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1.
J Org Chem ; 2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38741558

RESUMO

A novel three-component cyclization carbonylation reaction of iodoarene-tethered propargyl ethers with amine and CO is reported. This palladium-catalyzed cascade reaction undergoes a sequence of oxidative addition, unsaturated bond migration, carbonyl insertion, and nucleophilic attack to deliver the benzofuran skeleton. Both aromatic amines and aliphatic amines could proceed smoothly in this transformation under one atm of CO.

2.
Environ Sci Pollut Res Int ; 30(22): 62104-62120, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36940032

RESUMO

Using rhizobacteria as biological fertilizer is gradually expanding in agriculture as excellent substitutes for chemical fertilizers. Bacillus subtilis SL-44 is a plant growth-promoting rhizobacteria screened from the severely salinized cotton rhizosphere soil in Xinjiang. Study showed that indole-3-acetic acid, organic acid production, nitrogen fixation, and other beneficial secondary metabolite secretion can be synthesized by stain SL-44. At the same time, fencyclin, lipopeptide, chitinase, and other antifungal substances were also detected from the secretion of Bacillus subtilis SL-44, which can effectively control plant diseases. Siderophore separated from SL-44 was verified by HPLC, and results showed it was likely bacillibactin. This study also verified that SL-44 has high antifungal activity against Rhizoctonia solani through in vitro antifungal experiments. The B. subtilis SL-44 whole genome was sequenced and annotated to further explore the biotechnological potential of SL-44. And a large number of genes involved in the synthesis of anti-oxidative stress, antibiotic, and toxins were found. Genome-wide analysis provides clear evidence to support the great potential of B. subtilis SL-44 strain to produce multiple bioantagonistic natural products and growth-promoting metabolites, which may facilitate further research into effective therapies for harmful diseases.


Assuntos
Antifúngicos , Bacillus subtilis , Antifúngicos/metabolismo , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Desenvolvimento Vegetal , Antibacterianos/metabolismo , Agricultura , Doenças das Plantas/microbiologia
3.
Environ Sci Pollut Res Int ; 30(8): 21548-21564, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36272007

RESUMO

The genus Enterobacter is widely recognized for its biotechnology potential in improving soil environment and crop growth promotion. To further explore these biotechnological potentials, we sequenced and analyzed the whole genome of Enterobacter cloacae Rs-2. The analysis showed that the total length of the Rs-2 genome was 6,965,070,514 bp, and GC content was 55.80%; the annotation results of GO and COG databases showed that the genome contains a variety of growth-promoting genes, such as iscU, glnA, glnB (nitrogen fixation); iucABCD (siderophore synthesis) and fepA, fcuA, fhuA, and pfeA, etc. (siderophore transport); ipdC (secreted IAA) and gcd, pqqBCDEF (dissolved phosphorus), etc. No pathogenic factors such as virulence genes were found. The application of Rs-2 as a soil inoculant in pot experiments showed great potential for growth promotion. This study proved the plant growth-promoting ability of Rs-2 at the molecular level through genetic screening and analysis, which provided guidance for the further improvement of the strain and laid a foundation for its application in agricultural production.


Assuntos
Enterobacter cloacae , Sideróforos , Enterobacter cloacae/genética , Enterobacter/genética , Fósforo , Desenvolvimento Vegetal , Solo
5.
Biochem Biophys Res Commun ; 523(4): 947-953, 2020 03 19.
Artigo em Inglês | MEDLINE | ID: mdl-31964531

RESUMO

Cancer stem-like cells are rare immortal cells within tumor, which are thought to play important roles in ionizing radiation (IR) therapy-resistance. Quercetin is a natural flavonoid with potential anti-cancer properties without significant cytotoxicity in normal tissues. In this study, we demonstrated that quercetin-IR combination treatment exhibited more dramatic anti-cancer effect than either quercetin or IR treatment alone via targeting colon cancer stem cells (CSCs) and inhibiting the Notch-1 signaling. These effects were further verified by in vivo studies which showed remarkable decrease of the CSCs markers and the expression of Notch-1 signaling proteins in human colon cancer xenografts in nude mice. Co-treatment with quercetin and low dose of radiation significantly reduced the expressions of all five proteins of γ-secretase complex in HT-29 and DLD-1 cells. In addition, ectopic expression of the Notch intracellular domain (NICD) partly reversed the inhibition effects by the combination therapy. In conclusion, our results indicated that the combination of quercetin (20 µM) and IR (5Gy) might be a promising therapeutic strategy for colon cancer treatment by targeting colon cancer stem-like cells and inhibiting the Notch-1 signaling. In future studies, we intend to further explore the potential therapeutic efficacy of the quercetin-radiation combination treatment in clinical trials.


Assuntos
Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/radioterapia , Quercetina/uso terapêutico , Tolerância a Radiação/efeitos dos fármacos , Receptores Notch/metabolismo , Transdução de Sinais , Secretases da Proteína Precursora do Amiloide/metabolismo , Animais , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/efeitos da radiação , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Células-Tronco Neoplásicas/efeitos da radiação , Quercetina/farmacologia , Tolerância a Radiação/efeitos da radiação , Radiação Ionizante , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/efeitos da radiação , Ensaios Antitumorais Modelo de Xenoenxerto
6.
J Cardiovasc Pharmacol ; 75(1): 45-53, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31895879

RESUMO

Atherosclerosis is a chronic inflammation condition resulting from the interaction between lipoproteins, monocyte-derived macrophages, T lymphocytes, and other cellular elements in the arterial wall. Macrophage-derived foam cells play a key role in both early and advanced stage of atherosclerosis. Previous studies have shown that berberine could inhibit foam cell formation and prevent experimental atherosclerosis. However, its underlying molecular mechanisms have not been fully clarified. In this study, we explored the cholesterol-lowering effects of berberine in macrophage-derived foam cells and investigated its possible mechanisms in prevention and treatment of atherosclerosis. Here, we demonstrated that berberine could inhibit atherosclerosis in apolipoprotein E-deficient mice and induce cholesterol reduction as well as decrease the content of macrophages. Berberine can regulate oxLDL uptake and cholesterol efflux, thus suppresses foam cell formation. Mechanisms study showed that berberine can suppress scavenger receptor expression via inhibiting the activity of AP-1 and upregulate ATP-binding cassette transporter via activating Nrf2/HO-1 signaling in human macrophage. In summary, berberine significantly inhibits atherosclerotic disease development by regulating lipid homeostasis and suppressing macrophage foam cell formation.


Assuntos
Aterosclerose/prevenção & controle , Berberina/farmacologia , Colesterol/metabolismo , Antagonistas Colinérgicos/farmacologia , Células Espumosas/efeitos dos fármacos , Heme Oxigenase-1/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Fator de Transcrição AP-1/metabolismo , Transportador 1 de Cassete de Ligação de ATP/metabolismo , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Aterosclerose/enzimologia , Aterosclerose/genética , Aterosclerose/patologia , Antígenos CD36/metabolismo , Modelos Animais de Doenças , Células Espumosas/enzimologia , Células Espumosas/patologia , Heme Oxigenase-1/genética , Humanos , Lipoproteínas LDL/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Knockout para ApoE , Fator 2 Relacionado a NF-E2/genética , Receptores Depuradores Classe A/metabolismo , Células THP-1
7.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-823060

RESUMO

@#With the continuous development of maxillary sinus floor elevation technology, the osteogenesis mechanism of maxillary sinus floor elevation has always been a concern of scholars. The membrane of the maxillary sinus is an indispensable physiological structure in the process of space osteogenesis under the sinus floor after elevation of the sinus floor. In recent years, the role of the maxillary sinus floor mucosa in sinus floor space osteogenesis has been a research hotspot. Recent studies have found that the maxillary sinus floor membrane plays a role as a natural biological barrier membrane in the process of sinus floor space osteogenesis after maxillary sinus floor elevation; in addition, it has the ability to undergo osteogenesis. It has also been found that maxillary sinus membrane stem cells (MSMSCs) derived from the maxillary sinus floor membrane have characteristics of mesenchymal stem cells, which can differentiate into osteoblasts and participate in sinus floor space osteogenesis after maxillary sinus floor elevation. New studies have also found that small RNAs such as microRNAs, long noncoding RNAs and circular RNAs can regulate the osteogenic differentiation of MSMSCs, which may be important biological targets for promoting osteogenesis in the sinus floor space. In this paper, the relationship between the maxillary sinus floor mucosa and bone formation after maxillary sinus floor elevation, the barrier and osteogenic function of the maxillary sinus floor mucosa, the sources of osteoblasts involved in osteogenesis of the sinus floor space, and the molecular regulatory mechanisms of stem cells derived from maxillary sinus mucosa will be elucidated step by step.

8.
Oral Health Prev Dent ; 17(6): 505-514, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31825023

RESUMO

PURPOSE: Matrix metalloproteinases (MMPs) are closely associated with the progression of oral cancer. This study is a meta-analysis of the published literature on MMPs and oral cancer prognosis. MATERIALS AND METHODS: A comprehensive search of the literature on the prognosis of MMPs and oral cancer in PubMed, Embase, Web of science databases and the Cochrane Library was performed. All eligible studies were included in the final study and HR data were extracted. Pooled hazard ratio (HR) was analysed. RESULTS: A total of 15 studies concerning 1266 patients were included for the meta-analysis. Pooled HR showed that overexpression of MMPs negatively influenced overall survival (OS) (HR = 2.18; 95% CI: 1.80-2.65; heterogeneity test, p = 0.080; I2 = 35.5%). In the subgroup analysis, the HR of MMP-2 was positive and the OS was poor (HR = 3.93; 95% CI: 2.19-7.07; weight 10.94%; heterogeneity test, p = 0.481; I2 = 0.0%). The HR of MMP-9 was positive and the OS was poor (HR = 1.96; 95% CI: 1.55-2.48; weight 68.12%; heterogeneity test, p = 0.115; I2 = 15.3%). A few articles showed that MMP-11 and -13 overexpression is associated with differences in OS. The combined HR showed that the overexpression of MMPs negatively afftected the disease-free time (DFS) (HR = 2.45; 95% CI: 1.49-4.05; heterogeneity test, p = 0.061; I2 = 50.2%). Overexpression of MMPs tends to worsen DFS. No publication bias was found in the current meta-analysis. CONCLUSION: MMP-9 and -2 overexpression was associated with poor prognostes in oral cancer patients. Therefore, early and accurate detection of MMP-9 and -2 levels can help improve the prognosis of MMP-9- and MMP-2-positive oral cancer patients.


Assuntos
Metaloproteinases da Matriz , Neoplasias Bucais , Biomarcadores , Progressão da Doença , Humanos , Prognóstico
9.
Oncol Rep ; 42(6): 2738-2749, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31638251

RESUMO

Circular RNAs (circRNAs) have emerged as important regulators of carcinogenesis. However, the role of circRNAs in oral squamous cell carcinoma (OSCC) remains limited. Here, total RNAs were extracted from three pairs of OSCC and adjacent normal tissues and subjected to circRNA microarrays to detect the differentially expressed circRNAs. Gene Ontology (GO) and functional category analyses were used to identify circRNAs associated with tumor cell proliferation pathways. Then, gain­of­function assays or loss­of­function assays were conducted to investigate the functions of the most upregulated and downregulated circRNAs on TSCC1 cell proliferation, cell cycle and apoptosis using CCK­8 and EdU assays, flow cytometry and Hoechst 33258 staining, respectively. The results revealed that hsa_circRNA_102459 was significantly downregulated and hsa_circRNA_043621 was significantly upregulated in OSCC tissues. Clinical stage, tumor differentiation, lymph node metastasis presented significant difference in regards to the expression of circRNA_043621 and circRNA_102459. The in vitro experiments further demonstrated that upregulation of circRNA_102459 or downregulation of circRNA_043621 significantly suppressed TSCC1 cell proliferation, induced cell cycle G0/G1 phase arrest and promoted apoptosis. Furthermore, the MAPK and PI3K/Akt pathways were suppressed, while Bcl­2 family members were activated by circRNA_102459 overexpression and circRNA_043621 knockdown. Taken together, our study indicates that differentially expressed circRNAs are closely related to the carcinogenesis of OSCC. Among these, circRNA_102459 and circRNA_043621 may function as a tumor­suppressor and promoter, respectively, of OSCC carcinogenesis, and thus may be valuable diagnostic biomarkers of OSCC.


Assuntos
Carcinoma de Células Escamosas/genética , Neoplasias Bucais/genética , RNA Circular/genética , Idoso , Carcinoma de Células Escamosas/epidemiologia , Carcinoma de Células Escamosas/patologia , Proliferação de Células/genética , Intervalo Livre de Doença , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Sistema de Sinalização das MAP Quinases/genética , Masculino , MicroRNAs/genética , Análise em Microsséries , Pessoa de Meia-Idade , Neoplasias Bucais/epidemiologia , Neoplasias Bucais/patologia , Fosfatidilinositol 3-Quinases/genética
10.
Biomed Pharmacother ; 109: 1709-1717, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30551425

RESUMO

BACKGROUND: Circular RNAs (circRNAs) comprise a novel class of noncoding RNAs that play important roles in a variety of diseases. However, the mechanism by which circRNAs regulate the osteogenic differentiation of maxillary sinus membrane stem cells (MSMSCs) remains largely unclear. METHODS: Microarray analysis was used to explore the expression profiles of circRNAs during the osteogenic differentiation of normal and BMP2 induced-MSMSCs. CircRNA_33287 was identified by agarose electrophoresis, quantitative real-time PCR (qRT-PCR), and western blotting. The function of circRNA_33287 was assessed by loss- and gain-of-function techniques and Alizarin red staining. Potential miRNA binding sites for circRNA_33287, and the target genes of miR-214-3p, were predicted by using online bioinformatics analysis tools. The relationships among the regulatory roles played by circRNA_33287, miR-214-3p, and Runt-related transcription factor 3 (Runx3), during the osteogenic differentiation of MSMSCs were verified by use of the dual luciferase reporter assay, qRT-PCR, and western blotting techniques, respectively. In addition, the molecular sponge potential of circRNA_33287 for miRNA was assessed via in vivo ectopic bone formation and a histological analysis performed after hematoxylin and eosin staining. RESULTS: Expression of circRNA_33287 was confirmed to be up-regulated during the osteogenic differentiation of MSMSCS. Overexpression and silencing of circRNA_33287 increased and decreased the expression levels of key markers of osteogenesis, respectively, including Runx2, OSX, and ALP. Furthermore, circRNA_33287 acted as a molecular sponge for miR-214-3p, which regulated Runx3 expression by targeting its 3'UTR. Moreover, circRNA_33287 protected Runx3 from miR-214-3p-mediated suppression. In addition, circRNA_33287 was shown to increase ectopic bone formation in vivo and displayed the strongest ability to stimulate bone formation when co-transfected with a miR-214-3p inhibitor. CONCLUSION: The novel pathway circRNA_33287/miR-214-3p/Runx3 was found to play a role in regulating the osteoblastic differentiation of MSMSCs in the posterior maxilla.


Assuntos
Subunidade alfa 3 de Fator de Ligação ao Core/biossíntese , Seio Maxilar/metabolismo , MicroRNAs/biossíntese , Osteogênese/fisiologia , RNA/biossíntese , Células-Tronco/metabolismo , Animais , Diferenciação Celular/fisiologia , Células Cultivadas , Células HEK293 , Humanos , Masculino , Seio Maxilar/citologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , RNA Circular , RNA Longo não Codificante/biossíntese
11.
Biochem Biophys Res Commun ; 503(2): 815-821, 2018 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-29913147

RESUMO

Long noncoding RNAs (lncRNAs) were proposed to be important regulators influencing various differentiation processes. Yet, the molecular mechanisms of lncRNAs governing osteogenic differentiation of Periodontal Ligament Stem Cells (PDLSCs) remain unclear. Here, PDLSCs were isolated from normal periodontal ligament of human (PDL) whereas P-PDLSCs were isolated from periodontitis affected PDL. Quantitative real-time PCR (qRT-PCR) was performed to examine the relative expression level of lncRNA-ANCR and of Osterix (OSX), Alkaline Phosphatase (ALP) as well as Runt-related transcription factor 2 (RUNX2) in PDLSCs. Gain- and loss-of- function experiments was performed to study the role of lncRNA-ANCR. Alizarin Red staining was used to evaluate the function of lncRNA-ANCR and miRNA-758 on osteogenic differentiation. In addition, via dual luciferase reporter assay and RNA immunoprecipitation the microRNA sponge potential of lncRNA-ANCR was assessed. A luciferase reporter assay identified the correlation between miR-758 and Notch2. Our results showed that the expression of ALP, RUNX2 and OSX were increased whereas lncRNA-ANCR was decreased during the process of differentiation in PDLSCs. Overexpression of lncRNA-ANCR decreased the expression of ALP, RUNX2 and OSX as confirmed by Alizarin red staining. Overexpression of lncRNA-ANCR resulted in reduction of the miR-758 expression level. Furthermore, RNA immunoprecipitation proved that lncRNA-ANCR targets miR-758 directly. The results of dual luciferase reporter assay also demonstrated that miR-758 regulated Notch2 expression by targeting 3'-UTR of Notch2. In conclusion, the novel pathway lncRNA-ANCR/miR-758/Notch2 plays an important role in the process of regulating osteogenic differentiation of PDLSCs.


Assuntos
Regulação da Expressão Gênica , MicroRNAs/genética , Osteogênese/genética , Ligamento Periodontal/citologia , RNA Longo não Codificante/genética , Células-Tronco/metabolismo , Regiões 3' não Traduzidas/genética , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Sequência de Bases , Diferenciação Celular/genética , Humanos , Receptor Notch2/genética , Receptor Notch2/metabolismo , Homologia de Sequência do Ácido Nucleico , Fator de Transcrição Sp7/genética , Fator de Transcrição Sp7/metabolismo , Via de Sinalização Wnt/genética
12.
Clin Implant Dent Relat Res ; 20(2): 110-121, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29106055

RESUMO

BACKGROUND: The function and the mechanism of long non-coding RNAs (lncRNAs) on the osteogenic differentiation of maxillary sinus membrane stem cells (MSMSCs) remain largely unknown. MATERIALS AND METHODS: The expression of lnc-NTF3-5 and Runt-related transcription factor 2 (RUNX2), Osterix (OSX), and Alkaline Phosphatase (ALP) was examined by quantitative real-time PCR (qRT-PCR) in MSMSCs during the process osteogenic differentiation. Then the function of lnc-NTF3-5 was evaluated by loss- and gain-of-function techniques, as well as qRT-PCR, western blot, and Alizarin Red staining. In addition, the microRNAs (miRNAs) sponge potential of lnc-NTF3-5 was assessed through RNA immunoprecipitation, dual luciferase reporter assay, and in vivo ectopic bone formation. RESULTS: Lnc-NTF3-5, RUNX2, OSX, and ALP increased alone with the differentiation. Inhibition of lnc-NTF3-5 decreased the expression of RUNX2, OSX, and ALP both at mRNA and protein levels. Alizarin red staining showed similar trend. In contrast, overexpression of lnc-NTF3-5 presented totally opposite effects. Besides, overexpression of lnc-NTF3-5 could decrease the expression of microRNA-93-3p (miR-93-3p). Enhance miR-93-3p could also inhibit the expression level of lnc-NTF3-5. RNA immunoprecipitation demonstrated that lnc-NTF3-5 is directly bound to miR-93-3p and dual luciferase reporter assay proved that miR-93-3p targets 3' UTR of RUNX2 to regulate its expression. Ultimately, in vivo bone formation study showed that lnc-NTF3-5 and miR-93-3p inhibitor co-transfection group displayed the strongest bone formation. CONCLUSIONS: The novel pathway lnc-NTF3-5/miR-93-3p/RUNX2 could regulate osteogenic differentiation of MSMSCs and might serve as a therapeutic target for bone regeneration in the posterior maxilla.


Assuntos
Diferenciação Celular/fisiologia , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Seio Maxilar/citologia , MicroRNAs/metabolismo , Osteoblastos/citologia , Fosfatase Alcalina/metabolismo , Células Cultivadas , Humanos , Redes e Vias Metabólicas , Células-Tronco Multipotentes/citologia , Osteogênese , RNA Longo não Codificante/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Fator de Transcrição Sp7/metabolismo
13.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-777769

RESUMO

Objective@#To investigate the osteogenic properties of maxillary sinus membrane stem cells (MSMSCs). @*Methods @#Beagle maxillary sinus mucosa was collected, immunomagnetic bead method was applied for isolation of CD146+ cells, and MSMSCs were harvested and cultured from the canine maxillary sinus floor mucosa. The levels of the cell surface antigens CD44, CD146, and CD34 were determined at passage one by flow cytometry. Cells at passage one were cultured in basal medium and osteogenic inductive medium. Real-time PCR, immunohistochemical staining, alkaline phosphatase activity, alizarin red staining and Von Kossa staining were used to investigate the osteogenic properties in vitro. @*Results@#The canine MSMSCs were cultured successfully. The results of flow cytometry were positive for CD146 and CD44 expression but negative for CD34 expression. The relative mRNA expression of runt-related transcription factor 2 (RUNX2) (t = 14.44,P < 0.001), osteopontin (OPN) (t = 7.85,P = 0.001) and alkaline phosphatase alkaline phosphatase (t = 14.27,P < 0.001) was apparently higher in the osteoinductive medium group than in the basal medium group, the differences in relative mRNA expression between the groups were significant. The protein levels of RUNX2 and OPN increased in the osteoinductive medium group. The alkaline phosphatase activity of the MSMSCs increased when the cells were cultured in osteoinductive medium; the activity increased to a level that was significantly higher than that in basal medium, particularly at days 3 (t = 8.79, P < 0.001), 7 (t = 9.75,P < 0.001), 14 (t = 12.14,P < 0.001), 21 (t = 19.62,P < 0.001) and 28 (t = 17.53,P < 0.001). Obvious mineralized nodules were observed by alizarin red staining or Von Kossa staining.@*Conclusion @#Maxillary sinus membrane stem cells exhibit osteogenic ability.

14.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-777739

RESUMO

Objective@#To detect the expression level of miR-27a during the osteogenic differentiation of beagle maxillary sinus membrane stem cells (MSMSCs) and explore the role of miR-27a in the osteogenic differentiation of MSMSCs.@*Methods@#Beagle MSMSCs were cultured in vitro. The expression level of miR-27a was detected via RT-PCR after an osteogenic inductive culture was prepared. The mRNA expression levels of Runx2 and OPN were examined via RT-PCR, and the protein expression levels of Runx2 and OPN were examined via Western blot after the cells were transfected with pre-miR-27a or anti-miR-27a. Finally, osteoprogenitor cells transfected with pre-miR-27a were composited with Bio-Oss particles and subcutaneously implanted into nude mice to form ectopic bone formation models, and then the inhibition of bone formation from miR-27a was observed in vivo. @*Results@#The expression level of miR-27a in the beagle MSMSCs decreased after osteogenic inductive culturing. The relative miR-27a levels were significantly decreased at day 1 (t=3.795, P=0.023), day 3 (t=4.493, P=0.011), day 7 (t=11.591, P < 0.001), day 14 (t=12.542, P < 0.001), and day 21 (t=5.621, P=0.008) compared with day 0. In addition, the expression levels of Runx2 mRNA (t=4.923, P=0.007) and protein (t=4.425, P=0.008) were reduced after the cells were transfected with pre-miR-27a. The expression levels OPN mRNA (t=5.253, P=0.006) and protein (t=5.132, P=0.006) were also reduced. In contrast, the mRNA expression levels of Runx2 (t=3.925, P=0.013) and OPN (t=3.712, P=0.019) were increased after the cells were transfected with anti-miR-27a, and bone formation was observed after the subcutaneous implantation of beagle MSMSCs composited with Bio-Oss in nude mice. Nevertheless, ectopic bone formation was inhibited by pre-miR-27a-transfected beagle MSMSCs composited with Bio-Oss (t=7.219, P=0.0020). @* Conclusion @# MiR-27a negatively regulates the osteogenic differentiation of MSMSCs.

15.
Implant Dent ; 26(4): 492-499, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28719571

RESUMO

OBJECTIVE: To explore the critical role and function of miRNAs in the regulation of development and physiology of maxillary sinus membrane stem cell (MSMSC) osteogenesis. METHODS: Microarray analysis was performed to screen the miRNAs expression profiles during the process of MSMSC osteogenic differentiation. Quantitative real-time polymerase chain reaction was applied to verify the miRNAs expression profiles. Gain- and loss-of-function experiments were used to demonstrate that miR-27b-3p inhibited MSMSC osteoblastic differentiation. Bioinformatic analysis was performed to predict the potential target of miR-27b-3p and then demonstrated by luciferase reporter assay and western blot. The negative regulation between miR-27b-3p and Sp7 was further confirmed using mimic and inhibitor of miR-27b-3p in vitro. Xenograft mice model was generated to confirm the relationship between miR-27b-3p and Sp7 using recombinant adenoviruses in vivo. RESULTS: MiR-27b-3p was downregulated during osteogenic differentiation of MSMSCs. The expression of Sp7, alkaline phosphatase, and osteocalcin decreased when transfected with miR-27b-3p-mimic in MSMSCs after osteogenic differentiation. MiR-27b-3p directly targeted Sp7 and inhibited the MSMSC osteogenesis in vivo. CONCLUSION: MiR-27b-3p suppressed the osteogenic differentiation of MSMSCs by directly inhibiting Sp7.


Assuntos
Diferenciação Celular/fisiologia , Seio Maxilar/citologia , MicroRNAs/metabolismo , Osteogênese/fisiologia , Fator de Transcrição Sp7/metabolismo , Células-Tronco/citologia , Fosfatase Alcalina/metabolismo , Animais , Western Blotting , Proliferação de Células , Regulação para Baixo , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Análise em Microsséries , Osteocalcina/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
16.
J Craniomaxillofac Surg ; 45(9): 1531-1534, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28687469

RESUMO

OBJECTIVE: To explore endoscope-assisted surgical treatment for idiopathic trigeminal neuralgia (TN), with neurectomy and avulsion of the inferior alveolar nerve (IAN), when patients are not willing to undergo intracranial procedures. CLINICAL MATERIALS AND METHODS: Five TN patients with inferior alveolar neuralgia in the mandibular branch were admitted; endoscope-assisted inferior alveolar neurectomy and avulsion was performed for TN treatment. Clinical observation and postoperative follow-up were conducted. RESULTS: After the operation, all patients had complete relief of pain, with mild postoperative swelling, limited mouth opening, and fast recovery. Patients were followed up for 3-24 months. Facial numbness and paresthesia were felt in the IAN distribution area of the operated side, with no recurrence. CONCLUSION: Endoscope-assisted neurectomy and avulsion of the IAN is a safe surgical method to treat TN in the mandibular branch when craniotomy is contraindicated or when patients decline neurosurgical options. It is effective in pain relief, with limited invasiveness and speedy recovery.


Assuntos
Denervação/métodos , Nervo Mandibular/cirurgia , Neuralgia do Trigêmeo/cirurgia , Idoso , Denervação/efeitos adversos , Endoscopia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Parestesia/etiologia , Complicações Pós-Operatórias
17.
Sci Rep ; 7: 46136, 2017 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-28387248

RESUMO

We recently reported that maxillary sinus membrane stem cells (MSMSCs) have osteogenic potential. However, the biological mechanisms of bone formation remain unclear. In this study, we investigated the role and mechanisms of microRNAs (miRNAs) in the osteogenic differentiation of MSMSCs. The expression of miRNAs was determined in differentiated MSMSCs by comprehensive miRNA microarray analysis and quantitative RT-PCR (qRT-PCR). We selected miR-1827 for functional follow-up studies to explore its significance in MSMSCs. Here, miR-1827 was found to be up-regulated during osteogenic differentiation of MSMSCs. Over expression of miR-1827 inhibited osteogenic differentiation of MSMSCs in vitro, whereas the repression of miR-1827 greatly promoted cell differentiation. Further experiments confirmed that insulin-like growth factor 1 (IGF1) is a direct target of miR-1827. miR-1827 inhibited osteogenic differentiation partially via IGF1, which in turn is a positive regulator of osteogenic differentiation. Moreover, miR-1827 suppressed ectopic bone formation and silencing of miR-1827 led to increased bone formation in vivo. In summary, this study is the first to demonstrate that miR-1827 can regulate osteogenic differentiation. The increase in miR-1827 expression observed during osteogenesis is likely a negative feedback mechanism, thus offering a potential therapeutic target to address inadequate bone volume for dental implantation through inhibiting miR-1827.


Assuntos
Diferenciação Celular/genética , Fator de Crescimento Insulin-Like I/metabolismo , Seio Maxilar/citologia , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/metabolismo , Osteogênese/genética , Animais , Sequência de Bases , Coristoma , Perfilação da Expressão Gênica , Inativação Gênica , Humanos , Fator de Crescimento Insulin-Like I/genética , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , Regulação para Cima/genética
18.
Implant Dent ; 26(2): 178-186, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28301382

RESUMO

OBJECTIVE: Although increasing evidence has shown that long noncoding RNAs (lncRNAs) play an important regulatory role in pluripotency and differentiation of mesenchymal stem cells, little is known about the role of lncRNA in maxillary sinus membrane stem cells (MSMSCs). The goal of this study was to investigate the expression profile and function of lncRNAs on osteogenic differentiation of MSMSCs. MATERIALS AND METHODS: By using lncRNA microarray, we identify a novel osteogenesis differentiation-related lncRNA of MSMSCs (lncRNA-MODR). The functional role of lncRNA-MODR in regulating osteogenesis was evaluated by quantitative real-time polymerase chain reaction, western blot, and alizarin red staining. Bioinformatic analyses of the predicted target genes (gene ontology, pathway, and network analysis) were applied for further study of lncRNA-MODR. RESULTS: We show that lncRNA-MODR is gradually upregulated during osteogenic differentiation. lncRNA-MODR overexpression upregulated, whereas lncRNA-MODR silencing decreased the expression of the osteogenic key marker, runt-related transcription factor 2 (RUNX2). In-depth analyses showed that lncRNA-MODR acts as a molecular sponge for microRNA-454 (miR-454) and that prevents RUNX2 from mi-454-mediated suppression. CONCLUSION: The lncRNAs act as a competing endogenous RNA to sequester microRNA-454 (miR-454), leading to heightened RUNX2 expression and thus promotes osteogenesis of MSMSCs.


Assuntos
Seio Maxilar/efeitos dos fármacos , MicroRNAs/metabolismo , Osteogênese/fisiologia , RNA Longo não Codificante/metabolismo , Células-Tronco/efeitos dos fármacos , Western Blotting , Diferenciação Celular/efeitos dos fármacos , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Humanos , Seio Maxilar/citologia , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase em Tempo Real , Células-Tronco/fisiologia
19.
Sci Rep ; 5: 11660, 2015 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-26119339

RESUMO

Maxillary sinus membrane (MSM) elevation is a common surgical technique for increasing bone height in the posterior maxilla prior to dental implant placement. However, the biological nature of bone regeneration in MSM remains largely unidentified. In this study, MSM tissue was obtained from 16 individuals during orthognathic surgery and used to isolate MSM stem cells (MSMSCs) by single-colony selection and STRO-1 cell sorting. The cell characteristics in terms of colony-forming ability, cell surface antigens, multi-differentiation potential and in vivo implantation were all evaluated. It was found that MSMSCs were of mesenchymal origin and positive for mesenchymal stem cell (MSC) markers such as STRO-1, CD146, CD29 and CD44; furthermore, under defined culture conditions, MSMSCs were able to form mineral deposits and differentiate into adipocytes and chondrocytes. When transplanted into immunocompromised rodents, MSMSCs showed the capacity to generate bone-like tissue and, importantly, maintain their MSC characteristics after in vivo implantation. These findings provide cellular and molecular evidence that MSM contains stem cells that show functional potential in bone regeneration for dental implant.


Assuntos
Seio Maxilar/citologia , Células-Tronco Multipotentes/citologia , Adipogenia , Adolescente , Células-Tronco Adultas/citologia , Células da Medula Óssea/citologia , Separação Celular , Condrogênese , Células Clonais , Polpa Dentária/citologia , Humanos , Imuno-Histoquímica , Imunofenotipagem , Seio Maxilar/diagnóstico por imagem , Células-Tronco Multipotentes/transplante , Osteogênese , Ligamento Periodontal/citologia , Radiografia , Transplante de Células-Tronco , Adulto Jovem
20.
J Craniomaxillofac Surg ; 43(1): 97-101, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25465489

RESUMO

PURPOSE: To discuss an effective surgical treatment of a subtype of trigeminal neuralgia with descending palatine neuralgia of the maxillary division. METHOD: Nine patients, who suffered from trigeminal neuralgia with descending palatine neuralgia of the maxillary division, received neurotomy and avulsion of the descending palatine nerve in the pterygopalatine fossa via the greater palatine foramen-pterygopalatine canal approach. Seven of the patients had a recurrence of descending palatine neuralgia after they received treatment of maxillary neuralgia with neurotomy and avulsion of the infraorbital nerve; two patients were diagnosed with descending palatine neuralgia of the maxillary division in our department. Postoperative follow-up was conducted. RESULTS: Pain in the palate disappeared; all patients felt numb and paresthetic in the area innervated by the trigeminal nerve, with no pain. During the 3-36 months of follow-up, no recurrence occurred. CONCLUSIONS: Descending palatine neurotomy in the pterygopalatine fossa via the greater palatine foramen-pterygopalatine canal approach is a simple, safe and effective way to treat a subtype of trigeminal neuralgia--descending palatine neuralgia.


Assuntos
Palato/inervação , Fossa Pterigopalatina/inervação , Neuralgia do Trigêmeo/cirurgia , Adulto , Idoso , Doenças dos Nervos Cranianos/cirurgia , Eletrocoagulação/métodos , Feminino , Seguimentos , Humanos , Masculino , Nervo Maxilar/cirurgia , Seio Maxilar/cirurgia , Pessoa de Meia-Idade , Neuralgia/cirurgia , Procedimentos Neurocirúrgicos/métodos , Órbita/inervação , Osteotomia/instrumentação , Palato/cirurgia , Palato Duro/inervação , Palato Duro/cirurgia , Fossa Pterigopalatina/cirurgia , Recidiva
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