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1.
Artigo em Inglês | MEDLINE | ID: mdl-33166473

RESUMO

Rationale Cross-sectional human data suggest that enrichment of oral anaerobic bacteria in the lung is associated with increased Th17 inflammatory phenotype. In this study we evaluated the microbial and host immune response dynamics after aspiration with a oral commensals using a preclinical mouse model. Methods Aspiration with a mixture of human oral commensals (MOC; Prevotella melaninogenica, Veillonella parvula, and Streptococcus mitis) was modeled in mice followed by variable time of sacrifice. Genetic background of mice included WT, MyD88 knock out and STAT3C. Measurements 16S rRNA gene sequencing characterized changes in microbiota. Flow cytometry, cytokine measurement via Luminex and RNA host transcriptome sequencing was used to characterize host immune phenotype. Main Results While MOC aspiration correlated with lower airway dysbiosis that resolved within five days, it induced an extended inflammatory response associated with IL17-producing T-cells lasting at least 14 days. MyD88 expression was required for the IL-17 response to MOC aspiration, but not for T-cell activation or IFN-γ expression. MOC aspiration prior to a respiratory challenge with S. pneumoniae led to a decreased in host's susceptibility to this pathogen. Conclusions Thus, in otherwise healthy mice, a single aspiration event with oral commensals are rapidly cleared from the lower airways, but induce a prolonged Th17 response that secondarily decreased susceptibility to respiratory pathogens. Translationally, these data implicate an immuno-protective role of episodic microaspiration of oral microbes in the regulation of the lung immune phenotype and mitigation of host susceptibility to infection with lower airway pathogens.

2.
J Int Med Res ; 48(10): 300060520962388, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33032480

RESUMO

OBJECTIVE: This study aimed to examine the characteristics of blood lactate in neonates undergoing mechanical ventilation in Tibet. METHODS: We recruited 67 neonates undergoing mechanical ventilation in Naqu People's Hospital as the plateau observation group and 94 neonates undergoing mechanical ventilation in Shengjing Hospital as the control group. We analyzed the differences in lactate levels between the two groups. RESULTS: The lactate clearance rates of neonates with asphyxia and those with respiratory distress syndrome were significantly lower in the plateau group than in the control group. Lactate levels in neonates who died in the plateau group were significantly higher and the lactate clearance rate was significantly lower than those in neonates who survived. The cut-off point for the lactate clearance rate at 6 hours for predicting mortality was 6.09% in the plateau group. CONCLUSION: The lactate clearance rate of neonates on mechanical ventilation in the plateau area is lower than that in neonates in the non-plateau area. The lactate clearance rate at 6 hours is important for evaluating the prognoses of critical neonates in plateau areas.

3.
Cancer Sci ; 111(11): 4288-4302, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32945042

RESUMO

Non-alcoholic fatty liver disease (NAFLD) is one of the most common causes of hepatocellular carcinoma (HCC), but the underlying mechanisms behind the correlation of NAFLD with HCC are unclear. We aimed to uncover the genes and potential mechanisms that drive this progression. This study uncovered the genes and potential mechanisms through a multiple 'omics integration approach. Quantitative proteomics combined with phenotype-association analysis was performed. To investigate the potential mechanisms, a comprehensive transcriptome/lipidome/phenome-wide association analysis was performed in genetic reference panel BXD mice strains. The quantitative proteomics combined with phenotype-association results showed that VDAC1 was significantly increased in tumor tissues and correlated with NAFLD-related traits. Gene co-expression network analysis indicated that VDAC1 is involved in mitochondria dysfunction in the tumorigenic/tumor progression. The association between VDAC1 and mitochondria dysfunction can be explained by the fact that VDAC1 was associated with mitochondria membrane lipids cardiolipin (CL) composition shift. VDAC1 was correlated with the suppression of mature specie CL(LLLL) and elevation level of nascent CL species. Such profiling shift was supported by the significant positive correlation between VDAC1 and PTPMT1, as well as negative correlation with CL remodeling enzyme Tafazzin (TAZ). This study confirmed that the expression of VADC1 was dysregulated in NAFLD-driven HCC and associated with NAFLD progression. The VDAC1-driven mitochondria dysfunction is associated with cardiolipin composition shift, which causes alteration of mitochondria membrane properties.


Assuntos
Carcinoma Hepatocelular/etiologia , Neoplasias Hepáticas/etiologia , Mitocôndrias/genética , Mitocôndrias/metabolismo , Hepatopatia Gordurosa não Alcoólica/complicações , Hepatopatia Gordurosa não Alcoólica/genética , Canais de Ânion Dependentes de Voltagem/genética , Idoso , Animais , Carcinoma Hepatocelular/diagnóstico , Modelos Animais de Doenças , Suscetibilidade a Doenças , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Variação Genética , Humanos , Neoplasias Hepáticas/diagnóstico , Masculino , Camundongos , Pessoa de Meia-Idade , Gradação de Tumores , Estadiamento de Neoplasias , Hepatopatia Gordurosa não Alcoólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , Proteoma , Proteômica/métodos , Locos de Características Quantitativas , Transdução de Sinais , Canais de Ânion Dependentes de Voltagem/metabolismo
4.
Cancer Manag Res ; 12: 5893-5902, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32765090

RESUMO

Purpose: The expression of programmed death-ligand 1 (PD-L1) is common in various solid human cancers and it is an important therapeutic target. However, the expression pattern, clinical significance and potential mechanism of PD-L1 in hypopharyngeal squamous cell carcinoma (HSCC) are still lacking. Methods: PD-L1 expression in HSCC tumor tissues and paired adjacent hypopharyngeal mucosal tissues was detected using immunohistochemistry assay, and the clinical significance of PD-L1 in HSCC was characterized. In vitro assays including cell viability assays, migration assays, invasion assays as well as Western blot assays were performed to illuminate the biological functions and underlying molecular mechanisms of PD-L1 in HSCC development. Results: PD-L1 expression was detected in HSCC samples but we found no positive expression in matched normal hypopharyngeal mucosal tissues. The levels of PD-L1 expression were significantly correlated with advanced clinical progression and poor patient survival. Multivariable analysis of Cox model showed that PD-L1 expression was an independent predictor for the prognosis of HSCC patients. Functional experiments showed that the ectopic expression of PD-L1 markedly influenced the proliferation, migration and invasion of FaDu cells in vitro. Mechanistically, investigations demonstrated that PD-L1 could promote the epithelial-mesenchymal transition of FaDu cells. Meanwhile, PD-L1 knockdown inhibited, while PD-L1 overexpression activated the Akt-mTOR signaling pathway in FaDu cells. The EMT induced by PD-L1 overexpression could be reversed by the Akt inhibitor. Conclusion: In summary, the expression of PD-L1 can act as a significant biomarker for the adverse clinicopathological features and poor prognosis of patients with HSCC. PD-L1 can promote the proliferation, migration and invasion of FaDu cells and consequently enhance the aggressiveness. Moreover, PD-L1 induces EMT through AKT-mTOR signaling pathway. These suggest that PD-L1 has important tumor-intrinsic functions independent of its immunopathogenic effects.

5.
Exp Cell Res ; 395(2): 112234, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32822723

RESUMO

Skeletal muscle preservation is a dynamic process that involves constant repair and regeneration. However, the regenerative capacity of muscle cells declines in hyperglycemia. This study aimed to explore the molecular mechanisms underlying this glucotoxicity during myoblast differentiation. C2C12 cells were exposed to different concentrations of glucose, to recapitulate the development of skeletal muscles in vivo in normo- and hyperglycemic conditions. In high glucose conditions, we found significant increases in levels of total cellular reactive oxygen species (ROS) and a reorganization of SUMO enzyme transcripts and SUMOylated proteins. Furthermore, in anticipation of the ROS-induced damage to newly formed myotubes, we observed acceleration of myogenesis. Interestingly, we found a tight relationship between SUMOylation of the Histone methyltransferase SET7/9 and the maintenance of sarcomeric structures of newly formed myotubes. Finally, treatment with the antioxidant anacardic acid preserved the function and activity of myotubes generated in high-glucose conditions by interfering with both ROS and SUMO pathways. Combined, these results suggest that increased oxidative stress and modulation of SUMO reactions are key mediators of glucotoxicity and inhibition of these perturbations using antioxidants might improve muscle regeneration in hyperglycemia.

6.
J Mol Cell Cardiol ; 146: 60-68, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32668281

RESUMO

Genetic lineage tracing has been widely used for studying in vivo cell fate plasticity during embryogenesis, tissue homeostasis, and disease development. Recent applications with multiple site-specific recombinases have been used in complex and sophisticated genetic fate mapping studies. However, the previous multicolor reporters for dual recombinases had limitations of precise in situ quantification of cell number, which is mainly due to the intermingling of cells in condensed tissues. Here, we generated a dual recombinase-mediated nuclear-localized GFP and tdTomato reporter line, which enables clear, simultaneous quantification of two distinct cell lineages in vivo. Combining this dual genetic reporter with Tbx18-Cre and Cdh5-Dre lines, which genetically trace epicardial and endothelial cells, respectively, we obtained high-resolution images for the anatomic distribution of the descendants of these two distinct cell lineages in the valve mesenchyme during development, remodeling, and maturation stages. This new dual genetic reporter is expected to facilitate fate tracing of two cell lineages and their objective quantification in vivo.

7.
Biosci Rep ; 40(6)2020 06 26.
Artigo em Inglês | MEDLINE | ID: mdl-32490526

RESUMO

Chronic intermittent hypoxia (CIH) induced by sleep-disordered breathing (SDB) is a key factor involved in cognitive dysfunction (CD). Increasing evidence has shown that atractylon (ATR) has anti-inflammatory effects. However, it remains unclear if ATR has a protective effect against SDB-induced nerve cell injury and CD. So, in the present study, CIH-exposed mice and CIH-induced BV2 cells were used to mimic SDB. The results showed that ATR treatment decreased CIH-induced CD and the expression of inflammatory factors in the hippocampal region by suppression of M1 microglial activation and promotion of M2 microglial activation. Also, ATR treatment promoted sirtuin 3 (SIRT3) expression. Down-regulation of SIRT3 decreased the protective effect of ATR against CIH-induced microglial cell injury. Furthermore, in vitro detection found that SIRT3 silencing suppressed ATR-induced M2 microglial activation after exposure to CIH conditions. Taken together, these results indicate that ATR treatment prevents SDB-induced CD by inhibiting CIH-induced M1 microglial activation, which is mediated by SIRT3 activation.

8.
Bioorg Chem ; 97: 103670, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32088417

RESUMO

Selective COX-2 inhibitor celecoxib was found directly inhibiting the growth of tested phytopathogenic fungi with the inhibitory rate ranging from 30 to 40% at 100 µg/ml. Lead optimization of celecoxib led to the identification of compound 12 among its derivatives as the most active antifungal candidate. The antifungal effect of compound 12 was supposed to be independent of COX-2 inhibition. Transcriptome profiling analysis of Fusarium graminearium (PH-1) treated with compound 12 brought about 406 up-regulated and 572 down-regulated differentially express genes (DEGs) respectively.

9.
EMBO J ; 39(4): e102675, 2020 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-31943281

RESUMO

Site-specific recombinase-mediated genetic technology, such as inducible Cre-loxP recombination (CreER), is widely used for in vivo genetic manipulation with temporal control. The Cre-loxP technology improves our understanding on the in vivo function of specific genes in organ development, tissue regeneration, and disease progression. However, inducible CreER often remains inefficient in gene deletion. In order to improve the efficiency of gene manipulation, we generated a self-cleaved inducible CreER (sCreER) that switches inducible CreER into a constitutively active Cre by itself. We generated endocardial driver Npr3-sCreER and fibroblast driver Col1a2-sCreER, and compared them with conventional Npr3-CreER and Col1a2-CreER, respectively. For easy-to-recombine alleles such as R26-tdTomato, there was no significant difference in recombination efficiency between sCreER and the conventional CreER. However, for alleles that were relatively inert for recombination such as R26-Confetti, R26-LZLT, R26-GFP, or VEGFR2flox/flox alleles, sCreER showed a significantly higher efficiency in recombination compared with conventional CreER in endocardial cells or fibroblasts. Compared with conventional CreER, sCreER significantly enhances the efficiency of recombination to induce gene expression or gene deletion, allowing temporal yet effective in vivo genomic modification for studying gene function in specific cell lineages.


Assuntos
Integrases/genética , Recombinação Genética , Alelos , Animais , Linhagem da Célula , Feminino , Fibroblastos , Deleção de Genes , Expressão Gênica , Integrases/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos
10.
J Biol Chem ; 295(3): 690-700, 2020 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-31771978

RESUMO

Genetic lineage tracing is widely used to study organ development and tissue regeneration. Multicolor reporters are a powerful platform for simultaneously tracking discrete cell populations. Here, combining Dre-rox and Cre-loxP systems, we generated a new dual-recombinase reporter system, called Rosa26 traffic light reporter (R26-TLR), to monitor red, green, and yellow fluorescence. Using this new reporter system with the three distinct fluorescent reporters combined on one allele, we found that the readouts of the two recombinases Cre and Dre simultaneously reflect Cre+Dre-, Cre-Dre+, and Cre+Dre+ cell lineages. As proof of principle, we show specific labeling in three distinct progenitor/stem cell populations, including club cells, AT2 cells, and bronchoalveolar stem cells, in Sftpc-DreER;Scgb1a1-CreER;R26-TLR mice. By using this new dual-recombinase reporter system, we simultaneously traced the cell fate of these three distinct cell populations during lung repair and regeneration, providing a more comprehensive picture of stem cell function in distal airway repair and regeneration. We propose that this new reporter system will advance developmental and regenerative research by facilitating a more sophisticated genetic approach to studying in vivo cell fate plasticity.

11.
Exp Cell Res ; 386(2): 111746, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31778670

RESUMO

Hypopharyngeal squamous cell carcinoma (HSCC) is a relatively rare malignancy and has the worst prognosis among head and neck cancer. Metastasis is the major cause of poor prognosis in HSCC patients. In this study, we found that 3-phosphoinositide-dependent protein kinase 1 (PDK1 or PDPK1) was overexpressed in HSCC. The overexpression was positively correlated lymph node metastasis, clinical stage, and distant metastasis and indicated poor outcome. Loss and gain-of-function revealed that PDK1 increased cell proliferation, migration and invasion in vitro as well as tumor growth and metastasis in vivo. Mechanically, PDK1 induced epithelial-mesenchymal transition and promoted metastasis by activating the Notch1 signaling pathway. We further illustrated that PDK1 bound with the Notch1 intracellular domain, thereby inhibiting its ubiquitin-mediated degradation in a protein kinase B (Akt-) independent manner. In summary, PDK1/Notch1 axis played an important role in HSCC metastasis, and this investigation provided a new perspective on potential therapeutic targets for HSCC.

12.
Arch Biochem Biophys ; 680: 108239, 2020 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-31881189

RESUMO

c-Met receptor is frequently overexpressed in hepatocellular carcinoma and thus considered as an attractive target for pharmacological intervention with small molecule tyrosine kinase inhibitors. Albeit with the development of multiple c-Met inhibitors, none reached clinical application in the treatment of hepatoma so far. To improve the efficacy of c-Met inhibitors towards hepatocellular carcinoma, we investigated the combined effects of the dynamin inhibitor dynasore with several c-Met inhibitors, including tivantinib, PHA-665752, and JNJ-38877605. We provide several lines of evidence that dynasore enhanced the inhibitory effects of these inhibitors on hepatoma cell proliferation and migration, accompanied with increased cell cycle arrest and apoptosis. Mechanically, the combinatorial treatments decreased c-Met levels and hence markedly disrupted downstream signaling, as revealed by the dramatically declined phosphorylation of AKT and MEK. Taken together, our findings demonstrate that the candidate agent dynasore potentiated the inhibitory effects of c-Met inhibitors against hepatoma cells and will shed light on the development of novel therapeutic strategies to target c-Met in the clinical management of hepatocellular carcinoma patients.


Assuntos
Antineoplásicos/farmacologia , Carcinoma Hepatocelular/tratamento farmacológico , Hidrazonas/farmacologia , Neoplasias Hepáticas/tratamento farmacológico , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-met/antagonistas & inibidores , Carcinoma Hepatocelular/metabolismo , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Hep G2 , Humanos , Neoplasias Hepáticas/metabolismo , Proteínas Proto-Oncogênicas c-met/metabolismo
13.
J Exp Med ; 217(1)2020 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-31699822

RESUMO

Blood-brain barrier (BBB) dysfunction has been suggested to play an important role in epilepsy. However, the mechanism mediating the transition from cerebrovascular damage to epilepsy remains unknown. Here, we report that endothelial cyclin-dependent kinase 5 (CDK5) is a central regulator of neuronal excitability. Endothelial-specific Cdk5 knockout led to spontaneous seizures in mice. Knockout mice showed increased endothelial chemokine (C-X-C motif) ligand 1 (Cxcl1) expression, decreased astrocytic glutamate reuptake through the glutamate transporter 1 (GLT1), and increased glutamate synaptic function. Ceftriaxone restored astrocytic GLT1 function and inhibited seizures in endothelial Cdk5-deficient mice, and these effects were also reversed after silencing Cxcl1 in endothelial cells and its receptor chemokine (C-X-C motif) receptor 2 (Cxcr2) in astrocytes, respectively, in the CA1 by AAV transfection. These results reveal a previously unknown link between cerebrovascular factors and epileptogenesis and provide a rationale for targeting endothelial signaling as a potential treatment for epilepsy.


Assuntos
Quimiocina CXCL1/metabolismo , Quinase 5 Dependente de Ciclina/metabolismo , Células Endoteliais/metabolismo , Epilepsia/metabolismo , Gliose/metabolismo , Receptores de Interleucina-8B/metabolismo , Animais , Astrócitos/metabolismo , Astrócitos/patologia , Barreira Hematoencefálica/metabolismo , Células Cultivadas , Células Endoteliais/patologia , Epilepsia/patologia , Gliose/patologia , Ácido Glutâmico/metabolismo , Camundongos , Camundongos Knockout , Neurônios/metabolismo , Neurônios/patologia , Convulsões/metabolismo , Convulsões/patologia , Transdução de Sinais/fisiologia
14.
Biomed Phys Eng Express ; 6(2): 027002, 2020 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-33438644

RESUMO

OBJECTIVE: Parkinson's disease (PD) is a neurodegenerative disease for which there is no known cure. Deep brain stimulation (DBS) is a surgical treatment effective in reducing motor symptoms for PD patients. Previous work implicates DBS may directly influence motor cortex through stochastic antidromic spikes originating from the site of stimulation. Here we tested the hypothesis that direct randomized cortical stimulation is therapeutically effective in PD animal models. APPROACH: As a proof-of-principle study, we utilized a multi-channel stimulating system to mimic the effects of stochastic antidromic activation on the motor cortex of rat, by delivering microcurrents randomized temporally and spatially, and assessed the efficacy in ameliorating motor symptoms in a rat PD model. MAIN RESULTS: We found that different combinations of frequency, amplitude and pulse width of randomized electrical currents delivered to the motor cortex exerted different effects on Parkinsonian rats. Among these, some stimulus patterns, defined by specific ranges of pulse width and stimulation frequencies, were able to produce transient beneficial effect on locomotive ability assessed by open-field locomotor activities. These results indicate that, in principle, cortical stimulation could achieve therapeutic outcome in PD. SIGNIFICANCE: Direct cortical simulation based on a randomized protocol could be a less invasive approach than standard DBS in treating Parkinsonism. More refined mode of stimulation to achieve long-lasting and more robust effect should be explored.

15.
Front Pediatr ; 7: 476, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31803698

RESUMO

Objective: To explore the characteristics of mechanical ventilation parameters and the arterial partial pressure of oxygen in neonatal respiratory distress syndrome (RDS) at high altitude. Methods: From the 1st May 2017 to the 31st December 2018, we recruited 33 neonates with severe RDS who were undergoing mechanical ventilation in the NICU of Naqu People's Hospital in Tibet (4,580 m above sea level); these neonates formed a plateau observation group. We also recruited a non-plateau control group: 66 neonates with severe RDS undergoing mechanical ventilation of Shengjing Hospital in Liaoning (51 m above sea level). Various ventilation parameters and the arterial partial pressure of oxygen were then compared between the two groups, between the survivors of the two groups, and between those who died and survived in the plateau group. Results: In terms of initial ventilator parameters, peak inspiratory pressure (PIP), positive end expiratory pressure (PEEP), and the fraction of inspired oxygen (FiO2) in the plateau group were significantly higher than those in the non-plateau group (P < 0.01). PIP, PEEP, and FiO2 in the survivors from the plateau group were also significantly higher than those in the non-plateau group (P < 0.01). In addition, the arterial partial pressure of oxygen in the non-plateau group was higher (P < 0.05) than that in the plateau group during the early postnatal period, and the arterial partial pressure of oxygen at 6 and 12 h was lower than that in the plateau group (P < 0.05). Conclusion: Mechanical ventilation can effectively improve the arterial oxygen partial pressure and reduce the mortality of newborns with RDS in a plateau environment. It was clearly evident that ventilation parameters are closely related to altitude. It is therefore not advisable to apply mechanical ventilation parameters used in a non-plateau area as a guide for the treatment of newborns with RDS in plateau areas.

16.
Int J Biochem Cell Biol ; 117: 105640, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31689531

RESUMO

The tyrosine kinase receptor ErbB2 is frequently found to be overexpressed in multiple cancer types. Targeted therapeutic approaches against ErbB2 have shown promising results and received FDA approvals in the treatment of breast cancer. However, this approach has not been granted in ovarian cancers till now. In order to assess the validity of ErbB2-targeted therapy in ovarian cancer, we investigated the effectiveness of two FDA-approved tyrosine kinase inhibitors of ErbB2, lapatinib and neratinib, on the growth of ovarian cancers. We observed that both lapatinib and neratinib displayed inhibitory effects towards the proliferation and migration of ErbB2-positive ovarian cancer cells in vitro, with neratinib showing stronger suppression in general. Neratinib treatment led to the reduction of ErbB2 protein levels, with concomitant attenuation of the phosphorylation of AKT, MEK, and ERK1/2. Immunofluorescence assays revealed that neratinib induced the internalization and lysosomal degradation of ErbB2, which was accompanied by its hyperubiquitylation. Lapatinib and neratinib also repressed the in vivo growth of SKOV3 cells, and neratinib downregulated ErbB2 levels in xenograft tumors to cause potent inhibition. Therefore, the ubiquitylation-mediated endocytic degradation of ErbB2 incurred by neratinib treatment conferred potent inhibition of ovarian cancer growth. Clinical investigations of neratinib in ErbB2-positive ovarian cancer are warranted.

17.
J Neurosci Methods ; 328: 108441, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31574288

RESUMO

BACKGROUND: Recombinant adeno-associated virus (rAAV) is increasingly applied in neuroscience research or gene therapy. However, there is no simple and efficient tool for specific transfection of rAAV into cerebrovascular tissues. It has been reported that fluorescent tracers or beta-amyloid protein can enter the brain through perivascular spaces, named as "glymphatic system". The purpose of this study was to explore whether rAAV could transduce the cerebral vasculature through the glymphatic pathway. NEW METHOD: An AAV1-GFP vector suspension (15 µL) was injected into the intracisternal space of anesthetized mice (n = 2) and 5 µl was injected into the bulbus medullae (n = 2). As controls, 15 µl of artificial cerebrospinal fluid (aCSF) was injected into the cisterna magna. The endothelial specific transduction was verified by Glut1 or PDGFRß immunofluorescent staining. Immunofluorescence images for all groups were captured with a laser microscope. RESULTS: It was observed that infection with rAAV1 vectors encoding green fluorescence protein resulted in a successful cerebrovascular transduction when injected into cisterna magna, compared to aCSF or intra-parenchymal injection at 30 days post-transduction in adult mice. In addition, GFP was co-localized with Glut1 based on immuno-fluorescence. These results indicate that glymphatic system delivery enhances the transduction efficiency of AAV1 to brain endothelial cells. COMPARISON WITH EXISTING METHODS: The AAV1 vector can simply and efficiently transduce the cerebral endothelial cells through the glymphatic pathway. CONCLUSION: The findings of this study reveal that rAAV1-based vectors have high application potential for endothelial-targeted neurologic disease research or gene-based therapies.

20.
Int J Oncol ; 55(2): 488-498, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31268159

RESUMO

Fascin­1 is an actin­bundling protein, which specifically interacts with F­actin to form parallel actin bundles, and participates in the regulation of cell adhesion, interactions and migration. However, the expression and regulatory mechanisms of fascin­1 in hypopharyngeal squamous cell carcinoma (HSCC) remain poorly understood. The present study investigated the effects and underlying molecular mechanism of fascin­1 on the invasion and metastasis of HSCC. The results demonstrated that fascin­1 was overexpressed and correlated with lymph node metastasis and tumor­node­metastasis stage in HSCC tissues. Further in vitro study revealed that fascin­1 promoted cell morphology polarization to increase the motility of FaDu cells. In addition, fascin­1 significantly promoted the migration and invasion of FaDu cells. At the molecular level, fascin­1 promoted cell invasion and migration by upregulating matrix metalloproteinase­2 (MMP­2) expression in FaDu cells. Immunohistochemical analysis revealed that a correlation existed between hypoxia inducible factor (HIF)­1α and fascin­1 expression in the HSCC tissues. Furthermore, the results from a cobalt chloride­induced hypoxia model demonstrated that fascin­1 may be upregulated by HIF­1α in FaDu cells. Further analysis revealed that fascin­1 knockdown significantly decreased the invasion of cells under hypoxia and partially reversed hypoxia­induced MMP­2 expression under hypoxia in FaDu cells. In conclusion, fascin­1 was upregulated by HIF­1α, and promoted the invasion and migration of HSCC cells; therefore, fascin­1 may provide a potential target for the treatment of invasion and metastasis in HSCC.


Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/patologia , Proteínas de Transporte/metabolismo , Movimento Celular , Neoplasias Hipofaríngeas/patologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Neoplasias Pulmonares/secundário , Proteínas dos Microfilamentos/metabolismo , Apoptose , Carcinoma de Células Escamosas/metabolismo , Proliferação de Células , Feminino , Seguimentos , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hipofaríngeas/metabolismo , Neoplasias Pulmonares/metabolismo , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Prognóstico , Transdução de Sinais , Células Tumorais Cultivadas
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