T cells modified with chimeric antigen receptor are an attractive strategy to treat Epstein-Barr virus（EBV） associated malignancies.The EBV latent membrane protein 1（LMP1） is a 66-KD integral membrane protein enco...T cells modified with chimeric antigen receptor are an attractive strategy to treat Epstein-Barr virus（EBV） associated malignancies.The EBV latent membrane protein 1（LMP1） is a 66-KD integral membrane protein encoded by EBV that consists of transmembrane-spanning loops.Previously,we have identified a functional signal chain variable fragment（scFv） that specifically recognizes LMP1 through phage library screening.Here,we constructed a LMP1 specific chimeric antigen receptor containing anti-LMP1 scFv,the CD28 signalling domain,and the CD3ζchain（HELA/CAR）.We tested its functional ability to target LMP1 positive nasopharyngeal carcinoma cells.HELA/CAR cells were efficiently generated using lentivirus vector encoding the LMP1-specific chimeric antigen receptor to infect activated human CD3＋ T cells.The HELA/CAR T cells displayed LMP1 specific cytolytic action and produced IFN-γ and IL-2 in response to nasopharyngeal carcinoma cells overexpressing LMP1.To demonstrate in vivo anti-tumor activity,we tested the HELA/CAR T cells in a xenograft model using an LMP1 overexpressing tumor.Intratumoral injection of anti-LMP1 HELA/CAR-T cells significantly reduced tumor growth in vivo.These results show that targeting LMP1 using HELA/CAR cells could represent an alternative therapeutic approach for patients with EBV-positive cancers.展开更多
Ischemia-reperfusion(I/R)injury induces region-specific neuronal vulnerability within the hippocampus,with the cornu ammonis 1(CA1)subfield particularly prone to delayed neuronal death.While intrinsic neuronal factors...Ischemia-reperfusion(I/R)injury induces region-specific neuronal vulnerability within the hippocampus,with the cornu ammonis 1(CA1)subfield particularly prone to delayed neuronal death.While intrinsic neuronal factors have been implicated,emerging evidence highlights the decisive contribution of astrocyte endfeet(AEF)—specialized perivascular structures that regulate ion and water homeostasis,glutamate clearance,and blood–brain barrier(BBB)stability.This review synthesizes structural and molecular alterations of AEF across the CA1-CA3 subfields following I/R and their correlation with neuronal fate.In CA1,AEF undergo early-onset swelling and detachment from the vascular basal lamina due to dysfunction of critical proteins such as aquaporin-4(AQP4)and Kir4.1.These changes impair glutamate uptake,metabolic support,and potassium buffering,contributing to neuronal hyperexcitability and degeneration.In contrast,AEF in CA3 preserves polarity and functional coupling of AQP4 and Kir4.1,conferring regional resilience.At the signaling level,AEF disruption activates mitogen-activated protein kinase(MAPK)/c-Jun N-terminal kinase(JNK)pathways,promotes reactive oxygen species(ROS)accumulation,and induces inducible nitric oxide synthase(iNOS)-mediated inflammation,amplifying neurotoxicity.Furthermore,subfield-specific astrocytic transcriptional profiles modulate inflammatory responses and gliovascular interactions.By reframing AEF not as passive scaffolds but as active regulators of neuronal survival,this review provides novel insight into the astrocyte-dependent mechanisms of hippocampal vulnerability.Therapeutic strategies that preserve AEF structure and function may offer targeted protection against delayed neuronal death in ischemic brain injury.展开更多
Objective:Small cell lung cancer(SCLC)is commonly recognized as the most fatal lung cancer type.Despite substantial advances in immune checkpoint blockade therapies for treating solid cancers,their benefits are limite...Objective:Small cell lung cancer(SCLC)is commonly recognized as the most fatal lung cancer type.Despite substantial advances in immune checkpoint blockade therapies for treating solid cancers,their benefits are limited to a minority of patients with SCLC.In the present study,novel indicators for predicting the outcomes and molecular targets for SCLC treatment were elucidated.Methods:We conducted bioinformatics analysis to identify the key genes associated with tumor-infiltrating lymphocytes in SCLC.The functional role of the key gene identified in SCLC was determined both in vitro and in vivo.Results:A significant correlation was observed between patient survival and CD56dim natural killer(NK)cell proportion.Furthermore,we noted that the hub gene ubiquitin-specific protease 1(USP1)is closely correlated with both CD56dim NK cells and overall survival in SCLC.Bioinformatics analysis revealed that USP1 is upregulated in SCLC.In addition,gene set enrichment analysis revealed that USP1 overexpression hinders NK cell-mediated immune responses.By co-cultivating NK-92 cells with SCLC cells,we demonstrated that NK cell cytotoxicity against SCLC could be improved either via USP1 knock-down or pharmacological inhibition.Furthermore,using a nude-mice xenograft tumor model,we noted that USP1 inhibition effectively suppressed tumor proliferation and increased the expression of NK cell-associated markers.Conclusions:Our study findings highlight the importance of NK cells in regulating SCLC.USP1 overexpression can inhibit NK cell-mediated immunity;therefore,USP1 may serve not only as a prognostic biomarker but also as a potential molecular target of SCLC therapy.展开更多
Protein aggregation drives proteinopathies ranging from ALS to systemic amyloidosis,yet the multiscale determinants bridging sequence,structure,and kinetics remain elusive.We present SKALE,an interpretable machine lea...Protein aggregation drives proteinopathies ranging from ALS to systemic amyloidosis,yet the multiscale determinants bridging sequence,structure,and kinetics remain elusive.We present SKALE,an interpretable machine learning framework that integrates sequence motifs,AlphaFold-derived structural descriptors,and experimental kinetics to decode aggregation mechanisms.SKALE identifies latent hotspots that evade conventional tools and matches high-performing neural baselines while preserving computational efficiency.In ALS-linked SOD1 G86R,the model isolates a risk region at residues 72-91 where preserved β-sheet geometry coincides with weakened hydrogen bonding to drive nucleation.Similarly,analysis of TDP-43 S332N reveals that a locally unwound helix increases surface exposure,a prediction validated by showing that targeted deletion of model-identified regions significantly reduces cellular aggregation.The framework generalizes to Tau P301L and PRNP variants where it uncovers distal aggregation-prone regions to discriminate pathogenic drivers from neutral mutations.Interpretability analysis further disentangles global from mutation-local mechanisms to reveal that β-sheet propensity acts as a shared determinant while hydrogen bond dynamics define specific routes to nucleation.These findings establish SKALE as a scalable,disease-agnostic engine that combines high-fidelity prediction with biophysical resolution to decode the molecular logic of misfolding and guide therapeutic design.展开更多
BACKGROUND Irreversible electroporation(IRE)is a novel local tumor ablation approach with the potential to activate the host’s immune system.However,this approach is insufficient to prevent cancer progression,and com...BACKGROUND Irreversible electroporation(IRE)is a novel local tumor ablation approach with the potential to activate the host’s immune system.However,this approach is insufficient to prevent cancer progression,and complementary approaches are required for effective immunotherapy.AIM To assess the immunomodulatory effects and mechanism of IRE combined antiprogrammed cell death protein 1(PD-1)treatment in subcutaneous pancreatic cancer models.METHODS C57BL-6 tumor-bearing mice were randomly divided into four groups:Control group;IRE group;anti-PD-1 group;and IRE+anti-PD-1 group.Tumor-infiltrating T,B,and natural killer cell levels and plasma concentrations of T helper type 1 cytokines(interleukin-2,interferon-γ,and tumor necrosis factor-α)were evaluated.Real-time PCR was used to determine the expression of CD8(marker of CD8+T cells)in tumor tissues of the mice of all groups at different points of time.The growth curves of tumors were drawn.RESULTS The results demonstrated that the IRE+anti-PD-1 group exhibited significantly higher percentages of T lymphocyte infiltration,including CD4+and CD8+T cells compared with the control group.Additionally,the IRE+anti-PD-1 group showed increased infiltration of natural killer and B cells,elevated cytokine levels,and higher CD8 mRNA expression.Tumor volume was significantly reduced in the IRE+anti-PD-1 group,indicating a more pronounced therapeutic effect.CONCLUSION The combination of IRE and anti-PD-1 therapy promotes CD8+T cell immunity responses,leading to a more effective reduction in tumor volume and improved therapeutic outcomes,which provides a new direction for ablation and immunotherapy of pancreatic cancer.展开更多
Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis(Bt)have proven to be highly effective in managing some key pests.However,the evolution of resistance by the target pests threa...Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis(Bt)have proven to be highly effective in managing some key pests.However,the evolution of resistance by the target pests threatens the sustainability of Bt crops.The L31S mutation in a tetraspanin encoded by Harm TspC5(previously known as Ha TSPAN1)has been shown to confer dominant resistance to the Bt protein Cry1Ac in Helicoverpa armigera,a globally damaging lepidopteran pest.However,the broader implications of the L31S mutation in the tetraspanins of other lepidopteran species remain unclear.The evolutionary analyses in this study indicate that TspC5s have evolved in a species-specific manner among the lepidopteran insects.To investigate the role of TspC5s in conferring dominant resistance to Cry1Ac,we used the piggyBac-based transformation system to generate four transgenic H.armigera strains that express exogenous TspC5 variants from three phylogenetically close species(Helicoverpa zea,Helicoverpa assulta and Heliothis virescens)and one phylogenetically distant species(Plutella xylostella).In comparison with the background SCD strain of H.armigera,the transgenic strains expressing HzeaTspC5-L31S,HassTspC5-L31S,or HvirTspC5-L31S exhibited significant resistance to Cry1Ac(10.0-,21.4-,and 81.1-fold,respectively),whereas the strain expressing PxylTspC5-L27S remained susceptible.Furthermore,the Cry1Ac resistant phenotypes followed an autosomal dominant inheritance pattern and were closely linked to the introduced mutant TspC5s.These findings reveal the conserved role of TspC5s from Helicoverpa and Heliothis species in mediating the dominant resistance to Cry1Ac,and they provide crucial insights for assessing resistance risks related to mutant tetraspanins and devising adaptive resistance management strategies for these major lepidopteran pests.展开更多
AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiati...AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un⁃explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1^(fl/fl))mice and Treg-specific AMPKα1 knockout mice(AMPKα1^(fl/fl)Foxp3^(cre)mice).RESULTS:Compared to AMPKα1^(fl/fl)mice,the percentage of eosinophils in the bone marrow of AMPKα1^(fl/fl)Foxp3^(cre)mice was significant⁃ly reduced.Additionally,while the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice exhibited normal structure,both its size and the ratio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduction in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4^(+)T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice compared to AMPKα1^(fl/fl)mice.Importantly,the proportion of Siglec-F+CD11b^(+)eosinophils in the thymus was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4^(+)T cells in peripheral blood,alongside a decrease in the proportion of mature CD8^(+)T cells.Similarly,the proportion of CD4^(+)T cells in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice was elevated compared to AMPKα1^(fl/fl)mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice.In lymph nodes,the medullary boundaries in AMPKα1^(fl/fl)Foxp3^(cre)mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1^(fl/fl)mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3^(+)T cell population,particularly the CD8^(+)T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice,the percentage of CD4^(+)T cells was markedly in⁃creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1^(fl/fl)Foxp3^(cre)and AMPKα1^(fl/fl)mice.CONCLUSION:The populations of mature Tregs,CD8^(+)T cells and eosinophils in various im⁃mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.展开更多
Mutations in PTEN-induced putative kinase 1(PINK1) are implicated in early-onset Parkinson's disease(PD).Despite various in vitro studies indicating the importance of PINK1 in mitophagy,its physiological function ...Mutations in PTEN-induced putative kinase 1(PINK1) are implicated in early-onset Parkinson's disease(PD).Despite various in vitro studies indicating the importance of PINK1 in mitophagy,its physiological function in the brain remains poorly defined due to undetectable protein levels in rodents and cultured cells under basal conditions.Here,PINK1 was found to be selectively expressed in the primate brain,enabling exploration of its endogenous role in vivo.Proteomic profiling via mass spectrometry identified the ubiquitin-conjugating enzyme E2M(UBC12)as a PINK1-interacting partner,with strong colocalization in the monkey brain.Knockdown of PINK1 in monkeys resulted in marked reductions in UBC12 protein abundance and global neddylation,effects not observed in brain tissues from PINK1 knockout mice or pigs.These findings reveal a primate-specific PINK1-UBC12 axis and uncover a previously unrecognized role for PINK1 in protein neddylation,distinct from its established mitophagy function.展开更多
Flight feathers represent a hallmark innovation of avian evolution.Recent comparative genomic analyses identified a 284 bp avian-specific highly conserved element(ASHCE)located within the eighth intron of the SIM bHLH...Flight feathers represent a hallmark innovation of avian evolution.Recent comparative genomic analyses identified a 284 bp avian-specific highly conserved element(ASHCE)located within the eighth intron of the SIM bHLH transcription factor 1(Sim1)gene,postulated to act as a cis-regulatory element governing flight feather morphogenesis.To investigate its functional significance,genome-edited(GE)primordial germ cell(PGC)lines carrying targeted ASHCE deletions were generated using CRISPR/Cas9-mediated editing,with germline chimeric males subsequently mated with wild-type(WT)hens to obtain GE progeny.The resulting GE chickens harbored 257-260 bp deletions,excising approximately half of the Sim1-ASHCE sequence.Reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR)analysis showed an average 0.32-fold reduction in Sim1 expression in the forelimbs of GE embryos at day 8(E8)compared to WT counterparts.Despite this,GE chickens developed structurally normal flight and tail feathers.In situ hybridization localized Sim1 expression to the posterior mesenchyme surrounding flight feather buds in E8 WT embryos,but not within the buds themselves.These results suggest that partial deletion of Sim1-ASHCE,despite diminishing Sim1 expression,does not disrupt flight feather formation.The excised region appears to possess enhancer activity toward Sim1 but is dispensable for flight feather development.Complete ablation of the ASHCE will be necessary to fully resolve the regulatory role of Sim1 in avian feather morphogenesis.展开更多
Infiltration and activation of peripheral immune cells are critical in the progression of multiple sclerosis and its experimental animal model,experimental autoimmune encephalomyelitis(EAE).This study investigates the...Infiltration and activation of peripheral immune cells are critical in the progression of multiple sclerosis and its experimental animal model,experimental autoimmune encephalomyelitis(EAE).This study investigates the role of high mobility group box 1(HMGB1)in oligodendrocyte precursor cells(OPCs)in modulating pathogenic T cells infiltrating the central nervous system through the blood-brain barrier(BBB)by using OPC-specific HMGB1 knockout(KO)mice.We found that HMGB1 released from OPCs promotes BBB disruption,subsequently allowing increased immune cell infiltration.The migration of CD4+T cells isolated from EAE-induced mice was enhanced when co-cultured with OPCs compared to oligodendrocytes(OLs).OPC-specific HMGB1 KO mice exhibited lower BBB permeability and reduced immune cell infiltration into the CNS,leading to less damage to the myelin sheath and mitigated EAE progression.CD4+T cell migration was also reduced when co-cultured with HMGB1 knock-out OPCs.Our findings reveal that HMGB1 secretion from OPCs is crucial for regulating immune cell infiltration and provides insights into the immunomodulatory function of OPCs in autoimmune diseases.展开更多
BACKGROUND Type 1 diabetes(T1D)results from the autoimmune-mediated loss of pancreatic β-cells.Current insulin therapies offer symptomatic relief but fall short of providing a definitive cure.Islet cell transplantati...BACKGROUND Type 1 diabetes(T1D)results from the autoimmune-mediated loss of pancreatic β-cells.Current insulin therapies offer symptomatic relief but fall short of providing a definitive cure.Islet cell transplantation,while promising,faces limitations related to donor scarcity,procedural complexities,and the necessity for long-term immunosuppression.Consequently,there is an urgent need for innovative strategies aimed at β-cell regeneration.Patient-derived induced pluripotent stem cells(iPSCs),obtained from peripheral blood mononuclear cells(PBMCs)of T1D patients,hold great potential as a source of cells for therapeutic purposes.Therefore,the differentiation of T1D-iPSCs into functional pancreatic β-cells is a critical step toward effective β-cell replacement therapy.AIM To assess the potential of patient-derived T1D-β-like cells(differentiated from T1D-iPSCs reprogrammed from T1D-PBMCs)for restoring β-cell function in T1D.METHODS T1D-iPSCs were reprogrammed from T1D-PBMCs using an episomal vectorbased approach.Pluripotency was confirmed by flow cytometry(FCM),quantitative real-time polymerase chain reaction,genomic stability analysis,and teratoma formation assays.Differentiation into pancreatic β-cells was optimized using triiodothyronine(T3),vitamin C(Vc),and an adenovirus(M3C)encoding pancreatic duodenal homeobox-1,neurogenin 3(Ngn3),and MAF bZIP transcription factor A(MafA).Following characterization of β-cell features by immunofluorescence,quantitative real-time polymerase chain reaction,and flow cytometry,therapeutic efficacy was assessed through blood glucose monitoring after transplantation under the renal capsule of streptozotocin-induced diabetic mice.RESULTS T1D-iPSCs were successfully generated from T1D-PBMCs.These cells exhibited the hallmark characteristics of pluripotent stem cells,including appropriate morphology,differentiation potential,genomic integrity,and expression of pluripotency-associated genes.Differentiation yielded insulin-positive(insulin+)pancreatic β-like cells that,at the mRNA level,expressed key β-cell markers such as pancreatic duodenal homeobox-1,Ngn3,MafA,NeuroD,glucagon-like peptide-1 receptor,Nkx6.1,glucose transporter 2,and Kir6.2.Notably,the T3+Vc group displayed the lowest Ngn3 expression(1.31±0.38 vs 1.96±0.25 vs 2.51±0.24,P<0.01),while the M3C+T3+Vc group exhibited the highest MafA expression(0.49±0.11 vs 0.32±0.06 vs 0.29±0.08,P<0.05).Both in vitro and in vivo assessments confirmed the insulin secretion ability of the generated β-like cells;however,they did not demonstrate appropriate modulation of insulin release in response to variations in extracellular glucose concentrations.CONCLUSION T1D-iPSCs derived from T1D-PBMCs can be differentiated into insulin+β-like cells,albeit with functional immaturity.These cells represent a potential source of seed cells for β-cell replacement therapy in T1D.展开更多
Introducing the 1S^1 chromosome of Aegilops longissima into wheat genome can significantly improve wheat grain quality and contents of iron and zinc. Therefore, the development of molecular markers specific to 1S^1 ch...Introducing the 1S^1 chromosome of Aegilops longissima into wheat genome can significantly improve wheat grain quality and contents of iron and zinc. Therefore, the development of molecular markers specific to 1S^1 chromosome of A. longissima is of important significance for breeding high-quality wheat with high contents of iron and zinc in grains. In this study, nine molecular markers specific to 1S^1 chromosome of A. longissima were developed, including two 1S^1S specific markers,six 1S^1L specific markers and one 1S^1 specific marker which was located on both short and long arms. The practicability of these molecular markers were verified using hybrid population as materials. The results showed that hybrid population could be effectively screened and identified, which indicated that the developed 1S^1 chromosome-specific molecular markers could be used for screening and identification of hybrid population and could be used in marker-assisted breeding of high-quality wheat with high contents of Fe and Zn in grains.展开更多
基金supported in part by grants from the Special Fund of Clinical Medicine in Jiangsu Province(BL2013038)the Graduate Student Innovation Fund(CXZZ12_0563)
文摘T cells modified with chimeric antigen receptor are an attractive strategy to treat Epstein-Barr virus（EBV） associated malignancies.The EBV latent membrane protein 1（LMP1） is a 66-KD integral membrane protein encoded by EBV that consists of transmembrane-spanning loops.Previously,we have identified a functional signal chain variable fragment（scFv） that specifically recognizes LMP1 through phage library screening.Here,we constructed a LMP1 specific chimeric antigen receptor containing anti-LMP1 scFv,the CD28 signalling domain,and the CD3ζchain（HELA/CAR）.We tested its functional ability to target LMP1 positive nasopharyngeal carcinoma cells.HELA/CAR cells were efficiently generated using lentivirus vector encoding the LMP1-specific chimeric antigen receptor to infect activated human CD3＋ T cells.The HELA/CAR T cells displayed LMP1 specific cytolytic action and produced IFN-γ and IL-2 in response to nasopharyngeal carcinoma cells overexpressing LMP1.To demonstrate in vivo anti-tumor activity,we tested the HELA/CAR T cells in a xenograft model using an LMP1 overexpressing tumor.Intratumoral injection of anti-LMP1 HELA/CAR-T cells significantly reduced tumor growth in vivo.These results show that targeting LMP1 using HELA/CAR cells could represent an alternative therapeutic approach for patients with EBV-positive cancers.
文摘Ischemia-reperfusion(I/R)injury induces region-specific neuronal vulnerability within the hippocampus,with the cornu ammonis 1(CA1)subfield particularly prone to delayed neuronal death.While intrinsic neuronal factors have been implicated,emerging evidence highlights the decisive contribution of astrocyte endfeet(AEF)—specialized perivascular structures that regulate ion and water homeostasis,glutamate clearance,and blood–brain barrier(BBB)stability.This review synthesizes structural and molecular alterations of AEF across the CA1-CA3 subfields following I/R and their correlation with neuronal fate.In CA1,AEF undergo early-onset swelling and detachment from the vascular basal lamina due to dysfunction of critical proteins such as aquaporin-4(AQP4)and Kir4.1.These changes impair glutamate uptake,metabolic support,and potassium buffering,contributing to neuronal hyperexcitability and degeneration.In contrast,AEF in CA3 preserves polarity and functional coupling of AQP4 and Kir4.1,conferring regional resilience.At the signaling level,AEF disruption activates mitogen-activated protein kinase(MAPK)/c-Jun N-terminal kinase(JNK)pathways,promotes reactive oxygen species(ROS)accumulation,and induces inducible nitric oxide synthase(iNOS)-mediated inflammation,amplifying neurotoxicity.Furthermore,subfield-specific astrocytic transcriptional profiles modulate inflammatory responses and gliovascular interactions.By reframing AEF not as passive scaffolds but as active regulators of neuronal survival,this review provides novel insight into the astrocyte-dependent mechanisms of hippocampal vulnerability.Therapeutic strategies that preserve AEF structure and function may offer targeted protection against delayed neuronal death in ischemic brain injury.
基金supported by grants from the Dongguan Science and Technology of Social Development Program(No.20231800940192)the Talent Development Foundation of the First Dongguan Affiliated Hospital of Guangdong Medical University(No.PU2023002).
文摘Objective:Small cell lung cancer(SCLC)is commonly recognized as the most fatal lung cancer type.Despite substantial advances in immune checkpoint blockade therapies for treating solid cancers,their benefits are limited to a minority of patients with SCLC.In the present study,novel indicators for predicting the outcomes and molecular targets for SCLC treatment were elucidated.Methods:We conducted bioinformatics analysis to identify the key genes associated with tumor-infiltrating lymphocytes in SCLC.The functional role of the key gene identified in SCLC was determined both in vitro and in vivo.Results:A significant correlation was observed between patient survival and CD56dim natural killer(NK)cell proportion.Furthermore,we noted that the hub gene ubiquitin-specific protease 1(USP1)is closely correlated with both CD56dim NK cells and overall survival in SCLC.Bioinformatics analysis revealed that USP1 is upregulated in SCLC.In addition,gene set enrichment analysis revealed that USP1 overexpression hinders NK cell-mediated immune responses.By co-cultivating NK-92 cells with SCLC cells,we demonstrated that NK cell cytotoxicity against SCLC could be improved either via USP1 knock-down or pharmacological inhibition.Furthermore,using a nude-mice xenograft tumor model,we noted that USP1 inhibition effectively suppressed tumor proliferation and increased the expression of NK cell-associated markers.Conclusions:Our study findings highlight the importance of NK cells in regulating SCLC.USP1 overexpression can inhibit NK cell-mediated immunity;therefore,USP1 may serve not only as a prognostic biomarker but also as a potential molecular target of SCLC therapy.
基金International Brain Research Organization(IBRO)Rising Star Awardee and received an IBRO Early Career Principal Investigator Grant(No.PM010CNI000148)supported by Sunway University internal grant(No.GRTIN-IGS[02]-CVVR-11-2023)+2 种基金supported by the Fundamental Research Funds from the Central of Public Welfare Research Institute,China Rehabilitation Institutesupported by the research initiation funding scheme provided by Henan University of Technology(No.0004/31401568)Shenzhen Vaccine Biopharmaceuticals Limited(No.0004/51100292).
文摘Protein aggregation drives proteinopathies ranging from ALS to systemic amyloidosis,yet the multiscale determinants bridging sequence,structure,and kinetics remain elusive.We present SKALE,an interpretable machine learning framework that integrates sequence motifs,AlphaFold-derived structural descriptors,and experimental kinetics to decode aggregation mechanisms.SKALE identifies latent hotspots that evade conventional tools and matches high-performing neural baselines while preserving computational efficiency.In ALS-linked SOD1 G86R,the model isolates a risk region at residues 72-91 where preserved β-sheet geometry coincides with weakened hydrogen bonding to drive nucleation.Similarly,analysis of TDP-43 S332N reveals that a locally unwound helix increases surface exposure,a prediction validated by showing that targeted deletion of model-identified regions significantly reduces cellular aggregation.The framework generalizes to Tau P301L and PRNP variants where it uncovers distal aggregation-prone regions to discriminate pathogenic drivers from neutral mutations.Interpretability analysis further disentangles global from mutation-local mechanisms to reveal that β-sheet propensity acts as a shared determinant while hydrogen bond dynamics define specific routes to nucleation.These findings establish SKALE as a scalable,disease-agnostic engine that combines high-fidelity prediction with biophysical resolution to decode the molecular logic of misfolding and guide therapeutic design.
基金Science and Technology Program of Guangzhou,No.202102010077International Science Foundation of Guangzhou Fuda Cancer Hospital,No.Y2020-ZD-03.
文摘BACKGROUND Irreversible electroporation(IRE)is a novel local tumor ablation approach with the potential to activate the host’s immune system.However,this approach is insufficient to prevent cancer progression,and complementary approaches are required for effective immunotherapy.AIM To assess the immunomodulatory effects and mechanism of IRE combined antiprogrammed cell death protein 1(PD-1)treatment in subcutaneous pancreatic cancer models.METHODS C57BL-6 tumor-bearing mice were randomly divided into four groups:Control group;IRE group;anti-PD-1 group;and IRE+anti-PD-1 group.Tumor-infiltrating T,B,and natural killer cell levels and plasma concentrations of T helper type 1 cytokines(interleukin-2,interferon-γ,and tumor necrosis factor-α)were evaluated.Real-time PCR was used to determine the expression of CD8(marker of CD8+T cells)in tumor tissues of the mice of all groups at different points of time.The growth curves of tumors were drawn.RESULTS The results demonstrated that the IRE+anti-PD-1 group exhibited significantly higher percentages of T lymphocyte infiltration,including CD4+and CD8+T cells compared with the control group.Additionally,the IRE+anti-PD-1 group showed increased infiltration of natural killer and B cells,elevated cytokine levels,and higher CD8 mRNA expression.Tumor volume was significantly reduced in the IRE+anti-PD-1 group,indicating a more pronounced therapeutic effect.CONCLUSION The combination of IRE and anti-PD-1 therapy promotes CD8+T cell immunity responses,leading to a more effective reduction in tumor volume and improved therapeutic outcomes,which provides a new direction for ablation and immunotherapy of pancreatic cancer.
基金primarily funded by a grant from the National Natural Science Foundation of China(31930093)Additional support was provided by the Natural Science Foundation of Jiangsu Province,China(BK20230983)the Project of Fund for Stable Support to Agricultural Sci-Tech Renovation,China(xjnkywdzc-2022004)。
文摘Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis(Bt)have proven to be highly effective in managing some key pests.However,the evolution of resistance by the target pests threatens the sustainability of Bt crops.The L31S mutation in a tetraspanin encoded by Harm TspC5(previously known as Ha TSPAN1)has been shown to confer dominant resistance to the Bt protein Cry1Ac in Helicoverpa armigera,a globally damaging lepidopteran pest.However,the broader implications of the L31S mutation in the tetraspanins of other lepidopteran species remain unclear.The evolutionary analyses in this study indicate that TspC5s have evolved in a species-specific manner among the lepidopteran insects.To investigate the role of TspC5s in conferring dominant resistance to Cry1Ac,we used the piggyBac-based transformation system to generate four transgenic H.armigera strains that express exogenous TspC5 variants from three phylogenetically close species(Helicoverpa zea,Helicoverpa assulta and Heliothis virescens)and one phylogenetically distant species(Plutella xylostella).In comparison with the background SCD strain of H.armigera,the transgenic strains expressing HzeaTspC5-L31S,HassTspC5-L31S,or HvirTspC5-L31S exhibited significant resistance to Cry1Ac(10.0-,21.4-,and 81.1-fold,respectively),whereas the strain expressing PxylTspC5-L27S remained susceptible.Furthermore,the Cry1Ac resistant phenotypes followed an autosomal dominant inheritance pattern and were closely linked to the introduced mutant TspC5s.These findings reveal the conserved role of TspC5s from Helicoverpa and Heliothis species in mediating the dominant resistance to Cry1Ac,and they provide crucial insights for assessing resistance risks related to mutant tetraspanins and devising adaptive resistance management strategies for these major lepidopteran pests.
基金Supported by the National Natural Science Foundation of China(No.81800423)the Guangdong Medical Science and Technology Research project(No.B2022102)。
文摘AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un⁃explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1^(fl/fl))mice and Treg-specific AMPKα1 knockout mice(AMPKα1^(fl/fl)Foxp3^(cre)mice).RESULTS:Compared to AMPKα1^(fl/fl)mice,the percentage of eosinophils in the bone marrow of AMPKα1^(fl/fl)Foxp3^(cre)mice was significant⁃ly reduced.Additionally,while the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice exhibited normal structure,both its size and the ratio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduction in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4^(+)T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice compared to AMPKα1^(fl/fl)mice.Importantly,the proportion of Siglec-F+CD11b^(+)eosinophils in the thymus was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4^(+)T cells in peripheral blood,alongside a decrease in the proportion of mature CD8^(+)T cells.Similarly,the proportion of CD4^(+)T cells in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice was elevated compared to AMPKα1^(fl/fl)mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice.In lymph nodes,the medullary boundaries in AMPKα1^(fl/fl)Foxp3^(cre)mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1^(fl/fl)mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3^(+)T cell population,particularly the CD8^(+)T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice,the percentage of CD4^(+)T cells was markedly in⁃creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1^(fl/fl)Foxp3^(cre)and AMPKα1^(fl/fl)mice.CONCLUSION:The populations of mature Tregs,CD8^(+)T cells and eosinophils in various im⁃mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.
基金supported by the National Key Research and Development Program of China (2021YFF0702201)National Natural Science Foundation of China (32070534, 32370567, 81830032, 31872779,82071421, 81873736)+4 种基金Department of Science and Technology of Guangdong Province (2021ZT09Y007, 2020B121201006)Guangdong Basic and Applied Basic Research Foundation (2023B1515020031,2022A1515012301)K.C. Wong Education FoundationFundamental Research Funds for the Central Universities (Jinan University, 11620358)Hubei Topgene Biotechnology Co.Ltd。
文摘Mutations in PTEN-induced putative kinase 1(PINK1) are implicated in early-onset Parkinson's disease(PD).Despite various in vitro studies indicating the importance of PINK1 in mitophagy,its physiological function in the brain remains poorly defined due to undetectable protein levels in rodents and cultured cells under basal conditions.Here,PINK1 was found to be selectively expressed in the primate brain,enabling exploration of its endogenous role in vivo.Proteomic profiling via mass spectrometry identified the ubiquitin-conjugating enzyme E2M(UBC12)as a PINK1-interacting partner,with strong colocalization in the monkey brain.Knockdown of PINK1 in monkeys resulted in marked reductions in UBC12 protein abundance and global neddylation,effects not observed in brain tissues from PINK1 knockout mice or pigs.These findings reveal a primate-specific PINK1-UBC12 axis and uncover a previously unrecognized role for PINK1 in protein neddylation,distinct from its established mitophagy function.
基金supported by the Ministry of Agriculture and Rural Affairs of the People's Republic of China(125A0607)Department of Science and Technology of Yunnan Province(XDYC-KJLJ-2022-0004)。
文摘Flight feathers represent a hallmark innovation of avian evolution.Recent comparative genomic analyses identified a 284 bp avian-specific highly conserved element(ASHCE)located within the eighth intron of the SIM bHLH transcription factor 1(Sim1)gene,postulated to act as a cis-regulatory element governing flight feather morphogenesis.To investigate its functional significance,genome-edited(GE)primordial germ cell(PGC)lines carrying targeted ASHCE deletions were generated using CRISPR/Cas9-mediated editing,with germline chimeric males subsequently mated with wild-type(WT)hens to obtain GE progeny.The resulting GE chickens harbored 257-260 bp deletions,excising approximately half of the Sim1-ASHCE sequence.Reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR)analysis showed an average 0.32-fold reduction in Sim1 expression in the forelimbs of GE embryos at day 8(E8)compared to WT counterparts.Despite this,GE chickens developed structurally normal flight and tail feathers.In situ hybridization localized Sim1 expression to the posterior mesenchyme surrounding flight feather buds in E8 WT embryos,but not within the buds themselves.These results suggest that partial deletion of Sim1-ASHCE,despite diminishing Sim1 expression,does not disrupt flight feather formation.The excised region appears to possess enhancer activity toward Sim1 but is dispensable for flight feather development.Complete ablation of the ASHCE will be necessary to fully resolve the regulatory role of Sim1 in avian feather morphogenesis.
基金supported by the National Research Foundation of Korea(NRF)funded by the government of the Republic of Korea[2023R1A2C1004955]the Technology Innovation Program funded by the Ministry of Trade,Industry&Energy(Korea)(20009707)+1 种基金the Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education(2020R1A6A3A01099056)the Korea Institute for Advancement of Technology funded by the Ministry of Trade,Industry and Energy(P0025489).
文摘Infiltration and activation of peripheral immune cells are critical in the progression of multiple sclerosis and its experimental animal model,experimental autoimmune encephalomyelitis(EAE).This study investigates the role of high mobility group box 1(HMGB1)in oligodendrocyte precursor cells(OPCs)in modulating pathogenic T cells infiltrating the central nervous system through the blood-brain barrier(BBB)by using OPC-specific HMGB1 knockout(KO)mice.We found that HMGB1 released from OPCs promotes BBB disruption,subsequently allowing increased immune cell infiltration.The migration of CD4+T cells isolated from EAE-induced mice was enhanced when co-cultured with OPCs compared to oligodendrocytes(OLs).OPC-specific HMGB1 KO mice exhibited lower BBB permeability and reduced immune cell infiltration into the CNS,leading to less damage to the myelin sheath and mitigated EAE progression.CD4+T cell migration was also reduced when co-cultured with HMGB1 knock-out OPCs.Our findings reveal that HMGB1 secretion from OPCs is crucial for regulating immune cell infiltration and provides insights into the immunomodulatory function of OPCs in autoimmune diseases.
基金Supported by the Nonprofit Research Institutes Foundation of Fujian Province,China,No.2020R1011003 and No.2022R1012001the Talents Training Project for the Key Young Scholars of Fujian Provincial Health Commission,China,No.2021GGA056.
文摘BACKGROUND Type 1 diabetes(T1D)results from the autoimmune-mediated loss of pancreatic β-cells.Current insulin therapies offer symptomatic relief but fall short of providing a definitive cure.Islet cell transplantation,while promising,faces limitations related to donor scarcity,procedural complexities,and the necessity for long-term immunosuppression.Consequently,there is an urgent need for innovative strategies aimed at β-cell regeneration.Patient-derived induced pluripotent stem cells(iPSCs),obtained from peripheral blood mononuclear cells(PBMCs)of T1D patients,hold great potential as a source of cells for therapeutic purposes.Therefore,the differentiation of T1D-iPSCs into functional pancreatic β-cells is a critical step toward effective β-cell replacement therapy.AIM To assess the potential of patient-derived T1D-β-like cells(differentiated from T1D-iPSCs reprogrammed from T1D-PBMCs)for restoring β-cell function in T1D.METHODS T1D-iPSCs were reprogrammed from T1D-PBMCs using an episomal vectorbased approach.Pluripotency was confirmed by flow cytometry(FCM),quantitative real-time polymerase chain reaction,genomic stability analysis,and teratoma formation assays.Differentiation into pancreatic β-cells was optimized using triiodothyronine(T3),vitamin C(Vc),and an adenovirus(M3C)encoding pancreatic duodenal homeobox-1,neurogenin 3(Ngn3),and MAF bZIP transcription factor A(MafA).Following characterization of β-cell features by immunofluorescence,quantitative real-time polymerase chain reaction,and flow cytometry,therapeutic efficacy was assessed through blood glucose monitoring after transplantation under the renal capsule of streptozotocin-induced diabetic mice.RESULTS T1D-iPSCs were successfully generated from T1D-PBMCs.These cells exhibited the hallmark characteristics of pluripotent stem cells,including appropriate morphology,differentiation potential,genomic integrity,and expression of pluripotency-associated genes.Differentiation yielded insulin-positive(insulin+)pancreatic β-like cells that,at the mRNA level,expressed key β-cell markers such as pancreatic duodenal homeobox-1,Ngn3,MafA,NeuroD,glucagon-like peptide-1 receptor,Nkx6.1,glucose transporter 2,and Kir6.2.Notably,the T3+Vc group displayed the lowest Ngn3 expression(1.31±0.38 vs 1.96±0.25 vs 2.51±0.24,P<0.01),while the M3C+T3+Vc group exhibited the highest MafA expression(0.49±0.11 vs 0.32±0.06 vs 0.29±0.08,P<0.05).Both in vitro and in vivo assessments confirmed the insulin secretion ability of the generated β-like cells;however,they did not demonstrate appropriate modulation of insulin release in response to variations in extracellular glucose concentrations.CONCLUSION T1D-iPSCs derived from T1D-PBMCs can be differentiated into insulin+β-like cells,albeit with functional immaturity.These cells represent a potential source of seed cells for β-cell replacement therapy in T1D.
基金Supported by National Natural Science Foundation of China(31201203)Earmarked Fund for Modern Agro-industry Technology Research System(CARS-03-1-8)+3 种基金China Postdoctoral Science Foundation(2013T60850)Program for Youth Talent of Shandong Academy of Agricultural Sciences(1-18-024)Seed Industry Foundation Grant to Taishan ScholarAgricultural Improved Variety Industrialization Project of Shandong Province(2-B-08)~~
文摘Introducing the 1S^1 chromosome of Aegilops longissima into wheat genome can significantly improve wheat grain quality and contents of iron and zinc. Therefore, the development of molecular markers specific to 1S^1 chromosome of A. longissima is of important significance for breeding high-quality wheat with high contents of iron and zinc in grains. In this study, nine molecular markers specific to 1S^1 chromosome of A. longissima were developed, including two 1S^1S specific markers,six 1S^1L specific markers and one 1S^1 specific marker which was located on both short and long arms. The practicability of these molecular markers were verified using hybrid population as materials. The results showed that hybrid population could be effectively screened and identified, which indicated that the developed 1S^1 chromosome-specific molecular markers could be used for screening and identification of hybrid population and could be used in marker-assisted breeding of high-quality wheat with high contents of Fe and Zn in grains.
