Osteoporosis is a known risk factor for rotator cuff tears(RCTs),but the causal correlation and underlying mechanisms remain unclear.This study aims to evaluate the impact of osteoporosis on RCT risk and investigate t...Osteoporosis is a known risk factor for rotator cuff tears(RCTs),but the causal correlation and underlying mechanisms remain unclear.This study aims to evaluate the impact of osteoporosis on RCT risk and investigate their genetic associations.Using data from the UK Biobank(n=457871),cross-sectional analyses demonstrated that osteoporosis was significantly associated with an increased risk of RCTs(adjusted OR[95%CI]=1.38[1.25–1.52]).A longitudinal analysis of a subset of patients(n=268117)over 11 years revealed that osteoporosis increased the risk of RCTs(adjusted HR[95%CI]=1.56[1.29–1.87]),which is notably varied between sexes in sex-stratified analysis.Causal inference methods,including propensity score matching,inverse probability weighting,causal random forest and survival random forest models further confirmed the causal effect,both from cross-sectional and longitudinal perspectives.展开更多
Background:Diabetic retinopathy(DR)urgently needs novel and effective therapeutic targets.Integrated analyses of plasma proteomic and genetic markers can clarify the causal relevance of proteins and discover novel tar...Background:Diabetic retinopathy(DR)urgently needs novel and effective therapeutic targets.Integrated analyses of plasma proteomic and genetic markers can clarify the causal relevance of proteins and discover novel targets for diseases,but no systematic screening for DR has been performed.Methods:Summary statistics of plasma protein quantitative trait loci(pQTL)were derived from two extensive genome-wide analysis study(GWAS)datasets and one systematic review,with over 100 thousand participants covering thousands of plasma proteins.DR data were sourced from the largest FinnGen study,comprising 10,413 DR cases and 308,633 European controls.Genetic instrumental variables were identified using multiple filters.In the two-sample MR analysis,Wald ratio and inverse variance-weighted(IVW)MR were utilized to investigate the causality of plasma proteins with DR.Bidirectional MR,Bayesian Co-localization,and phenotype scanning were employed to test for potential reverse causality and confounding factors in the main MR analyses.By systemically searching druggable gene lists,the ChEMBL database,DrugBank,and Gene Ontology database,the druggability and relevant functional pathways of the identified proteins were systematically evaluated.Results:Genetically predicted levels of 24 proteins were significantly associated with DR risk at a false discovery rate<0.05 including 11 with positive associations and 13 with negative associations.For each standard deviation increase in plasm protein levels,the odds ratios(ORs)for DR varied from 0.51(95%CI:0.36-0.73;P=2.22×10-5)for tubulin polymerization-promoting protein family member 3(TPPP3)to 2.02(95%CI:1.44-2.83;P=5.01×10-5)for olfactomedin like 3(OLFML3).Bidirectional MR indicated there was no reverse causality that interfered with the results of the main MR analyses.Four proteins exhibited strong co-localization evidence(PH4≥0.8):cytoplasmic tRNA synthetase(WARS),acrosin binding protein(ACRBP),and intercellular adhesion molecule 1(ICAM1)were negatively associated with DR risk,while neurogenic locus notch homolog protein 2(NOTCH2)showed a positive association.No confounding factors were detected between pQTLs and DR according to the phenotypic scan.Drugability assessments highlighted 6 proteins already in drug development endeavor and 18 novel drug targets,with metalloproteinase inhibitor 3(TIMP)currently in phase I clinical trials for DR.GO analysis identified 18 of 24 plasma proteins enriching 22 pathways related to cell differentiation and proliferation regulation.Conclusions:Twenty-four promising drug targets for DR were identified,including four plasma proteins with particular co-localization evidence.These findings offer new insights into DR's etiology and therapeutic targeting,exemplifying the value of genomic and proteomic data in drug target discovery.展开更多
Background Lung squamous cell carcinoma(LUSC)is a major subtype of non-small cell lung cancer with a high mortality rate.Identifying causal plasma proteins associated with LUSC could provide new insights into the path...Background Lung squamous cell carcinoma(LUSC)is a major subtype of non-small cell lung cancer with a high mortality rate.Identifying causal plasma proteins associated with LUSC could provide new insights into the pathophysiology of the disease and potential therapeutic targets.This study aimed to identify plasma proteins causally linked to LUSC risk using proteome-wide Mendelian randomization(MR)and colocalization analyses.Methods Proteome-wide MR analysis was conducted using data from the UK Biobank Pharma Proteomics Project and deCODE genetics.Summary-level data for LUSC were obtained from the ILCCO Consortium,the FinnGen study,and a separate GWAS study.A total of 1,046 shared protein quantitative trait loci(pQTLs)were analyzed.Sensitivity analyses included the HEIDI test for horizontal pleiotropy and colocalization analysis to validate the causal associations.Results MR analysis identified six plasma proteins associated with LUSC risk:HSPA1L,PCSK7,POLI,SPINK2,TCL1A,and VARS.HSPA1L(OR=0.47;95%CI:0.34–0.65;P=4.89×10^(–6)),SPINK2(OR=0.68;95%CI:0.58–0.80;P=3.17×10^(–6)),and VARS(OR=0.44;95%CI:0.31–0.63;P=5.94×10^(–6))were associated with a decreased risk of LUSC.Conversely,PCSK7(OR=1.37;95%CI:1.21–1.56;P=1.40×10^(–6)),POLI(OR=4.50;95%CI:2.25–9.00;P=2.13×10–5),and TCL1A(OR=1.72;95%CI:1.34–2.21;P=1.89×10–5)were associated with an increased risk.The SMR analysis and HEIDI test confirmed the robustness of these associations.HSPA1L,SPINK2,and VARS showed significant inverse associations,with strong colocalization evidence for TCL1A(PPH4=0.817).Conclusions This study identified six plasma proteins potentially causal for LUSC risk.HSPA1L,SPINK2,and VARS are associated with decreased risk,while PCSK7,POLI,and TCL1A are linked to increased risk.These findings provide new insights into LUSC pathogenesis and highlight potential targets for therapeutic intervention.展开更多
Background A detailed understanding of genetic variants that affect beef merit helps maximize the efficiency of breeding for improved production merit in beef cattle.To prioritize the putative variants and genes,we ra...Background A detailed understanding of genetic variants that affect beef merit helps maximize the efficiency of breeding for improved production merit in beef cattle.To prioritize the putative variants and genes,we ran a com-prehensive genome-wide association studies(GWAS)analysis for 21 agronomic traits using imputed whole-genome variants in Simmental beef cattle.Then,we applied expression quantitative trait loci(eQTL)mapping between the genotype variants and transcriptome of three tissues(longissimus dorsi muscle,backfat,and liver)in 120 cattle.Results We identified 1,580 association signals for 21 beef agronomic traits using GWAS.We then illuminated 854,498 cis-eQTLs for 6,017 genes and 46,970 trans-eQTLs for 1,903 genes in three tissues and built a synergistic network by integrating transcriptomics with agronomic traits.These cis-eQTLs were preferentially close to the transcription start site and enriched in functional regulatory regions.We observed an average of 43.5%improvement in cis-eQTL discovery using multi-tissue eQTL mapping.Fine-mapping analysis revealed that 111,192,and 194 variants were most likely to be causative to regulate gene expression in backfat,liver,and muscle,respectively.The transcriptome-wide association studies identified 722 genes significantly associated with 11 agronomic traits.Via the colocalization and Mendelian randomization analyses,we found that eQTLs of several genes were associated with the GWAS signals of agronomic traits in three tissues,which included genes,such as NADSYN1,NDUFS3,LTF and KIFC2 in liver,GRAMD1C,TMTC2 and ZNF613 in backfat,as well as TIGAR,NDUFS3 and L3HYPDH in muscle that could serve as the candidate genes for economic traits.Conclusions The extensive atlas of GWAS,eQTL,fine-mapping,and transcriptome-wide association studies aid in the suggestion of potentially functional variants and genes in cattle agronomic traits and will be an invaluable source for genomics and breeding in beef cattle.展开更多
Objective: To investigate the colocalization of Somatostatin (SOM ) and NADPH--diaphorase (NADPHd ) of the neurons in raphe nuclei innervating pharyngeal muscles in the rats. Methods: After PRV was injected into the p...Objective: To investigate the colocalization of Somatostatin (SOM ) and NADPH--diaphorase (NADPHd ) of the neurons in raphe nuclei innervating pharyngeal muscles in the rats. Methods: After PRV was injected into the pharyngeal muscles. PRV and SOM immunofluorescence double labeling procedure was completed at first,then proceeded NADPH -d histochemistry. Results: PRV and SOM double--labled cells were present mainly in the nucleus raphe magnus. but some PRV and SOM double labled neurons were found in the other raphe nuclei as well, such as nucleus raphe pallidus. nucleus raphe obsurus, median raphe nucleus and dorsal raphe nucleus.NADPH -d positive neurons were also observed in the raphe nuclei. PRV. SOM and NADPH-d triple labeling neurons were found in the nucleus raphe magnus. Conclusion: It is suggested that the colocalization of SOM and NADPH--d of the neurons in the raphe nuclei innervating pharyngeal muscles may play an important role in the coordination of the pharyngeal motility.展开更多
To the Editor:Atopic dermatitis(AD)is a chronic inflammatory skin condition marked by recurrent eczematous lesions and intense pruritus,affecting approximately 15–20%of children and up to 10%of adults worldwide.[1]Th...To the Editor:Atopic dermatitis(AD)is a chronic inflammatory skin condition marked by recurrent eczematous lesions and intense pruritus,affecting approximately 15–20%of children and up to 10%of adults worldwide.[1]The presence of AD significantly impairs patient quality of life and imposes substantial economic burdens on society.In recent years,the potential link between AD and lymphoma has attracted considerable attention from researchers and clinicians.Lymphoma,a malignancy of the lymphatic system,poses significant health risks due to its high mortality potential.[2]However,observational studies investigating the potential correlation between AD and lymphoma have yielded inconsistent results.Consequently,the association between AD and lymphoma remains controversial and warrants further investigation.展开更多
Background Semen quality is one of the most important indicators of boar reproductive performance.In the past,boar breeding has mostly emphasized characteristics such as lean meat percentage,feed conversion efficiency...Background Semen quality is one of the most important indicators of boar reproductive performance.In the past,boar breeding has mostly emphasized characteristics such as lean meat percentage,feed conversion efficiency,and growth rate,while overlooking the genetic improvement of reproductive traits.This study employs advanced multi-omics approaches,such as transcriptome-wide association studies(TWAS)and colocalization between genome-wide association studies(GWAS)and expression quantitative trait loci(eQTLs),to provide a comprehensive understanding of the genetic mechanisms governing semen quality traits in boars.Results Here,we collected 190,000 ejaculate records across 11 semen quality traits from 3,604 Duroc boars.The heritability of semen quality traits ranged from 0.095 to 0.343.Genetic correlations between semen quality traits varied from−0.802 to 0.661,and phenotypic correlations ranged from−0.833 to 0.776.Single-trait GWAS identified 19 independent variants,corresponding to 13 quantitative trait loci(QTLs).By integrating PigGTEx and FAANG resources,we combined TWAS and colocalization analyses to reveal genetic regulation of semen quality traits.Notably,both GWAS and colocalization analyses pinpointed the DCAF12 as a crucial gene associated with multiple semen quality traits.Additionally,the ZSCAN9 gene and the variant rs322211455 were found to significantly affect sperm motility(SPMOT),possibly through hypothalamic-pituitary-gonadal axis.PheWAS further highlighted an association between rs322211455 and sperm abnormality rate,demonstrating the crucial role of ZSCAN9 in male fertility.Conclusion This study reveals the genetic basis and regulatory mechanisms underlying semen quality traits in Duroc boars,identifying key candidate genes such as DCAF12 and ZSCAN9.These findings provide important insight into the genetic regulation of semen quality in boars.展开更多
Although the spatial characteristics within the tumor microenvironment of lung adenocarcinoma(LUAD)have been identified,the mechanisms by which these factors promote LUAD progression and immune evasion remain unclear....Although the spatial characteristics within the tumor microenvironment of lung adenocarcinoma(LUAD)have been identified,the mechanisms by which these factors promote LUAD progression and immune evasion remain unclear.Using spatial transcriptomics and single-cell RNA-sequencing data from multi-regional LUAD biopsies consisting of tumor core,tumor edge,and normal area,we sought to delineate the spatial heterogeneity and driving factors of cell colocalization.Two cancer cell sub-clusters(Cancer_c1 and Cancer_c2),associated with LUAD initiation and metastasis,respectively,exhibit distinct spatial distributions and immune cell colocalizations.In particular,Cancer_c1,enriched within the tumor core,could directly interact with B cells or indirectly recruit B cells through macrophages.Conversely,Cancer_c2 enriched within the tumor edge exhibits colocalization with CD8^(+)T cells.Collectively,our work elucidates the spatial distribution of cancer cell subtypes and their interaction with immune cells in the core and edge of LUAD,providing insights for developing therapeutic strategies for cancer intervention.展开更多
Genome-wide association studies(GWAS)have identified thousands of genomic loci associated with complex diseases and traits,including cancer.The vast majority of common traitassociated variants identified via GWAS fall...Genome-wide association studies(GWAS)have identified thousands of genomic loci associated with complex diseases and traits,including cancer.The vast majority of common traitassociated variants identified via GWAS fall in non-coding regions of the genome,posing a challenge in elucidating the causal variants,genes,and mechanisms involved.Expression quantitative trait locus(eQTL)and other molecular QTL studies have been valuable resources in identifying candidate causal genes from GWAS loci through statistical colocalization methods.While QTL colocalization is becoming a standard analysis in post-GWAS investigation,an easy web tool for users to perform formal colocalization analyses with either user-provided or public GWAS and eQTL datasets has been lacking.Here,we present ezQTL,a web-based bioinformatic application to interactively visualize and analyze genetic association data such as GWAS loci and molecular QTLs under different linkage disequilibrium(LD)patterns(1000 Genomes Project,UK Biobank,or user-provided data).This application allows users to perform data quality control for variants matched between different datasets,LD visualization,and two-trait colocalization analyses using two state-of-the-art methodologies(eCAVIAR and HyPrColoc),including batch processing.ezQTL is a free and publicly available cross-platform web tool,which can be accessed online at https://analysistools.cancer.gov/ezqtl.展开更多
AIM: To investigate the mechanism of endoplasmic reticulum(ER) stress induction by an occult infection related hepatitis B virus S surface antigen(HBsAg)variant.METHODS: We used an HBsAg variant with lower secretion c...AIM: To investigate the mechanism of endoplasmic reticulum(ER) stress induction by an occult infection related hepatitis B virus S surface antigen(HBsAg)variant.METHODS: We used an HBsAg variant with lower secretion capacity, which was a KD variant from a Korean subject who was occultly infected with the genotype C. We compared the expression profiles of ER stress-related proteins between HuH-7 cells transfected with HBsAg plasmids of a wild-type and a KD variant using Western blot.RESULTS: Confocal microscopy indicated that the KD variant had higher levels of co-localization with ER than the wild-type HBsAg. The KD variant upregulated ER stress-related proteins and induced reactive oxygen species(ROS) compared to the wildtype via an increase in calcium. The KD variant also down-regulated anti-oxidant proteins(HO-1, catalase and SOD) compared to the wild-type, which indicates positive amplification loops of the ER-ROS axis. The KD variant also induced apoptotic cell death via the upregulation of caspase proteins(caspase 6, 9 and 12).Furthermore, the KD variant induced a higher level of nitric oxide than wild-type HBsAg via the up-regulation of the iNOS protein.CONCLUSION: Our data indicate that occult infection related HBsAg variants can lead to ER-derived oxidative stress and liver cell death in HuH-7 cells.展开更多
High-grade colonic neuroendocrine carcinomas (NECs) are uncommon but extremely aggressive. Their co-existence with tubular adenoma (TA) has rarely been reported. We present a 68-year-old man who was found on routine c...High-grade colonic neuroendocrine carcinomas (NECs) are uncommon but extremely aggressive. Their co-existence with tubular adenoma (TA) has rarely been reported. We present a 68-year-old man who was found on routine colonoscopy to have multiple colorectal TAs and an ulcerated lesion in the ascending colon. Microscopically, a poorly-differentiated invasive carcinoma juxtaposed with a TA was identified. Differential diagnosis included a poorly-differentiated adenocarcinoma, medullary carcinoma, high-grade NEC and lymphoma. The immunohistochemical profile showed positive staining for keratins, synaptophysin and chromogranin but negative for LCA, CDX2, CK7, CK20, TTF-1 and PSA, supporting the NEC diagnosis. Upon subsequent laparoscopic right hemicolectomy, the tumor was identified as a 3.0 cm umbilicated and ulcerated mass with an adjacent TA. Both TA and NEC showed positive staining for β-catenin indicating a shared colonic origin. The mitotic counts (77/10 high power fields) and a high proliferation rate (75% by Ki-67) corroborated a high-grade stratification. Mutational analysis indicated a wild-type BRAF and KRAS with mismatch repair proficiency. The AJCC (7<sup>th</sup> edition) pathologic stage is pT3, pN0, pMx. The patient received adjuvant chemotherapy with cisplatin/etoposides for three cycles and will be followed up for a year to detect recurrence. In conclusion, the co-existence of TA with high grade-NEC in our case allowed early identification and intervention of the otherwise asymptomatic but aggressive tumor. In addition, the finding of a high-grade NEC within a large TA in this case suggests a link between the two lesions and could represent a shared stem cell origin.展开更多
Mitochondrial division inhibitor 1（Mdivi-1） is a selective cell-permeable inhibitor of dynamin-related protein-1（Drp1） and mitochondrial division.To investigate the effect of Mdivi-1 on cells treated with glutamat...Mitochondrial division inhibitor 1（Mdivi-1） is a selective cell-permeable inhibitor of dynamin-related protein-1（Drp1） and mitochondrial division.To investigate the effect of Mdivi-1 on cells treated with glutamate,cerebral cortex neurons isolated from neonatal rats were treated with 10 m M glutamate for 24 hours.Normal cultured cells and dimethyl sulfoxide-cultured cells were considered as controls.Apoptotic cells were detected by flow cytometry.Changes in mitochondrial morphology were examined by electron microscopy.Drp1,Bax,and casp ase-3 expression was evaluated by western blot assays and immunocytochemistry.Mitochondrial membrane potential was detected using the JC-1 probe.Twenty-four hours after 10 m M glutamate treatment,Drp1,Bax and caspase-3 expression was upregulated,Drp1 and Bax were translocated to mitochondria,mitochondrial membrane potential was decreased and the rate of apoptosis was increased.These effects were inhibited by treatment with 50 μM Mdivi-1 for 2 hours.This finding indicates that Mdivi-1 is a candidate neuroprotective drug that can potentially mitigate against neuronal injury caused by glutamate-induced excitotoxicity.展开更多
Adolescent alcohol abuse is a substantive public health problem that has been the subject of intensive study in recent years.Despite reports of a wide range of effects of adolescent intermittent ethanol(AIE)exposure o...Adolescent alcohol abuse is a substantive public health problem that has been the subject of intensive study in recent years.Despite reports of a wide range of effects of adolescent intermittent ethanol(AIE)exposure on brain and behavior,little is known about the mechanisms that may underlie those effects,and even less about treatments that might reverse them.Recent studies from our laboratory have indicated that AIE produced enduring changes in astrocyte function and synaptic activity in the hippocampal formation,suggesting the possibility of an alteration in astrocyte-neuronal connectivity and function.We utilized astrocyte-specific,membrane restricted viral labeling paired with immunohistochemistry to perform confocal single cell astrocyte imaging,three-dimensional reconstruction,and quantification of astrocyte morphology in hippocampal area CA1 from adult rats after AIE.Additionally,we assessed the colocalization of astrocyte plasma membrane labeling with immunoreactivity for AMPA-(α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid)glutamate receptor 1,an AMPA receptor subunit and established neuronal marker of excitatory synapses,as a metric of astrocyte-synapse proximity.AIE significantly reduced the colocalization of the astrocyte plasma membrane with synaptic marker puncta in adulthood.This is striking in that it suggests not only an alteration of the physical association of astrocytes with synapses by AIE,but one that lasts into adulthood-well after the termination of alcohol exposure.Perhaps even more notable,the AIE-induced reduction of astrocyte-synapse interaction was reversed by sub-chronic treatment with the clinically used agent,gabapentin(Neurontin),in adulthood.This suggests that a medication in common clinical use may have the potential to reverse some of the enduring effects of adolescent alcohol exposure on brain function.All animal experiments conducted were approved by the Duke University Institutional Animal Care and Use Committee(Protocol Registry Number A159-18-07)on July 27,2018.展开更多
Rabies virus(RABV) is a highly neurotropic virus that follows clathrin-mediated endocytosis and p H-dependent pathway for trafficking and invasion into endothelial cells. Early(Rab5, EEA1) and late(Rab7, LAMP1) endoso...Rabies virus(RABV) is a highly neurotropic virus that follows clathrin-mediated endocytosis and p H-dependent pathway for trafficking and invasion into endothelial cells. Early(Rab5, EEA1) and late(Rab7, LAMP1) endosomal proteins play critical roles in endosomal sorting, maturity and targeting various molecular cargoes, but their precise functions in the early stage of RABV neuronal infection remain elusive. In this study, the relationship between enigmatic entry of RABV with these endosomal proteins into neuronal and SH-SY5 Y cells was investigated.Immunofluorescence, TCID_(50) titers, electron microscopy and western blotting were carried out to determine the molecular interaction of the nucleoprotein(N) of RABV with early or late endosomal proteins in these cell lines. The expression of N was also determined by down-regulating Rab5 and Rab7 in both cell lines through RNA interference. The results were indicative that N proficiently colocalized with Rab5/EEA1 and Rab7/LAMP1 in both cell lines at 24 and 48 h post-infection, while N titers significantly decreased in early infection of RABV. Down-regulation of Rab5 and Rab7 did not inhibit N expression, but it prevented productive infection via blocking the normal trafficking of RABV in a low pH environment. Ultrathin sections of cells studied by electron microscope also verified the close association of RABV with Rab5 and Rab7 in neurons. From the data it was concluded that primary entry of RABV strongly correlates with the kinetics of Rab-proteins present on early and late vesicles, which provides helpful clues to explain the early events of RABV in nerve cells.展开更多
Summary:Interleukin 17A(IL 17A)is reported to be involved in many inflammatory processes,but its role in aortic valve diseases remains unknown.We examined the role of IL17A based on an ApoE^-/-mouse model with strateg...Summary:Interleukin 17A(IL 17A)is reported to be involved in many inflammatory processes,but its role in aortic valve diseases remains unknown.We examined the role of IL17A based on an ApoE^-/-mouse model with strategies as fed with high-fat diet or treated with ILI7A monoclonal antibody(mAb).12 weeks of high-fat diet feeding can elevate cytokines secretion,inflammatory cells infiltration and myofibroblastic transition of valvular interstitial cells(VICs)in aortic valve.Moreover,diet-induction accelerated interleukin 17 receptor A(IL17RA)activation in VICs.In an IL17A inhibition model,the treatment group was intra-peritoneally injected with anti-IL17A mAb while controls received irrelevant antibody.Functional blockade of IL17A markedly reduced cellular infiltration and transition in aortic valve.To investigate potential mechanisms,NF-kB was co-stained in IL17RA^+VICs and IL17RA macrophages,and further confirmed by Western blotting in VICs.High-fat diet could activate NF-kB nuclear translocation in IL17RA^+VICs and IL17RA^+macrophages and this process was depressed after IL17A mAb-treatment.In conclusion,high-fat diet can lead to IL17A upregulation,VICs myofibroblastic transition and inflammatory cells infiltration in the aortic value of ApoE^-/-mice.Blocking IL17A with IL17A mAb can alleviate aortic valve inflammatory states.展开更多
The development of long-term imaging agents and subcellular imaging materials is of great importance in the research of cancer cell behaviors. In this work, a cationic poly（p-phenylenevinylene） derivative（PPV） is ...The development of long-term imaging agents and subcellular imaging materials is of great importance in the research of cancer cell behaviors. In this work, a cationic poly（p-phenylenevinylene） derivative（PPV） is designed and synthesized to link quaternized N-methyl-imidazole groups as pendants which endow the polymer to bear positive charges. Absorption and fluorescence emission spectra of PPV display a large Stokes shift of 102 nm which is much larger than the commercial cell dyes. Positively charged polymer could adsorb onto the surface of cells via electrostatic interactions followed by cell endocytosis process to enter cells. Importantly, PPV barely has influence on the cell viability through cytotoxicity analysis. The colocalization data demonstrates that PPV and commercial lysosome-specific dye are highly colocalized in the same region, indicating that the green fluorescent PPV mainly distributes in the lysosomes. Moreover, the continuous imaging investigation shows that PPV could stay in cells for more than seven days while the commercial Lyso-Tracker would be extruded by cells after three days. PPV exhibits superior capabilities including strong fluorescence, large Stokes shift, good biocompatibility and high photostablity, which has great potential in the applications of cellular process monitoring.展开更多
基金the Scientific Research Innovation Capability Support Project for Young Faculty(ZYGXQNJSKYCXNLZCXM-H8)Fundamental Research Funds for the Central Universities(2024ZYGXZR077)+3 种基金Guangdong Basic and Applied Basic Research Foundation(2023B1515120006)Guangzhou Basic and Applied Basic Research Foundation(2024A04J5776)the Research Fund(2023QN10Y421)Guangzhou Talent Recruitment Team Program(2024D03J0004),all related to this study.
文摘Osteoporosis is a known risk factor for rotator cuff tears(RCTs),but the causal correlation and underlying mechanisms remain unclear.This study aims to evaluate the impact of osteoporosis on RCT risk and investigate their genetic associations.Using data from the UK Biobank(n=457871),cross-sectional analyses demonstrated that osteoporosis was significantly associated with an increased risk of RCTs(adjusted OR[95%CI]=1.38[1.25–1.52]).A longitudinal analysis of a subset of patients(n=268117)over 11 years revealed that osteoporosis increased the risk of RCTs(adjusted HR[95%CI]=1.56[1.29–1.87]),which is notably varied between sexes in sex-stratified analysis.Causal inference methods,including propensity score matching,inverse probability weighting,causal random forest and survival random forest models further confirmed the causal effect,both from cross-sectional and longitudinal perspectives.
基金funded by the Hainan Province Clinical Medical Center(82171084)the National Natural Science Foundation of China(82371086).
文摘Background:Diabetic retinopathy(DR)urgently needs novel and effective therapeutic targets.Integrated analyses of plasma proteomic and genetic markers can clarify the causal relevance of proteins and discover novel targets for diseases,but no systematic screening for DR has been performed.Methods:Summary statistics of plasma protein quantitative trait loci(pQTL)were derived from two extensive genome-wide analysis study(GWAS)datasets and one systematic review,with over 100 thousand participants covering thousands of plasma proteins.DR data were sourced from the largest FinnGen study,comprising 10,413 DR cases and 308,633 European controls.Genetic instrumental variables were identified using multiple filters.In the two-sample MR analysis,Wald ratio and inverse variance-weighted(IVW)MR were utilized to investigate the causality of plasma proteins with DR.Bidirectional MR,Bayesian Co-localization,and phenotype scanning were employed to test for potential reverse causality and confounding factors in the main MR analyses.By systemically searching druggable gene lists,the ChEMBL database,DrugBank,and Gene Ontology database,the druggability and relevant functional pathways of the identified proteins were systematically evaluated.Results:Genetically predicted levels of 24 proteins were significantly associated with DR risk at a false discovery rate<0.05 including 11 with positive associations and 13 with negative associations.For each standard deviation increase in plasm protein levels,the odds ratios(ORs)for DR varied from 0.51(95%CI:0.36-0.73;P=2.22×10-5)for tubulin polymerization-promoting protein family member 3(TPPP3)to 2.02(95%CI:1.44-2.83;P=5.01×10-5)for olfactomedin like 3(OLFML3).Bidirectional MR indicated there was no reverse causality that interfered with the results of the main MR analyses.Four proteins exhibited strong co-localization evidence(PH4≥0.8):cytoplasmic tRNA synthetase(WARS),acrosin binding protein(ACRBP),and intercellular adhesion molecule 1(ICAM1)were negatively associated with DR risk,while neurogenic locus notch homolog protein 2(NOTCH2)showed a positive association.No confounding factors were detected between pQTLs and DR according to the phenotypic scan.Drugability assessments highlighted 6 proteins already in drug development endeavor and 18 novel drug targets,with metalloproteinase inhibitor 3(TIMP)currently in phase I clinical trials for DR.GO analysis identified 18 of 24 plasma proteins enriching 22 pathways related to cell differentiation and proliferation regulation.Conclusions:Twenty-four promising drug targets for DR were identified,including four plasma proteins with particular co-localization evidence.These findings offer new insights into DR's etiology and therapeutic targeting,exemplifying the value of genomic and proteomic data in drug target discovery.
基金supported by The Medical Engineering Cross Research Funding of Shanghai Jiaotong University"Star of Jiaotong University"Program(24X010301595).
文摘Background Lung squamous cell carcinoma(LUSC)is a major subtype of non-small cell lung cancer with a high mortality rate.Identifying causal plasma proteins associated with LUSC could provide new insights into the pathophysiology of the disease and potential therapeutic targets.This study aimed to identify plasma proteins causally linked to LUSC risk using proteome-wide Mendelian randomization(MR)and colocalization analyses.Methods Proteome-wide MR analysis was conducted using data from the UK Biobank Pharma Proteomics Project and deCODE genetics.Summary-level data for LUSC were obtained from the ILCCO Consortium,the FinnGen study,and a separate GWAS study.A total of 1,046 shared protein quantitative trait loci(pQTLs)were analyzed.Sensitivity analyses included the HEIDI test for horizontal pleiotropy and colocalization analysis to validate the causal associations.Results MR analysis identified six plasma proteins associated with LUSC risk:HSPA1L,PCSK7,POLI,SPINK2,TCL1A,and VARS.HSPA1L(OR=0.47;95%CI:0.34–0.65;P=4.89×10^(–6)),SPINK2(OR=0.68;95%CI:0.58–0.80;P=3.17×10^(–6)),and VARS(OR=0.44;95%CI:0.31–0.63;P=5.94×10^(–6))were associated with a decreased risk of LUSC.Conversely,PCSK7(OR=1.37;95%CI:1.21–1.56;P=1.40×10^(–6)),POLI(OR=4.50;95%CI:2.25–9.00;P=2.13×10–5),and TCL1A(OR=1.72;95%CI:1.34–2.21;P=1.89×10–5)were associated with an increased risk.The SMR analysis and HEIDI test confirmed the robustness of these associations.HSPA1L,SPINK2,and VARS showed significant inverse associations,with strong colocalization evidence for TCL1A(PPH4=0.817).Conclusions This study identified six plasma proteins potentially causal for LUSC risk.HSPA1L,SPINK2,and VARS are associated with decreased risk,while PCSK7,POLI,and TCL1A are linked to increased risk.These findings provide new insights into LUSC pathogenesis and highlight potential targets for therapeutic intervention.
基金supported by grants from the Central Public-interest Scientific Institution Basal Research Fund(2020-YWF-YB-02)the Young Scientists Fund of the National Natural Science Foundation of China(32202652)+1 种基金China Agriculture Research System of MOF and MARA(CARS-37)the Science and Technology Project of Inner Mongolia Autonomous Region(2020GG0210).
文摘Background A detailed understanding of genetic variants that affect beef merit helps maximize the efficiency of breeding for improved production merit in beef cattle.To prioritize the putative variants and genes,we ran a com-prehensive genome-wide association studies(GWAS)analysis for 21 agronomic traits using imputed whole-genome variants in Simmental beef cattle.Then,we applied expression quantitative trait loci(eQTL)mapping between the genotype variants and transcriptome of three tissues(longissimus dorsi muscle,backfat,and liver)in 120 cattle.Results We identified 1,580 association signals for 21 beef agronomic traits using GWAS.We then illuminated 854,498 cis-eQTLs for 6,017 genes and 46,970 trans-eQTLs for 1,903 genes in three tissues and built a synergistic network by integrating transcriptomics with agronomic traits.These cis-eQTLs were preferentially close to the transcription start site and enriched in functional regulatory regions.We observed an average of 43.5%improvement in cis-eQTL discovery using multi-tissue eQTL mapping.Fine-mapping analysis revealed that 111,192,and 194 variants were most likely to be causative to regulate gene expression in backfat,liver,and muscle,respectively.The transcriptome-wide association studies identified 722 genes significantly associated with 11 agronomic traits.Via the colocalization and Mendelian randomization analyses,we found that eQTLs of several genes were associated with the GWAS signals of agronomic traits in three tissues,which included genes,such as NADSYN1,NDUFS3,LTF and KIFC2 in liver,GRAMD1C,TMTC2 and ZNF613 in backfat,as well as TIGAR,NDUFS3 and L3HYPDH in muscle that could serve as the candidate genes for economic traits.Conclusions The extensive atlas of GWAS,eQTL,fine-mapping,and transcriptome-wide association studies aid in the suggestion of potentially functional variants and genes in cattle agronomic traits and will be an invaluable source for genomics and breeding in beef cattle.
文摘Objective: To investigate the colocalization of Somatostatin (SOM ) and NADPH--diaphorase (NADPHd ) of the neurons in raphe nuclei innervating pharyngeal muscles in the rats. Methods: After PRV was injected into the pharyngeal muscles. PRV and SOM immunofluorescence double labeling procedure was completed at first,then proceeded NADPH -d histochemistry. Results: PRV and SOM double--labled cells were present mainly in the nucleus raphe magnus. but some PRV and SOM double labled neurons were found in the other raphe nuclei as well, such as nucleus raphe pallidus. nucleus raphe obsurus, median raphe nucleus and dorsal raphe nucleus.NADPH -d positive neurons were also observed in the raphe nuclei. PRV. SOM and NADPH-d triple labeling neurons were found in the nucleus raphe magnus. Conclusion: It is suggested that the colocalization of SOM and NADPH--d of the neurons in the raphe nuclei innervating pharyngeal muscles may play an important role in the coordination of the pharyngeal motility.
基金supported by grants from the National Key R&D Program of China(Nos.2022YFC3602002 and 2024YFF0507404)National High Level Hospital Clinical Research Funding(Nos.2022-NHLHCRF-LX-02-03 and 2023-NHLHCRF-YXHZ-ZRZD-06).
文摘To the Editor:Atopic dermatitis(AD)is a chronic inflammatory skin condition marked by recurrent eczematous lesions and intense pruritus,affecting approximately 15–20%of children and up to 10%of adults worldwide.[1]The presence of AD significantly impairs patient quality of life and imposes substantial economic burdens on society.In recent years,the potential link between AD and lymphoma has attracted considerable attention from researchers and clinicians.Lymphoma,a malignancy of the lymphatic system,poses significant health risks due to its high mortality potential.[2]However,observational studies investigating the potential correlation between AD and lymphoma have yielded inconsistent results.Consequently,the association between AD and lymphoma remains controversial and warrants further investigation.
基金supported by grants from The Sci-Tech Innovation 2030 Agenda(2023ZD04045)Guangxi Key Research and Development Program(AB241484033).
文摘Background Semen quality is one of the most important indicators of boar reproductive performance.In the past,boar breeding has mostly emphasized characteristics such as lean meat percentage,feed conversion efficiency,and growth rate,while overlooking the genetic improvement of reproductive traits.This study employs advanced multi-omics approaches,such as transcriptome-wide association studies(TWAS)and colocalization between genome-wide association studies(GWAS)and expression quantitative trait loci(eQTLs),to provide a comprehensive understanding of the genetic mechanisms governing semen quality traits in boars.Results Here,we collected 190,000 ejaculate records across 11 semen quality traits from 3,604 Duroc boars.The heritability of semen quality traits ranged from 0.095 to 0.343.Genetic correlations between semen quality traits varied from−0.802 to 0.661,and phenotypic correlations ranged from−0.833 to 0.776.Single-trait GWAS identified 19 independent variants,corresponding to 13 quantitative trait loci(QTLs).By integrating PigGTEx and FAANG resources,we combined TWAS and colocalization analyses to reveal genetic regulation of semen quality traits.Notably,both GWAS and colocalization analyses pinpointed the DCAF12 as a crucial gene associated with multiple semen quality traits.Additionally,the ZSCAN9 gene and the variant rs322211455 were found to significantly affect sperm motility(SPMOT),possibly through hypothalamic-pituitary-gonadal axis.PheWAS further highlighted an association between rs322211455 and sperm abnormality rate,demonstrating the crucial role of ZSCAN9 in male fertility.Conclusion This study reveals the genetic basis and regulatory mechanisms underlying semen quality traits in Duroc boars,identifying key candidate genes such as DCAF12 and ZSCAN9.These findings provide important insight into the genetic regulation of semen quality in boars.
基金supported by the National Natural Science Foundation of China(82002432 to J.W.,82302068 to M.Z.,and 32300568 to T.W.)the Natural Science Foundation of Shandong Province(ZR2024MH159 to Y.Z.,ZR2020QH179 to J.W.,ZR2022QH057 to M.Z.,and ZR2021QH005 to T.W.)the China Postdoctoral Science Foundation(2024M752006 to S.M.)。
文摘Although the spatial characteristics within the tumor microenvironment of lung adenocarcinoma(LUAD)have been identified,the mechanisms by which these factors promote LUAD progression and immune evasion remain unclear.Using spatial transcriptomics and single-cell RNA-sequencing data from multi-regional LUAD biopsies consisting of tumor core,tumor edge,and normal area,we sought to delineate the spatial heterogeneity and driving factors of cell colocalization.Two cancer cell sub-clusters(Cancer_c1 and Cancer_c2),associated with LUAD initiation and metastasis,respectively,exhibit distinct spatial distributions and immune cell colocalizations.In particular,Cancer_c1,enriched within the tumor core,could directly interact with B cells or indirectly recruit B cells through macrophages.Conversely,Cancer_c2 enriched within the tumor edge exhibits colocalization with CD8^(+)T cells.Collectively,our work elucidates the spatial distribution of cancer cell subtypes and their interaction with immune cells in the core and edge of LUAD,providing insights for developing therapeutic strategies for cancer intervention.
基金the Intramural Research Program(Grant No.1ZIACP010201)the Division of Cancer Epidemiology and Genetics Informatics Tool Challenge of NCI.
文摘Genome-wide association studies(GWAS)have identified thousands of genomic loci associated with complex diseases and traits,including cancer.The vast majority of common traitassociated variants identified via GWAS fall in non-coding regions of the genome,posing a challenge in elucidating the causal variants,genes,and mechanisms involved.Expression quantitative trait locus(eQTL)and other molecular QTL studies have been valuable resources in identifying candidate causal genes from GWAS loci through statistical colocalization methods.While QTL colocalization is becoming a standard analysis in post-GWAS investigation,an easy web tool for users to perform formal colocalization analyses with either user-provided or public GWAS and eQTL datasets has been lacking.Here,we present ezQTL,a web-based bioinformatic application to interactively visualize and analyze genetic association data such as GWAS loci and molecular QTLs under different linkage disequilibrium(LD)patterns(1000 Genomes Project,UK Biobank,or user-provided data).This application allows users to perform data quality control for variants matched between different datasets,LD visualization,and two-trait colocalization analyses using two state-of-the-art methodologies(eCAVIAR and HyPrColoc),including batch processing.ezQTL is a free and publicly available cross-platform web tool,which can be accessed online at https://analysistools.cancer.gov/ezqtl.
基金Supported by National Research Foundation of Korea grant funded by the Korea government(MEST),No.2013-005810
文摘AIM: To investigate the mechanism of endoplasmic reticulum(ER) stress induction by an occult infection related hepatitis B virus S surface antigen(HBsAg)variant.METHODS: We used an HBsAg variant with lower secretion capacity, which was a KD variant from a Korean subject who was occultly infected with the genotype C. We compared the expression profiles of ER stress-related proteins between HuH-7 cells transfected with HBsAg plasmids of a wild-type and a KD variant using Western blot.RESULTS: Confocal microscopy indicated that the KD variant had higher levels of co-localization with ER than the wild-type HBsAg. The KD variant upregulated ER stress-related proteins and induced reactive oxygen species(ROS) compared to the wildtype via an increase in calcium. The KD variant also down-regulated anti-oxidant proteins(HO-1, catalase and SOD) compared to the wild-type, which indicates positive amplification loops of the ER-ROS axis. The KD variant also induced apoptotic cell death via the upregulation of caspase proteins(caspase 6, 9 and 12).Furthermore, the KD variant induced a higher level of nitric oxide than wild-type HBsAg via the up-regulation of the iNOS protein.CONCLUSION: Our data indicate that occult infection related HBsAg variants can lead to ER-derived oxidative stress and liver cell death in HuH-7 cells.
文摘High-grade colonic neuroendocrine carcinomas (NECs) are uncommon but extremely aggressive. Their co-existence with tubular adenoma (TA) has rarely been reported. We present a 68-year-old man who was found on routine colonoscopy to have multiple colorectal TAs and an ulcerated lesion in the ascending colon. Microscopically, a poorly-differentiated invasive carcinoma juxtaposed with a TA was identified. Differential diagnosis included a poorly-differentiated adenocarcinoma, medullary carcinoma, high-grade NEC and lymphoma. The immunohistochemical profile showed positive staining for keratins, synaptophysin and chromogranin but negative for LCA, CDX2, CK7, CK20, TTF-1 and PSA, supporting the NEC diagnosis. Upon subsequent laparoscopic right hemicolectomy, the tumor was identified as a 3.0 cm umbilicated and ulcerated mass with an adjacent TA. Both TA and NEC showed positive staining for β-catenin indicating a shared colonic origin. The mitotic counts (77/10 high power fields) and a high proliferation rate (75% by Ki-67) corroborated a high-grade stratification. Mutational analysis indicated a wild-type BRAF and KRAS with mismatch repair proficiency. The AJCC (7<sup>th</sup> edition) pathologic stage is pT3, pN0, pMx. The patient received adjuvant chemotherapy with cisplatin/etoposides for three cycles and will be followed up for a year to detect recurrence. In conclusion, the co-existence of TA with high grade-NEC in our case allowed early identification and intervention of the otherwise asymptomatic but aggressive tumor. In addition, the finding of a high-grade NEC within a large TA in this case suggests a link between the two lesions and could represent a shared stem cell origin.
基金supported by the National Natural Science Foundation of China,No.81371967 and 81401807a grant from the 5th Phase of "Project 333"of Jiangsu Province of China,No.BRA2016512a grant from the Six Talent Peaks Project of Jiangsu Province of China,No.2014-WSN-012
文摘Mitochondrial division inhibitor 1（Mdivi-1） is a selective cell-permeable inhibitor of dynamin-related protein-1（Drp1） and mitochondrial division.To investigate the effect of Mdivi-1 on cells treated with glutamate,cerebral cortex neurons isolated from neonatal rats were treated with 10 m M glutamate for 24 hours.Normal cultured cells and dimethyl sulfoxide-cultured cells were considered as controls.Apoptotic cells were detected by flow cytometry.Changes in mitochondrial morphology were examined by electron microscopy.Drp1,Bax,and casp ase-3 expression was evaluated by western blot assays and immunocytochemistry.Mitochondrial membrane potential was detected using the JC-1 probe.Twenty-four hours after 10 m M glutamate treatment,Drp1,Bax and caspase-3 expression was upregulated,Drp1 and Bax were translocated to mitochondria,mitochondrial membrane potential was decreased and the rate of apoptosis was increased.These effects were inhibited by treatment with 50 μM Mdivi-1 for 2 hours.This finding indicates that Mdivi-1 is a candidate neuroprotective drug that can potentially mitigate against neuronal injury caused by glutamate-induced excitotoxicity.
基金supported by the National Institute on Alcohol Abuse and Alcoholism(NIAAA)Neurobiology of Adolescent Drinking In Adulthood(NADIA)Grant#2U01AA019925(to HSS)the National Institute on Alcohol Abuse and Alcoholism(NIAAA)R00AA022651(to TAW)the National Institute on Drug Abuse(NIDA)R01DA041455(to KJR)
文摘Adolescent alcohol abuse is a substantive public health problem that has been the subject of intensive study in recent years.Despite reports of a wide range of effects of adolescent intermittent ethanol(AIE)exposure on brain and behavior,little is known about the mechanisms that may underlie those effects,and even less about treatments that might reverse them.Recent studies from our laboratory have indicated that AIE produced enduring changes in astrocyte function and synaptic activity in the hippocampal formation,suggesting the possibility of an alteration in astrocyte-neuronal connectivity and function.We utilized astrocyte-specific,membrane restricted viral labeling paired with immunohistochemistry to perform confocal single cell astrocyte imaging,three-dimensional reconstruction,and quantification of astrocyte morphology in hippocampal area CA1 from adult rats after AIE.Additionally,we assessed the colocalization of astrocyte plasma membrane labeling with immunoreactivity for AMPA-(α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid)glutamate receptor 1,an AMPA receptor subunit and established neuronal marker of excitatory synapses,as a metric of astrocyte-synapse proximity.AIE significantly reduced the colocalization of the astrocyte plasma membrane with synaptic marker puncta in adulthood.This is striking in that it suggests not only an alteration of the physical association of astrocytes with synapses by AIE,but one that lasts into adulthood-well after the termination of alcohol exposure.Perhaps even more notable,the AIE-induced reduction of astrocyte-synapse interaction was reversed by sub-chronic treatment with the clinically used agent,gabapentin(Neurontin),in adulthood.This suggests that a medication in common clinical use may have the potential to reverse some of the enduring effects of adolescent alcohol exposure on brain function.All animal experiments conducted were approved by the Duke University Institutional Animal Care and Use Committee(Protocol Registry Number A159-18-07)on July 27,2018.
基金supported by the National Key Research and Development Program of China(Grant No.216YFD0500402)Natural Science Foundation of China(Grants No.31272579 and 31472208)
文摘Rabies virus(RABV) is a highly neurotropic virus that follows clathrin-mediated endocytosis and p H-dependent pathway for trafficking and invasion into endothelial cells. Early(Rab5, EEA1) and late(Rab7, LAMP1) endosomal proteins play critical roles in endosomal sorting, maturity and targeting various molecular cargoes, but their precise functions in the early stage of RABV neuronal infection remain elusive. In this study, the relationship between enigmatic entry of RABV with these endosomal proteins into neuronal and SH-SY5 Y cells was investigated.Immunofluorescence, TCID_(50) titers, electron microscopy and western blotting were carried out to determine the molecular interaction of the nucleoprotein(N) of RABV with early or late endosomal proteins in these cell lines. The expression of N was also determined by down-regulating Rab5 and Rab7 in both cell lines through RNA interference. The results were indicative that N proficiently colocalized with Rab5/EEA1 and Rab7/LAMP1 in both cell lines at 24 and 48 h post-infection, while N titers significantly decreased in early infection of RABV. Down-regulation of Rab5 and Rab7 did not inhibit N expression, but it prevented productive infection via blocking the normal trafficking of RABV in a low pH environment. Ultrathin sections of cells studied by electron microscope also verified the close association of RABV with Rab5 and Rab7 in neurons. From the data it was concluded that primary entry of RABV strongly correlates with the kinetics of Rab-proteins present on early and late vesicles, which provides helpful clues to explain the early events of RABV in nerve cells.
基金This project was supported by grants from the National Key Research and Development Program of China(No.2016YFA0101100)National Natural Science Foundation of China(No.81700339 and No.31330029)Scientific Research Training Program for Young Talents sponsored by Union Hospital,Tongji Medical College,Huazhong University of Science and Technology。
文摘Summary:Interleukin 17A(IL 17A)is reported to be involved in many inflammatory processes,but its role in aortic valve diseases remains unknown.We examined the role of IL17A based on an ApoE^-/-mouse model with strategies as fed with high-fat diet or treated with ILI7A monoclonal antibody(mAb).12 weeks of high-fat diet feeding can elevate cytokines secretion,inflammatory cells infiltration and myofibroblastic transition of valvular interstitial cells(VICs)in aortic valve.Moreover,diet-induction accelerated interleukin 17 receptor A(IL17RA)activation in VICs.In an IL17A inhibition model,the treatment group was intra-peritoneally injected with anti-IL17A mAb while controls received irrelevant antibody.Functional blockade of IL17A markedly reduced cellular infiltration and transition in aortic valve.To investigate potential mechanisms,NF-kB was co-stained in IL17RA^+VICs and IL17RA macrophages,and further confirmed by Western blotting in VICs.High-fat diet could activate NF-kB nuclear translocation in IL17RA^+VICs and IL17RA^+macrophages and this process was depressed after IL17A mAb-treatment.In conclusion,high-fat diet can lead to IL17A upregulation,VICs myofibroblastic transition and inflammatory cells infiltration in the aortic value of ApoE^-/-mice.Blocking IL17A with IL17A mAb can alleviate aortic valve inflammatory states.
基金the National Natural Science Foundation of China(Nos.21504002,21401008)Beijing National Laboratory for Molecular Sciences
文摘The development of long-term imaging agents and subcellular imaging materials is of great importance in the research of cancer cell behaviors. In this work, a cationic poly（p-phenylenevinylene） derivative（PPV） is designed and synthesized to link quaternized N-methyl-imidazole groups as pendants which endow the polymer to bear positive charges. Absorption and fluorescence emission spectra of PPV display a large Stokes shift of 102 nm which is much larger than the commercial cell dyes. Positively charged polymer could adsorb onto the surface of cells via electrostatic interactions followed by cell endocytosis process to enter cells. Importantly, PPV barely has influence on the cell viability through cytotoxicity analysis. The colocalization data demonstrates that PPV and commercial lysosome-specific dye are highly colocalized in the same region, indicating that the green fluorescent PPV mainly distributes in the lysosomes. Moreover, the continuous imaging investigation shows that PPV could stay in cells for more than seven days while the commercial Lyso-Tracker would be extruded by cells after three days. PPV exhibits superior capabilities including strong fluorescence, large Stokes shift, good biocompatibility and high photostablity, which has great potential in the applications of cellular process monitoring.
