Background:The efficacy of standard 5-fluorouracil(5-FU)chemotherapy for colorectal cancer is limited by drug resistance and adverse effects,prompting research into esketamine,a potent ketamine variant with analgesic,...Background:The efficacy of standard 5-fluorouracil(5-FU)chemotherapy for colorectal cancer is limited by drug resistance and adverse effects,prompting research into esketamine,a potent ketamine variant with analgesic,antidepressant,and recently discovered anti-tumor properties,to determine if it can enhance 5-FU’s chemosensitivity.This study investigates whether esketamine synergizes with 5-FU to enhance therapeutic efficacy in colorectal adenocarcinoma cell models.Methods:We performed functional assays to evaluate proliferation(CCK-8),migration(wound healing),invasion(Transwell),and apoptosis(flow cytometry)in colorectal adenocarcinoma cell lines treated with 5-FU alone or in combination with esketamine.Transcriptomic profiling was conducted using RNA sequencing,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis was employed to identify critical molecular targets and signaling networks.Protein-level validation of key pathway components was performed via western blotting.Results:Combination therapy with esketamine and 5-FU synergistically inhibited cellular proliferation,migration,and invasion while significantly inducing apoptosis compared to monotherapy.Mechanistically,esketamine potentiated 5-FU-driven AMP-activated protein kinase(AMPK)phosphorylation,leading to inhibition of both mammalian target of rapamycin(mTOR)and hyaluronan-mediated motility receptor(HMMR).Conclusion:Esketamine enhances 5-FU chemosensitivity in colorectal adenocarcinoma by activating the AMPK/mTOR/HMMR signaling axis,thereby suppressing tumor progression and metastatic potential.These findings position esketamine as a potential adjunctive therapy for 5-FU-based regimens,offering the dual benefit of enhancing chemotherapeutic efficacy while addressing cancer-associated comorbidities including pain and depression.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
Promotion of angiogenesis is crucial for bone tissue repair,and the poor activity of angiogenic cells and growth factors is the main problem in angiogenesis.New proangiogenic nanomaterials are urgently needed to be a ...Promotion of angiogenesis is crucial for bone tissue repair,and the poor activity of angiogenic cells and growth factors is the main problem in angiogenesis.New proangiogenic nanomaterials are urgently needed to be a promising strategy for this issue.Nb promotes bone formation and fracture healing,possibly by increasing vascular endothelial growth factor(VEGF)production.Nanoniobium particles(nNb)may promote angiogenesis.However,the effect of nNb on angiogenesis is unclear,limiting its application.This study confirmed that nNb significantly promoted angiogenesis.nNb increased and Ras-related C3 botulinum toxin substrate(Rac)family small guanosine triphosphatase(GTPase)1(Rac1)expression,inducing F-actin aggregation at the front edge of cells and the formation of pseudopodia to mediate cell migration,further promoting angiogenesis.We discovered that cyclin-dependent kinase-like 5(CDKL5)is a new signaling molecule that activates Rac1.V-ets erythroblastosis virus E26 oncogene homolog(ETS)domain-containing protein(ELK1),regulating CDKL5 and Rac1,plays an upstream regulatory role.When ELK1 was inhibited,CDKL5 and Rac1 levels were decreased.ELK1,CDKL5 or Rac1 are effective regulatory targets of angiogenesis.Inhibiting expression of ELK1,CDKL5 or Rac1 decreased angiogenesis.Thus,nNb has good angiogenic effects.The ELK1-CDKL5-Rac1 signaling pathway regulates the migration of endothelial cells to promote angiogenesis.nNb can be used in bone tissue engineering as a new nanomaterial,and it will promote the development of a new strategy for tissue engineering.展开更多
Ulcerative colitis(UC)is an idiopathic,relapsing,and etiologically complicated chronic inflammatory bowel disease.Despite substantial progress in the management of UC,the outcomes of mucosal barrier repair are unsatis...Ulcerative colitis(UC)is an idiopathic,relapsing,and etiologically complicated chronic inflammatory bowel disease.Despite substantial progress in the management of UC,the outcomes of mucosal barrier repair are unsatisfactory.In this study,phillygenin(PHI)treatment alleviated the symptoms of chronic colitis in mice,including body weight loss,severe disease activity index scores,colon shortening,splenomegaly,oxidative stress,and inflammatory response.In particular,PHI treatment ameliorated the tight junction proteins(TJs)reduction,fibrosis,apoptosis,and intestinal stem cell activity,indicating that PHI exerted beneficial effects on the intestinal mucosal barrier in mice with chronic colitis.In the NCM460 cells damage model,dextran sulfate sodium triggered the sequential induction of TJs reduction,fibrosis,and apoptosis.Takeda G protein-coupled receptor-5(TGR5)dysfunction mediated NCM460 cell injury.Moreover,PHI treatment enhanced TJs and suppressed fibrosis and apoptosis to maintain NCM460 cell function,depending on TGR5 activation.PHI promoted TGR5 activation and elevated intracellular cyclic adenosine monophosphate levels in HEK 293T cells transfected with TGR5 expression plasmids.Cellular thermal shift assay and molecular docking studies confirmed that PHI directly binds to TGR5,indicating that PHI is an agonist of TGR5.The process of PERK-eIF2α pathway-mediated endoplasmic reticulum Ca^(2+) release was involved in NCM460 cell injury as well,which was associated with TGR5 dysfunction.When NCM460 cells were pretreated with PHI,the PERK-eIF2α pathway and elevated Ca^(2+) levels were blocked.In conclusion,our study demonstrated a novel mechanism that PHI inhibited the PERK-eIF2α-Ca^(2+) pathway through TGR5 activation to against DSS-induced TJs reduction,fibrosis,and apoptosis.展开更多
Background Uterine aging is a key factor contributing to the deterioration of egg quality and reproductive performance in laying hens.Despite its importance,the molecular mechanisms underlying uterine aging remain poo...Background Uterine aging is a key factor contributing to the deterioration of egg quality and reproductive performance in laying hens.Despite its importance,the molecular mechanisms underlying uterine aging remain poorly defined.This study aimed to characterize gene expression and regulatory changes associated with uterine aging in hens at different life stages.Results Transcriptomic Analysis of uterine tissue from hens aged 350,500,And 700 d revealed dynamic changes in gene expression patterns during aging.A significant upregulation of genes involved in cellular senescence was observed,including increased expression of the p53 signaling pathway And markers associated with inflammation And cell cycle arrest.The most notable changes occurred between 350 And 500 d of age,suggesting this as a critical window for the onset of uterine aging.MicroRNA sequencing identified miR-210a-5p as significantly reduced with age.Target prediction and experimental validation showed that miR-210a-5p directly suppresses the expression of RASL11B,a Ras-like small GTPase that activates the MAPK signaling pathway.In primary uterine epithelial cells,reduced miR-210a-5p levels led to elevated RASL11B expression,increased activation of B-Raf,MEK,and ERK proteins,and enhanced expression of aging-related genes and inflammatory factors.In contrast,overexpression of miR-210a-5p or inhibition of the MAPK pathway delayed senescence and reduced inflammatory signaling.RASL11B overexpression was sufficient to induce aging phenotypes,confirming its central role in promoting uterine cellular aging.Conclusions This study identifies a novel regulatory pathway in which miR-210a-5p modulates uterine aging through the RASL11B-MAPK signaling cascade.The findings provide mechanistic insight into age-related reproductive decline in hens and suggest that targeting this pathway may offer new strategies for maintaining uterine function and extending reproductive lifespan in poultry.展开更多
Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of th...Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of the neuroinflammation regulated by the FKBP prolyl isomerase 5(FKBP5)-IκB kinase alpha(IKKα)-nuclear factor kappa-B(NF-κB)-NOD-like receptor thermal protein domain-associated protein 3(NLRP3)signaling pathway in PTSD.Methods:The primary components of ADP,including ginsenosides Rg1 and Rb1,were quantified using ultra-performance liquid chromatography.Twelve C57BL/6 mice were allocated to control(D0)and experimental groups on days one,seven,and 14 of single prolonged stress(SPS).Eighteen C57BL/6 mice were allocated to control,SPS,and MCC950,an NLRP3 inhibitor(5 mg/kg)groups.Finally,24 C57BL/6 mice were allocated to control,SPS,paroxetine hydrochloride(PRX),or ADP(18.4 and 36.8 mg/kg)groups.Mice were administered MCC950,PRX,or ADP for 14 days.The open field test and elevated plus maze were used to evaluate anxiety-like behaviors,whereas fear memory extinction was evaluated using the fear memory test.Western blotting was employed to evaluate the expression levels of the FKBP5-IKKα-NF-κB-NLRP3 signaling pathway,tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β.The expression of FKBP5 and NLRP3 was further confirmed by immunofluorescence staining.Results:The amounts of ginsenosides Rg1 and Rb1 in ADP were(96.85±1.14)and(9.04±0.22)μg/g,respectively.Compared with the D0 group,the levels of the inflammatory cytokine proteins,TNF-α,IL-6,and IL-1β were elevated 1.33-to 1.51-fold and those of FKBP5-IKKα-NF-κB-NLRP3 signaling pathway were increased 1.16-to 1.41-fold in the hippocampus of the D14 group(P<0.05);the fluorescence intensity of FKBP5 and NLRP3 was also markedly increased(1.33-1.79-fold)in the hippocampus of the D14 group(P<0.5).Notably,injection of MCC950(5 mg/kg)reduced the levels of FKBP5-IKKα-NF-κB-NLRP3(0.80-0.88-fold)and inflammatory cytokines(0.74-0.83-fold),thereby improving the PTSD-like behaviors induced by SPS(P<0.05).In addition,ADP(36.8 g/kg)significantly improved PTSD-like behaviors and reduced levels of hippocampal inflammatory cytokines(0.70-0.79-fold)and FKBP5-IKKα-NF-κB-NLRP3(0.50-0.79-fold)(P<0.05)in SPS mice.Conclusion:The results suggest a potential therapeutic benefit of ADP in PTSD due to the inhibition of the FKBP5-IKKα-NF-κBNLRP3 signaling pathway.展开更多
BACKGROUND Gastric cancer(GC)is a widespread malignancy and associated with high rates of morbidity and mortality worldwide.AIM To examine the functional role of long non-coding RNAs small nucleolar RNA host gene 5(SN...BACKGROUND Gastric cancer(GC)is a widespread malignancy and associated with high rates of morbidity and mortality worldwide.AIM To examine the functional role of long non-coding RNAs small nucleolar RNA host gene 5(SNHG5)and its regulation of miR-92a-3p and B-cell translocation gene 2(BTG2)in GC progression.METHODS Quantitative reverse transcription PCR and western blot analysis determined the expression of SNHG5,miR-92a-3p,and BTG2 in GC and adjacent non-neoplastic mucosa.Dual-luciferase assays demonstrated interactions of SNHG5 with miR-92a-3p and BTG2.AGS cells were transfected with SNHG5 overexpression and miR-92a-3p knockdown models.Various assays,including CCK-8,colony formation,scratch wound healing,and Transwell assays,were used to determine cell proliferation and migration.An experimental model of a xenograft mouse was used to determine in vivo tumor growth.At the same time histological changes were evaluated by hematoxylin and eosin staining,with western blot analysis used to evaluate signaling pathway protein expression.RESULTS BTG2 and SNHG5 were downregulated in GC tissues,and miR-92a-3p was upregulated.Overexpression of SNHG5 or knockdown of miR-92a-3p reduced GC cell proliferation and migration,and increased BTG2 expression while decreasing PI3K/AKT signaling activity.The dual-luciferase assays demonstrated direct binding of miR-92a-3p to SNHG5 and BTG2.Tumor volume and weight were significantly reduced in mice transplanted with AGS cells treated with miR-92a-3p inhibitor or SNHG5 overexpression compared with control AGS cells.Hematoxylin and eosin staining revealed that treated tumors exhibited degenerative characteristics,including irregular morphology and nucleolysis.CONCLUSION LncRNA SNHG5 inhibited GC cell growth and migration by modulating the PI3K/AKT pathway via the miR-92a-3p/BTG2 axis.展开更多
Piezocatalytic hydrogen peroxide(H_(2)O_(2))generation is a promising synthesis method that has received increasing attention;however,the reaction pathway requires further investigation.Here,Bi_(5)Ti_(3)FeO_(15)nanofi...Piezocatalytic hydrogen peroxide(H_(2)O_(2))generation is a promising synthesis method that has received increasing attention;however,the reaction pathway requires further investigation.Here,Bi_(5)Ti_(3)FeO_(15)nanofibers are used to generate H_(2)O_(2)by harvesting mechanical energy,and the reaction pathways are investigated.The H_(2)O_(2)yield over Bi_(5)Ti_(3)FeO_(15)nanofibers steadily increases from 331μmol g1 h1 in the first cycle to 746μmol g1 h1 in the tenth cycle in pure water without a sacrificial agent.Reliable reaction pathways are revealed by monitoring the pH value changes in the reaction solution during the H_(2)O_(2)generation process.In the H_(2)O_(2)generation process,the water oxidation reaction(WOR)provides a large amount of H+in the reaction solution,which promotes the oxygen reduction reaction(ORR)for H_(2)O_(2)generation.Therefore,an efficient synergistic effect between ORR and WOR achieves dual-pathway H_(2)O_(2)generation,contributing to the excellent piezocatalytic performance of Bi_(5)Ti_(3)FeO_(15)nanofibers.Furthermore,mechanistic studies indicate that the piezocatalytic H_(2)O_(2)generation follows the energy band theory.This work not only demonstrates Bi_(5)Ti_(3)FeO_(15)nanofibers as efficient piezocatalysts for H_(2)O_(2)generation but also provides a simple and effective approach to elucidate reaction pathways.This approach can be applied in photocatalytic,tribocatalytic,and electrocatalytic H_(2)O_(2)generation.展开更多
Background Inflammatory bowel disease causes intestinal structural damage,impairs gut function,hinders animal growth and development,and reduces farming efficiency.Previous studies demonstrated that lactate alleviates...Background Inflammatory bowel disease causes intestinal structural damage,impairs gut function,hinders animal growth and development,and reduces farming efficiency.Previous studies demonstrated that lactate alleviates dextran sulfate sodium(DSS)-induced inflammation and mitigates weight loss by enhancing intestinal barrier functions.However,the mechanisms underlying lactate-mediated protection of the intestinal epithelial barrier remain unclear.This study aimed to explore the protective effect of lactate on intestinal barrier damage in colitis piglets and the possible underlying mechanisms through in vivo and in vitro experiments.Methods A total of 6021-day-old weaned female piglets were randomly assigned into three groups based on weight:the control group(basal diet with physiological saline gavage),the DSS group(basal diet with 5%DSS gavage),and the DSS+LA group(2%lactate diet with 5%DSS gavage).There were 10 replicates per treatment,with 2 piglets per replicate.Jejunal morphology was assessed via hematoxylin and eosin staining,while Western blotting quantified the protein levels of proliferation markers,including cluster of differentiation 24(CD24),cyclin D1,and wingless/integrated(Wnt)/β-catenin signaling components.In vitro,0.08%DSS and 2–32 mmol/L sodium lactate-treated intestinal porcine epithelial cell line-J2(IPEC-J2)cells(n=4)were assessed for viability(Cell Counting Kit-8 assay),apoptosis(flow cytometry),and proliferation parameters,including cell cycle analysis and Leucine-rich repeat-containing G-protein coupled receptor 5(Lgr5+)stem cell quantification.Results In vivo,DSS administration induced jejunal villus shortening(P<0.05),downregulated protein levels of CD24,cyclin D1,casein kinase 1(CK1),and dishevelled-2(DVL2)(P<0.05).In vitro,DSS promoted apoptosis,inhibited proliferation,diminished the Lgr5+cell populations(P<0.05),and reduced S-phase cell proportions(P<0.05).Conversely,lactate supplementation ameliorated DSS-induced villus atrophy(P<0.05),restored CD24,cyclin D1,CK1,and DVL2 protein levels(P<0.05).Furthermore,in vitro,sodium lactate attenuated DSS-induced apoptosis(P<0.05),enhanced IPEC-J2 proliferation(P<0.05),expanded Lgr5+cells(P<0.05),and increased S-phase progression(P<0.05).Conclusions In summary,lactate ameliorated intestinal barrier damage in DSS-induced colitis by activating the Wnt/β-catenin pathway and restoring the balance between epithelial cell proliferation and apoptosis.This study provides novel mechanistic evidence supporting lactate's therapeutic potential for IBD management.展开更多
Peripheral nerve defect repair is a complex process that involves multiple cell types;perineurial cells play a pivotal role.Hair follicle neural crest stem cells promote perineurial cell proliferation and migration vi...Peripheral nerve defect repair is a complex process that involves multiple cell types;perineurial cells play a pivotal role.Hair follicle neural crest stem cells promote perineurial cell proliferation and migration via paracrine signaling;however,their clinical applications are limited by potential risks such as tumorigenesis and xenogeneic immune rejection,which are similar to the risks associated with other stem cell transplantations.The present study therefore focuses on small extracellular vesicles derived from hair follicle neural crest stem cells,which preserve the bioactive properties of the parent cells while avoiding the transplantation-associated risks.In vitro,small extracellular vesicles derived from hair follicle neural crest stem cells significantly enhanced the proliferation,migration,tube formation,and barrier function of perineurial cells,and subsequently upregulated the expression of tight junction proteins.Furthermore,in a rat model of sciatic nerve defects bridged with silicon tubes,treatment with small extracellular vesicles derived from hair follicle neural crest stem cells resulted in higher tight junction protein expression in perineurial cells,thus facilitating neural tissue regeneration.At 10 weeks post-surgery,rats treated with small extracellular vesicles derived from hair follicle neural crest stem cells exhibited improved nerve function recovery and reduced muscle atrophy.Transcriptomic and micro RNA analyses revealed that small extracellular vesicles derived from hair follicle neural crest stem cells deliver mi R-21-5p,which inhibits mothers against decapentaplegic homolog 7 expression,thereby activating the transforming growth factor-β/mothers against decapentaplegic homolog signaling pathway and upregulating hyaluronan synthase 2 expression,and further enhancing tight junction protein expression.Together,our findings indicate that small extracellular vesicles derived from hair follicle neural crest stem cells promote the proliferation,migration,and tight junction protein formation of perineurial cells.These results provide new insights into peripheral nerve regeneration from the perspective of perineurial cells,and present a novel approach for the clinical treatment of peripheral nerve defects.展开更多
目的:通过观察醒鼻凝胶滴鼻剂干预变应性鼻炎(AR)豚鼠鼻黏膜成纤维细胞对Fyn-STAT5信号通路的影响,进一步阐明醒鼻凝胶剂治疗AR可能的作用机制。方法:体外分离培养AR豚鼠鼻黏膜成纤维细胞并进行鉴定;将成纤维细胞分为正常组、模型组、T...目的:通过观察醒鼻凝胶滴鼻剂干预变应性鼻炎(AR)豚鼠鼻黏膜成纤维细胞对Fyn-STAT5信号通路的影响,进一步阐明醒鼻凝胶剂治疗AR可能的作用机制。方法:体外分离培养AR豚鼠鼻黏膜成纤维细胞并进行鉴定;将成纤维细胞分为正常组、模型组、TGF-β1组、醒鼻剂组和雷诺考特组,分别检测各组成纤维细胞中Fyn-STAT5信号通路及相关因子的基因和蛋白变化。结果:细胞鉴定结果显示,F3代成纤维细胞波形蛋白表达呈阳性。MTT检测结果提示,经25ng/m L TGF-β1干预24h后,细胞活力增长最明显。Real-time PCR和Western blot检测结果显示,模型组中Fyn m RNA及蛋白和SCF m RNA高表达(P<0.01),IL-10 m RNA和STAT5蛋白呈现低表达(P<0.01);与模型组比较,3个给药组干预12、24h后,Fyn m RNA及蛋白、SCF m RNA的表达均显著降低,IL-10 m RNA、STAT5蛋白显著升高(P<0.01,P<0.05),干预48h后,酸鼻剂组及雷诺考特组IL-10 m RNA仍明显升高(P<0.01)。结论:醒鼻凝胶滴鼻剂可能通过降低Fyn和SCF表达,同时上调STAT5和IL-10,从而抑制Fyn-STAT5信号传导通路,减轻AR成纤维细胞免疫反应。展开更多
Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a nov...Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a novel miRNA implicated in porcine IMF adipogenesis was found, and its effect and regulatory mechanism were further explored with respect to intramuscular preadipocyte proliferation and differentiation.Results: By porcine adipose tissue miRNA sequencing analysis, we found that miR-146a-5p is a potential regulator of porcine IMF adipogenesis. Further studies showed that miR-146a-5p mimics inhibited porcine intramuscular preadipocyte proliferation and differentiation, while the miR-146a-5p inhibitor promoted cell proliferation and adipogenic differentiation. Mechanistically, miR-146a-5p suppressed cell proliferation by directly targeting SMAD family member 4(SMAD4) to attenuate TGF-β signaling. Moreover, miR-146a-5p inhibited the differentiation of intramuscular preadipocytes by targeting TNF receptor-associated factor 6(TRAF6) to weaken the AKT/mTORC1 signaling downstream of the TRAF6 pathway.Conclusions: MiR-146a-5p targets SMAD4 and TRAF6 to inhibit porcine intramuscular adipogenesis by attenuating TGF-β and AKT/mTORC1 signaling, respectively. These findings provide a novel miRNA biomarker for regulating intramuscular adipogenesis to promote pork quality.展开更多
We investigated the importance of HMGN5, a nuclear protein that binds to nucleosomes, unfolds chromatin, and affects transcription, in the LNCaP prostate cancer cell line. We also examined the molecular mechanisms tha...We investigated the importance of HMGN5, a nuclear protein that binds to nucleosomes, unfolds chromatin, and affects transcription, in the LNCaP prostate cancer cell line. We also examined the molecular mechanisms that promote apoptosis of LNCaP cells after infection with small interfering RNA （siRNA） targeting HMGN5 （siRNA-HMGN5）. The androgen-dependent LNCaP human prostate cancer cells were infected with siRNA-HMGN5. Apoptosis was detected using the Annexin V-PE/7-AAD double staining and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling （TUNEL） assays. Mitochondrial membrane potential was measured by JC-1 staining. HMGN5and GAPDHmRNA expression were determined using real-time PCR. Bcl-2 and other apoptosis-related protein levels were determined by Western blot analysis. Caspase activity was measured by cleavage of the caspase substrate. Infection with siRNA targeting HMGN5 efficiently and specifically reduced the HMGN5 expression in LNCaP cells. The downregulation of HMGN5 induced remarkable apoptosis of LNCaP cells and resulted in the reduction of mitochondrial membrane potential. The induction of cell apoptosis was accompanied by the upregulation of Bax, the Bax/Bcl-2 ratio and the activation of caspase3. The HMGN5-targeted siRNA was effective in downregulating the expression of HMGN5 in androgen-dependent prostate cancer cells and inducing cell apoptosis via the regulation of a caspase-related mitochondrial pathway and Bcl-2 family proteins. This study suggests that HMGN5 may be a potential molecular target with therapeutic relevance for the treatment of prostate cancer.展开更多
Hepatocellular carcinoma(HCC)is a malignant tumor with high morbidity and mortality globally.It accounts for the majority of primary liver cancer cases.Amyloid precursor protein(APP),a cell membrane protein,plays a vi...Hepatocellular carcinoma(HCC)is a malignant tumor with high morbidity and mortality globally.It accounts for the majority of primary liver cancer cases.Amyloid precursor protein(APP),a cell membrane protein,plays a vital role in the pathogenesis of Alzheimer’s disease,and has been found to be implicated in tumor growth and metastasis.Therefore,to understand the relationship between APP and 5-fluorouracil(5-FU)resistance in liver cancer,Cell Counting Kit-8,apoptosis and cell cycle assays,western blotting,and reverse transcription-quantitative polymerase chain reaction(q PCR)analysis were performed.The results demonstrated that APP expression in Bel7402-5-FU cells was significantly up-regulated,as compared with that in Bel7402 cells.Through successful construction of APP-silenced(si APP)and overexpressed(OE)Bel7402 cell lines,data revealed that the Bel7402-APP751-OE cell line was insensitive,while the Bel7402-si APP cell line was sensitive to 5-FU in comparison to the matched control group.Furthermore,APP overexpression decreased,while APP silencing increased 5-FU-induced apoptosis in Bel7402 cells.Mechanistically,APP overexpression and silencing can regulate the mitochondrial apoptotic pathway and the expression of apoptotic suppressor genes(B-cell lymphoma-2(Bcl-2)and B-cell lymphoma-extra large(Bcl-xl)).Taken together,these results preliminarily revealed that APP overexpression contributes to the resistance of liver cancer cells to 5-FU,providing a new perspective for drug resistance.展开更多
Background:Tauroursodeoxycholic acid(TUDCA),a hydrophilic bile acid,is the main medicinal component of bear bile and is commonly used to treat a variety of hepatobiliary diseases.Meanwhile,TUDCA has been shown to modu...Background:Tauroursodeoxycholic acid(TUDCA),a hydrophilic bile acid,is the main medicinal component of bear bile and is commonly used to treat a variety of hepatobiliary diseases.Meanwhile,TUDCA has been shown to modulate the intestinal barrier function and alleviate DSS-induced colitis in mice.However,the effect of TUDCA on the intestinal barrier of weaned piglets remains largely unclear.Methods:The weaned piglets and porcine IPEC-J2 intestinal epithelial cells were used to investigate the effects of TUDCA on intestinal barrier function in weaned piglets and explore the possible underlying mechanisms.In vivo,72 healthy weaned piglets were randomly allocated into 2 groups according to their gender and body weight,and piglets were fed the basal diet with 0(control,CON)and 200 mg/kg TUDCA for 30 d,respectively.Three female and three male piglets reflecting the average bodyweight were slaughtered in each group and samples were collected.In vitro,IPEC-J2 cells were subjected to 100μmol/L TUDCA to explore the possible underlying mechanisms.Results:Our results demonstrated that dietary TUDCA supplementation significantly reduced the diarrhea incidence of weaned piglets,possibly attributing to the TUDCA-enhanced intestinal barrier function and immunity.In addition,TUDCA supplementation altered serum metabolites and the relative abundance of certain gut bacteria,which might contribute to the improved intestinal barrier function.Furthermore,the in-vitro results showed that TUDCA improved the E.coli-induced epithelial barrier impairment of IPEC-J2 cells and increased Takeda G-coupled protein receptor 5(TGR5)protein expression.However,knockdown of TGR5 and inhibition of myosin light chain kinase(MLCK)pathway abolished the TUDCA-improved epithelial barrier impairment in E.coli-treated IPEC-J2 cells,indicating the involvement of TGR5-MLCK in this process.Conclusions:These findings showed that TUDCA improved intestinal barrier function associated with TGR5-MLCK pathway and the alteration of serum metabolites and gut bacteria in weaned piglets,suggesting the potential application of TUDCA in improving gut health in piglet production.展开更多
AIM To investigate the potential role of micro RNA-30 a(mi R-30 a) in esophageal squamous cell carcinoma(ESCC).METHODS Expression of mi R-30 a-3 p/5 p was analyzed using microarray data and fresh ESCC tissue samples. ...AIM To investigate the potential role of micro RNA-30 a(mi R-30 a) in esophageal squamous cell carcinoma(ESCC).METHODS Expression of mi R-30 a-3 p/5 p was analyzed using microarray data and fresh ESCC tissue samples. Both in vitro and in vivo assays were used to investigate the effects of mi R-30 a-3 p/5 p on ESCC cell proliferation. Furthermore,Kyoto Encyclopedia of Genes and Genomes analysis was performed to explore underlying mechanisms involved in ESCC,and then,assays were carried out to verify the potential molecular mechanism of mi R-30 a in ESCC.RESULTS Low expression of mi R-30 a-3 p/5 p was closely associated with advanced ESCC progression and poor prognosis of patients with ESCC. Knock-down of mi R-30 a-3 p/5 p promoted ESCC cell proliferation. Increased mi R-30 a-3 p/5 p expression inhibited the Wnt signaling pathway by targeting Wnt2 and Fzd2.CONCLUSION Down-regulation of mi R-30 a-3 p/5 p promotes ESCC cell proliferation by activating the Wnt signaling pathway through inhibition of Wnt2 and Fzd2.展开更多
Avian infectious bronchitis virus(IBV) is a Gammacoronavirus in the family Coronaviridae and causes highly contagious respiratory disease in chickens. Innate immunity plays significant roles in host defense against IB...Avian infectious bronchitis virus(IBV) is a Gammacoronavirus in the family Coronaviridae and causes highly contagious respiratory disease in chickens. Innate immunity plays significant roles in host defense against IBV. Here, we explored the interaction between IBV and the host innate immune system. Severe histopathological lesions were observed in the tracheal mucosa at 3–5days post inoculation(dpi) and in the kidney at 8 dpi, with heavy viral loads at 1–11 and 1–28 dpi,respectively. The expression of m RNAs encoding Toll-like receptor(TLR) 3 and TLR7 were upregulated at 3–8 dpi, and that of TIR-domain-containing adapter-inducing interferon(IFN) β(TRIF) was upregulated at 21 dpi in the trachea and kidney. Myeloid differentiation primary response protein 88(My D88) was upregulated in the trachea during early infection. Tumor necrosis factor receptor-associated factor(TRAF) 3 and TRAF6 were upregulated expression in both tissues.Moreover, melanoma differentiation-associated protein 5(MDA5), laboratory of genetics and physiology 2(LGP2), stimulator of IFN genes(STING), and mitochondrial antiviral signaling protein(MAVS), as well as TANK binding kinase 1(TBK1), inhibitor of kappa B kinase(IKK) ?, IKKα, IKKβ,IFN regulatory factor(IRF) 7, nuclear factor of kappa B(NF-κB), IFN-α, IFN-β, various interleukins(ILs), and macrophage inflammatory protein-1β(MIP-1β) were significantly upregulated in the trachea and downregulated in the kidney. These results suggested that the TLR and MDA5 signaling pathways and innate immune cytokine were induced after IBV infection. Additionally,consistent responses to IBV infection were observed during early infection, with differential and complicated responses in the kidney.展开更多
AIM To investigate the mechanism by which hepatitis C virus(HCV) core protein-induced mi R-93-5 p up-regulation regulates the interferon(IFN) signaling pathway.METHODS HCV-1 b core protein was exogenously expressed in...AIM To investigate the mechanism by which hepatitis C virus(HCV) core protein-induced mi R-93-5 p up-regulation regulates the interferon(IFN) signaling pathway.METHODS HCV-1 b core protein was exogenously expressed in Huh7 cells using pc DNA3.1(+) vector. The expression of mi R-93-5 p and interferon receptor 1(IFNAR1) was measured using quantitative reverse transcriptionpolymerase chain reaction and Western blot. The protein expression and phosphorylation level of STAT1 were evaluated by Western blot. The overexpression and silencing of mi R-93-5 p and IFNAR1 were performed using mi R-93-5 p agomir and antagomir, and pc DNA3.1-IFNAR1 and IFNAR1 si RNA, respectively. Luciferase assay was used to identify whether IFNAR1 is a target of mi R-93-5 p. Cellular experiments were also conducted.RESULTS Serum mi R-93-5 p level was increased in patients with HCV-1 b infection and decreased to normal level after HCV-1 b clearance, but persistently increased in those with pegylated interferon-α resistance, compared with healthy subjects. Serum mi R-93-5 p expression had an AUC value of 0.8359 in distinguishing patients with pegylated interferon-α resistance from those with pegylated interferon-α sensitivity. HCV-1 b core protein increased mi R-93-5 p expression and induced inactivation of the IFN signaling pathway in Huh7 cells. Furthermore, IFNAR1 was identified as a direct target of mi R-93-5 p, and IFNAR1 restore could rescue mi R-93-5 p-reduced STAT1 phosphorylation, suggesting that the mi R-93-5 p-IFNAR1 axis regulates the IFN signaling pathway.CONCLUSION HCV-1 b core protein-induced mi R-93-5 p up-regulation inhibits the IFN signaling pathway by directly targeting IFNAR1, and the mi R-93-5 p-IFNAR1 axis regulates STAT1 phosphorylation. This axis may be a potential therapeutic target for HCV-1 b infection.展开更多
基金funded by the Natural Science Foundation of Guangdong Provice(2022A1515012543).
文摘Background:The efficacy of standard 5-fluorouracil(5-FU)chemotherapy for colorectal cancer is limited by drug resistance and adverse effects,prompting research into esketamine,a potent ketamine variant with analgesic,antidepressant,and recently discovered anti-tumor properties,to determine if it can enhance 5-FU’s chemosensitivity.This study investigates whether esketamine synergizes with 5-FU to enhance therapeutic efficacy in colorectal adenocarcinoma cell models.Methods:We performed functional assays to evaluate proliferation(CCK-8),migration(wound healing),invasion(Transwell),and apoptosis(flow cytometry)in colorectal adenocarcinoma cell lines treated with 5-FU alone or in combination with esketamine.Transcriptomic profiling was conducted using RNA sequencing,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis was employed to identify critical molecular targets and signaling networks.Protein-level validation of key pathway components was performed via western blotting.Results:Combination therapy with esketamine and 5-FU synergistically inhibited cellular proliferation,migration,and invasion while significantly inducing apoptosis compared to monotherapy.Mechanistically,esketamine potentiated 5-FU-driven AMP-activated protein kinase(AMPK)phosphorylation,leading to inhibition of both mammalian target of rapamycin(mTOR)and hyaluronan-mediated motility receptor(HMMR).Conclusion:Esketamine enhances 5-FU chemosensitivity in colorectal adenocarcinoma by activating the AMPK/mTOR/HMMR signaling axis,thereby suppressing tumor progression and metastatic potential.These findings position esketamine as a potential adjunctive therapy for 5-FU-based regimens,offering the dual benefit of enhancing chemotherapeutic efficacy while addressing cancer-associated comorbidities including pain and depression.
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 Guangzhou Science and Technology Plan Project(No.2023A03J0328)the National Natural Science Foundation of China(No.81600904)。
文摘Promotion of angiogenesis is crucial for bone tissue repair,and the poor activity of angiogenic cells and growth factors is the main problem in angiogenesis.New proangiogenic nanomaterials are urgently needed to be a promising strategy for this issue.Nb promotes bone formation and fracture healing,possibly by increasing vascular endothelial growth factor(VEGF)production.Nanoniobium particles(nNb)may promote angiogenesis.However,the effect of nNb on angiogenesis is unclear,limiting its application.This study confirmed that nNb significantly promoted angiogenesis.nNb increased and Ras-related C3 botulinum toxin substrate(Rac)family small guanosine triphosphatase(GTPase)1(Rac1)expression,inducing F-actin aggregation at the front edge of cells and the formation of pseudopodia to mediate cell migration,further promoting angiogenesis.We discovered that cyclin-dependent kinase-like 5(CDKL5)is a new signaling molecule that activates Rac1.V-ets erythroblastosis virus E26 oncogene homolog(ETS)domain-containing protein(ELK1),regulating CDKL5 and Rac1,plays an upstream regulatory role.When ELK1 was inhibited,CDKL5 and Rac1 levels were decreased.ELK1,CDKL5 or Rac1 are effective regulatory targets of angiogenesis.Inhibiting expression of ELK1,CDKL5 or Rac1 decreased angiogenesis.Thus,nNb has good angiogenic effects.The ELK1-CDKL5-Rac1 signaling pathway regulates the migration of endothelial cells to promote angiogenesis.nNb can be used in bone tissue engineering as a new nanomaterial,and it will promote the development of a new strategy for tissue engineering.
基金supported by the National Natural Science Fund of China(Grant Nos.:31800293 and 32370422)Project of Standard for TCM(Grant No.:ZYBZH-Y-JIN-34).
文摘Ulcerative colitis(UC)is an idiopathic,relapsing,and etiologically complicated chronic inflammatory bowel disease.Despite substantial progress in the management of UC,the outcomes of mucosal barrier repair are unsatisfactory.In this study,phillygenin(PHI)treatment alleviated the symptoms of chronic colitis in mice,including body weight loss,severe disease activity index scores,colon shortening,splenomegaly,oxidative stress,and inflammatory response.In particular,PHI treatment ameliorated the tight junction proteins(TJs)reduction,fibrosis,apoptosis,and intestinal stem cell activity,indicating that PHI exerted beneficial effects on the intestinal mucosal barrier in mice with chronic colitis.In the NCM460 cells damage model,dextran sulfate sodium triggered the sequential induction of TJs reduction,fibrosis,and apoptosis.Takeda G protein-coupled receptor-5(TGR5)dysfunction mediated NCM460 cell injury.Moreover,PHI treatment enhanced TJs and suppressed fibrosis and apoptosis to maintain NCM460 cell function,depending on TGR5 activation.PHI promoted TGR5 activation and elevated intracellular cyclic adenosine monophosphate levels in HEK 293T cells transfected with TGR5 expression plasmids.Cellular thermal shift assay and molecular docking studies confirmed that PHI directly binds to TGR5,indicating that PHI is an agonist of TGR5.The process of PERK-eIF2α pathway-mediated endoplasmic reticulum Ca^(2+) release was involved in NCM460 cell injury as well,which was associated with TGR5 dysfunction.When NCM460 cells were pretreated with PHI,the PERK-eIF2α pathway and elevated Ca^(2+) levels were blocked.In conclusion,our study demonstrated a novel mechanism that PHI inhibited the PERK-eIF2α-Ca^(2+) pathway through TGR5 activation to against DSS-induced TJs reduction,fibrosis,and apoptosis.
基金funded by the National Key Research and Development Program of China,grant number 2021YFD1300600Sichuan Science and Technology Program(2024YFNH0025,2022YFYZ0005,2021YFYZ0031)China Agriculture Research System of MOF and MARA(CARS-40).
文摘Background Uterine aging is a key factor contributing to the deterioration of egg quality and reproductive performance in laying hens.Despite its importance,the molecular mechanisms underlying uterine aging remain poorly defined.This study aimed to characterize gene expression and regulatory changes associated with uterine aging in hens at different life stages.Results Transcriptomic Analysis of uterine tissue from hens aged 350,500,And 700 d revealed dynamic changes in gene expression patterns during aging.A significant upregulation of genes involved in cellular senescence was observed,including increased expression of the p53 signaling pathway And markers associated with inflammation And cell cycle arrest.The most notable changes occurred between 350 And 500 d of age,suggesting this as a critical window for the onset of uterine aging.MicroRNA sequencing identified miR-210a-5p as significantly reduced with age.Target prediction and experimental validation showed that miR-210a-5p directly suppresses the expression of RASL11B,a Ras-like small GTPase that activates the MAPK signaling pathway.In primary uterine epithelial cells,reduced miR-210a-5p levels led to elevated RASL11B expression,increased activation of B-Raf,MEK,and ERK proteins,and enhanced expression of aging-related genes and inflammatory factors.In contrast,overexpression of miR-210a-5p or inhibition of the MAPK pathway delayed senescence and reduced inflammatory signaling.RASL11B overexpression was sufficient to induce aging phenotypes,confirming its central role in promoting uterine cellular aging.Conclusions This study identifies a novel regulatory pathway in which miR-210a-5p modulates uterine aging through the RASL11B-MAPK signaling cascade.The findings provide mechanistic insight into age-related reproductive decline in hens and suggest that targeting this pathway may offer new strategies for maintaining uterine function and extending reproductive lifespan in poultry.
基金supported by the National Natural Science Foundation of China(82404995,82404890)Research Funds of Center for Xin’an Medicine and Modernization of Traditional Chinese Medicine of IHM(2023CXMMTCM013)+3 种基金Scientific Research Program of Anhui Provincial Department of Education(2024AH051036,2024AH040137,2024AH051044)Excellent Funding for Academic and Scientific Research Activities for Academic and Technological Leaders in Anhui Province(2022D317)Key Research and Development Plan of Anhui Province(202104j07020004)Anhui University of Chinese Medicine Talent Support Program(DT2300000173).
文摘Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of the neuroinflammation regulated by the FKBP prolyl isomerase 5(FKBP5)-IκB kinase alpha(IKKα)-nuclear factor kappa-B(NF-κB)-NOD-like receptor thermal protein domain-associated protein 3(NLRP3)signaling pathway in PTSD.Methods:The primary components of ADP,including ginsenosides Rg1 and Rb1,were quantified using ultra-performance liquid chromatography.Twelve C57BL/6 mice were allocated to control(D0)and experimental groups on days one,seven,and 14 of single prolonged stress(SPS).Eighteen C57BL/6 mice were allocated to control,SPS,and MCC950,an NLRP3 inhibitor(5 mg/kg)groups.Finally,24 C57BL/6 mice were allocated to control,SPS,paroxetine hydrochloride(PRX),or ADP(18.4 and 36.8 mg/kg)groups.Mice were administered MCC950,PRX,or ADP for 14 days.The open field test and elevated plus maze were used to evaluate anxiety-like behaviors,whereas fear memory extinction was evaluated using the fear memory test.Western blotting was employed to evaluate the expression levels of the FKBP5-IKKα-NF-κB-NLRP3 signaling pathway,tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β.The expression of FKBP5 and NLRP3 was further confirmed by immunofluorescence staining.Results:The amounts of ginsenosides Rg1 and Rb1 in ADP were(96.85±1.14)and(9.04±0.22)μg/g,respectively.Compared with the D0 group,the levels of the inflammatory cytokine proteins,TNF-α,IL-6,and IL-1β were elevated 1.33-to 1.51-fold and those of FKBP5-IKKα-NF-κB-NLRP3 signaling pathway were increased 1.16-to 1.41-fold in the hippocampus of the D14 group(P<0.05);the fluorescence intensity of FKBP5 and NLRP3 was also markedly increased(1.33-1.79-fold)in the hippocampus of the D14 group(P<0.5).Notably,injection of MCC950(5 mg/kg)reduced the levels of FKBP5-IKKα-NF-κB-NLRP3(0.80-0.88-fold)and inflammatory cytokines(0.74-0.83-fold),thereby improving the PTSD-like behaviors induced by SPS(P<0.05).In addition,ADP(36.8 g/kg)significantly improved PTSD-like behaviors and reduced levels of hippocampal inflammatory cytokines(0.70-0.79-fold)and FKBP5-IKKα-NF-κB-NLRP3(0.50-0.79-fold)(P<0.05)in SPS mice.Conclusion:The results suggest a potential therapeutic benefit of ADP in PTSD due to the inhibition of the FKBP5-IKKα-NF-κBNLRP3 signaling pathway.
文摘BACKGROUND Gastric cancer(GC)is a widespread malignancy and associated with high rates of morbidity and mortality worldwide.AIM To examine the functional role of long non-coding RNAs small nucleolar RNA host gene 5(SNHG5)and its regulation of miR-92a-3p and B-cell translocation gene 2(BTG2)in GC progression.METHODS Quantitative reverse transcription PCR and western blot analysis determined the expression of SNHG5,miR-92a-3p,and BTG2 in GC and adjacent non-neoplastic mucosa.Dual-luciferase assays demonstrated interactions of SNHG5 with miR-92a-3p and BTG2.AGS cells were transfected with SNHG5 overexpression and miR-92a-3p knockdown models.Various assays,including CCK-8,colony formation,scratch wound healing,and Transwell assays,were used to determine cell proliferation and migration.An experimental model of a xenograft mouse was used to determine in vivo tumor growth.At the same time histological changes were evaluated by hematoxylin and eosin staining,with western blot analysis used to evaluate signaling pathway protein expression.RESULTS BTG2 and SNHG5 were downregulated in GC tissues,and miR-92a-3p was upregulated.Overexpression of SNHG5 or knockdown of miR-92a-3p reduced GC cell proliferation and migration,and increased BTG2 expression while decreasing PI3K/AKT signaling activity.The dual-luciferase assays demonstrated direct binding of miR-92a-3p to SNHG5 and BTG2.Tumor volume and weight were significantly reduced in mice transplanted with AGS cells treated with miR-92a-3p inhibitor or SNHG5 overexpression compared with control AGS cells.Hematoxylin and eosin staining revealed that treated tumors exhibited degenerative characteristics,including irregular morphology and nucleolysis.CONCLUSION LncRNA SNHG5 inhibited GC cell growth and migration by modulating the PI3K/AKT pathway via the miR-92a-3p/BTG2 axis.
文摘Piezocatalytic hydrogen peroxide(H_(2)O_(2))generation is a promising synthesis method that has received increasing attention;however,the reaction pathway requires further investigation.Here,Bi_(5)Ti_(3)FeO_(15)nanofibers are used to generate H_(2)O_(2)by harvesting mechanical energy,and the reaction pathways are investigated.The H_(2)O_(2)yield over Bi_(5)Ti_(3)FeO_(15)nanofibers steadily increases from 331μmol g1 h1 in the first cycle to 746μmol g1 h1 in the tenth cycle in pure water without a sacrificial agent.Reliable reaction pathways are revealed by monitoring the pH value changes in the reaction solution during the H_(2)O_(2)generation process.In the H_(2)O_(2)generation process,the water oxidation reaction(WOR)provides a large amount of H+in the reaction solution,which promotes the oxygen reduction reaction(ORR)for H_(2)O_(2)generation.Therefore,an efficient synergistic effect between ORR and WOR achieves dual-pathway H_(2)O_(2)generation,contributing to the excellent piezocatalytic performance of Bi_(5)Ti_(3)FeO_(15)nanofibers.Furthermore,mechanistic studies indicate that the piezocatalytic H_(2)O_(2)generation follows the energy band theory.This work not only demonstrates Bi_(5)Ti_(3)FeO_(15)nanofibers as efficient piezocatalysts for H_(2)O_(2)generation but also provides a simple and effective approach to elucidate reaction pathways.This approach can be applied in photocatalytic,tribocatalytic,and electrocatalytic H_(2)O_(2)generation.
基金funded by the Sichuan Science and Technology Program(2021ZDZX0009)the earmarked fund from the National Natural Science Foundation of China(31972577)。
文摘Background Inflammatory bowel disease causes intestinal structural damage,impairs gut function,hinders animal growth and development,and reduces farming efficiency.Previous studies demonstrated that lactate alleviates dextran sulfate sodium(DSS)-induced inflammation and mitigates weight loss by enhancing intestinal barrier functions.However,the mechanisms underlying lactate-mediated protection of the intestinal epithelial barrier remain unclear.This study aimed to explore the protective effect of lactate on intestinal barrier damage in colitis piglets and the possible underlying mechanisms through in vivo and in vitro experiments.Methods A total of 6021-day-old weaned female piglets were randomly assigned into three groups based on weight:the control group(basal diet with physiological saline gavage),the DSS group(basal diet with 5%DSS gavage),and the DSS+LA group(2%lactate diet with 5%DSS gavage).There were 10 replicates per treatment,with 2 piglets per replicate.Jejunal morphology was assessed via hematoxylin and eosin staining,while Western blotting quantified the protein levels of proliferation markers,including cluster of differentiation 24(CD24),cyclin D1,and wingless/integrated(Wnt)/β-catenin signaling components.In vitro,0.08%DSS and 2–32 mmol/L sodium lactate-treated intestinal porcine epithelial cell line-J2(IPEC-J2)cells(n=4)were assessed for viability(Cell Counting Kit-8 assay),apoptosis(flow cytometry),and proliferation parameters,including cell cycle analysis and Leucine-rich repeat-containing G-protein coupled receptor 5(Lgr5+)stem cell quantification.Results In vivo,DSS administration induced jejunal villus shortening(P<0.05),downregulated protein levels of CD24,cyclin D1,casein kinase 1(CK1),and dishevelled-2(DVL2)(P<0.05).In vitro,DSS promoted apoptosis,inhibited proliferation,diminished the Lgr5+cell populations(P<0.05),and reduced S-phase cell proportions(P<0.05).Conversely,lactate supplementation ameliorated DSS-induced villus atrophy(P<0.05),restored CD24,cyclin D1,CK1,and DVL2 protein levels(P<0.05).Furthermore,in vitro,sodium lactate attenuated DSS-induced apoptosis(P<0.05),enhanced IPEC-J2 proliferation(P<0.05),expanded Lgr5+cells(P<0.05),and increased S-phase progression(P<0.05).Conclusions In summary,lactate ameliorated intestinal barrier damage in DSS-induced colitis by activating the Wnt/β-catenin pathway and restoring the balance between epithelial cell proliferation and apoptosis.This study provides novel mechanistic evidence supporting lactate's therapeutic potential for IBD management.
基金supported by the National Natural Science Foundation of China,No.81571211(to FL)the Natural Science Foundation of Shanghai,No.22ZR1476800(to CH)。
文摘Peripheral nerve defect repair is a complex process that involves multiple cell types;perineurial cells play a pivotal role.Hair follicle neural crest stem cells promote perineurial cell proliferation and migration via paracrine signaling;however,their clinical applications are limited by potential risks such as tumorigenesis and xenogeneic immune rejection,which are similar to the risks associated with other stem cell transplantations.The present study therefore focuses on small extracellular vesicles derived from hair follicle neural crest stem cells,which preserve the bioactive properties of the parent cells while avoiding the transplantation-associated risks.In vitro,small extracellular vesicles derived from hair follicle neural crest stem cells significantly enhanced the proliferation,migration,tube formation,and barrier function of perineurial cells,and subsequently upregulated the expression of tight junction proteins.Furthermore,in a rat model of sciatic nerve defects bridged with silicon tubes,treatment with small extracellular vesicles derived from hair follicle neural crest stem cells resulted in higher tight junction protein expression in perineurial cells,thus facilitating neural tissue regeneration.At 10 weeks post-surgery,rats treated with small extracellular vesicles derived from hair follicle neural crest stem cells exhibited improved nerve function recovery and reduced muscle atrophy.Transcriptomic and micro RNA analyses revealed that small extracellular vesicles derived from hair follicle neural crest stem cells deliver mi R-21-5p,which inhibits mothers against decapentaplegic homolog 7 expression,thereby activating the transforming growth factor-β/mothers against decapentaplegic homolog signaling pathway and upregulating hyaluronan synthase 2 expression,and further enhancing tight junction protein expression.Together,our findings indicate that small extracellular vesicles derived from hair follicle neural crest stem cells promote the proliferation,migration,and tight junction protein formation of perineurial cells.These results provide new insights into peripheral nerve regeneration from the perspective of perineurial cells,and present a novel approach for the clinical treatment of peripheral nerve defects.
文摘目的:通过观察醒鼻凝胶滴鼻剂干预变应性鼻炎(AR)豚鼠鼻黏膜成纤维细胞对Fyn-STAT5信号通路的影响,进一步阐明醒鼻凝胶剂治疗AR可能的作用机制。方法:体外分离培养AR豚鼠鼻黏膜成纤维细胞并进行鉴定;将成纤维细胞分为正常组、模型组、TGF-β1组、醒鼻剂组和雷诺考特组,分别检测各组成纤维细胞中Fyn-STAT5信号通路及相关因子的基因和蛋白变化。结果:细胞鉴定结果显示,F3代成纤维细胞波形蛋白表达呈阳性。MTT检测结果提示,经25ng/m L TGF-β1干预24h后,细胞活力增长最明显。Real-time PCR和Western blot检测结果显示,模型组中Fyn m RNA及蛋白和SCF m RNA高表达(P<0.01),IL-10 m RNA和STAT5蛋白呈现低表达(P<0.01);与模型组比较,3个给药组干预12、24h后,Fyn m RNA及蛋白、SCF m RNA的表达均显著降低,IL-10 m RNA、STAT5蛋白显著升高(P<0.01,P<0.05),干预48h后,酸鼻剂组及雷诺考特组IL-10 m RNA仍明显升高(P<0.01)。结论:醒鼻凝胶滴鼻剂可能通过降低Fyn和SCF表达,同时上调STAT5和IL-10,从而抑制Fyn-STAT5信号传导通路,减轻AR成纤维细胞免疫反应。
基金supported by grants from the National Natural Science Foundation (31872979, 31572366)the National Key Research and Development Program of China (2017YFD0502002)the National Basic Research Programs of China (2015CB943102)。
文摘Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a novel miRNA implicated in porcine IMF adipogenesis was found, and its effect and regulatory mechanism were further explored with respect to intramuscular preadipocyte proliferation and differentiation.Results: By porcine adipose tissue miRNA sequencing analysis, we found that miR-146a-5p is a potential regulator of porcine IMF adipogenesis. Further studies showed that miR-146a-5p mimics inhibited porcine intramuscular preadipocyte proliferation and differentiation, while the miR-146a-5p inhibitor promoted cell proliferation and adipogenic differentiation. Mechanistically, miR-146a-5p suppressed cell proliferation by directly targeting SMAD family member 4(SMAD4) to attenuate TGF-β signaling. Moreover, miR-146a-5p inhibited the differentiation of intramuscular preadipocytes by targeting TNF receptor-associated factor 6(TRAF6) to weaken the AKT/mTORC1 signaling downstream of the TRAF6 pathway.Conclusions: MiR-146a-5p targets SMAD4 and TRAF6 to inhibit porcine intramuscular adipogenesis by attenuating TGF-β and AKT/mTORC1 signaling, respectively. These findings provide a novel miRNA biomarker for regulating intramuscular adipogenesis to promote pork quality.
文摘We investigated the importance of HMGN5, a nuclear protein that binds to nucleosomes, unfolds chromatin, and affects transcription, in the LNCaP prostate cancer cell line. We also examined the molecular mechanisms that promote apoptosis of LNCaP cells after infection with small interfering RNA （siRNA） targeting HMGN5 （siRNA-HMGN5）. The androgen-dependent LNCaP human prostate cancer cells were infected with siRNA-HMGN5. Apoptosis was detected using the Annexin V-PE/7-AAD double staining and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling （TUNEL） assays. Mitochondrial membrane potential was measured by JC-1 staining. HMGN5and GAPDHmRNA expression were determined using real-time PCR. Bcl-2 and other apoptosis-related protein levels were determined by Western blot analysis. Caspase activity was measured by cleavage of the caspase substrate. Infection with siRNA targeting HMGN5 efficiently and specifically reduced the HMGN5 expression in LNCaP cells. The downregulation of HMGN5 induced remarkable apoptosis of LNCaP cells and resulted in the reduction of mitochondrial membrane potential. The induction of cell apoptosis was accompanied by the upregulation of Bax, the Bax/Bcl-2 ratio and the activation of caspase3. The HMGN5-targeted siRNA was effective in downregulating the expression of HMGN5 in androgen-dependent prostate cancer cells and inducing cell apoptosis via the regulation of a caspase-related mitochondrial pathway and Bcl-2 family proteins. This study suggests that HMGN5 may be a potential molecular target with therapeutic relevance for the treatment of prostate cancer.
基金Project supported by the International Science&Technology Cooperation Program of China(No.2016G02).
文摘Hepatocellular carcinoma(HCC)is a malignant tumor with high morbidity and mortality globally.It accounts for the majority of primary liver cancer cases.Amyloid precursor protein(APP),a cell membrane protein,plays a vital role in the pathogenesis of Alzheimer’s disease,and has been found to be implicated in tumor growth and metastasis.Therefore,to understand the relationship between APP and 5-fluorouracil(5-FU)resistance in liver cancer,Cell Counting Kit-8,apoptosis and cell cycle assays,western blotting,and reverse transcription-quantitative polymerase chain reaction(q PCR)analysis were performed.The results demonstrated that APP expression in Bel7402-5-FU cells was significantly up-regulated,as compared with that in Bel7402 cells.Through successful construction of APP-silenced(si APP)and overexpressed(OE)Bel7402 cell lines,data revealed that the Bel7402-APP751-OE cell line was insensitive,while the Bel7402-si APP cell line was sensitive to 5-FU in comparison to the matched control group.Furthermore,APP overexpression decreased,while APP silencing increased 5-FU-induced apoptosis in Bel7402 cells.Mechanistically,APP overexpression and silencing can regulate the mitochondrial apoptotic pathway and the expression of apoptotic suppressor genes(B-cell lymphoma-2(Bcl-2)and B-cell lymphoma-extra large(Bcl-xl)).Taken together,these results preliminarily revealed that APP overexpression contributes to the resistance of liver cancer cells to 5-FU,providing a new perspective for drug resistance.
基金supported by the National Natural Science Foundation of China(31972636,31672508,31790411 and 31802103)the National Key Research and Development Program of China(2017YFD0500501)+1 种基金the Guangdong Key Areas Research and Development Project(2019B020218001)the Provincial Agricultural Science and Technology Innovation Promotion and Agricultural Resources and Ecological Environmental Protection Construction Project(2021KJ266).
文摘Background:Tauroursodeoxycholic acid(TUDCA),a hydrophilic bile acid,is the main medicinal component of bear bile and is commonly used to treat a variety of hepatobiliary diseases.Meanwhile,TUDCA has been shown to modulate the intestinal barrier function and alleviate DSS-induced colitis in mice.However,the effect of TUDCA on the intestinal barrier of weaned piglets remains largely unclear.Methods:The weaned piglets and porcine IPEC-J2 intestinal epithelial cells were used to investigate the effects of TUDCA on intestinal barrier function in weaned piglets and explore the possible underlying mechanisms.In vivo,72 healthy weaned piglets were randomly allocated into 2 groups according to their gender and body weight,and piglets were fed the basal diet with 0(control,CON)and 200 mg/kg TUDCA for 30 d,respectively.Three female and three male piglets reflecting the average bodyweight were slaughtered in each group and samples were collected.In vitro,IPEC-J2 cells were subjected to 100μmol/L TUDCA to explore the possible underlying mechanisms.Results:Our results demonstrated that dietary TUDCA supplementation significantly reduced the diarrhea incidence of weaned piglets,possibly attributing to the TUDCA-enhanced intestinal barrier function and immunity.In addition,TUDCA supplementation altered serum metabolites and the relative abundance of certain gut bacteria,which might contribute to the improved intestinal barrier function.Furthermore,the in-vitro results showed that TUDCA improved the E.coli-induced epithelial barrier impairment of IPEC-J2 cells and increased Takeda G-coupled protein receptor 5(TGR5)protein expression.However,knockdown of TGR5 and inhibition of myosin light chain kinase(MLCK)pathway abolished the TUDCA-improved epithelial barrier impairment in E.coli-treated IPEC-J2 cells,indicating the involvement of TGR5-MLCK in this process.Conclusions:These findings showed that TUDCA improved intestinal barrier function associated with TGR5-MLCK pathway and the alteration of serum metabolites and gut bacteria in weaned piglets,suggesting the potential application of TUDCA in improving gut health in piglet production.
基金Supported by the Youth Fund of the First Affiliated Hospital of Xinxiang Medical University(Type A-4)
文摘AIM To investigate the potential role of micro RNA-30 a(mi R-30 a) in esophageal squamous cell carcinoma(ESCC).METHODS Expression of mi R-30 a-3 p/5 p was analyzed using microarray data and fresh ESCC tissue samples. Both in vitro and in vivo assays were used to investigate the effects of mi R-30 a-3 p/5 p on ESCC cell proliferation. Furthermore,Kyoto Encyclopedia of Genes and Genomes analysis was performed to explore underlying mechanisms involved in ESCC,and then,assays were carried out to verify the potential molecular mechanism of mi R-30 a in ESCC.RESULTS Low expression of mi R-30 a-3 p/5 p was closely associated with advanced ESCC progression and poor prognosis of patients with ESCC. Knock-down of mi R-30 a-3 p/5 p promoted ESCC cell proliferation. Increased mi R-30 a-3 p/5 p expression inhibited the Wnt signaling pathway by targeting Wnt2 and Fzd2.CONCLUSION Down-regulation of mi R-30 a-3 p/5 p promotes ESCC cell proliferation by activating the Wnt signaling pathway through inhibition of Wnt2 and Fzd2.
基金supported by grants from the Natural Science Foundation of China (31360611 and 31160516)Guangxi Natural Science Foundation (2013GXNSFCA01 9010 and 2014GXNSFDA118011)
文摘Avian infectious bronchitis virus(IBV) is a Gammacoronavirus in the family Coronaviridae and causes highly contagious respiratory disease in chickens. Innate immunity plays significant roles in host defense against IBV. Here, we explored the interaction between IBV and the host innate immune system. Severe histopathological lesions were observed in the tracheal mucosa at 3–5days post inoculation(dpi) and in the kidney at 8 dpi, with heavy viral loads at 1–11 and 1–28 dpi,respectively. The expression of m RNAs encoding Toll-like receptor(TLR) 3 and TLR7 were upregulated at 3–8 dpi, and that of TIR-domain-containing adapter-inducing interferon(IFN) β(TRIF) was upregulated at 21 dpi in the trachea and kidney. Myeloid differentiation primary response protein 88(My D88) was upregulated in the trachea during early infection. Tumor necrosis factor receptor-associated factor(TRAF) 3 and TRAF6 were upregulated expression in both tissues.Moreover, melanoma differentiation-associated protein 5(MDA5), laboratory of genetics and physiology 2(LGP2), stimulator of IFN genes(STING), and mitochondrial antiviral signaling protein(MAVS), as well as TANK binding kinase 1(TBK1), inhibitor of kappa B kinase(IKK) ?, IKKα, IKKβ,IFN regulatory factor(IRF) 7, nuclear factor of kappa B(NF-κB), IFN-α, IFN-β, various interleukins(ILs), and macrophage inflammatory protein-1β(MIP-1β) were significantly upregulated in the trachea and downregulated in the kidney. These results suggested that the TLR and MDA5 signaling pathways and innate immune cytokine were induced after IBV infection. Additionally,consistent responses to IBV infection were observed during early infection, with differential and complicated responses in the kidney.
基金Supported by National Natural Science Foundation of China,No.81371849the TMMU Key Project for Clinical Research,No.2012XLC05
文摘AIM To investigate the mechanism by which hepatitis C virus(HCV) core protein-induced mi R-93-5 p up-regulation regulates the interferon(IFN) signaling pathway.METHODS HCV-1 b core protein was exogenously expressed in Huh7 cells using pc DNA3.1(+) vector. The expression of mi R-93-5 p and interferon receptor 1(IFNAR1) was measured using quantitative reverse transcriptionpolymerase chain reaction and Western blot. The protein expression and phosphorylation level of STAT1 were evaluated by Western blot. The overexpression and silencing of mi R-93-5 p and IFNAR1 were performed using mi R-93-5 p agomir and antagomir, and pc DNA3.1-IFNAR1 and IFNAR1 si RNA, respectively. Luciferase assay was used to identify whether IFNAR1 is a target of mi R-93-5 p. Cellular experiments were also conducted.RESULTS Serum mi R-93-5 p level was increased in patients with HCV-1 b infection and decreased to normal level after HCV-1 b clearance, but persistently increased in those with pegylated interferon-α resistance, compared with healthy subjects. Serum mi R-93-5 p expression had an AUC value of 0.8359 in distinguishing patients with pegylated interferon-α resistance from those with pegylated interferon-α sensitivity. HCV-1 b core protein increased mi R-93-5 p expression and induced inactivation of the IFN signaling pathway in Huh7 cells. Furthermore, IFNAR1 was identified as a direct target of mi R-93-5 p, and IFNAR1 restore could rescue mi R-93-5 p-reduced STAT1 phosphorylation, suggesting that the mi R-93-5 p-IFNAR1 axis regulates the IFN signaling pathway.CONCLUSION HCV-1 b core protein-induced mi R-93-5 p up-regulation inhibits the IFN signaling pathway by directly targeting IFNAR1, and the mi R-93-5 p-IFNAR1 axis regulates STAT1 phosphorylation. This axis may be a potential therapeutic target for HCV-1 b infection.
