Topographical properties,such as pattern and diameter,of biomaterials play important roles in influencing cell activities and manipulating the related immune response during wound healing.We prepared aligned electrosp...Topographical properties,such as pattern and diameter,of biomaterials play important roles in influencing cell activities and manipulating the related immune response during wound healing.We prepared aligned electrospinning membranes with different fiber diameters,including 319±100 nm(A300),588±132 nm(A600),and 1048±130 nm(A1000),by adjusting the distance from the tip to the collector,the injection rate,and the concentration of the solution.The A300 membranes significantly improved cell proliferation and spreading and facilitated wound healing(epithelization and vascularization)with the regeneration of immature hair follicles compared to the other membranes.Transcriptomics revealed the underlying molecular mechanism that A300 could promote immune-related processes towards a pro-healing direction,significantly promoting keratinocyte migration and skin wound healing.All the results indicated that wound healing requires the active participation of the immune process,and that A300 was a potential candidate for guided skin regeneration applications.展开更多
Dear Editor,We present this case report which discusses a patient who underwent flap amputation after repeated attempts to resolve persistent and severe epithelial ingrowth following laser-assisted in situ keratomileu...Dear Editor,We present this case report which discusses a patient who underwent flap amputation after repeated attempts to resolve persistent and severe epithelial ingrowth following laser-assisted in situ keratomileusis(LASIK)surgery.Epithelial ingrowth is a known complication of LASIK surgery,typically manageable with minimal measures.However,severe cases may necessitate more aggressive interventions,such as flap amputation[1].LASIK is a widely performed refractive surgery with high success rates and excellent visual outcome[2].展开更多
A recently published prospective study marks a breakthrough for congenital olfactory disorders in children.The study provides the first long-term,three-year follow-up data,robustly demonstrating the durable efficacy a...A recently published prospective study marks a breakthrough for congenital olfactory disorders in children.The study provides the first long-term,three-year follow-up data,robustly demonstrating the durable efficacy and safety of autologous nasal epithelial stem cell transplantation.This work reveals immense therapeutic potential for a condition traditionally considered untreatable.However,this milestone achievement also presents new challenges.To translate this pioneering therapy from a single-center success to a global standard,multicenter,controlled clinical trials must be initiated immediately.Only through rigorous validation can we ensure its widespread adoption and ultimately bring hope to millions of children worldwide.展开更多
AIM:To investigate whether vaccinia-related kinase 1(VRK1)mediates transforming growth factor-beta2(TGF-β2)-caused epithelial-mesenchymal transition(EMT)and inflammatory responses in retinal pigment epithelial(RPE)ce...AIM:To investigate whether vaccinia-related kinase 1(VRK1)mediates transforming growth factor-beta2(TGF-β2)-caused epithelial-mesenchymal transition(EMT)and inflammatory responses in retinal pigment epithelial(RPE)cells through regulating snail family transcriptional repressor 1(SNAI1),and to validate its role in a proliferative vitreoretinopathy(PVR)mouse model.METHODS:Human RPE cell line ARPE-19 cells were treated with TGF-β2 to construct an EMT model.Western blot detected VRK1 level.The effects of VRK1 on SNAI1 expression and biological behavior of ARPE-19 cells were detected by immunofluorescence,ELISA,Transwell,and scratch assay,and the interaction between VRK1 and SNAI1 was confirmed through immunoprecipitation.A PVR mouse model was constructed,and the effects of VRK1 or/and SNAI1 on retinal damage were assessed by pathologic staining.Inflammatory factors and EMT-related proteins were assessed with ELISA and Western blot.RESULTS:VRK1 was upregulated in ARPE-19 cells after TGF-β2 treatment.Overexpression of VRK1 increased cell viability,promoted cell migration and EMT,and the levels of inflammatory factors.Silencing of VRK1 reversed the above indexes.There was a direct interaction between VRK1 and SNAI1,and overexpresssion SNAI1 weakened the impacts of silencing of VRK1.In PVR mice,silencing of VRK1 ameliorated retinal structural damage,decreased proinflammatory factor levels,and suppressed SNAI1 and mesenchymal marker expression.SNAI1 overexpression antagonized the protective effects of silencing VRK1 and exacerbated EMT and inflammatory responses.CONCLUSION:VRK1 plays a key role in retinal structural and inflammatory damage in PVR mice by regulating SNAI1 and mediating TGF-β2-caused EMT and inflammatory responses in RPE cells.展开更多
AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)...AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.展开更多
Background:EMP2 is a tumor-associated membrane protein belonging to the GAS-3/PMP22 gene family.EMP2 expression demonstrates significant tissue specificity and heterogeneity in various human tissues and tumor tissues,...Background:EMP2 is a tumor-associated membrane protein belonging to the GAS-3/PMP22 gene family.EMP2 expression demonstrates significant tissue specificity and heterogeneity in various human tissues and tumor tissues,where it may play a role in either promoting or inhibiting tumor growth.This study aimed to investigate the expression level,biological functions,and molecular mechanisms of EMP2 in liver cancer.Methods:we analyzed the mRNA expression levels of EMPs family genes in hepatocellular carcinoma(HCC)tissues and normal liver tissues based on the TCGA database and immunohistochemical analysis of tissue microarrays.Subsequently,we constructed HCC cell lines with either knockdown or overexpression of EMP2 to examine the biological functions and molecular mechanisms of EMP2 in tumorigenesis in vivo and in vitro.Results:Bioinformatic and immunohistochemical analysis of tissue microarrays have confirmed the significant upregulation of EMP2 in HCC tissues.In vitro and in vivo studies have shown that downregulation of EMP2 results in a moderate reduction in the proliferation and invasive capacity of HCC cells.Conversely,overexpression of EMP2 enhances the invasive capacity of HCC cells and induces autophagy.Initial investigations into the molecular mechanisms underlying EMP2-mediated enhancement of HCC cell invasion have revealed the dual regulation of EMP2-induced autophagy and the integrin pathway,which synergistically influence the invasive and metastatic potential of HCC cells.Conclusion:EMP2 holds promise as a diagnostic marker for HCC metastasis and a potential target for targeted therapy.展开更多
Objectives:Apatinib has been reported to be a promising treatment for sorafenib-resistant hepatocellular carcinoma(HCC)patients.However,the underlying mechanism remains ambiguous.The study aimed to explore the efficac...Objectives:Apatinib has been reported to be a promising treatment for sorafenib-resistant hepatocellular carcinoma(HCC)patients.However,the underlying mechanism remains ambiguous.The study aimed to explore the efficacy of apatinib in sorafenib-resistant HCC and the underlying mechanism both in vitro and in vivo.Methods:After observing epithelial-mesenchymal transformation(EMT)changes in HepG2 and HepG2/Sorafenib cells,we treated them with varying concentrations of apatinib to assess its impact on sorafenib-resistant HCC.Subsequently,specific inhibitors of c-Jun N-terminal kinase(JNK,SP600125)and extracellular signal-regulated kinase(ERK,PD98059)were introduced to investigate whether apatinib influenced sorafenib-resistant HCC via modulation of the epidermal growth factor receptor(EGFR)/JNK/ERK signaling pathway in vitro and in vivo.Biological behavior changes were assessed through cell counting kit-8(CCK-8),colony formation,transwell,and immunofluorescence tests.Simultaneously,Western blot analysis was conducted to elucidate the expression of proteins associated with EMT and the EGFR/JNK/ERK signaling pathway.Results:The HepG2/Sorafenib cells exhibited greater resistance to sorafenib compared to HepG2 cells,and sorafenib-resistant HCC was characterized by EMT changes.Apatinib demonstrated concentration-dependent inhibition of biological behaviors in HepG2/Sorafenib cells,with minimal impact on HepG2 cells.Additionally,apatinib had a pronounced effect on the expression of EMT-related proteins in sorafenib-resistant cells similar to that in sorafenib-sensitive cells.Furthermore,there was a dose-dependent reduction in the expression of proteins associated with the EGFR/JNK/ERK pathway in apatinib-treated groups.Notably,SP600125 and PD98059 contributed to the inhibition of EMT and EGFR/JNK/ERK pathway-related proteins by apatinib in sorafenib-resistant HCC.Conclusion:Apatinib potentially hindered the progression of sorafenib-resistant HCC by suppressing both EMT and the EGFR/JNK/ERK pathway.展开更多
Epithelial-mesenchymal transition(EMT)plays an irreplaceable role in the development of silicosis.However,molecular mechanisms of EMT induced by silica exposure still remain to be addressed.Herein,metabolic profiles o...Epithelial-mesenchymal transition(EMT)plays an irreplaceable role in the development of silicosis.However,molecular mechanisms of EMT induced by silica exposure still remain to be addressed.Herein,metabolic profiles of human alveolar type II epithelial cells(A549 cells)exposed directly to silica were characterized using non-targeted metabolomic approaches.A total of 84 differential metabolites(DMs)were identified in silica-treated A549 cells undergoing EMT,which were mainly enriched in metabolisms of amino acids(e.g.,glutamate,alanine,aspartate),purine metabolism,glycolysis,etc.The number of DMs identified in the A549 cells obviously increased with the elevated exposure concentration of silica.Remarkably,glutamine catabolism was significantly promoted in the silica-treated A549 cells,and the levels of related metabolites(e.g.,succinate)and enzymes(e.g.,α-ketoglutarate(α-KG)dehydrogenase)were substantially up-regulated,with a preference toα-KG pathway.Supplementation of glutamine into the cell culture could substantially enhance the expression levels of both EMT-related markers and Snail(zinc finger transcription factor).Our results suggest that the EMT of human alveolar epithelial cells directly induced by silica can be essential to the development of silicosis.展开更多
Background:Dry eye disease(DED)predominantly results from elevated tear film os-molarity,which can not only cause ocular inconvenience but may lead to visual impair-ments,severely compromising patient well-being and e...Background:Dry eye disease(DED)predominantly results from elevated tear film os-molarity,which can not only cause ocular inconvenience but may lead to visual impair-ments,severely compromising patient well-being and exerting substantial economic burdens as well.Astaxanthin(AST),a member of the xanthophylls and recognized for its robust abilities to combat inflammation and oxidation,is a common dietary sup-plement.Nonetheless,the precise molecular pathways through which AST influences DED are still poorly understood.Methods:Therapeutic targets for AST were identified using data from the GeneCards,PharmMapper,and Swiss Target Prediction databases,and STITCH datasets.Similarly,targets for dry eye disease(DED)were delineated leveraging resources such as the Therapeutic Target Database(TTD),DisGeNET,GeneCards,and OMIM databases,and DrugBank datasets.Interactions among shared targets were charted and dis-played using CytoScape 3.9.0.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses were conducted to elucidate the functions of pivotal tar-gets within the protein-protein interaction network.Molecular interactions between AST and key targets were confirmed through molecular docking using AutoDock and PyMOL.Molecular dynamics simulations were performed using GROMACS 2022.3.Viability of human corneal epithelial cells(hCEC)was assessed across varying concen-trations of AST.A mouse model of experimental DED was developed using 0.1%ben-zalkonium chloride(BAC),and the animals were administered 100 mg/kg/day of AST orally for 7 days.The efficacy of the treatments was assessed through a series of di-agnostic tests to evaluate the condition of the ocular surface after the interventions.The levels of inflammation and oxidative stress were quantitatively assessed using methods such as reverse transcription-polymerase chain reaction(RT-PCR),Western blot,and immunofluorescence staining.Results:Network pharmacology suggests that AST may alleviate DED by influenc-ing oxidation-reduction signaling pathways and reducing oxidative stress provoked by BAC.In vivo experiments demonstrated an improved overall condition in AST-administered mice in contrast to the control group.Immunofluorescence staining analyses indicated a decrease in Keap1 protein in the corneal tissues of AST-treated mice and a significant increase in Nrf2 and HO-1 protein.In vitro studies demon-strated that AST significantly enhanced cell viability and suppressed reactive oxy-gen species expression under hyperosmotic(HS)conditions,thereby protecting the human corneal epithelium.Conclusion:AST is capable of shielding mice from BAC-induced DED,decelerating the progression of DED,and mitigating oxidative stress damage under HS conditions in hCEC cells.The protective impact of AST on DED may operate through stimulating the Keap1-Nrf2/HO-1 signaling pathway.Our research findings indicate that AST may be a promising treatment for DED,offering new insights into DED treatment.展开更多
BACKGROUND Oral squamous cell carcinoma(OSCC)is a frequent cancer affecting the oral cavity.OSCC is usually preceded by occurrence of precursor lesions,which demonstrate a varying degree of malignant transformation po...BACKGROUND Oral squamous cell carcinoma(OSCC)is a frequent cancer affecting the oral cavity.OSCC is usually preceded by occurrence of precursor lesions,which demonstrate a varying degree of malignant transformation potential.Epithelialmesenchymal transition is a significant biological phenomenon that facilitates tumor growth and metastasis through the involvement of diverse epithelial and mesenchymal proteins.E-cadherin down regulation and over-expression of zincfinger E-box-binding homeobox 1(ZEB1)has been reported in several cancers.AIM To evaluate the role of E-cadherin and ZEB1 in oral leukoplakia and OSCC.METHODS A total of 60 cases i.e.,oral leukoplakia/oral epithelial dysplasia(OED)(n=30)and OSCC(n=30)were included.Immunohistochemistry was performed utilizing two markers:E-cadherin and ZEB1.The Mann-Whitney U test was employed to compare individual markers across the study groups.Spearman’s correlation was done followed by discriminant function analysis.RESULTS Reduction in the expression E-cadherin and altered localization were noted from OED to OSCC.Overexpression of ZEB1 along with cytoplasmic accumulation was noted in OED,with marked expression in OSCC.ZEB1 epithelium intensity and ZEB1 connective tissue percentage contributed to the discriminant function with an accuracy of 85%for the classification of OED from OSCC.CONCLUSION Loss of E-cadherin and up-regulation of ZEB1 from OED to OSCC,predisposes to induction of epithelial-mesenchymal transition.Discriminant formulas are developed to classify cases of OED and OSCC with 85%accuracy.展开更多
The dark blue pigment(DBP)is a health ingredient from Vaccinium bracteatum Thunb.leaves used as a functional food supplement.However,the details of transepithelial absorption on the intestinal epithelial cells are bar...The dark blue pigment(DBP)is a health ingredient from Vaccinium bracteatum Thunb.leaves used as a functional food supplement.However,the details of transepithelial absorption on the intestinal epithelial cells are barely understood.This study aimed to clarify the absorption properties of DBP in the Caco-2 cell monolayer model and evaluate the effect on the endo-metabolism and barrier function of Caco-2 cells.The results showed that the DBP did not show the dose-dependent toxic effect to Caco-2 cells between 0.25 and 1.5 mg/mL,which did not cause disorder in the normal cell metabolism and absorption activity.The Caco-2 cell monolayer model could absorb DBP by passive and active transport,and the absorptive pattern was dosedependent when the concentration was more than 0.25 mg/mL.During DBP absorption,an increase in m RNA and protein expressions of glucose transporters demonstrated that the glucose transporters were the potential transporter of DBP.But the glucose transport amounts were significantly lowered after 30 min of DBP treatment,indicating that DBP owned the inhibitory effect on glucose transportation.Furthermore,DBP also owned protective effects on the barrier function of intestinal epithelial cells.展开更多
Background Using yeast culture as additives in ruminant feed prevents rumen microbial dysbiosis,enhances performance,and regulates rumen pH.The yeast culture used in this study was developed in-house,and has been show...Background Using yeast culture as additives in ruminant feed prevents rumen microbial dysbiosis,enhances performance,and regulates rumen pH.The yeast culture used in this study was developed in-house,and has been shown to promote rumen epithelial growth in several sheep trials.Changes in protein expression associated with the promotion of rumen epithelial development following the addition of yeast culture,along with the associated molecular mechanisms,remain unknown.We used 2045-day-old weaned lambs to investigate the specific proteins and molecular mechanisms involved in these processes.Half of the lambs were fed yeast culture,and the other half were used as controls.Results Yeast culture enhanced growth performance,facilitated rumen fermentation,and promoted rumen papilla development in weaned lambs.Proteomics data identified 4,831 proteins in the rumen epithelial tissue of lambs,comprising 87 upregulated and 425 downregulated proteins.Administration of yeast culture activated multiple molecular functions within rumen epithelial cells,including oxidative phosphorylation,glutathione metabolism,apoptosis,cell cycle,and vitamin digestion and absorption.The expression of proteins associated with cell cycle regulation increased,whereas those associated with apoptosis decreased.Administration of yeast culture also reduced the duration of the G0/G1 phase of rumen epithelial cells and accelerated the cell cycle.Furthermore,yeast culture showed increased cyclin D1,cyclin-dependent kinase(CDK)2,CDK4,CDK6,and cyclin E1 expressions and decreased cytochrome C(Cyto-c),Bcl-2-related X protein(Bax),cleaved caspase 3(C-caspase 3),caspase 3,and cleaved caspase 7(C-caspase 7)protein expressions.Yeast culture upregulated the insulin-like growth factor-1 receptor(IGF-1R)and insulin-like growth factor-binding protein 5(IGFBP-5)mRNA expressions in rumen epithelial cells.Conclusions Yeast culture facilitates rumen epithelial development by regulating the cell cycle and IGF-1 signaling and reducing the expression of proteins associated with apoptosis in rumen epithelial cells.The findings of this study provide novel insights into the molecular mechanisms through which yeast culture promotes rumen epithelial development in weaned lambs.展开更多
Background Clostridium perfringens is a pathogen that secretes multiple toxins,impacting humans and animals.It can cause intestinal diseases such as necrotic enteritis.Although tannins inhibit C.perfringens proliferat...Background Clostridium perfringens is a pathogen that secretes multiple toxins,impacting humans and animals.It can cause intestinal diseases such as necrotic enteritis.Although tannins inhibit C.perfringens proliferation,the precise underlying mechanisms are unclear.Objective This study integrated transcriptomics and metabolomics to systematically investigate the mechanism by which tannins,specifically pentagalloylglucose(PGG)and tannic acid(TA),inhibit C.perfringens and potential pathways to alleviate infection in vivo.Results Ion concentration measurements,flow cytometric analysis,and transmission electron microscopy revealed that PGG and TA damaged the cell membrane structure of C.perfringens,triggering cytoplasmic content leakage.Additionally,PGG and TA significantly affected C.perfringens at the transcriptional and metabolic levels.Bioinformatics analysis revealed that PGG and TA induced amino acid restriction,disrupted energy metabolism,and impeded the ability of C.perfringens to sense and respond to the external environment.In an in vitro C.perfringens-infected intestinal cell model,PGG and TA boundαtoxin,significantly reduced the mRNA expression of inflammatory factors,and improved intestinal barrier function and cell viability.Compared to PGG,TA exhibited stronger inhibitory activity against C.perfringens and binding toαtoxin.In vivo,PGG and TA alleviated C.perfringens-induced weight loss in mice,improved intestinal villi morphology,and reduced intestinal inflammation and tight junction gene dysregulation.Conclusion These findings indicate that tannins inhibit C.perfringens,improve gut tissue integrity and reduce inflammation,demonstrating their multi-target effects of resisting intestinal diseases caused by harmful bacteria.This offers new insights for plant polyphenol-based strategies against necrotic enteritis.展开更多
A lactating dairy cow mammary epithelial cell(DCMEC)model is useful for studying milk synthesis and lactation in the dairy cow mammary gland.However,the cells can only maintain their original characteristics for a lim...A lactating dairy cow mammary epithelial cell(DCMEC)model is useful for studying milk synthesis and lactation in the dairy cow mammary gland.However,the cells can only maintain their original characteristics for a limited period in vitro.Thus,the proliferative capacity and lactation pattern of subcultured DCMECs need to be characterized.In the present study,subcultured DCMECs appeared to proliferate without changes in morphology or growth pattern up to the 12th passage.Subculturing had no obvious effect on the lactation capacity of the subcultured DCMEC up to the 10th passage in vitro.These results indicated that the DCMEC line could be subpassaged up to 10 times and still maintain its original milk synthesis characteristics.Moreover,DCMECs were frozen using different methods,and the method of placing the vials in the foam box was determined after cryopreservation with the best adherence of 91.2%.This method was recommended for long-term cryopreservation for DCMECs.展开更多
The cornea is a transparent tissue that serves as the main refractive element of the eye ball.Limbal epithelial stem cells(LESCs),residing in the basal epithelial layer of the Palisades of Vogt located in the corneal ...The cornea is a transparent tissue that serves as the main refractive element of the eye ball.Limbal epithelial stem cells(LESCs),residing in the basal epithelial layer of the Palisades of Vogt located in the corneal limbus between cornea and scleral,are believed to be crucial for the continuous turnover of the corneal epithelium.The proliferation,migration,and differentiation of the LESCs are modulated by unique physical and chemical futures contained within the microenvironment known as the limbal niche.This niche,composed of nerve terminals,cells,extracellular matrix,vasculature,and signaling molecules,is the home for processes such as proliferation,migration and differentiation.Corneal nerve terminals possess special anatomical structures in the limbal region and basal epithelial cells,and they demonstrate pivotal biological effects in the regulation of the LESC function and corneal epithelium homeostasis.Biological molecules such as neuropeptides,neurotransmitters,and neurotrophic factors play a crucial role in modulating the LESCs phenotype responsible for corneal epithelium homeostasis.This paper will review recent studies on how these nerve derived molecules function in this process and provide clear orientations for future research.展开更多
Adenoid cystic carcinoma of the lacrimal gland(LGACC)is the most common type of malignant epithelial tumor of the lacrimal gland,which is characterized by a high recurrence rate,perineural invasion,and a propensity to...Adenoid cystic carcinoma of the lacrimal gland(LGACC)is the most common type of malignant epithelial tumor of the lacrimal gland,which is characterized by a high recurrence rate,perineural invasion,and a propensity to metastasize to distant sites.Due to its unclear pathogenesis,LGACC has a poor prognosis and a high mortality rate.In recent years,a range of radiotherapy and chemotherapy have been clinically applied,leading to a shift in the treatment approach for LGACC.This article discussed the advances being made in the treatment of LGACC and provides readers with an overview of the impact of LGACC treatment modalities on patient survival and prognostic levels.展开更多
Background Mastitis caused by Staphylococcus aureus(S.aureus)is one of the most intractable problems for the dairy industry,causing significantly reduced milk yields and early slaughter of cows worldwide.MicroRNAs(miR...Background Mastitis caused by Staphylococcus aureus(S.aureus)is one of the most intractable problems for the dairy industry,causing significantly reduced milk yields and early slaughter of cows worldwide.MicroRNAs(miRNAs)can post-transcriptionally regulate gene expression and studies in recent years have shown the importance of miRNA-associated gene regulation in S.aureus-induced mastitis.Results In this study,to investigate the role of miR-223 in mastitis,we performed experiments to overexpress and suppress miR-223 in an immortalized bovine mammary epithelial cell line(MAC-T)infected with S.aureus.Overexpression of miR-223 in MAC-T cells repressed cell apoptosis and necrosis induced by S.aureus infection,whereas suppression of miR-223 had the opposite effect.Transcriptome expression profiling with weighted gene co-expression network analysis(WGCNA)and gene set variation analysis(GSVA)showed that miR-223 affects apoptosis and inflammation-related pathways.Furthermore,differentially expressed(DE)genes were evaluated,and genes exhibiting contrasting expression trends in the miR-223 overexpressed and suppressed groups were assessed as potential target genes of miR-223.Potential target genes,including CDC25B,PTPRF,DCTN1,and DPP9,were observed to be associated with apoptosis and necroptosis.Finally,through integrative analysis of genome-wide association study(GWAS)data and the animal quantitative trait loci(QTL)database,we determined that target genes of miR-223 were significantly enriched in single-nucleotide polymorphisms(SNP)and QTLs related to somatic cell count(SCC)and mastitis.Conclusion In summary,miR-223 has an inhibitory effect on S.aureus-induced cell apoptosis and necrosis by regulating PTPRF,DCTN1,and DPP9.These genes were significantly enriched in QTL regions associated with bovine mastitis resistance,underscoring their relevance in genetic regulation of disease resilience.Our findings provide critical genetic markers for enhancing mastitis resistance,particularly S.aureus-induced mastitis,through selective breeding.This work offers valuable insights for developing cattle with improved resistance to mastitis via targeted genetic selection.展开更多
The oviduct epithelium is the initial maternal contact site for embryos after fertilization,offering the microenviron-ment before implantation.This early gestation period is particularly sensitive to stress,which can ...The oviduct epithelium is the initial maternal contact site for embryos after fertilization,offering the microenviron-ment before implantation.This early gestation period is particularly sensitive to stress,which can cause reduced fertil-ity and reproductive disorders in mammals.Nevertheless,the local impact of elevated stress hormones on the ovi-duct epithelium has received limited attention to date,except for a few reports on polyovulatory species like mice and pigs.In this study,we focused on the effects of chronic maternal stress on cattle,given its association with infertil-ity issues in this monoovulatory species.Bovine oviduct epithelial cells(BOEC)differentiated at the air–liquid interface(ALI)were stimulated with 250 nmol/L cortisol for 1 or 3 weeks.Subsequently,they were assessed for morphology,bioelectrical properties,and gene expression related to oviduct function,glucocorticoid pathway,cortisol metabo-lism,inflammation,and apoptosis.Results revealed adverse effects of cortisol on epithelium structure,featured by deciliation,vacuole formation,and multilayering.Additionally,cortisol exposure led to an increase in transepithelial potential difference,downregulated mRNA expression of the major glucocorticoid receptor(NR3C1),upregulated the expression of cortisol-responsive genes(FKBP5,TSC22D3),and significant downregulation of oviductal glycopro-tein 1(OVGP1)and steroid receptors PGR and ESR1.The systematic comparison to a similar experiment previously performed by us in porcine oviduct epithelial cells,indicated that bovine cultures were more susceptible to elevated cortisol levels than porcine.The distinct responses between both species are likely linked to their divergence in the cortisol-induced expression changes of HSD11B2,an enzyme controlling the cellular capacity to metabolise cortisol.These findings provide insights into the species-specific reactions and reproductive consequences triggered by maternal stress.展开更多
Background Ferroptosis is characterized by increased production of reactive oxygen species(ROS)and membrane lipid peroxidation that can exacerbate inflammatory damage.Extracellular vesicles(EVs)isolated from bovine mi...Background Ferroptosis is characterized by increased production of reactive oxygen species(ROS)and membrane lipid peroxidation that can exacerbate inflammatory damage.Extracellular vesicles(EVs)isolated from bovine milk have many biological functions,including antioxidant properties.However,the role of EVs on Klebsiella pneumoniaeinduced ferroptosis and oxidative stress in bovine mammary epithelial cells(bMECs)and murine mammary tissue is unclear.In this study,EVs were isolated from bovine colostrum,mature milk and clinical mastitis milk(defined as C-EVs,M-EVs and CM-EVs,respectively)and assessed by transmission electron microscopy,Western blot and transcriptome sequencing.Effects of EVs on K.pneumoniae-induced ferroptosis and oxidative stress in bMECs were evaluated with immunofluorescence and Western blot.Results In bMECs,infection with K.pneumoniae induced oxidative stress,decreasing protein expression of Nrf2,Keap1 and HO-1 plus SOD activity,and increasing ROS concentrations.However,protein expression of GPX4,ACSL4 and S100A4 in bMECs,all factors that regulate ferroptosis,was downregulated by K.pneumoniae.Furthermore,this bacterium compromised tight junctions in murine mammary tissue,with low expression of ZO-1 and Occludin,whereas protein expression of Nrf2 and GPX4 was also decreased in mammary tissue.Adding C-EVs,M-EVs or CMEVs reduced oxidative stress and ferroptosis in K.pneumoniae-infected bMECs in vitro and murine mammary tissues in vivo.Conclusion In conclusion,all 3 sources of milk-derived EVs alleviated oxidative stress and ferroptosis in K.pneumoniae-infected bMECs and mammary tissues.展开更多
Pigeons and certain other avian species produce a milklike secretion in their crop sacs to nourish offspring,yet the detailed processes involved are not fully elucidated.This study investigated the crop sacs of 225-da...Pigeons and certain other avian species produce a milklike secretion in their crop sacs to nourish offspring,yet the detailed processes involved are not fully elucidated.This study investigated the crop sacs of 225-day-old unpaired non-lactating male pigeons(MN)and males initiating lactation on the first day after incubation(ML).Using RNA sequencing,ribosomeprofiling,andsingle-cell transcriptome sequencing(scRNA-seq),we identified a significant up-regulation of genes associated with ribosome assembly and protein synthesis in ML compared to MN.Results from scRNA-seq analysis identified 12distinct cell types and 22 clusters,with secretory epithelial cells(SECs)exhibiting marked expression of plasma cell markers,including IGLL1 and MZB1.RNA fluorescence in situ hybridization(RNA FISH)and IgY quantification confirmed the critical role of SECs in producing endogenous IgY during lactation.We propose that fibroblast-derived BAFF signals activate SECs,mimicking B cell transformation and enhancing protein production through the unfolded protein response(UPR).These findings shed light on the cellular dynamics of pigeon milk production and contribute to a broader understanding of avian biology.展开更多
基金supported by Key R&D Projects of Sichuan Science and Technology Plan(2021YFS0030)Interdisciplinary Innovation Project,West China Hospital of Stomatology Sichuan University(RD-03-202006)Research and Develop Program,West China Hospital of Stomatology Sichuan University(No.LCYJ2019-19).
文摘Topographical properties,such as pattern and diameter,of biomaterials play important roles in influencing cell activities and manipulating the related immune response during wound healing.We prepared aligned electrospinning membranes with different fiber diameters,including 319±100 nm(A300),588±132 nm(A600),and 1048±130 nm(A1000),by adjusting the distance from the tip to the collector,the injection rate,and the concentration of the solution.The A300 membranes significantly improved cell proliferation and spreading and facilitated wound healing(epithelization and vascularization)with the regeneration of immature hair follicles compared to the other membranes.Transcriptomics revealed the underlying molecular mechanism that A300 could promote immune-related processes towards a pro-healing direction,significantly promoting keratinocyte migration and skin wound healing.All the results indicated that wound healing requires the active participation of the immune process,and that A300 was a potential candidate for guided skin regeneration applications.
文摘Dear Editor,We present this case report which discusses a patient who underwent flap amputation after repeated attempts to resolve persistent and severe epithelial ingrowth following laser-assisted in situ keratomileusis(LASIK)surgery.Epithelial ingrowth is a known complication of LASIK surgery,typically manageable with minimal measures.However,severe cases may necessitate more aggressive interventions,such as flap amputation[1].LASIK is a widely performed refractive surgery with high success rates and excellent visual outcome[2].
文摘A recently published prospective study marks a breakthrough for congenital olfactory disorders in children.The study provides the first long-term,three-year follow-up data,robustly demonstrating the durable efficacy and safety of autologous nasal epithelial stem cell transplantation.This work reveals immense therapeutic potential for a condition traditionally considered untreatable.However,this milestone achievement also presents new challenges.To translate this pioneering therapy from a single-center success to a global standard,multicenter,controlled clinical trials must be initiated immediately.Only through rigorous validation can we ensure its widespread adoption and ultimately bring hope to millions of children worldwide.
文摘AIM:To investigate whether vaccinia-related kinase 1(VRK1)mediates transforming growth factor-beta2(TGF-β2)-caused epithelial-mesenchymal transition(EMT)and inflammatory responses in retinal pigment epithelial(RPE)cells through regulating snail family transcriptional repressor 1(SNAI1),and to validate its role in a proliferative vitreoretinopathy(PVR)mouse model.METHODS:Human RPE cell line ARPE-19 cells were treated with TGF-β2 to construct an EMT model.Western blot detected VRK1 level.The effects of VRK1 on SNAI1 expression and biological behavior of ARPE-19 cells were detected by immunofluorescence,ELISA,Transwell,and scratch assay,and the interaction between VRK1 and SNAI1 was confirmed through immunoprecipitation.A PVR mouse model was constructed,and the effects of VRK1 or/and SNAI1 on retinal damage were assessed by pathologic staining.Inflammatory factors and EMT-related proteins were assessed with ELISA and Western blot.RESULTS:VRK1 was upregulated in ARPE-19 cells after TGF-β2 treatment.Overexpression of VRK1 increased cell viability,promoted cell migration and EMT,and the levels of inflammatory factors.Silencing of VRK1 reversed the above indexes.There was a direct interaction between VRK1 and SNAI1,and overexpresssion SNAI1 weakened the impacts of silencing of VRK1.In PVR mice,silencing of VRK1 ameliorated retinal structural damage,decreased proinflammatory factor levels,and suppressed SNAI1 and mesenchymal marker expression.SNAI1 overexpression antagonized the protective effects of silencing VRK1 and exacerbated EMT and inflammatory responses.CONCLUSION:VRK1 plays a key role in retinal structural and inflammatory damage in PVR mice by regulating SNAI1 and mediating TGF-β2-caused EMT and inflammatory responses in RPE cells.
文摘AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.
基金supported by the Fundamental Research Funds for the National Natural Science Foundation of China(Nos.22177084 and 82104373)the Fundamental Research Funds of Science&Technology Department of Sichuan Province(No.2022YFQ0054).
文摘Background:EMP2 is a tumor-associated membrane protein belonging to the GAS-3/PMP22 gene family.EMP2 expression demonstrates significant tissue specificity and heterogeneity in various human tissues and tumor tissues,where it may play a role in either promoting or inhibiting tumor growth.This study aimed to investigate the expression level,biological functions,and molecular mechanisms of EMP2 in liver cancer.Methods:we analyzed the mRNA expression levels of EMPs family genes in hepatocellular carcinoma(HCC)tissues and normal liver tissues based on the TCGA database and immunohistochemical analysis of tissue microarrays.Subsequently,we constructed HCC cell lines with either knockdown or overexpression of EMP2 to examine the biological functions and molecular mechanisms of EMP2 in tumorigenesis in vivo and in vitro.Results:Bioinformatic and immunohistochemical analysis of tissue microarrays have confirmed the significant upregulation of EMP2 in HCC tissues.In vitro and in vivo studies have shown that downregulation of EMP2 results in a moderate reduction in the proliferation and invasive capacity of HCC cells.Conversely,overexpression of EMP2 enhances the invasive capacity of HCC cells and induces autophagy.Initial investigations into the molecular mechanisms underlying EMP2-mediated enhancement of HCC cell invasion have revealed the dual regulation of EMP2-induced autophagy and the integrin pathway,which synergistically influence the invasive and metastatic potential of HCC cells.Conclusion:EMP2 holds promise as a diagnostic marker for HCC metastasis and a potential target for targeted therapy.
基金supported by Natural Science Foundation of Shandong Province,No.ZR2021QH186National Natural Science Foundation of China,No.81870205+3 种基金Foundation Research Project of Qinghai Province,2021-ZJ-719National Natural Science Foundation of China,No.82000579Qinghai Provincial Health System Key Project No.2021-wjzd-06Foundation Research Project of Qinghai Province,2023-ZJ-786.
文摘Objectives:Apatinib has been reported to be a promising treatment for sorafenib-resistant hepatocellular carcinoma(HCC)patients.However,the underlying mechanism remains ambiguous.The study aimed to explore the efficacy of apatinib in sorafenib-resistant HCC and the underlying mechanism both in vitro and in vivo.Methods:After observing epithelial-mesenchymal transformation(EMT)changes in HepG2 and HepG2/Sorafenib cells,we treated them with varying concentrations of apatinib to assess its impact on sorafenib-resistant HCC.Subsequently,specific inhibitors of c-Jun N-terminal kinase(JNK,SP600125)and extracellular signal-regulated kinase(ERK,PD98059)were introduced to investigate whether apatinib influenced sorafenib-resistant HCC via modulation of the epidermal growth factor receptor(EGFR)/JNK/ERK signaling pathway in vitro and in vivo.Biological behavior changes were assessed through cell counting kit-8(CCK-8),colony formation,transwell,and immunofluorescence tests.Simultaneously,Western blot analysis was conducted to elucidate the expression of proteins associated with EMT and the EGFR/JNK/ERK signaling pathway.Results:The HepG2/Sorafenib cells exhibited greater resistance to sorafenib compared to HepG2 cells,and sorafenib-resistant HCC was characterized by EMT changes.Apatinib demonstrated concentration-dependent inhibition of biological behaviors in HepG2/Sorafenib cells,with minimal impact on HepG2 cells.Additionally,apatinib had a pronounced effect on the expression of EMT-related proteins in sorafenib-resistant cells similar to that in sorafenib-sensitive cells.Furthermore,there was a dose-dependent reduction in the expression of proteins associated with the EGFR/JNK/ERK pathway in apatinib-treated groups.Notably,SP600125 and PD98059 contributed to the inhibition of EMT and EGFR/JNK/ERK pathway-related proteins by apatinib in sorafenib-resistant HCC.Conclusion:Apatinib potentially hindered the progression of sorafenib-resistant HCC by suppressing both EMT and the EGFR/JNK/ERK pathway.
基金supported by the National Natural Science Foundation of China(Nos.22206207,22127810,and 22276224)the Natural Science Foundation of Guangdong Province(Nos.2021A1515011546 and 2023A1515010085)the Science and Technology Planning Project of Guangzhou(No.202102080005)。
文摘Epithelial-mesenchymal transition(EMT)plays an irreplaceable role in the development of silicosis.However,molecular mechanisms of EMT induced by silica exposure still remain to be addressed.Herein,metabolic profiles of human alveolar type II epithelial cells(A549 cells)exposed directly to silica were characterized using non-targeted metabolomic approaches.A total of 84 differential metabolites(DMs)were identified in silica-treated A549 cells undergoing EMT,which were mainly enriched in metabolisms of amino acids(e.g.,glutamate,alanine,aspartate),purine metabolism,glycolysis,etc.The number of DMs identified in the A549 cells obviously increased with the elevated exposure concentration of silica.Remarkably,glutamine catabolism was significantly promoted in the silica-treated A549 cells,and the levels of related metabolites(e.g.,succinate)and enzymes(e.g.,α-ketoglutarate(α-KG)dehydrogenase)were substantially up-regulated,with a preference toα-KG pathway.Supplementation of glutamine into the cell culture could substantially enhance the expression levels of both EMT-related markers and Snail(zinc finger transcription factor).Our results suggest that the EMT of human alveolar epithelial cells directly induced by silica can be essential to the development of silicosis.
基金supported by grants from the Beijing Municipal Public Welfare Development and Reform Pilot Project for Medical Research Institutes(PWD&RPP-MRI,JYY2023-6)the R&D Program of Beijing Municipal Education Commission(KZ20231002543).
文摘Background:Dry eye disease(DED)predominantly results from elevated tear film os-molarity,which can not only cause ocular inconvenience but may lead to visual impair-ments,severely compromising patient well-being and exerting substantial economic burdens as well.Astaxanthin(AST),a member of the xanthophylls and recognized for its robust abilities to combat inflammation and oxidation,is a common dietary sup-plement.Nonetheless,the precise molecular pathways through which AST influences DED are still poorly understood.Methods:Therapeutic targets for AST were identified using data from the GeneCards,PharmMapper,and Swiss Target Prediction databases,and STITCH datasets.Similarly,targets for dry eye disease(DED)were delineated leveraging resources such as the Therapeutic Target Database(TTD),DisGeNET,GeneCards,and OMIM databases,and DrugBank datasets.Interactions among shared targets were charted and dis-played using CytoScape 3.9.0.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses were conducted to elucidate the functions of pivotal tar-gets within the protein-protein interaction network.Molecular interactions between AST and key targets were confirmed through molecular docking using AutoDock and PyMOL.Molecular dynamics simulations were performed using GROMACS 2022.3.Viability of human corneal epithelial cells(hCEC)was assessed across varying concen-trations of AST.A mouse model of experimental DED was developed using 0.1%ben-zalkonium chloride(BAC),and the animals were administered 100 mg/kg/day of AST orally for 7 days.The efficacy of the treatments was assessed through a series of di-agnostic tests to evaluate the condition of the ocular surface after the interventions.The levels of inflammation and oxidative stress were quantitatively assessed using methods such as reverse transcription-polymerase chain reaction(RT-PCR),Western blot,and immunofluorescence staining.Results:Network pharmacology suggests that AST may alleviate DED by influenc-ing oxidation-reduction signaling pathways and reducing oxidative stress provoked by BAC.In vivo experiments demonstrated an improved overall condition in AST-administered mice in contrast to the control group.Immunofluorescence staining analyses indicated a decrease in Keap1 protein in the corneal tissues of AST-treated mice and a significant increase in Nrf2 and HO-1 protein.In vitro studies demon-strated that AST significantly enhanced cell viability and suppressed reactive oxy-gen species expression under hyperosmotic(HS)conditions,thereby protecting the human corneal epithelium.Conclusion:AST is capable of shielding mice from BAC-induced DED,decelerating the progression of DED,and mitigating oxidative stress damage under HS conditions in hCEC cells.The protective impact of AST on DED may operate through stimulating the Keap1-Nrf2/HO-1 signaling pathway.Our research findings indicate that AST may be a promising treatment for DED,offering new insights into DED treatment.
文摘BACKGROUND Oral squamous cell carcinoma(OSCC)is a frequent cancer affecting the oral cavity.OSCC is usually preceded by occurrence of precursor lesions,which demonstrate a varying degree of malignant transformation potential.Epithelialmesenchymal transition is a significant biological phenomenon that facilitates tumor growth and metastasis through the involvement of diverse epithelial and mesenchymal proteins.E-cadherin down regulation and over-expression of zincfinger E-box-binding homeobox 1(ZEB1)has been reported in several cancers.AIM To evaluate the role of E-cadherin and ZEB1 in oral leukoplakia and OSCC.METHODS A total of 60 cases i.e.,oral leukoplakia/oral epithelial dysplasia(OED)(n=30)and OSCC(n=30)were included.Immunohistochemistry was performed utilizing two markers:E-cadherin and ZEB1.The Mann-Whitney U test was employed to compare individual markers across the study groups.Spearman’s correlation was done followed by discriminant function analysis.RESULTS Reduction in the expression E-cadherin and altered localization were noted from OED to OSCC.Overexpression of ZEB1 along with cytoplasmic accumulation was noted in OED,with marked expression in OSCC.ZEB1 epithelium intensity and ZEB1 connective tissue percentage contributed to the discriminant function with an accuracy of 85%for the classification of OED from OSCC.CONCLUSION Loss of E-cadherin and up-regulation of ZEB1 from OED to OSCC,predisposes to induction of epithelial-mesenchymal transition.Discriminant formulas are developed to classify cases of OED and OSCC with 85%accuracy.
基金supported by the China Postdoctoral Science Foundation(2021M701462)the National Natural Science Foundation of China(32072254)+2 种基金the Postdoctoral Research Funding Program of Jiangsu Province(2021K097A)the Fundamental Research Funds for the Central Universities(JUSRP121106)the“Qing Lan Project”of Jiangsu Province。
文摘The dark blue pigment(DBP)is a health ingredient from Vaccinium bracteatum Thunb.leaves used as a functional food supplement.However,the details of transepithelial absorption on the intestinal epithelial cells are barely understood.This study aimed to clarify the absorption properties of DBP in the Caco-2 cell monolayer model and evaluate the effect on the endo-metabolism and barrier function of Caco-2 cells.The results showed that the DBP did not show the dose-dependent toxic effect to Caco-2 cells between 0.25 and 1.5 mg/mL,which did not cause disorder in the normal cell metabolism and absorption activity.The Caco-2 cell monolayer model could absorb DBP by passive and active transport,and the absorptive pattern was dosedependent when the concentration was more than 0.25 mg/mL.During DBP absorption,an increase in m RNA and protein expressions of glucose transporters demonstrated that the glucose transporters were the potential transporter of DBP.But the glucose transport amounts were significantly lowered after 30 min of DBP treatment,indicating that DBP owned the inhibitory effect on glucose transportation.Furthermore,DBP also owned protective effects on the barrier function of intestinal epithelial cells.
基金supported by the National Key Research and Development Program of China(2023YFE0100400)Science and Technology Project of Inner Mongolia Autonomous Region(2020GG0036)+2 种基金Basic Scientific Research Business Project of Universities directly under the Inner Mongolia Autonomous Region(BR22-11-17)National Center of Technology Innovation for Dairy(2023-JSGG-5)the Special Project for Improving the Research Ability of Young Teachers of Inner Mongolia Agricultural University(BR220133).
文摘Background Using yeast culture as additives in ruminant feed prevents rumen microbial dysbiosis,enhances performance,and regulates rumen pH.The yeast culture used in this study was developed in-house,and has been shown to promote rumen epithelial growth in several sheep trials.Changes in protein expression associated with the promotion of rumen epithelial development following the addition of yeast culture,along with the associated molecular mechanisms,remain unknown.We used 2045-day-old weaned lambs to investigate the specific proteins and molecular mechanisms involved in these processes.Half of the lambs were fed yeast culture,and the other half were used as controls.Results Yeast culture enhanced growth performance,facilitated rumen fermentation,and promoted rumen papilla development in weaned lambs.Proteomics data identified 4,831 proteins in the rumen epithelial tissue of lambs,comprising 87 upregulated and 425 downregulated proteins.Administration of yeast culture activated multiple molecular functions within rumen epithelial cells,including oxidative phosphorylation,glutathione metabolism,apoptosis,cell cycle,and vitamin digestion and absorption.The expression of proteins associated with cell cycle regulation increased,whereas those associated with apoptosis decreased.Administration of yeast culture also reduced the duration of the G0/G1 phase of rumen epithelial cells and accelerated the cell cycle.Furthermore,yeast culture showed increased cyclin D1,cyclin-dependent kinase(CDK)2,CDK4,CDK6,and cyclin E1 expressions and decreased cytochrome C(Cyto-c),Bcl-2-related X protein(Bax),cleaved caspase 3(C-caspase 3),caspase 3,and cleaved caspase 7(C-caspase 7)protein expressions.Yeast culture upregulated the insulin-like growth factor-1 receptor(IGF-1R)and insulin-like growth factor-binding protein 5(IGFBP-5)mRNA expressions in rumen epithelial cells.Conclusions Yeast culture facilitates rumen epithelial development by regulating the cell cycle and IGF-1 signaling and reducing the expression of proteins associated with apoptosis in rumen epithelial cells.The findings of this study provide novel insights into the molecular mechanisms through which yeast culture promotes rumen epithelial development in weaned lambs.
基金The China Agriculture Research System Program(Project No.CARS-41-G04)Shenyang Governmental Science and Technology Program(Project No.22316-2-02)supported this work.
文摘Background Clostridium perfringens is a pathogen that secretes multiple toxins,impacting humans and animals.It can cause intestinal diseases such as necrotic enteritis.Although tannins inhibit C.perfringens proliferation,the precise underlying mechanisms are unclear.Objective This study integrated transcriptomics and metabolomics to systematically investigate the mechanism by which tannins,specifically pentagalloylglucose(PGG)and tannic acid(TA),inhibit C.perfringens and potential pathways to alleviate infection in vivo.Results Ion concentration measurements,flow cytometric analysis,and transmission electron microscopy revealed that PGG and TA damaged the cell membrane structure of C.perfringens,triggering cytoplasmic content leakage.Additionally,PGG and TA significantly affected C.perfringens at the transcriptional and metabolic levels.Bioinformatics analysis revealed that PGG and TA induced amino acid restriction,disrupted energy metabolism,and impeded the ability of C.perfringens to sense and respond to the external environment.In an in vitro C.perfringens-infected intestinal cell model,PGG and TA boundαtoxin,significantly reduced the mRNA expression of inflammatory factors,and improved intestinal barrier function and cell viability.Compared to PGG,TA exhibited stronger inhibitory activity against C.perfringens and binding toαtoxin.In vivo,PGG and TA alleviated C.perfringens-induced weight loss in mice,improved intestinal villi morphology,and reduced intestinal inflammation and tight junction gene dysregulation.Conclusion These findings indicate that tannins inhibit C.perfringens,improve gut tissue integrity and reduce inflammation,demonstrating their multi-target effects of resisting intestinal diseases caused by harmful bacteria.This offers new insights for plant polyphenol-based strategies against necrotic enteritis.
基金Supported by the National Natural Science Foundation of China(31601141)。
文摘A lactating dairy cow mammary epithelial cell(DCMEC)model is useful for studying milk synthesis and lactation in the dairy cow mammary gland.However,the cells can only maintain their original characteristics for a limited period in vitro.Thus,the proliferative capacity and lactation pattern of subcultured DCMECs need to be characterized.In the present study,subcultured DCMECs appeared to proliferate without changes in morphology or growth pattern up to the 12th passage.Subculturing had no obvious effect on the lactation capacity of the subcultured DCMEC up to the 10th passage in vitro.These results indicated that the DCMEC line could be subpassaged up to 10 times and still maintain its original milk synthesis characteristics.Moreover,DCMECs were frozen using different methods,and the method of placing the vials in the foam box was determined after cryopreservation with the best adherence of 91.2%.This method was recommended for long-term cryopreservation for DCMECs.
文摘The cornea is a transparent tissue that serves as the main refractive element of the eye ball.Limbal epithelial stem cells(LESCs),residing in the basal epithelial layer of the Palisades of Vogt located in the corneal limbus between cornea and scleral,are believed to be crucial for the continuous turnover of the corneal epithelium.The proliferation,migration,and differentiation of the LESCs are modulated by unique physical and chemical futures contained within the microenvironment known as the limbal niche.This niche,composed of nerve terminals,cells,extracellular matrix,vasculature,and signaling molecules,is the home for processes such as proliferation,migration and differentiation.Corneal nerve terminals possess special anatomical structures in the limbal region and basal epithelial cells,and they demonstrate pivotal biological effects in the regulation of the LESC function and corneal epithelium homeostasis.Biological molecules such as neuropeptides,neurotransmitters,and neurotrophic factors play a crucial role in modulating the LESCs phenotype responsible for corneal epithelium homeostasis.This paper will review recent studies on how these nerve derived molecules function in this process and provide clear orientations for future research.
基金Supported by National Key R&D Program of China(No.2023YFC2410203)Beijing Hospitals Authority Clinical medicine Development of Special Funding Support(No.ZLRK202503).
文摘Adenoid cystic carcinoma of the lacrimal gland(LGACC)is the most common type of malignant epithelial tumor of the lacrimal gland,which is characterized by a high recurrence rate,perineural invasion,and a propensity to metastasize to distant sites.Due to its unclear pathogenesis,LGACC has a poor prognosis and a high mortality rate.In recent years,a range of radiotherapy and chemotherapy have been clinically applied,leading to a shift in the treatment approach for LGACC.This article discussed the advances being made in the treatment of LGACC and provides readers with an overview of the impact of LGACC treatment modalities on patient survival and prognostic levels.
基金supported by the National Key Research and Development Program of China(Grant No.2023YFF1000902,2021YFD1200903)the National Science Foundation for Young Scientists of China(Grant No.32302706)the Beijing Dairy Industry Innovation Team(Grant No.BAIC06).
文摘Background Mastitis caused by Staphylococcus aureus(S.aureus)is one of the most intractable problems for the dairy industry,causing significantly reduced milk yields and early slaughter of cows worldwide.MicroRNAs(miRNAs)can post-transcriptionally regulate gene expression and studies in recent years have shown the importance of miRNA-associated gene regulation in S.aureus-induced mastitis.Results In this study,to investigate the role of miR-223 in mastitis,we performed experiments to overexpress and suppress miR-223 in an immortalized bovine mammary epithelial cell line(MAC-T)infected with S.aureus.Overexpression of miR-223 in MAC-T cells repressed cell apoptosis and necrosis induced by S.aureus infection,whereas suppression of miR-223 had the opposite effect.Transcriptome expression profiling with weighted gene co-expression network analysis(WGCNA)and gene set variation analysis(GSVA)showed that miR-223 affects apoptosis and inflammation-related pathways.Furthermore,differentially expressed(DE)genes were evaluated,and genes exhibiting contrasting expression trends in the miR-223 overexpressed and suppressed groups were assessed as potential target genes of miR-223.Potential target genes,including CDC25B,PTPRF,DCTN1,and DPP9,were observed to be associated with apoptosis and necroptosis.Finally,through integrative analysis of genome-wide association study(GWAS)data and the animal quantitative trait loci(QTL)database,we determined that target genes of miR-223 were significantly enriched in single-nucleotide polymorphisms(SNP)and QTLs related to somatic cell count(SCC)and mastitis.Conclusion In summary,miR-223 has an inhibitory effect on S.aureus-induced cell apoptosis and necrosis by regulating PTPRF,DCTN1,and DPP9.These genes were significantly enriched in QTL regions associated with bovine mastitis resistance,underscoring their relevance in genetic regulation of disease resilience.Our findings provide critical genetic markers for enhancing mastitis resistance,particularly S.aureus-induced mastitis,through selective breeding.This work offers valuable insights for developing cattle with improved resistance to mastitis via targeted genetic selection.
基金German research Foundation(DFG,grant numbers:CH2321/1–1 and SCHO1231/7–1)JH has received a scholarship from the Chinese Scholarship Council(CSC No.:201908350115).
文摘The oviduct epithelium is the initial maternal contact site for embryos after fertilization,offering the microenviron-ment before implantation.This early gestation period is particularly sensitive to stress,which can cause reduced fertil-ity and reproductive disorders in mammals.Nevertheless,the local impact of elevated stress hormones on the ovi-duct epithelium has received limited attention to date,except for a few reports on polyovulatory species like mice and pigs.In this study,we focused on the effects of chronic maternal stress on cattle,given its association with infertil-ity issues in this monoovulatory species.Bovine oviduct epithelial cells(BOEC)differentiated at the air–liquid interface(ALI)were stimulated with 250 nmol/L cortisol for 1 or 3 weeks.Subsequently,they were assessed for morphology,bioelectrical properties,and gene expression related to oviduct function,glucocorticoid pathway,cortisol metabo-lism,inflammation,and apoptosis.Results revealed adverse effects of cortisol on epithelium structure,featured by deciliation,vacuole formation,and multilayering.Additionally,cortisol exposure led to an increase in transepithelial potential difference,downregulated mRNA expression of the major glucocorticoid receptor(NR3C1),upregulated the expression of cortisol-responsive genes(FKBP5,TSC22D3),and significant downregulation of oviductal glycopro-tein 1(OVGP1)and steroid receptors PGR and ESR1.The systematic comparison to a similar experiment previously performed by us in porcine oviduct epithelial cells,indicated that bovine cultures were more susceptible to elevated cortisol levels than porcine.The distinct responses between both species are likely linked to their divergence in the cortisol-induced expression changes of HSD11B2,an enzyme controlling the cellular capacity to metabolise cortisol.These findings provide insights into the species-specific reactions and reproductive consequences triggered by maternal stress.
基金supported by the National Natural Science Foundation of China(32273082 and U21A20262).
文摘Background Ferroptosis is characterized by increased production of reactive oxygen species(ROS)and membrane lipid peroxidation that can exacerbate inflammatory damage.Extracellular vesicles(EVs)isolated from bovine milk have many biological functions,including antioxidant properties.However,the role of EVs on Klebsiella pneumoniaeinduced ferroptosis and oxidative stress in bovine mammary epithelial cells(bMECs)and murine mammary tissue is unclear.In this study,EVs were isolated from bovine colostrum,mature milk and clinical mastitis milk(defined as C-EVs,M-EVs and CM-EVs,respectively)and assessed by transmission electron microscopy,Western blot and transcriptome sequencing.Effects of EVs on K.pneumoniae-induced ferroptosis and oxidative stress in bMECs were evaluated with immunofluorescence and Western blot.Results In bMECs,infection with K.pneumoniae induced oxidative stress,decreasing protein expression of Nrf2,Keap1 and HO-1 plus SOD activity,and increasing ROS concentrations.However,protein expression of GPX4,ACSL4 and S100A4 in bMECs,all factors that regulate ferroptosis,was downregulated by K.pneumoniae.Furthermore,this bacterium compromised tight junctions in murine mammary tissue,with low expression of ZO-1 and Occludin,whereas protein expression of Nrf2 and GPX4 was also decreased in mammary tissue.Adding C-EVs,M-EVs or CMEVs reduced oxidative stress and ferroptosis in K.pneumoniae-infected bMECs in vitro and murine mammary tissues in vivo.Conclusion In conclusion,all 3 sources of milk-derived EVs alleviated oxidative stress and ferroptosis in K.pneumoniae-infected bMECs and mammary tissues.
基金supported by the Department of Agriculture and Rural Affairs of Jiangxi Province,China (JXARS-09)Science and Technology Program of Guangdong Province,China (2020B1212060060)。
文摘Pigeons and certain other avian species produce a milklike secretion in their crop sacs to nourish offspring,yet the detailed processes involved are not fully elucidated.This study investigated the crop sacs of 225-day-old unpaired non-lactating male pigeons(MN)and males initiating lactation on the first day after incubation(ML).Using RNA sequencing,ribosomeprofiling,andsingle-cell transcriptome sequencing(scRNA-seq),we identified a significant up-regulation of genes associated with ribosome assembly and protein synthesis in ML compared to MN.Results from scRNA-seq analysis identified 12distinct cell types and 22 clusters,with secretory epithelial cells(SECs)exhibiting marked expression of plasma cell markers,including IGLL1 and MZB1.RNA fluorescence in situ hybridization(RNA FISH)and IgY quantification confirmed the critical role of SECs in producing endogenous IgY during lactation.We propose that fibroblast-derived BAFF signals activate SECs,mimicking B cell transformation and enhancing protein production through the unfolded protein response(UPR).These findings shed light on the cellular dynamics of pigeon milk production and contribute to a broader understanding of avian biology.
