inhibited

摘要： A gain-of-function mutation in the fibroblast growth factor receptor 3 gene(FGFR3)results in achondroplasia(ACH),the most frequent form of dwarfism.Constitutive activation of FGFR3 impairs bone formation and elongation and many signal transduction pathways.Identification of new and relevant compounds targeting the FGFR3 signaling pathway is of broad importance for the treatment of ACH,and natural plant compounds are prime drug candidate sources.Here,we found that the phenolic compound(-)-epicatechin,isolated from Theobroma cacao,effectively inhibited FGFR3’s downstream signaling pathways.Transcriptomic analysis in an Fgfr3 mouse model showed that ciliary mRNA expression was modified and influenced significantly by the Indian hedgehog and PKA pathways.(-)-Epicatechin is able to rescue mRNA expression impairments that control both the structural organization of the primary cilium and ciliogenesis-related genes.In femurs isolated from a mouse model(Fgfr3^(Y367C/+))of ACH,we showed that(-)-epicatechin eliminated bone growth impairment during 6 days of ex vivo culture.In vivo,we confirmed that daily subcutaneous injections of(-)-epicatechin to Fgfr3^(Y367C/+) mice increased bone elongation and rescued the primary cilium defects observed in chondrocytes.This modification to the primary cilia promoted the typical columnar arrangement of flat proliferative chondrocytes and thus enhanced bone elongation.The results of the present proof-of-principle study support(-)-epicatechin as a potential drug for the treatment of ACH.

摘要： Multiple signaling pathways are involved in the regulation of cell proliferation and differentiation in odontogenesis and dental tissue renewal,but the details of these mechanisms remain unknown.Here,we investigated the expression patterns of a transcription factor,Krüppel-like factor 6(KLF6),during the development of murine tooth germ and its function in odontoblastic differentiation.KLF6 was almost ubiquitously expressed in odontoblasts at various stages,and it was co-expressed with P21(to varying degrees)in mouse dental germ.To determine the function of Klf6,overexpression and knockdown experiments were performed in a mouse dental papilla cell line(iMDP-3).Klf6 functioned as a promoter of odontoblastic differentiation and inhibited the proliferation and cell cycle progression of i MDP-3 through p21 upregulation.Dual-luciferase reporter assay and chromatin immunoprecipitation showed that Klf6 directly activates p21 transcription.Additionally,the in vivo study showed that KLF6 and P21were also co-expressed in odontoblasts around the reparative dentin.In conclusion,Klf6 regulates the transcriptional activity of p21,thus promoting the cell proliferation to odontoblastic differentiation transition in vitro.This study provides a theoretical basis for odontoblast differentiation and the formation of reparative dentine regeneration.

摘要： Structure-assisted reverse chemical discovery approaches are becoming attractive due to target based screening.However,Small molecules(SMs)identified by reverse chemical genetic approach require robust phenotyping assays to study the plant responses.We optimized a screening method in rice seedlings using methyl viologen(MV)induced oxidative stress under high light conditions.This method was suitable for studying the efficacy of SM targeting oxidative stress related genes.Structure-assisted drug designing approach was used to identify SM targeting DREB2A transcription factor(TF).Rice seedlings treated with pronetalol showed susceptible phenotype when exposed to oxidative stress.Pronetalol inhibited the DREB2A activity and suppressed the expression of its target LEA7 and HSFA3 genes under oxidative stress.The assay is quite simple and robust and can be expanded to high throughput screening method for functional validation of TF.

摘要： BACKGROUND Pathological cardiac hypertrophy is a compensated response to various stimuli and is considered a key risk factor for heart failure.7,8-Dihydroxyflavone(7,8-DHF)is a flavonoid derivative that acts as a small-molecule brain-derived neurotrophic factor mimetic.The present study aimed to explore the potential role of 7,8-DHF in cardiac hypertrophy.METHODS Kunming mice and H9c2 cells were exposed to transverse aortic constriction or isoproterenol(ISO)with or without 7,8-DHF,respectively.F-actin staining was performed to calculate the cell area.Transcriptional levels of hypertrophic markers,including ANP,BNP,andβ-MHC,were detected.Echocardiography,hematoxylin-eosin staining,and transmission electron microscopy were used to examine the cardiac function,histology,and ultrastructure of ventricles.Protein levels of mitochondria-related factors,such as adenosine monophosphate-activated protein kinase(AMPK),and peroxisome proliferator-activated receptorγcoactivator-1α(PGC-1α),were detected.RESULTS 7,8-DHF inhibited compensated and decompensated cardiac hypertrophy,diminished the cross-sectional area,and alleviated the mitochondrial disorders of cardiomyocytes.Meanwhile,7,8-DHF reduced the cell size and repressed the mRNA levels of the hypertrophic markers of ISO-treated cardiomyocytes.In addition,7,8-DHF activated AMPK and PGC-1αsignals without affecting the protein levels of mitochondrial dynamics-related molecules.The effects of 7,8-DHF were eliminanted by Compound C,an AMPK inhibitor.CONCLUSIONS These findings suggest that 7,8-DHF inhibited cardiac hypertrophy and mitochondrial dysfunction by activating AMPK signaling,providing a potential agent for the treatment of pathological cardiac hypertrophy.

摘要： Oral squamous cell carcinoma(OSCC)has a high incidence of metastasis.Tumour immunotherapy targeting PD-L1 or PD-1 has been revolutionary;however,only a few patients with OSCC respond to this treatment.Therefore,it is essential to gain insights into the molecular mechanisms underlying the growth and metastasis of OSCC.In this study,we analysed the expression levels of protein kinase D3(PKD3)and PD-L1 and their correlation with the expression of mesenchymal and epithelial markers.We found that the expression of PKD3 and PD-L1 in OSCC cells and tissues was significantly increased,which correlated positively with that of mesenchymal markers but negatively with that of epithelial markers.Silencing PKD3 significantly inhibited the growth,metastasis and invasion of OSCC cells,while its overexpression promoted these processes.Our further analyses revealed that there was positive feedback regulation between PKD3 and PD-L1,which could drive EMT of OSCC cells via the ERK/STAT1/3 pathway,thereby promoting tumour growth and metastasis.Furthermore,silencing PKD3 significantly inhibited the expression of PD-L1,and lymph node metastasis of OSCC was investigated with a mouse footpad xenograft model.Thus,our findings provide a theoretical basis for targeting PKD3 as an alternative method to block EMT for regulating PD-L1 expression and inhibiting OSCC growth and metastasis.

摘要： Dental pulp can initiate its damage repair after an injury of the pulp–dentin complex by rearrangement of odontoblasts and formation of newly differentiated odontoblast-like cells.Connexin 43(Cx43)is one of the gap junction proteins that participates in multiple tissue repair processes.However,the role of Cx43 in the repair of the dental pulp remains unclear.This study aimed to determine the function of Cx43 in the odontoblast arrangement patterns and odontoblastic differentiation.Human teeth for in vitro experiments were acquired,and a pulp injury model in Sprague-Dawley rats was used for in vivo analysis.The odontoblast arrangement pattern and the expression of Cx43 and dentin sialophosphoprotein(DSPP)were assessed.To investigate the function of Cx43 in odontoblastic differentiation,we overexpressed or inhibited Cx43.The results indicated that polarized odontoblasts were arranged along the pulp–dentin interface and had high levels of Cx43 expression in the healthy teeth;however,the odontoblast arrangement pattern was slightly changed concomitant to an increase in the Cx43 expression in the carious teeth.Regularly arranged odontoblast-like cells had high levels of the Cx43 expression during the formation of mature dentin,but the odontoblastlike cells were not regularly arranged beneath immature osteodentin in the pulp injury models.Subsequent in vitro experiments demonstrated that Cx43 is upregulated during odontoblastic differentiation of the dental pulp cells,and inhibition or overexpression of Cx43 influence the odontoblastic differentiation.Thus,Cx43 may be involved in the maintenance of odontoblast arrangement patterns,and influence the pulp repair outcomes by the regulation of odontoblastic differentiation.

摘要： Specificity protein 1(Sp1)is a ubiquitously expressed transcription factor involved in the regulation of a large number of genes including housekeeping genes as well as actively regulated genes.Increasing evidences have indicated that Sp1 is closely correlated with the occurrence and progression of many tumors.The plasmid expression vector of short hairpin RNA(shRNA)targeting Sp1 was constructed and transfected into HepG2 cells by Lipo2000 transfection reagent.Immunocytochemistry,Western blot and quantitative real-time PCR were applied to determine Sp1 expression,and MTT and Transwell assay were used to analyze the cell proliferation and migration,respectively.The expressions of Sp1 protein and mRNA decreased significantly in HepG2 cell after transfection with Sp1 shRNA,and reduced Sp1 expression inhibited proliferation and migration of HepG2 cells.

摘要： Clinical efforts to repair damaged articular cartilage(AC)currently face major obstacles due to the limited intrinsic repair capacity of the cartilage tissue.Thus,it is essential to understand the molecular mechanisms involved in the chondrogenic differentiation.It is known that miRNAs play a significant role in chondrogenic differentiation.In this study,the expression of miR-222-3p and the level of autophagic activity were investigated during chondrogenic induction in ATDC5 cells.The results showed that miR-222-3p was down-regulated significantly and the level of autophagic activity was enhanced during this process,and there was a negative correlation between them.Moreover,inhibition of miR 222-3p promoted chondrogenic differentiation and inhibited hypertrophic differentiation of ATDC5 cells,as supported by enhancing the gene and protein expression of chondrocyte specific markers of SOX9,aggrecan and collagen type II(Col2a1),but reduced the expression of hypertrophic differentiation marker collagen type X(Col10a1).In contrast,inhibiting autophagic activity decreased chondrogenic differentiation and promoted hypertrophic differentiation of ATDC5 cells.

摘要： Methamphetamine(METH)is one of the most widely abused synthetic drugs in the world,with the abuser present hyperthermia(HT)and psychiatric symptoms.However,the mechanism involved in METH/HT-induced neurotoxicity remains elusive.Here,we investigated the role of heat shock protein 90 alpha(HSP90a)in METH/HT(39.5C)-induced necroptosis in rat striatum neurons and rat in vivo model.Western blot showed that the expression of HSP90a increased,and HSP90a specific inhibitor geldanamycin(GA)and shRNA of HSP90a could attenuate METH/HT-induced upregulation of receptor interacting protein 3(RIP3),pRIP3,mixed lineage kinase domain-like protein(MLKL)and pMLKL in cultured striatum neurons and in vivo model.Propidium iodide staining and lactate dehydrogenase assay results showed that METH/HT-induced necroptosis is partially inhibited by both necrostain-1,GA and shRNA of HSP90a in vitro and in vivo model.

摘要： Circadian rhythm is involved in the development and diseases of many tissues.However,as an essential environmental regulating factor,its effect on amelogenesis has not been fully elucidated.The present study aims to investigate the correlation between circadian rhythm and ameloblast differentiation and to explore the mechanism by which circadian genes regulate ameloblast differentiation.Circadian disruption models were constructed in mice for in vivo experiments.An ameloblast-lineage cell(ALC)line was used for in vitro studies.As essential molecules of the circadian system,Bmal1 and Per2 exhibited circadian expression in ALCs.Circadian disruption mice showed reduced amelogenin(AMELX)expression and enamel matrix secretion and downregulated expression of BMAL1,PER2,PPARγ,phosphorylated AKT1 andβ-catenin,cytokeratin-14 and F-actin in ameloblasts.According to previous findings and our study,BMAL1 positively regulated PER2.Therefore,the present study focused on PER2-mediated ameloblast differentiation and enamel formation.Per2 knockdown decreased the expression of AMELX,PPARγ,phosphorylated AKT1 andβ-catenin,promoted nuclearβ-catenin accumulation,inhibited mineralization and altered the subcellular localization of E-cadherin in ALCs.Overexpression of PPARγpartially reversed the above results in Per2-knockdown ALCs.Furthermore,in in vivo experiments,the length of incisor eruption was significantly decreased in the circadian disturbance group compared to that in the control group,which was rescued by using a PPARγagonist in circadian disturbance mice.In conclusion,through regulation of the PPARγ/AKT1/β-catenin signalling axis,PER2 played roles in amelogenin expression,cell junctions and arrangement,enamel matrix secretion and mineralization during ameloblast differentiation,which exert effects on enamel formation.