The overdiagnosis of prostate cancer(PCa)caused by nonspecific elevation serum prostate-specific antigen(PSA)and the overtreatment of indolent PCa have become a global problem that needs to be solved urgently.We aimed...The overdiagnosis of prostate cancer(PCa)caused by nonspecific elevation serum prostate-specific antigen(PSA)and the overtreatment of indolent PCa have become a global problem that needs to be solved urgently.We aimed to construct a prediction model and provide a risk stratification system to reduce unnecessary biopsies.In this retrospective study,clinical data of 1807 patients from three Chinese hospitals were used.The final model was built using stepwise logistic regression analysis.The apparent performance of the model was assessed by receiver operating characteristic curves,calibration plots,and decision curve analysis.Finally,a risk stratification system of clinically significant prostate cancer(csPCa)was created,and diagnosis-free survival analyses were performed.Following multivariable screening and evaluation of the diagnostic performances,a final diagnostic model comprised of the PSA density and Prostate Imaging-Reporting and Data System(PI-RADS)score was established.Model validation in the development cohort and two external cohorts showed excellent discrimination and calibration.Finally,we created a risk stratification system using risk thresholds of 0.05 and 0.60 as the cut-off values.The follow-up results indicated that the diagnosis-free survival rate for csPCa at 12 months and 24 months postoperatively was 99.7%and 99.4%,respectively,for patients with a risk threshold below O.05 after the initial negative prostate biopsy,which was significantly better than patients with higher risk.Our diagnostic model and risk stratification system can achieve a personalized risk calculation of csPCa.It provides a standardized tool for Chinese patients and physicians when considering thenecessity of prostatebiopsy.展开更多
Although per-and polyfluoroalkyl substances(PFAS)have been frequently linked to cardiovascular and renal disease separately,evidence remains scarce regarding their systematic effect.Therefore,we recruited 546 newly di...Although per-and polyfluoroalkyl substances(PFAS)have been frequently linked to cardiovascular and renal disease separately,evidence remains scarce regarding their systematic effect.Therefore,we recruited 546 newly diagnosed acute coronary syndrome(ACS)patients and detected seven myocardial enzymes and six kidney function biomarkers.Twelve PFASwere also assessedwith ultra-high-performance liquid chromatography-tandem mass spectrometry.Generalized linear model and restricted cubic spline model were applied to single pollutant analysis.Quantile g-computation was used for mixture analysis.Network model was utilized to identify central and bridge nodes of pollutants and phenotypes.In the present study,perfluorohexane sulfonic acid was positively associated with uric acid(UA)(β=0.04,95%confidence interval(CI):0.01,0.07),and perfluorobutanoic acid was negatively associated with estimated glomerular filtration rate(β=-0.04,95%CI:-0.07,-0.01)but positively associated with UA(β=0.03,95%CI:0.01,0.06).In mixture analysis,each quantile increase in the PFAS mixture was significantly associated with UA(β=0.08,95%CI:0.04,0.11).Network analysis revealed that perfluorooctanoate,UA,and myoglobin were denoted as bridge nodes,and the first principal component of lactate dehydrogenase and creatine kinase-myocardial band was identified as the node with the highest strength and expected influence.This study investigates the systematic impact of PFAS exposure through cardiorenal interaction network,which highlights that PFAS may serve as an upstream approach in UA-modulated cardiorenal network to affect cardiorenal system comprehensively.展开更多
Objective To investigate the role and molecular mechanism of membrane-associated guanylate kinase inverted 3 （MAGI3） in glioma cell proliferation. Methods The expression levels of MAGI3 and PTEN were assessed in gli...Objective To investigate the role and molecular mechanism of membrane-associated guanylate kinase inverted 3 （MAGI3） in glioma cell proliferation. Methods The expression levels of MAGI3 and PTEN were assessed in glioma samples by Western blotting. MAGI3 was stably transfected into C6 glioma cells to obtain C6-MAGI3 cells. Then, the proliferation, the expression levels of MAGI3 and PTEN, and Akt phosphorylation were evaluated in C6 and C6-MAGI3 cells. Xenograft tumor models were established by subcutaneous injection of C6 and C6-MAGI3 cells into nude mice, and the growth rates of xenografts in the mice were compared. The potential role of MAGI3 expression in PI3K/Akt signaling activation was further investigated by examining the correlation between MAGI3 expression and the expression of PI3K/Akt signaling downstream target genes in a glioma dataset using gene set enrichment analysis （GSEA）. Results Expression levels of MAGI3 and PTEN were significantly downregulated in gliomas. Overexpression of MAGI3 in the glioma C6 cell line upregulated PTEN protein expression, inhibited the phosphorylation of Akt, and suppressed cell proliferation. MAGI3 overexpression also inhibited the growth of C6 glioma tumor xenografts in nude mice. Analysis based on the GEO database confirmed the negative correlation between activation of PI3K/Akt pathway and MAGI3 mRNA levels in human glioma samples. Conclusion The loss of MAGI3 expression in downregulation of PTEN expression, leading potential glioma suppressor. glioma may enhance the proliferation of glioma cells via to the activation of the PI3K/Akt pathway. MAGI3 is a展开更多
Calcium influx into neurons triggers neuronal death during cerebral ischemia/reperfusion injury.Various calcium channels are involved in cerebral ischemia/reperfusion injury.Cav3.2 channel is a main subtype of T-type ...Calcium influx into neurons triggers neuronal death during cerebral ischemia/reperfusion injury.Various calcium channels are involved in cerebral ischemia/reperfusion injury.Cav3.2 channel is a main subtype of T-type calcium channels.T-type calcium channel blockers,such as pimozide and mibefradil,have been shown to prevent cerebral ischemia/reperfusion injury-induced brain injury.However,the role of Cav3.2 channels in cerebral ischemia/reperfusion injury remains unclear.Here,in vitro and in vivo models of cerebral ischemia/reperfusion injury were established using middle cerebral artery occlusion in mice and high glucose hypoxia/reoxygenation exposure in primary hippocampal neurons.The results showed that Cav3.2 expression was significantly upregulated in injured hippocampal tissue and primary hippocampal neurons.We further established a Cav3.2 gene-knockout mouse model of cerebral ischemia/reperfusion injury.Cav3.2 knockout markedly reduced infarct volume and brain water content,and alleviated neurological dysfunction after cerebral ischemia/reperfusion injury.Additionally,Cav3.2 knockout attenuated cerebral ischemia/reperfusion injury-induced oxidative stress,inflammatory response,and neuronal apoptosis.In the hippocampus of Cav3.2-knockout mice,calcineurin overexpression offset the beneficial effect of Cav3.2 knockout after cerebral ischemia/reperfusion injury.These findings suggest that the neuroprotective function of Cav3.2 knockout is mediated by calcineurin/nuclear factor of activated T cells 3 signaling.Findings from this study suggest that Cav3.2 could be a promising target for treatment of cerebral ischemia/reperfusion injury.展开更多
Leptin,an adipocyte-derived peptide hormone,has been shown to facilitate breathing.However,the central sites and circuit mechanisms underlying the respiratory effects of leptin remain incompletely understood.The prese...Leptin,an adipocyte-derived peptide hormone,has been shown to facilitate breathing.However,the central sites and circuit mechanisms underlying the respiratory effects of leptin remain incompletely understood.The present study aimed to address whether neurons expressing leptin receptor b(LepRb)in the nucleus tractus solitarii(NTS)contribute to respiratory control.Both chemogenetic and optogenetic stimulation of LepRb-ex-pressing NTS(NTS^(LepRb))neurons notably activated breathing.Moreover,stimulation of NTS^(LepRb) neurons projecting to the lateral parabrachial nucleus(LPBN)not only remarkably increased basal ventilation to a level similar to that of the stimulation of all NTS^(LepRb) neurons,but also activated LPBN neurons projecting to the preBotzinger complex(preBotC).By contrast,ablation of NTS^pRb neurons projecting to the LPBN notably eliminated the enhanced respiratory effect induced by NTSLepRb neuron stimulation.In brainstem slices,bath application of leptin rapidly depolarized the membrane potential,increased the spontaneous firing rate,and accelerated the Ca2+transients in most NTSLepRb neurons.Therefore,leptin potentiates breathing in the NTS most likely via an NTS-LPBN-preBdtC circuit.展开更多
The frontal pole cortex(FPC)plays key roles in various higher-order functions and is highly developed in non-human primates.An essential missing piece of information is the detailed anatomical connections for finer pa...The frontal pole cortex(FPC)plays key roles in various higher-order functions and is highly developed in non-human primates.An essential missing piece of information is the detailed anatomical connections for finer parcellation of the macaque FPC than provided by the previous tracer results.This is important for understanding the functional architecture of the cerebral cortex.Here,combining cross-validation and principal component analysis,we formed a tractography-based parcellation scheme that applied a machine learning algorithm to divide the macaque FPC(2 males and 6 females)into eight subareas using high-resolution diffusion magnetic resonance imaging with the 9.4 T Bruker system,and then revealed their subregional connections.Furthermore,we applied improved hierarchical clustering to the obtained parcels to probe the modular structure of the subregions,and found that the dorsolateral FPC,which contains an extension to the medial FPC,was mainly connected to regions of the default-mode network.The ventral FPC was mainly involved in the social-interaction network and the dorsal FPC in the metacognitive network.These results enhance our understanding of the anatomy and circuitry of the macaque brain,and contribute to FPC-related clinical research.展开更多
The three-phase Enriched Environment(EE)paradigm has been shown to promote post-stroke functional improvement,but the neuronal mechanisms are still unclear.In this study,we applied a multimodal neuroimaging protocol c...The three-phase Enriched Environment(EE)paradigm has been shown to promote post-stroke functional improvement,but the neuronal mechanisms are still unclear.In this study,we applied a multimodal neuroimaging protocol combining magnetic resonance imaging(MRI)and positron emission tomography(PET)to examine the effects of post-ischemic EE treatment on structural and functional neuroplasticity in the bilateral sensorimotor cortex.Rats were subjected to permanent middle cerebral artery occlusion.The motor function of the rats was examined using the DigiGait test.MRI was applied to investigate the EE-induced structural modifications of the bilateral sensorimotor cortex.[^(18)F]-fluorodeoxyglucose PET was used to detect glucose metabolism.Blood oxygen level-dependent(BOLD)-functional MRI(fMRI)was used to identify the regional brain activity and functional connectivity(FC).In addition,the expression of neuroplasticity-related signaling pathways including neurotrophic factors(BDNF/CREB),axonal guidance proteins(Robo1/Slit2),and axonal growth-inhibitory proteins(NogoA/NgR)as well as downstream proteins(RhoA/ROCK)in the bilateral sensorimotor cortex were measured by Western blots.Our results showed the three-phase EE improved the walking ability.Structural T2 mapping imaging and diffusion tensor imaging demonstrated that EE benefited structure integrity in the bilateral sensorimotor cortex.PET-MRI fused images showed improved glucose metabolism in the corresponding regions after EE intervention.Specifically,the BOLD-based amplitude of low-frequency fluctuations showed that EE increased spontaneous activity in the bilateral motor cortex and ipsilateral sensory cortex.In addition,FC results showed increased sensorimotor connectivity in the ipsilateral hemisphere and increased interhemispheric motor cortical connectivity and motor cortical-thalamic connectivity following EE intervention.In addition,a strong correlation was found between increased functional connectivity and improved motor performance of limbs.Specifically,EE regulated the expression of neuroplasticity-related signaling,involving BDNF/CREB,Slit2/Robo1,as well as the axonal growth–inhibitory pathways Nogo-A/Nogo receptor and RhoA/ROCK in the bilateral sensorimotor cortex.Our results indicated that the three-phase enriched environment paradigm enhances neuronal plasticity of the bilateral sensorimotor cortex and consequently ameliorates post-stroke gait deficits.These findings might provide some new clues for the development of EE and thus facilitate the clinical translation of EE.展开更多
We investigated the role of metal atomization and solvent decomposition into reductive species and carbon clusters in the phase formation of transition-metal carbides(TMCs;namely,Co_(3)C,Fe_(3)C,TiC,and MoC)by pulsed ...We investigated the role of metal atomization and solvent decomposition into reductive species and carbon clusters in the phase formation of transition-metal carbides(TMCs;namely,Co_(3)C,Fe_(3)C,TiC,and MoC)by pulsed laser ablation of Co,Fe,Ti,and Mo metals in acetone.The interaction between carbon s-p-orbitals and metal d-orbitals causes a redistribution of valence structure through charge transfer,leading to the formation of surface defects as observed by X-ray photoelectron spectroscopy.These defects influence the evolved TMCs,making them effective for hydrogen and oxygen evolution reactions(HER and OER)in an alkaline medium.Co_(3)C with more oxygen affinity promoted CoO(OH)intermediates,and the electrochemical surface oxidation to Co_(3)O_(4)was captured via in situ/operando electrochemical Raman probes,increasing the number of active sites for OER activity.MoC with more d-vacancies exhibits strong hydrogen binding,promoting HER kinetics,whereas Fe_(3)C and TiC with more defect states to trap charge carriers may hinder both OER and HER activities.The results show that the assembled membrane-less electrolyzer with Co_(3)C∥Co_(3)C and MoC∥MoC electrodes requires~2.01 and 1.99 V,respectively,to deliver a 10 mA cm−2 with excellent electrochemical and structural stability.In addition,the ascertained pulsed laser synthesis mechanism and unit-cell packing relations will open up sustainable pathways for obtaining highly stable electrocatalysts for electrolyzers.展开更多
The nucleus tractus solitarii(NTS)is one of the morphologically and functionally defined centers that engage in the autonomic regulation of cardiovascular activity.Phenotypically-characterized NTS neurons have been im...The nucleus tractus solitarii(NTS)is one of the morphologically and functionally defined centers that engage in the autonomic regulation of cardiovascular activity.Phenotypically-characterized NTS neurons have been implicated in the differential regulation of blood pressure(BP).Here,we investigated whether phenylethanolamine N-methyltransferase(PNMT)-expressing NTS(NTS^(PNMT))neurons contribute to the control of BP.We demonstrate that photostimulation of NTS^(PNMT)neurons has variable effects on BP.A depressor response was produced during optogenetic stimulation of NTS^(PNMT)neurons projecting to the paraventricular nucleus of the hypothalamus,lateral parabrachial nucleus,and caudal ventrolateral medulla.Conversely,photostimulation of NTS^(PNMT)neurons projecting to the rostral ventrolateral medulla produced a robust pressor response and bradycardia.In addition,genetic ablation of both NTS^(PNMT)neurons and those projecting to the rostral ventrolateral medulla impaired the arterial baroreflex.Overall,we revealed the neuronal phenotype-and circuit-specific mechanisms underlying the contribution of NTS^(PNMT)neurons to the regulation of BP.展开更多
The combination of nucleic acid and small-molecule drugs in tumor treatment holds significant promise;however,the precise delivery and controlled release of drugs within the cytoplasm encounter substantial obstacles,i...The combination of nucleic acid and small-molecule drugs in tumor treatment holds significant promise;however,the precise delivery and controlled release of drugs within the cytoplasm encounter substantial obstacles,impeding the advancement of formulations.To surmount the challenges associated with precise drug delivery and controlled release,we have developed a multi-level p H-responsive co-loaded drug lipid nanoplatform.This platform first employs cyclic cell-penetrating peptides to exert a multi-level pH response,thereby enhancing the uptake efficiency of tumor cells and endow the nanosystem with effective endosomal/lysosomal escape.Subsequently,small interferring RNA(siRNA)complexes are formed by compacting siRNA with stearic acid octahistidine,which is capable of responding to the lysosome-tocytoplasm pH gradient and facilitate siRNA release.The siRNA complexes and docetaxel are simultaneously encapsulated into liposomes,thereby creating a lipid nanoplatform capable of co-delivering nucleic acid and small-molecule drugs.The efficacy of this platform has been validated through both in vitro and in vivo experiments,affirming its significant potential for practical applications in the co-delivery of nucleic acids and small-molecule drugs.展开更多
The gut microbiota plays a crucial role in maintaining host health and modulating disease progression.Microbiota-derived metabolites(MDMs)form a complex and diverse repertoire of molecules that interact uniquely with ...The gut microbiota plays a crucial role in maintaining host health and modulating disease progression.Microbiota-derived metabolites(MDMs)form a complex and diverse repertoire of molecules that interact uniquely with the host and regulate epigenetics.In this review,we provide a comprehensive overview by examining the current evidence on how MDMs reshape host epigenetic landscape,and we highlight the innovative concept of the“MDMs-epigenetic(MDME)axis”as a potential framework for exploring and understanding microbiota-host interactions.Next,we underscore the significance of the MDME axis in driving mechanistic studies and elucidating the underlying biological processes and pathophysiological pathways involved in various diseases.Finally,we discuss the impact of MDMs on epigenetic landscapes,outline future directions,and highlight their pivotal role in both mechanistic investigation and the development of clinical therapies.展开更多
Transcription activator-like effectors(TALEs) mimic eukaryotic transcriptional activators and translocate into host plant cells via the bacterial type Ⅲ secretion system(T3SS) during pathogenic interactions. They pla...Transcription activator-like effectors(TALEs) mimic eukaryotic transcriptional activators and translocate into host plant cells via the bacterial type Ⅲ secretion system(T3SS) during pathogenic interactions. They play a crucial role in disease development by regulating host genes. Despite this, the regulatory mechanisms by which TALEs control OsWRKY transcription factors(TFs) remain poorly understood. In this study, we show that two TALEs from Xanthomonas oryzae pv. oryzae(Xoo) individually modulate two OsWRKY TFs, resulting in increased susceptibility and reduced host defense.Specifically, Xoo1219 and Xoo2145 activate the expression of OsWRKY104 and OsWRKY55, respectively, through direct interactions. OsWRKY104 increases the susceptibility to Xoo by activating OsSWEET11 and OsSWEET14, while OsWRKY55suppresses host defense against Xoo by directly regulating OsWRKY62. These findings suggest that TALEs hijack the host's OsWRKY TFs to create a favorable environment for bacterial survival.展开更多
Background： In the current society, infertility related to age has become a social problem. The in vitro fertilization （IVF） success rate in women with poor ovarian response （POR） is very low. Dandelion extract T...Background： In the current society, infertility related to age has become a social problem. The in vitro fertilization （IVF） success rate in women with poor ovarian response （POR） is very low. Dandelion extract T-1 （DE-T1） is an effective component of the extract from the leaves and stems of Traxacum officinale, which is one of the medicines used in some patients with POR, but its molecular mechanism remains unclear. Methods： Following IVF, ovarian granulosa cells （GCs） of sixty patients were extracted and divided into normal ovarian response （NOR） and POR groups. GCs were cultured in a dose-dependent and time-dependent manner with DE-TI, proliferation of GCs was determined by Cell Counting Kit-8 assay, and mRNA levels of insulin-like growth factor 1 receptor （IGF-1R）, luteotropic hormone receptor （LHR）, follicle-stimulating hormone receptor （FSHR）, LHR, and CYP19A1 （aromatase） were determined by quantitative polymerase chain reaction. Progesterone and estradiol （E2） concentrations were determined by enzyme-linked immunosorbent assay. Results： The cell viability gradually increased with the progressive increase in the DE-T1 concentration. Compared with the control group （without DE-T1）, the mRNA expressions of FSHR, LHR, IGF-IR, and CYPIgAI were upregulated after the addition of DE-T l, especially in the 2.5% DE-T 1 group （P 〈 0.01 ）. The expression of IGF- 1R was upregulated approximately 25 times （24.97 ± 4.02 times） in the POR group with 2.5% DE-T1. E2 and progesterone levels increased with the increasing DE-T1 concentration. There were highly significant differences in the E2 and progesterone secretion between the NOR and POR groups （P 〈 0.01）. Conclusion： DE-TI may promote steroid hormone synthesis by promoting GC proliferation and upregulating GC receptor expression, thereby improving ovarian endocrine function.展开更多
Severe pandemic influenza A(H1N1)2009 virus(pH1N1)infection causes significant morbidity and mortality.We and other groups have reported that immunopathological damage induced by excessive pulmonary inflammation plays...Severe pandemic influenza A(H1N1)2009 virus(pH1N1)infection causes significant morbidity and mortality.We and other groups have reported that immunopathological damage induced by excessive pulmonary inflammation plays a critical role in the pathogenesis of severe pneumonia and provides novel strategies for the treatment of severe influenza infection[1–3].The complement system plays critical roles in both innate and adaptive immunity by activating the classical,alternative and lectin pathways[4].展开更多
Medicago truncatula is a model legume species that has been studied for decades to understand the symbiotic relationship between legumes and soil bacteria collectively named rhizobia.This symbiosis called nodulation i...Medicago truncatula is a model legume species that has been studied for decades to understand the symbiotic relationship between legumes and soil bacteria collectively named rhizobia.This symbiosis called nodulation is initiated in roots with the infection of root hair cells by the bacteria,as well as the initiation of nodule primordia from root cortical,endodermal,and pericycle cells,leading to the development of a new root organ,the nodule,where bacteria fix and assimilate the atmospheric dinitrogen for the benefit of the plant.Here,we report the isolation and use of the nuclei from mock and rhizobia-inoculated roots for the single nuclei RNA-seq(sNucRNA-seq)profiling to gain a deeper understanding of early responses to rhizobial infection in Medicago roots.A gene expression map of the Medicago root was generated,comprising 25 clusters,which were annotated as specific cell types using 119 Medicago marker genes and orthologs to Arabidopsis cell-type marker genes.A focus on root hair,cortex,endodermis,and pericycle cell types,showing the strongest differential regulation in response to a short-term(48 h)rhizobium inoculation,revealed not only known genes and functional pathways,validating the sNucRNA-seq approach,but also numerous novel genes and pathways,allowing a comprehensive analysis of early root symbiotic responses at a cell type-specific level.展开更多
Hemophilic articular cartilage damage presents a significant challenge for surgeons,characterized by recurrent intraarticular bleeding,a severe inflammatory microenvironment,and limited self-repair capability of carti...Hemophilic articular cartilage damage presents a significant challenge for surgeons,characterized by recurrent intraarticular bleeding,a severe inflammatory microenvironment,and limited self-repair capability of cartilage tissue.Currently,there is a lack of tissue engineering-based integrated therapies that address both early hemostasis,anti-inflammation,and long-lasting chondrogenesis for hemophilic articular cartilage defects.Herein,we developed an adhesive hydrogel using oxidized chondroitin sulfate and gelatin,loaded with exosomes derived from bone marrow stem cells(BMSCs)(Hydrogel-Exos).This hydrogel demonstrated favorable injectability,self-healing,biocompatibility,biodegradability,swelling,frictional and mechanical properties,providing a comprehensive approach to treating hemophilic articular cartilage defects.The adhesive hydrogel,featuring dynamic Schiff base bonds and hydrogen bonds,exhibited excellent wet tissue adhesiveness and hemostatic properties.In a pig model,the hydrogel could be smoothly injected into the knee joint cartilage defect site and gelled in situ under fluid-irrigated arthroscopic conditions.Our in vitro and in vivo experiments confirmed that the sustained release of exosomes yielded anti-inflammatory effects by modulating macrophage M2 polarization through the NF-κB pathway.This immunoregulatory effect,coupled with the extracellular matrix components provided by the adhesive hydrogel,enhanced chondrogenesis,promoted the cartilage repair and joint function restoration after hemophilic articular cartilage defects.In conclusion,our results highlight the significant application potential of Hydrogel-Exos for early hemostasis,immunoregulation,and long-term chondrogenesis in hemophilic patients with cartilage injuries.This innovative approach is well-suited for application during arthroscopic procedures,offering a promising solution for addressing the complex challenges associated with hemophilic articular cartilage damage.展开更多
Cancer stem cells (CSCs) are largely responsible for the formation of tumor heterogeneity and tumor’s resistance to traditional treatments such as chemotherapy due to its self-renew capability and multi-lineage diffe...Cancer stem cells (CSCs) are largely responsible for the formation of tumor heterogeneity and tumor’s resistance to traditional treatments such as chemotherapy due to its self-renew capability and multi-lineage differentiation potential (1)However, it remains elusive how the CSCs maintain their stemness, thus restricting the development of therapeutic treatments specifically targeting the CSCs.展开更多
Activation-induced cytidine deaminase(AID)is required for the generation of antibody diversity through initiat-ing both somatic hypermutation(SHM)and class switch recombination.A few research groups have success-fully...Activation-induced cytidine deaminase(AID)is required for the generation of antibody diversity through initiat-ing both somatic hypermutation(SHM)and class switch recombination.A few research groups have success-fully used the feature of AID for generating mutant li-braries in directed evolution of target proteins in B cells in vitro.B cells,cultured in suspension,are not con-venient for transfection and cloning.In this study,we established an AID-based mutant accumulation and sorting system in adherent human cells.Mouse AID gene was first transfected into the human non-small cell lung carcinoma H1299 cells,and a stable cell clone(H1299-AID)was selected.Afterwards,anti-hTNF-αscFv(ATscFv)was transfected into H1299-AID cells and ATscFv was displayed on the surface of H1299-AID cells.By 4-round amplification/flow cytometric sorting for cells with the highest affinities to hTNF-alpha,two ATscFv mutant gene clones were isolated.Compared with the wild type ATscFv,the two mutants were much more efficient in neutralizing cytotoxicity of hTNF-alpha.The results indicate that directed evolution by somatic hypermutation can be carried out in adherent non-B cells,which makes directed evolution in mammalian cells easier and more efficient.展开更多
D-xylose is an abundant sugar found in plant biomass and can be used as a renewable feedstock for the microbial production of diverse biofuels and bioproducts.However,D-xylose metabolism is slow in many industrial mic...D-xylose is an abundant sugar found in plant biomass and can be used as a renewable feedstock for the microbial production of diverse biofuels and bioproducts.However,D-xylose metabolism is slow in many industrial microorganisms,at least as compared to glucose metabolism.Not surprisingly,a number of approaches have been developed for improving D-xylose metabolism in diverse microorganisms.In this work,we applied a previously developed evolution strategy based on media-in-oil emulsions for improving the growth yield of Escherichia coli NCM3722 on D-xylose.After 30 rounds of evolutions,we isolated multiple mutants with increased growth yield on D-xylose.In addition,we also observed similar increases in the growth rate.Three mutants were selected for whole-genome sequencing.Two mutants had an amber stop mutation in adenylate cyclase,which truncates nearly 60%of the enzyme.However,the ability of this mutant to grow on xylose indicated that truncated enzyme,lacking the C-terminal regulatory domain,is still active.The other mutant had a point mutation in the cyclic AMP receptor protein(CRP),near the high affinity binding site for cyclic AMP.Both mutations,when intro-duced into wild type E.coli,were able to increase the growth yields at levels similar to the isolated mutants.In addition to D-xylose,these mutant strains and their genetic mimics also exhibited higher growth rates and yields on glucose,lactose,and L-arabinose.These results suggest that the improved growth rates and yields are due to changes in the production and sensing of intracellular cyclic AMP concentrations and also suggest native concentrations are suboptimal with respect to the growth rate and yield under the growth conditions tested.Collectively,these results may prove useful for engineering strains of E.coli for high-density fermentations or protein production.展开更多
Neutrophils are derived from bone marrow hematopoietic stem cells(HSCs)and are the largest population among circulating white blood cells in humans,acting as the first line of defense against invading pathogens.Whethe...Neutrophils are derived from bone marrow hematopoietic stem cells(HSCs)and are the largest population among circulating white blood cells in humans,acting as the first line of defense against invading pathogens.Whether neutrophils can be generated by transdifferentiation strategies is unknown.Here,we show that thymidine induces the conversion of mouse fibroblasts to neutrophils.Induced neutrophils(iNeus)showed antibacterial effects and did not undergo malignant transformation in vivo.Importantly,iNeu transplantation cured neutropenia in mice in vivo.Mechanistically,thymidine mediates iNeu conversion by enhancing Tet3 activity.Tet3 initiates the expression of the neutrophil fate decision factors Cebpδ and Rfx1 that drive the transdifferentiation of mouse fibroblasts to neutrophils.Therefore,the induction of functional neutrophils by chemicals may provide a potential therapeutic strategy for patients with neutropenia patients and infectious diseases.展开更多
基金This study was supported by the Key Research and Development Program of Anhui Province(No.202204295107020003)the National Natural Science Foundation of Anhui Province(No.2108085MH293)+1 种基金the Distinguished Young Scholars Fund of Anhui Province(No.2022AH020078)the Key health Project of Anhui Province(AHWJ2022a037).
文摘The overdiagnosis of prostate cancer(PCa)caused by nonspecific elevation serum prostate-specific antigen(PSA)and the overtreatment of indolent PCa have become a global problem that needs to be solved urgently.We aimed to construct a prediction model and provide a risk stratification system to reduce unnecessary biopsies.In this retrospective study,clinical data of 1807 patients from three Chinese hospitals were used.The final model was built using stepwise logistic regression analysis.The apparent performance of the model was assessed by receiver operating characteristic curves,calibration plots,and decision curve analysis.Finally,a risk stratification system of clinically significant prostate cancer(csPCa)was created,and diagnosis-free survival analyses were performed.Following multivariable screening and evaluation of the diagnostic performances,a final diagnostic model comprised of the PSA density and Prostate Imaging-Reporting and Data System(PI-RADS)score was established.Model validation in the development cohort and two external cohorts showed excellent discrimination and calibration.Finally,we created a risk stratification system using risk thresholds of 0.05 and 0.60 as the cut-off values.The follow-up results indicated that the diagnosis-free survival rate for csPCa at 12 months and 24 months postoperatively was 99.7%and 99.4%,respectively,for patients with a risk threshold below O.05 after the initial negative prostate biopsy,which was significantly better than patients with higher risk.Our diagnostic model and risk stratification system can achieve a personalized risk calculation of csPCa.It provides a standardized tool for Chinese patients and physicians when considering thenecessity of prostatebiopsy.
基金supported by the China Postdoctoral Science Foundation(Nos.2023M730317 and 2023T160066)the Fundamental Research Funds for the Central Universities(No.3332023042)+3 种基金the Open Project of Hebei Key Laboratory of Environment and Human Health(No.202301)the Programs of the National Natural Science Foundation of China(No.21976050)the Science and Technology Program of Hebei Province(No.21377779D)the Natural Science Foundation of Hebei Province,China(No.B2020206008).
文摘Although per-and polyfluoroalkyl substances(PFAS)have been frequently linked to cardiovascular and renal disease separately,evidence remains scarce regarding their systematic effect.Therefore,we recruited 546 newly diagnosed acute coronary syndrome(ACS)patients and detected seven myocardial enzymes and six kidney function biomarkers.Twelve PFASwere also assessedwith ultra-high-performance liquid chromatography-tandem mass spectrometry.Generalized linear model and restricted cubic spline model were applied to single pollutant analysis.Quantile g-computation was used for mixture analysis.Network model was utilized to identify central and bridge nodes of pollutants and phenotypes.In the present study,perfluorohexane sulfonic acid was positively associated with uric acid(UA)(β=0.04,95%confidence interval(CI):0.01,0.07),and perfluorobutanoic acid was negatively associated with estimated glomerular filtration rate(β=-0.04,95%CI:-0.07,-0.01)but positively associated with UA(β=0.03,95%CI:0.01,0.06).In mixture analysis,each quantile increase in the PFAS mixture was significantly associated with UA(β=0.08,95%CI:0.04,0.11).Network analysis revealed that perfluorooctanoate,UA,and myoglobin were denoted as bridge nodes,and the first principal component of lactate dehydrogenase and creatine kinase-myocardial band was identified as the node with the highest strength and expected influence.This study investigates the systematic impact of PFAS exposure through cardiorenal interaction network,which highlights that PFAS may serve as an upstream approach in UA-modulated cardiorenal network to affect cardiorenal system comprehensively.
基金supported by the National Natural Science Foundation of the People’s Republic of China(No.81272887 and 81141033)Beijing Municipal Natural Science Foundation(No.7131003)
文摘Objective To investigate the role and molecular mechanism of membrane-associated guanylate kinase inverted 3 （MAGI3） in glioma cell proliferation. Methods The expression levels of MAGI3 and PTEN were assessed in glioma samples by Western blotting. MAGI3 was stably transfected into C6 glioma cells to obtain C6-MAGI3 cells. Then, the proliferation, the expression levels of MAGI3 and PTEN, and Akt phosphorylation were evaluated in C6 and C6-MAGI3 cells. Xenograft tumor models were established by subcutaneous injection of C6 and C6-MAGI3 cells into nude mice, and the growth rates of xenografts in the mice were compared. The potential role of MAGI3 expression in PI3K/Akt signaling activation was further investigated by examining the correlation between MAGI3 expression and the expression of PI3K/Akt signaling downstream target genes in a glioma dataset using gene set enrichment analysis （GSEA）. Results Expression levels of MAGI3 and PTEN were significantly downregulated in gliomas. Overexpression of MAGI3 in the glioma C6 cell line upregulated PTEN protein expression, inhibited the phosphorylation of Akt, and suppressed cell proliferation. MAGI3 overexpression also inhibited the growth of C6 glioma tumor xenografts in nude mice. Analysis based on the GEO database confirmed the negative correlation between activation of PI3K/Akt pathway and MAGI3 mRNA levels in human glioma samples. Conclusion The loss of MAGI3 expression in downregulation of PTEN expression, leading potential glioma suppressor. glioma may enhance the proliferation of glioma cells via to the activation of the PI3K/Akt pathway. MAGI3 is a
基金supported by the Natural Science Foundation of Anhui Province of China,No.2208085Y32Scientific Research Plan Project of Anhui Province of China,No.2022AH020076the Chen Xiao-Ping Foundation for the Development of Science and Technology of Hubei Province,No.CXPJJH12000005-07-115(all to CT).
文摘Calcium influx into neurons triggers neuronal death during cerebral ischemia/reperfusion injury.Various calcium channels are involved in cerebral ischemia/reperfusion injury.Cav3.2 channel is a main subtype of T-type calcium channels.T-type calcium channel blockers,such as pimozide and mibefradil,have been shown to prevent cerebral ischemia/reperfusion injury-induced brain injury.However,the role of Cav3.2 channels in cerebral ischemia/reperfusion injury remains unclear.Here,in vitro and in vivo models of cerebral ischemia/reperfusion injury were established using middle cerebral artery occlusion in mice and high glucose hypoxia/reoxygenation exposure in primary hippocampal neurons.The results showed that Cav3.2 expression was significantly upregulated in injured hippocampal tissue and primary hippocampal neurons.We further established a Cav3.2 gene-knockout mouse model of cerebral ischemia/reperfusion injury.Cav3.2 knockout markedly reduced infarct volume and brain water content,and alleviated neurological dysfunction after cerebral ischemia/reperfusion injury.Additionally,Cav3.2 knockout attenuated cerebral ischemia/reperfusion injury-induced oxidative stress,inflammatory response,and neuronal apoptosis.In the hippocampus of Cav3.2-knockout mice,calcineurin overexpression offset the beneficial effect of Cav3.2 knockout after cerebral ischemia/reperfusion injury.These findings suggest that the neuroprotective function of Cav3.2 knockout is mediated by calcineurin/nuclear factor of activated T cells 3 signaling.Findings from this study suggest that Cav3.2 could be a promising target for treatment of cerebral ischemia/reperfusion injury.
基金supported by the National Natural Science Foundation of China(31800981 and 31971058)the Natural Science Foundation of Hebei Province for Distinguished Young Scholars(H2020206509)a Hebei Province Government Grant(CXZZBS2020119).
文摘Leptin,an adipocyte-derived peptide hormone,has been shown to facilitate breathing.However,the central sites and circuit mechanisms underlying the respiratory effects of leptin remain incompletely understood.The present study aimed to address whether neurons expressing leptin receptor b(LepRb)in the nucleus tractus solitarii(NTS)contribute to respiratory control.Both chemogenetic and optogenetic stimulation of LepRb-ex-pressing NTS(NTS^(LepRb))neurons notably activated breathing.Moreover,stimulation of NTS^(LepRb) neurons projecting to the lateral parabrachial nucleus(LPBN)not only remarkably increased basal ventilation to a level similar to that of the stimulation of all NTS^(LepRb) neurons,but also activated LPBN neurons projecting to the preBotzinger complex(preBotC).By contrast,ablation of NTS^pRb neurons projecting to the LPBN notably eliminated the enhanced respiratory effect induced by NTSLepRb neuron stimulation.In brainstem slices,bath application of leptin rapidly depolarized the membrane potential,increased the spontaneous firing rate,and accelerated the Ca2+transients in most NTSLepRb neurons.Therefore,leptin potentiates breathing in the NTS most likely via an NTS-LPBN-preBdtC circuit.
基金the National Natural Science Foundation of China(91432302 and 31620103905)the Science Frontier Program of the Chinese Academy of Sciences(QYZDJ-SSW-SMC019)+3 种基金the National Key R&D Program of China(2017YFA0105203)Beijing Municipal Science and Technology Commission(Z161100000216152,Z161100000216139,Z181100001518004and Z171100000117002)the Beijing Brain Initiative of Beijing Municipal Science and Technology Commission(Z181100001518004)the Guangdong Pearl River Talents Plan(2016ZT06S220)。
文摘The frontal pole cortex(FPC)plays key roles in various higher-order functions and is highly developed in non-human primates.An essential missing piece of information is the detailed anatomical connections for finer parcellation of the macaque FPC than provided by the previous tracer results.This is important for understanding the functional architecture of the cerebral cortex.Here,combining cross-validation and principal component analysis,we formed a tractography-based parcellation scheme that applied a machine learning algorithm to divide the macaque FPC(2 males and 6 females)into eight subareas using high-resolution diffusion magnetic resonance imaging with the 9.4 T Bruker system,and then revealed their subregional connections.Furthermore,we applied improved hierarchical clustering to the obtained parcels to probe the modular structure of the subregions,and found that the dorsolateral FPC,which contains an extension to the medial FPC,was mainly connected to regions of the default-mode network.The ventral FPC was mainly involved in the social-interaction network and the dorsal FPC in the metacognitive network.These results enhance our understanding of the anatomy and circuitry of the macaque brain,and contribute to FPC-related clinical research.
基金supported by the National Natural Science Foundation of China(82174471).
文摘The three-phase Enriched Environment(EE)paradigm has been shown to promote post-stroke functional improvement,but the neuronal mechanisms are still unclear.In this study,we applied a multimodal neuroimaging protocol combining magnetic resonance imaging(MRI)and positron emission tomography(PET)to examine the effects of post-ischemic EE treatment on structural and functional neuroplasticity in the bilateral sensorimotor cortex.Rats were subjected to permanent middle cerebral artery occlusion.The motor function of the rats was examined using the DigiGait test.MRI was applied to investigate the EE-induced structural modifications of the bilateral sensorimotor cortex.[^(18)F]-fluorodeoxyglucose PET was used to detect glucose metabolism.Blood oxygen level-dependent(BOLD)-functional MRI(fMRI)was used to identify the regional brain activity and functional connectivity(FC).In addition,the expression of neuroplasticity-related signaling pathways including neurotrophic factors(BDNF/CREB),axonal guidance proteins(Robo1/Slit2),and axonal growth-inhibitory proteins(NogoA/NgR)as well as downstream proteins(RhoA/ROCK)in the bilateral sensorimotor cortex were measured by Western blots.Our results showed the three-phase EE improved the walking ability.Structural T2 mapping imaging and diffusion tensor imaging demonstrated that EE benefited structure integrity in the bilateral sensorimotor cortex.PET-MRI fused images showed improved glucose metabolism in the corresponding regions after EE intervention.Specifically,the BOLD-based amplitude of low-frequency fluctuations showed that EE increased spontaneous activity in the bilateral motor cortex and ipsilateral sensory cortex.In addition,FC results showed increased sensorimotor connectivity in the ipsilateral hemisphere and increased interhemispheric motor cortical connectivity and motor cortical-thalamic connectivity following EE intervention.In addition,a strong correlation was found between increased functional connectivity and improved motor performance of limbs.Specifically,EE regulated the expression of neuroplasticity-related signaling,involving BDNF/CREB,Slit2/Robo1,as well as the axonal growth–inhibitory pathways Nogo-A/Nogo receptor and RhoA/ROCK in the bilateral sensorimotor cortex.Our results indicated that the three-phase enriched environment paradigm enhances neuronal plasticity of the bilateral sensorimotor cortex and consequently ameliorates post-stroke gait deficits.These findings might provide some new clues for the development of EE and thus facilitate the clinical translation of EE.
基金National Research Foundation of Korea,Grant/Award Numbers:2019H1D3A1A01071209,2021R1I1A1A01060380,2022R1A2C2010686,2022R1A4A3033528Korea Basic Science Institute,Grant/Award Numbers:2019R1A6C1010042,2021R1A6C103A427。
文摘We investigated the role of metal atomization and solvent decomposition into reductive species and carbon clusters in the phase formation of transition-metal carbides(TMCs;namely,Co_(3)C,Fe_(3)C,TiC,and MoC)by pulsed laser ablation of Co,Fe,Ti,and Mo metals in acetone.The interaction between carbon s-p-orbitals and metal d-orbitals causes a redistribution of valence structure through charge transfer,leading to the formation of surface defects as observed by X-ray photoelectron spectroscopy.These defects influence the evolved TMCs,making them effective for hydrogen and oxygen evolution reactions(HER and OER)in an alkaline medium.Co_(3)C with more oxygen affinity promoted CoO(OH)intermediates,and the electrochemical surface oxidation to Co_(3)O_(4)was captured via in situ/operando electrochemical Raman probes,increasing the number of active sites for OER activity.MoC with more d-vacancies exhibits strong hydrogen binding,promoting HER kinetics,whereas Fe_(3)C and TiC with more defect states to trap charge carriers may hinder both OER and HER activities.The results show that the assembled membrane-less electrolyzer with Co_(3)C∥Co_(3)C and MoC∥MoC electrodes requires~2.01 and 1.99 V,respectively,to deliver a 10 mA cm−2 with excellent electrochemical and structural stability.In addition,the ascertained pulsed laser synthesis mechanism and unit-cell packing relations will open up sustainable pathways for obtaining highly stable electrocatalysts for electrolyzers.
基金supported by the Natural Science Foundation of China(31971050)the Natural Science Foundation of Hebei Province for Distinguished Young Scholars(H2020206509)+1 种基金Hebei Province Innovative Research Project for Postgraduate(2021074)the Natural Science Foundation of Hebei Province for Innovative Research Group Project(H2021206203).
文摘The nucleus tractus solitarii(NTS)is one of the morphologically and functionally defined centers that engage in the autonomic regulation of cardiovascular activity.Phenotypically-characterized NTS neurons have been implicated in the differential regulation of blood pressure(BP).Here,we investigated whether phenylethanolamine N-methyltransferase(PNMT)-expressing NTS(NTS^(PNMT))neurons contribute to the control of BP.We demonstrate that photostimulation of NTS^(PNMT)neurons has variable effects on BP.A depressor response was produced during optogenetic stimulation of NTS^(PNMT)neurons projecting to the paraventricular nucleus of the hypothalamus,lateral parabrachial nucleus,and caudal ventrolateral medulla.Conversely,photostimulation of NTS^(PNMT)neurons projecting to the rostral ventrolateral medulla produced a robust pressor response and bradycardia.In addition,genetic ablation of both NTS^(PNMT)neurons and those projecting to the rostral ventrolateral medulla impaired the arterial baroreflex.Overall,we revealed the neuronal phenotype-and circuit-specific mechanisms underlying the contribution of NTS^(PNMT)neurons to the regulation of BP.
基金supported by the grants from the National Natural Science Foundation of China(Nos.81973251 and 81302725)Hebei Province Funding Project for Introduced Overseas Personnel(Nos.C20230351 and C20220345)+3 种基金Key Research and Development Program of Hebei Province(No.22372701D)Hebei Province Natural Science Fund(No.H2020206610)Hebei Provincial Health Commission Government-Funded Clinical Medicine Talent Program(No.ZF2024048)Hebei Medical University Undergraduate Innovative Experiment Program(No.USIP2023008)。
文摘The combination of nucleic acid and small-molecule drugs in tumor treatment holds significant promise;however,the precise delivery and controlled release of drugs within the cytoplasm encounter substantial obstacles,impeding the advancement of formulations.To surmount the challenges associated with precise drug delivery and controlled release,we have developed a multi-level p H-responsive co-loaded drug lipid nanoplatform.This platform first employs cyclic cell-penetrating peptides to exert a multi-level pH response,thereby enhancing the uptake efficiency of tumor cells and endow the nanosystem with effective endosomal/lysosomal escape.Subsequently,small interferring RNA(siRNA)complexes are formed by compacting siRNA with stearic acid octahistidine,which is capable of responding to the lysosome-tocytoplasm pH gradient and facilitate siRNA release.The siRNA complexes and docetaxel are simultaneously encapsulated into liposomes,thereby creating a lipid nanoplatform capable of co-delivering nucleic acid and small-molecule drugs.The efficacy of this platform has been validated through both in vitro and in vivo experiments,affirming its significant potential for practical applications in the co-delivery of nucleic acids and small-molecule drugs.
基金supported by the National Key Research and Development Program(2022YFA1303900)the National Natural Science Foundation of China(NSFC)(82172288).
文摘The gut microbiota plays a crucial role in maintaining host health and modulating disease progression.Microbiota-derived metabolites(MDMs)form a complex and diverse repertoire of molecules that interact uniquely with the host and regulate epigenetics.In this review,we provide a comprehensive overview by examining the current evidence on how MDMs reshape host epigenetic landscape,and we highlight the innovative concept of the“MDMs-epigenetic(MDME)axis”as a potential framework for exploring and understanding microbiota-host interactions.Next,we underscore the significance of the MDME axis in driving mechanistic studies and elucidating the underlying biological processes and pathophysiological pathways involved in various diseases.Finally,we discuss the impact of MDMs on epigenetic landscapes,outline future directions,and highlight their pivotal role in both mechanistic investigation and the development of clinical therapies.
基金support of the Next-Generation BioGreen 21 Program of the Rural Development Administration(PJ013269)the Brain Pool Program of the Ministry of Science and ICT through the National Research Foundation of Korea(RS-2023-00262576)M.-Y.J.was supported by an NRF grant funded by the Korean government(Ministry of Science and ICT)(RS-2025000518246).
文摘Transcription activator-like effectors(TALEs) mimic eukaryotic transcriptional activators and translocate into host plant cells via the bacterial type Ⅲ secretion system(T3SS) during pathogenic interactions. They play a crucial role in disease development by regulating host genes. Despite this, the regulatory mechanisms by which TALEs control OsWRKY transcription factors(TFs) remain poorly understood. In this study, we show that two TALEs from Xanthomonas oryzae pv. oryzae(Xoo) individually modulate two OsWRKY TFs, resulting in increased susceptibility and reduced host defense.Specifically, Xoo1219 and Xoo2145 activate the expression of OsWRKY104 and OsWRKY55, respectively, through direct interactions. OsWRKY104 increases the susceptibility to Xoo by activating OsSWEET11 and OsSWEET14, while OsWRKY55suppresses host defense against Xoo by directly regulating OsWRKY62. These findings suggest that TALEs hijack the host's OsWRKY TFs to create a favorable environment for bacterial survival.
文摘Background： In the current society, infertility related to age has become a social problem. The in vitro fertilization （IVF） success rate in women with poor ovarian response （POR） is very low. Dandelion extract T-1 （DE-T1） is an effective component of the extract from the leaves and stems of Traxacum officinale, which is one of the medicines used in some patients with POR, but its molecular mechanism remains unclear. Methods： Following IVF, ovarian granulosa cells （GCs） of sixty patients were extracted and divided into normal ovarian response （NOR） and POR groups. GCs were cultured in a dose-dependent and time-dependent manner with DE-TI, proliferation of GCs was determined by Cell Counting Kit-8 assay, and mRNA levels of insulin-like growth factor 1 receptor （IGF-1R）, luteotropic hormone receptor （LHR）, follicle-stimulating hormone receptor （FSHR）, LHR, and CYP19A1 （aromatase） were determined by quantitative polymerase chain reaction. Progesterone and estradiol （E2） concentrations were determined by enzyme-linked immunosorbent assay. Results： The cell viability gradually increased with the progressive increase in the DE-T1 concentration. Compared with the control group （without DE-T1）, the mRNA expressions of FSHR, LHR, IGF-IR, and CYPIgAI were upregulated after the addition of DE-T l, especially in the 2.5% DE-T 1 group （P 〈 0.01 ）. The expression of IGF- 1R was upregulated approximately 25 times （24.97 ± 4.02 times） in the POR group with 2.5% DE-T1. E2 and progesterone levels increased with the increasing DE-T1 concentration. There were highly significant differences in the E2 and progesterone secretion between the NOR and POR groups （P 〈 0.01）. Conclusion： DE-TI may promote steroid hormone synthesis by promoting GC proliferation and upregulating GC receptor expression, thereby improving ovarian endocrine function.
基金This work was supported by grants from National Natural Science Foundation of China(82071747,81373114)Beijing Municipal Natural Science Foundation,China(7182013).
文摘Severe pandemic influenza A(H1N1)2009 virus(pH1N1)infection causes significant morbidity and mortality.We and other groups have reported that immunopathological damage induced by excessive pulmonary inflammation plays a critical role in the pathogenesis of severe pneumonia and provides novel strategies for the treatment of severe influenza infection[1–3].The complement system plays critical roles in both innate and adaptive immunity by activating the classical,alternative and lectin pathways[4].
基金Supported by grants to M.L.from the U.S.National Sclence Foundation (I0S#1854326 and 2127485),USDA-NIFA(2022-67013-36144)by the Center for Plant Science Innovation,and by the Department of Agronomy and Horticulture at the University of Nebraska-Lincoln.Work in F.F.labo-ratory was supported by the"Ecole Universitaire de Recherche"Saclay Plant Sciences(EUR-SPS).
文摘Medicago truncatula is a model legume species that has been studied for decades to understand the symbiotic relationship between legumes and soil bacteria collectively named rhizobia.This symbiosis called nodulation is initiated in roots with the infection of root hair cells by the bacteria,as well as the initiation of nodule primordia from root cortical,endodermal,and pericycle cells,leading to the development of a new root organ,the nodule,where bacteria fix and assimilate the atmospheric dinitrogen for the benefit of the plant.Here,we report the isolation and use of the nuclei from mock and rhizobia-inoculated roots for the single nuclei RNA-seq(sNucRNA-seq)profiling to gain a deeper understanding of early responses to rhizobial infection in Medicago roots.A gene expression map of the Medicago root was generated,comprising 25 clusters,which were annotated as specific cell types using 119 Medicago marker genes and orthologs to Arabidopsis cell-type marker genes.A focus on root hair,cortex,endodermis,and pericycle cell types,showing the strongest differential regulation in response to a short-term(48 h)rhizobium inoculation,revealed not only known genes and functional pathways,validating the sNucRNA-seq approach,but also numerous novel genes and pathways,allowing a comprehensive analysis of early root symbiotic responses at a cell type-specific level.
基金supported by the National Natural Science Foundation of China Youth Fund(82202662)the Guangzhou Science and Technology Program(2023A04J2314)+11 种基金the National Natural Science Foundation of China(12,272,164)the China Postdoctoral Science Foundation(2023M741563)the Clinical Research Startup Program of Southern Medical University by High-level University Construction Funding of Guangdong Provincial Department of Education(LC2019ZD001)the Clinical Research Program of Nanfang Hospital,Southern Medical University(2019CR016)the Project of Drug Clinical Evaluate Research of Chinese Pharmaceutical Association(CPA-Z06-ZC-2021-004)the National Natural Science Foundation of China(82370497)the Medical Scientific Research Foundation of Guangdong(A2024366)Huizhou Science Technology Project Foundation(2022CZ010423)the Macao Science and Technology Development fund(FDCT(0012/2021/AMJ,003/2022/ALC,0092/2022/A2,0144/2022/A3))the Shenzhen-Hong Kong-Macao Science and Technology Fund(Category C:SGDX20220530111203020)the Foundation of Guangdong Basic and Applied Basic Research Foundation(2022A1515140151&2022A1515140189&2023A1515140045&2022A1515140071)the National Orthopaedics Key Clinical Specialty Construction Research Foundation of Huizhou Central People’s Hospital.
文摘Hemophilic articular cartilage damage presents a significant challenge for surgeons,characterized by recurrent intraarticular bleeding,a severe inflammatory microenvironment,and limited self-repair capability of cartilage tissue.Currently,there is a lack of tissue engineering-based integrated therapies that address both early hemostasis,anti-inflammation,and long-lasting chondrogenesis for hemophilic articular cartilage defects.Herein,we developed an adhesive hydrogel using oxidized chondroitin sulfate and gelatin,loaded with exosomes derived from bone marrow stem cells(BMSCs)(Hydrogel-Exos).This hydrogel demonstrated favorable injectability,self-healing,biocompatibility,biodegradability,swelling,frictional and mechanical properties,providing a comprehensive approach to treating hemophilic articular cartilage defects.The adhesive hydrogel,featuring dynamic Schiff base bonds and hydrogen bonds,exhibited excellent wet tissue adhesiveness and hemostatic properties.In a pig model,the hydrogel could be smoothly injected into the knee joint cartilage defect site and gelled in situ under fluid-irrigated arthroscopic conditions.Our in vitro and in vivo experiments confirmed that the sustained release of exosomes yielded anti-inflammatory effects by modulating macrophage M2 polarization through the NF-κB pathway.This immunoregulatory effect,coupled with the extracellular matrix components provided by the adhesive hydrogel,enhanced chondrogenesis,promoted the cartilage repair and joint function restoration after hemophilic articular cartilage defects.In conclusion,our results highlight the significant application potential of Hydrogel-Exos for early hemostasis,immunoregulation,and long-term chondrogenesis in hemophilic patients with cartilage injuries.This innovative approach is well-suited for application during arthroscopic procedures,offering a promising solution for addressing the complex challenges associated with hemophilic articular cartilage damage.
基金supported by National Key Scientific Program of China (2016YFA0100502 to X.S.)the National Natural Science Foundation of China (81701765 to J.C.)Anhui Provincial Natural Science Foundation (1808085MH295 to J.C.)
文摘Cancer stem cells (CSCs) are largely responsible for the formation of tumor heterogeneity and tumor’s resistance to traditional treatments such as chemotherapy due to its self-renew capability and multi-lineage differentiation potential (1)However, it remains elusive how the CSCs maintain their stemness, thus restricting the development of therapeutic treatments specifically targeting the CSCs.
基金funded by grants from the Ministry of Science and Technology of People’s Republic of China(Nos.2011CBA00906 and 2011YQ03013404).
文摘Activation-induced cytidine deaminase(AID)is required for the generation of antibody diversity through initiat-ing both somatic hypermutation(SHM)and class switch recombination.A few research groups have success-fully used the feature of AID for generating mutant li-braries in directed evolution of target proteins in B cells in vitro.B cells,cultured in suspension,are not con-venient for transfection and cloning.In this study,we established an AID-based mutant accumulation and sorting system in adherent human cells.Mouse AID gene was first transfected into the human non-small cell lung carcinoma H1299 cells,and a stable cell clone(H1299-AID)was selected.Afterwards,anti-hTNF-αscFv(ATscFv)was transfected into H1299-AID cells and ATscFv was displayed on the surface of H1299-AID cells.By 4-round amplification/flow cytometric sorting for cells with the highest affinities to hTNF-alpha,two ATscFv mutant gene clones were isolated.Compared with the wild type ATscFv,the two mutants were much more efficient in neutralizing cytotoxicity of hTNF-alpha.The results indicate that directed evolution by somatic hypermutation can be carried out in adherent non-B cells,which makes directed evolution in mammalian cells easier and more efficient.
基金supported by the Energy Biosciences Institute and by the DOE Center for Advanced Bioenergy and Bioproducts Innovation(U.S.Department of Energy,Office of Science,Office of Biological and Environmental Research under Award Number DE-SC0018420)US Department of Energy,DE-SC0018420,CV Rao.
文摘D-xylose is an abundant sugar found in plant biomass and can be used as a renewable feedstock for the microbial production of diverse biofuels and bioproducts.However,D-xylose metabolism is slow in many industrial microorganisms,at least as compared to glucose metabolism.Not surprisingly,a number of approaches have been developed for improving D-xylose metabolism in diverse microorganisms.In this work,we applied a previously developed evolution strategy based on media-in-oil emulsions for improving the growth yield of Escherichia coli NCM3722 on D-xylose.After 30 rounds of evolutions,we isolated multiple mutants with increased growth yield on D-xylose.In addition,we also observed similar increases in the growth rate.Three mutants were selected for whole-genome sequencing.Two mutants had an amber stop mutation in adenylate cyclase,which truncates nearly 60%of the enzyme.However,the ability of this mutant to grow on xylose indicated that truncated enzyme,lacking the C-terminal regulatory domain,is still active.The other mutant had a point mutation in the cyclic AMP receptor protein(CRP),near the high affinity binding site for cyclic AMP.Both mutations,when intro-duced into wild type E.coli,were able to increase the growth yields at levels similar to the isolated mutants.In addition to D-xylose,these mutant strains and their genetic mimics also exhibited higher growth rates and yields on glucose,lactose,and L-arabinose.These results suggest that the improved growth rates and yields are due to changes in the production and sensing of intracellular cyclic AMP concentrations and also suggest native concentrations are suboptimal with respect to the growth rate and yield under the growth conditions tested.Collectively,these results may prove useful for engineering strains of E.coli for high-density fermentations or protein production.
基金supported by the National Key R&D Program of China (2020YFA0803501,2019YFA0508501,2021YFF0702802)the National Natural Science Foundation of China (82130088,31930036,81921003,31871494,92042302,91940305,32070533,81772646)+2 种基金the Beijing Natural Science Foundation (5192018)the Biological Resources Program of Chinese Academy of Sciences (KFJ-BRP-017)the Strategic Priority Research Programs of the Chinese Academy of Sciences (XDB19030203).
文摘Neutrophils are derived from bone marrow hematopoietic stem cells(HSCs)and are the largest population among circulating white blood cells in humans,acting as the first line of defense against invading pathogens.Whether neutrophils can be generated by transdifferentiation strategies is unknown.Here,we show that thymidine induces the conversion of mouse fibroblasts to neutrophils.Induced neutrophils(iNeus)showed antibacterial effects and did not undergo malignant transformation in vivo.Importantly,iNeu transplantation cured neutropenia in mice in vivo.Mechanistically,thymidine mediates iNeu conversion by enhancing Tet3 activity.Tet3 initiates the expression of the neutrophil fate decision factors Cebpδ and Rfx1 that drive the transdifferentiation of mouse fibroblasts to neutrophils.Therefore,the induction of functional neutrophils by chemicals may provide a potential therapeutic strategy for patients with neutropenia patients and infectious diseases.
