catenin pathway stimulated by oxidative stress and assess its clinical correlation with psychiatric symptoms in patients with CVSD.AIM To explore whether exosomal miR-320e could suppress the Wnt/β-catenin pathway and play a protective role in CVSD progression,as well as examine its potential correlation with cognitive impairment and depression in patients with CVSD.METHODS Differentially expressed exosomal miRNAs were filtered by sequencing plasma exosomes from patients with CVSD and healthy controls.Bioinformatics and dual luciferase analyses were used to confirm the binding of miR-320e to Wnt2,and the mRNA and protein levels of downstream components in the Wnt/β-catenin pathway were evaluated when overexpressed or with knockdown of miR-320e under H2O2-induced oxidative stress.In addition,Wnt2-targeting siRNA was used to confirm the role of miR-320e in the Wnt2-mediated inhibition of the Wnt/β-catenin pathway.A retrospective analysis was conducted among patients with CVSD to confirm the correlation between miR-320e expression and the severity of cognitive impairment and depression,which were quantified using the Montreal Cognitive Assessment(MoCA)/Executive Function Assessment(EFA),and the Hamilton Depression Scale(HAMD)/Beck Depression Inventory(BDI),respectively.RESULTS High-throughput sequencing revealed that exosomal miR-320e was downregulated in patients with CVSD.Bioinformatics analysis and dual-luciferase reporter gene experiments showed that exosomal miR-320e inhibited the Wnt/β-catenin pathway in response to oxidative stress by targeting the 3'noncoding region of Wnt2.Uptake of exosomes carrying miR-320e into endothelial cells could also target Wnt2 and inhibit the Wnt2/β-catenin pathway.Elevated miR-320e expression may protect patients with CVSD from relatively severe cognitive impairment and depression,as it was found to have a positive correlation with the MoCA/EFA and HAMD/BDI scores.CONCLUSION Our results suggest that exosomal miR-320e suppresses the Wnt/β-catenin pathway and may play a protective role in CVSD progression.

Hes1 Knockdown Exacerbates Ischemic Stroke Following tMCAO by Increasing ER Stress-Dependent Apoptosis via the PERK/ eIF2a/ATF4/CHOP Signaling Pathway

Apoptosis induced by endoplasmic reticulum(ER)stress plays a crucial role in mediating brain damage after ischemic stroke.Recently,Hes1(hairy and enhancer of split 1)has been implicated in the regulation of ER stress,but whether it plays a functional role after ischemic stroke and the underlying mechanism remain unclear.In this study,using a mouse model of ischemic stroke via transient middle cerebral artery occlusion(tMCAO),we found that Hes1 was induced following brain injury,and that siRNA-mediated knockdown of Hes1 increased the cerebral infarction and worsened the neurological outcome,suggesting that Hes1 knockdown exacerbates ischemic stroke.In addition,mechanistically,Hes1 knockdown promoted apoptosis and activated the PERK/eIF2a/ATF4/CHOP signaling pathway after tMCAO.These results suggest that Hes1 knockdown promotes ER stress-induced apoptosis.Furthermore,inhibition of PERK with the specific inhibitor GSK2606414 markedly attenuated the Hes1 knockdown-induced apoptosis and the increased cerebral infarction as well as the worsened neurological outcome following tMCAO,implying that the protection of Hes1 against ischemic stroke is associated with the amelioration of ER stress via modulating the PERK/eIF2a/ATF4/CHOP signaling pathway.Taken together,these results unveil the detrimental role of Hes1 knockdown after ischemic stroke and further relate it to the regulation of ER stress-induced apoptosis,thus highlighting the importance of targeting ER stress in the treatment of ischemic stroke.

A de novo missense mutation in MPP2 confers an increased risk of Vogt–Koyanagi–Harada disease as shown by trio-based whole-exome sequencing

Vogt–Koyanagi–Harada(VKH)disease is a leading cause of blindness in young and middle-aged people.However,the etiology of VKH disease remains unclear.Here,we performed the first trio-based whole-exome sequencing study,which enrolled 25 VKH patients and 50 controls,followed by a study of 2081 VKH patients from a Han Chinese population to uncover detrimental mutations.A total of 15 de novo mutations in VKH patients were identified,with one of the most important being the membrane palmitoylated protein 2(MPP2)p.K315N(MPP2-N315)mutation.The MPP2-N315 mutation was highly deleterious according to bioinformatic predictions.Additionally,this mutation appears rare,being absent from the 1000 Genome Project and Genome Aggregation Database,and it is highly conserved in 10 species,including humans and mice.Subsequent studies showed that pathological phenotypes and retinal vascular leakage were aggravated in MPP2-N315 mutation knock-in or MPP2-N315 adeno-associated virus-treated mice with experimental autoimmune uveitis(EAU).In vitro,we used clustered regularly interspaced short palindromic repeats(CRISPR‒Cas9)gene editing technology to delete intrinsic MPP2 before overexpressing wild-type MPP2 or MPP2-N315.Levels of cytokines,such as IL-1β,IL-17E,and vascular endothelial growth factor A,were increased,and barrier function was destroyed in the MPP2-N315 mutant ARPE19 cells.Mechanistically,the MPP2-N315 mutation had a stronger ability to directly bind to ANXA2 than MPP2-K315,as shown by LC‒MS/MS and Co-IP,and resulted in activation of the ERK3/IL-17E pathway.Overall,our results demonstrated that the MPP2-K315N mutation may increase susceptibility to VKH disease.

斑块型银屑病患者外周血SLC35E2B基因的表达及其变异研究

目的通过检测斑块型银屑病患者外周血中SLC35E2B的表达情况,并利用数据库分析基因富集的功能和参与的通路,探讨SLC35E2B在银屑病发病机制中的作用。方法收集斑块型银屑病47例、正常对照14例的外周静脉血,提取样本基因组DNA,用外显子捕获-高通量测序(exon-capture high-throughput sequencing,EC-HTS)技术进行检测,数据经过Calling、数据库比对、软件分析得到最终SLC35E2B基因病理性突变位点,利用GO和KEGG pathway数据库分析基因富集的功能和参与的通路。结果对61例样本所得数据进行生物信息学分析得到致病性突变位点1个:SLC35E2Bc.833C>T(p.T278M),仅见于斑块型银屑病组,正常组未见;GO富集提示SLC35E2B生物学过程主要涉及调节IL-12家族细胞因子、IL-17家族细胞因子、调节对生物刺激的细胞应答、TNF的合成、调节Th1细胞相关的免疫应答以及抵抗病原微生物等多细胞生物过程;KEGG通路分析提示SLC35E2B基因主要与结核病、1型糖尿病、弓形虫病、哮喘、类风湿性关节炎等疾病相关。结论SLC35E2B可能参与银屑病发病的病理过程。

Therapeutic Role of Chinese Medicine Targeting Nrf2/HO-1 Signaling Pathway in Myocardial Ischemia/Reperfusion Injury

Acute myocardial infarction(AMI),characterized by high incidence and mortality rates,poses a significant public health threat.Reperfusion therapy,though the preferred treatment for AMI,often exacerbates cardiac damage,leading to myocardial ischemia/reperfusion injury(MI/RI).Consequently,the development of strategies to reduce MI/RI is an urgent priority in cardiovascular therapy.Chinese medicine,recognized for its multi-component,multi-pathway,and multi-target capabilities,provides a novel approach for alleviating MI/RI.A key area of interest is the nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)pathway.This pathway is instrumental in regulating inflammatory responses,oxidative stress,apoptosis,endoplasmic reticulum stress,and ferroptosis in MI/RI.This paper presents a comprehensive overview of the Nrf2/HO-1 signaling pathway's structure and its influence on MI/RI.Additionally,it reviews the latest research on leveraging Chinese medicine to modulate the Nrf2/HO-1 pathway in MI/RI treatment.

Efficient oxygen reduction reaction by a highly porous,nitrogen-doped carbon sphere electrocatalyst through space confinement effect in nanopores

A highly porous nitrogen-doped carbon sphere(NPC)electrocatalyst was prepared through the carbonization of biomass carbon spheres mixed with urea and zinc chloride in N_(2) atmosphere.The sample carbonized at.1000℃ demonstrates a superior oxygen reduction reaction(ORR)performance over the Pt/C electrocatalyst,while its contents of pyridinic nitrogen and graphitic nitrogen are the lowest among samples synthesized at the same or lower carbonization temperatures.This unusual result is explained by a space confinement effect from the microporous and mesoporous structures in the microflakes,which induces the further reduction of peroxide ions or other oxygen species produced in the first step reduction to water to have the preferred overall four electron reduction ORR process.This work demonstrates that in addition to the amount or species of its aptive sites,the space confinement can be a new approach to enhance the ORR performance of precious-metal-free,nitrogen-doped carbon electrocatalysts.

Mapping the Binding Site of P53 on UBC9 by NMR Spectroscopy

Human UBC9 is a member of the E2 family of proteins. However, instead of conjugating to ubiquitin, it conjugates to a ubiquitin homologue SUMO-1 (also known as UBL1, GMP1, SMTP3, PICT-1 and sentrin). The SUMO-1 conjugation pathway is very similar to that of ubiquitin with regard to the primary sequences of the ubiquitin activating enzymes (E1), the three-dimensional structures of the ubiquitin conjugating enzymes (E2), and the chemistry of the overall conjugation pathway. The interaction of p53 and UBC9, the E2 of the SUMO-1 pathway, has been studied by nuclear magnetic resonance spectroscopy. A peptide corresponding to the nuclear localization domain of p53 specifically interacts with UBC9 and this interaction is likely to be important for conjugation of p53 with SUMO-1. The largest chemical shift changes on UBC9 occur at residues 94 and 129-135. This region is adjacent to the active site and has significant dynamic behavior on the μs-ms and ps-ns timescales. Correlation of chemical shift changes and mobility of these residues further suggest the importance of these residues in substrate recognition.