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1.
Cell ; 158(2): 368-382, 2014 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-25036633

RESUMO

Adenomatous polyposis coli (APC) is a microtubule plus-end scaffolding protein important in biology and disease. APC is implicated in RNA localization, although the mechanisms and functional significance remain unclear. We show APC is an RNA-binding protein and identify an RNA interactome by HITS-CLIP. Targets were highly enriched for APC-related functions, including microtubule organization, cell motility, cancer, and neurologic disease. Among the targets is ß2B-tubulin, known to be required in human neuron and axon migration. We show ß2B-tubulin is synthesized in axons and localizes preferentially to dynamic microtubules in the growth cone periphery. APC binds the ß2B-tubulin 3' UTR; experiments interfering with this interaction reduced ß2B-tubulin mRNA axonal localization and expression, depleted dynamic microtubules and the growth cone periphery, and impaired neuron migration. These results identify APC as a platform binding functionally related protein and RNA networks, and suggest a self-organizing model for the microtubule to localize synthesis of its own subunits.


Assuntos
Proteína da Polipose Adenomatosa do Colo/metabolismo , Microtúbulos/metabolismo , Neurogênese , Proteínas de Ligação a RNA/metabolismo , Animais , Axônios/metabolismo , Sequência de Bases , Encéfalo/citologia , Encéfalo/metabolismo , Linhagem Celular , Movimento Celular , Gânglios Espinais/citologia , Estudo de Associação Genômica Ampla , Cones de Crescimento/metabolismo , Camundongos , Dados de Sequência Molecular , Neurônios/metabolismo , Mapas de Interação de Proteínas , RNA Mensageiro/metabolismo , Ratos , Alinhamento de Sequência , Tubulina (Proteína)/metabolismo
2.
Cell ; 153(6): 1185-7, 2013 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-23746834

RESUMO

The navigation of axons to their final destination can involve a sequence of steps that require different sets of guidance receptors. In this issue, Colak et al. show that regulated intra-axonal protein synthesis coupled to nonsense-mediated mRNA decay (NMD) controls a switch in Robo3.2 expression that is critical for navigation.


Assuntos
Axônios/metabolismo , Embrião de Mamíferos/metabolismo , Cones de Crescimento/metabolismo , Proteínas de Membrana/genética , Proteínas do Tecido Nervoso/genética , Degradação do RNAm Mediada por Códon sem Sentido , Medula Espinal/embriologia , Animais , Receptores de Superfície Celular
3.
Immunopharmacol Immunotoxicol ; 44(4): 574-585, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35485905

RESUMO

OBJECTIVE: This study aimed to explore the effect and mechanism of remifentanil on cardiopulmonary bypass (CPB)-induced cerebral nerve injury. METHODS: After pretreating with remifentanil, or dexmedetomidine (DEX), SD rats were subjected to the CPB for 2 h. The data of body temperature, blood gas and mean arterial pressure (MAP) and hematocrit (HCT) were recorded at different time points. The cerebral tissue water content of rats was determined and immunohistochemical (IHC) and H&E assays on the hippocampal CA1 region of rats was performed. The levels of interleukin (IL)-6, IL-10, soluble protein-100ß (S100ß) and neuron-specific enolase (NSE) were analyzed by ELISA, and those of the indexes for oxidative stress (malondialdehyde (MDA) and superoxide dismutase (SOD)) were detected by the commercial kits. Morris water maze was used to evaluate the learning and memory abilities. Western blot/qRT-PCR were used to detect the protein/mRNA expressions in hippocampus. RESULTS: CPB increased the levels/expressions of IL-6, IL-10, S100ß, NSE, MDA, cleaved caspase-3, Bax and decreased those of Bcl-2, SOD, p-AKT, HO-1, in serum and parietal cortex tissue, with increased brain water content, lesions in the hippocampal CA1 area, swimming distance, brain nerve injury and decreased escape latency, retention time on platform and times of crossing the platform of rats. The preconditioning of remifentanil or DEX partially attenuated CPB-induced injury and -decreased expressions on p-AKT and HO-1, while further promoting CPB-induced expression of nuclear Nrf2 expression and inhibiting that of cytoplasm Nrf2. CONCLUSION: This paper demonstrates that remifentanil preconditioning could partially attenuate CPB-induced brain nerve injury of rats.


Assuntos
Lesões Encefálicas , Fator 2 Relacionado a NF-E2 , Animais , Apoptose , Encéfalo/metabolismo , Ponte Cardiopulmonar/efeitos adversos , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos Sprague-Dawley , Remifentanil/farmacologia , Transdução de Sinais , Superóxido Dismutase/metabolismo
4.
BMC Surg ; 22(1): 221, 2022 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-35672718

RESUMO

BACKGROUND: The purpose of the present study was to evaluate the clinical effectiveness of ultrasonography-guided needle release of A1 pulley combined with corticosteroid injection by comparing it with ultrasound-guided needle release of the A1 pulley alone. METHODS: A total of 49 patients (55 fingers, thumb) with trigger fingers were included in this retrospective study. Twenty-seven fingers were treated with ultrasound-guided needle release of the A1 pulley alone (monotherapy group), and 28 fingers were treated with needle release of the A1 pulley combined with corticosteroid injection (combination group). Visual analog scale (VAS), Froimson scale, postoperative recurrence rate, and thickness of A1 pulley at baseline, Week-2, Week-12, and Month-6 were recorded. RESULTS: Higher clinical cure rates were observed in the combination group at Week-2 after treatment among patients with the Froimson scale Grade III and IV (p < 0.05). Among Froimson scale Grade IV patients, the combination group had a significantly thinner thickness of A1 pulley and better articular pain relief at Week-2 (all p < 0.05). No significant differences were found in the clinical cure rate, the thickness of the A1 pulley, articular pain relief, and recurrence rate between the two groups at Week-12 and Month-6 (all p > 0.05). CONCLUSIONS: Ultrasonography-guided needle release of A1 pulley plus corticosteroid injection was superior to ultrasonography-guided A1 pulley needle release alone during early-stage treatment of severe patients with trigger fingers. Moreover, ultrasonography-guided A1 pulley needle release combined with corticosteroid injection narrows the thickness of the A1 pulley. It is necessary to carry out preoperative evaluation and individualized treatment for patients of various severities.


Assuntos
Dedo em Gatilho , Corticosteroides/uso terapêutico , Humanos , Dor , Estudos Retrospectivos , Dedo em Gatilho/diagnóstico por imagem , Dedo em Gatilho/tratamento farmacológico , Dedo em Gatilho/cirurgia , Ultrassonografia , Ultrassonografia de Intervenção
5.
BMC Genomics ; 21(1): 64, 2020 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-31959126

RESUMO

BACKGROUND: The advent of Next Generation Sequencing has allowed transcriptomes to be profiled with unprecedented accuracy, but the high costs of full-length mRNA sequencing have posed a limit on the accessibility and scalability of the technology. To address this, we developed 3'Pool-seq: a simple, cost-effective, and scalable RNA-seq method that focuses sequencing to the 3'-end of mRNA. We drew from aspects of SMART-seq, Drop-seq, and TruSeq to implement an easy workflow, and optimized parameters such as input RNA concentrations, tagmentation conditions, and read depth specifically for bulk-RNA. RESULTS: Thorough optimization resulted in a protocol that takes less than 12 h to perform, does not require custom sequencing primers or instrumentation, and cuts over 90% of the costs associated with TruSeq, while still achieving accurate gene expression quantification (Pearson's correlation coefficient with ERCC theoretical concentration r = 0.96) and differential gene detection (ROC analysis of 3'Pool-seq compared to TruSeq AUC = 0.921). The 3'Pool-seq dual indexing scheme was further adapted for a 96-well plate format, and ERCC spike-ins were used to correct for potential row or column pooling effects. Transcriptional profiling of troglitazone and pioglitazone treatments at multiple doses and time points in HepG2 cells was then used to show how 3'Pool-seq could distinguish the two molecules based on their molecular signatures. CONCLUSIONS: 3'Pool-seq can accurately detect gene expression at a level that is on par with TruSeq, at one tenth of the total cost. Furthermore, its unprecedented TruSeq/Nextera hybrid indexing scheme and streamlined workflow can be applied in several different formats, including 96-well plates, which allows users to thoroughly evaluate biological systems under several conditions and timepoints. Care must be taken regarding experimental design and plate layout such that potential pooling effects can be accounted for and corrected. Lastly, further studies using multiple sets of ERCC spike-ins may be used to simulate differential gene expression in a system with known ground-state values.


Assuntos
RNA-Seq/métodos , Animais , Análise Custo-Benefício , Células Hep G2 , Humanos , Camundongos , Pioglitazona/farmacologia , RNA-Seq/economia , Transcriptoma/efeitos dos fármacos , Troglitazona/farmacologia
6.
J Virol ; 93(1)2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30333168

RESUMO

Type I interferon (IFN) inhibits viruses by inducing the expression of antiviral proteins. The IFN-induced myxovirus resistance B (MxB) protein has been reported to inhibit a limited number of viruses, including HIV-1 and herpesviruses, but its antiviral coverage remains to be explored further. Here we show that MxB interferes with RNA replication of hepatitis C virus (HCV) and significantly inhibits viral replication in a cyclophilin A (CypA)-dependent manner. Our data further show that MxB interacts with the HCV protein NS5A, thereby impairing NS5A interaction with CypA and NS5A localization to the endoplasmic reticulum, two events essential for HCV RNA replication. Interestingly, we found that MxB significantly inhibits two additional CypA-dependent viruses of the Flaviviridae family, namely, Japanese encephalitis virus and dengue virus, suggesting a potential link between virus dependence on CypA and virus susceptibility to MxB inhibition. Collectively, these data have identified MxB as a key factor behind IFN-mediated suppression of HCV infection, and they suggest that other CypA-dependent viruses may also be subjected to MxB restriction.IMPORTANCE Viruses of the Flaviviridae family cause major illness and death around the world and thus pose a great threat to human health. Here we show that IFN-inducible MxB restricts several members of the Flaviviridae, including HCV, Japanese encephalitis virus, and dengue virus. This finding not only suggests an active role of MxB in combating these major pathogenic human viruses but also significantly expands the antiviral spectrum of MxB. Our study further strengthens the link between virus dependence on CypA and susceptibility to MxB restriction and also suggests that MxB may employ a common mechanism to inhibit different viruses. Elucidating the antiviral functions of MxB advances our understanding of IFN-mediated host antiviral defense and may open new avenues to the development of novel antiviral therapeutics.


Assuntos
Ciclofilina A/farmacologia , Hepacivirus/fisiologia , Interferons/farmacologia , Proteínas de Resistência a Myxovirus/metabolismo , Proteínas não Estruturais Virais/metabolismo , Replicação Viral/efeitos dos fármacos , Animais , Linhagem Celular , Chlorocebus aethiops , Ciclosporina/farmacologia , Retículo Endoplasmático/metabolismo , Técnicas de Silenciamento de Genes , Células HEK293 , Hepacivirus/efeitos dos fármacos , Humanos , Proteínas de Resistência a Myxovirus/genética , Ligação Proteica/efeitos dos fármacos , Células Vero
7.
J Xray Sci Technol ; 28(3): 573-581, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32116288

RESUMO

OBJECTIVE: To compare the clinical effectiveness of ultrasound-guided corticosteroid injection with and without needle release of the A1 pulley in treating trigger finger. METHODS: A total of 60 patients with trigger finger were enrolled in this retrospective study. Among them, 30 patients were treated with ultrasound-guided needle release of the A1 pulley with corticosteroid injection (group A) and 30 patients were treated with single ultrasound-guided corticosteroids injection (group B). The following parameters were evaluated including clinical parameters (pain degree, function of joint, finger tendon function, postoperative satisfaction), and ultrasound parameter (thickness of A1 pulley). RESULTS: The postoperative visual analogue scale (VAS) and Quinnell scores in two groups were significantly lower than that before operation (p < 0.05). The postoperative Quinnell score of group A was significantly lower than that in group B (p < 0.05). The TAM results showed that the postoperative overall excellent and good rate of group A was significantly higher than that in group B (p < 0.05). The postoperative survey showed that more than 80% patients reported satisfaction in the two groups. The ultrasound imaging results showed that the postoperative thickness of A1 pulley in two groups were thinner than that before operation (p < 0.05). There were no adverse effects and complications in the two groups. CONCLUSIONS: Both approaches had treatment benefit in trigger finger. Ultrasound-guided needle release of the A1 pulley with corticosteroid injection had better treatment benefits than single ultrasound-guided corticosteroids injection in improving finger tendon function and joint function.


Assuntos
Corticosteroides , Dedo em Gatilho , Ultrassonografia de Intervenção/métodos , Corticosteroides/administração & dosagem , Corticosteroides/uso terapêutico , Idoso , Feminino , Dedos/diagnóstico por imagem , Dedos/cirurgia , Humanos , Injeções , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Resultado do Tratamento , Dedo em Gatilho/diagnóstico por imagem , Dedo em Gatilho/tratamento farmacológico , Dedo em Gatilho/cirurgia
8.
Development ; 143(15): 2753-9, 2016 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-27385015

RESUMO

RNA-based regulatory mechanisms play important roles in the development and plasticity of neural circuits and neurological disease. Developing axons provide a model well suited to the study of RNA-based regulation, and contain specific subsets of mRNAs that are locally translated and have roles in axon pathfinding. However, the RNA-binding proteins involved in axon pathfinding, and their corresponding mRNA targets, are still largely unknown. Here we find that the RNA-binding protein IMP2 (Igf2bp2) is strikingly enriched in developing axon tracts, including in spinal commissural axons. We used the HITS-CLIP approach to perform a genome-wide identification of RNAs that interact directly with IMP2 in the native context of developing mouse brain. This IMP2 interactome was highly enriched for mRNA targets related to axon guidance. Accordingly, IMP2 knockdown in the developing spinal cord led to strong defects in commissural axon trajectories at the midline intermediate target. These results reveal a highly distinctive axonal enrichment of IMP2, show that it interacts with a network of axon guidance-related mRNAs, and reveal that it is required for normal axon pathfinding during vertebrate development.


Assuntos
Axônios/metabolismo , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/metabolismo , Medula Espinal/citologia , Animais , Orientação de Axônios/genética , Orientação de Axônios/fisiologia , Axônios/fisiologia , Embrião de Galinha , Eletroporação , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Camundongos , RNA Mensageiro/genética , Proteínas de Ligação a RNA/genética
9.
PLoS Pathog ; 13(9): e1006625, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28938017

RESUMO

Enterovirus 71 (EV71) is the major causative agent of hand, foot and mouth disease (HFMD) in children, causing severe clinical outcomes and even death. Here, we report an important role of the highly conserved alanine residue at position 107 in the capsid protein VP1 (VP1A107) in the efficient replication of EV71. Substitutional mutations of VP1A107 significantly diminish viral growth kinetics without significant effect on viral entry, expression of viral genes and viral production. The results of mechanistic studies reveal that VP1A107 regulates the efficient cleavage of the VP0 precursor during EV71 assembly, which is required, in the next round of infection, for the transformation of the mature virion (160S) into an intermediate or A-particle (135S), a key step of virus uncoating. Furthermore, the results of molecular dynamic simulations and hydrogen-bond networks analysis of VP1A107 suggest that flexibility of the VP1 BC loop or the region surrounding the VP1107 residue directly correlates with viral infectivity. It is possible that sufficient flexibility of the region surrounding the VP1107 residue favors VP0 conformational change that is required for the efficient cleavage of VP0 as well as subsequent viral uncoating and viral replication. Taken together, our data reveal the structural role of the highly conserved VP1A107 in regulating EV71 maturation. Characterization of this novel determinant of EV71 virulence would promote the study on pathogenesis of Enteroviruses.


Assuntos
Enterovirus Humano A/fisiologia , Infecções por Enterovirus/virologia , Células Vero/virologia , Replicação Viral/genética , Aminoácidos/genética , Animais , Proteínas do Capsídeo/metabolismo , Chlorocebus aethiops , Mutação de Sentido Incorreto/genética , Internalização do Vírus
10.
J Neuroinflammation ; 15(1): 142, 2018 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-29759062

RESUMO

BACKGROUND: Acute neurological insults caused by infection, systemic inflammation, ischemia, or traumatic injury are often associated with breakdown of the blood-brain barrier (BBB) followed by infiltration of peripheral immune cells, cytotoxic proteins, and water. BBB breakdown and extravasation of these peripheral components into the brain parenchyma result in inflammation, oxidative stress, edema, excitotoxicity, and neurodegeneration. These downstream consequences of BBB dysfunction can drive pathophysiological processes and play a substantial role in the morbidity and mortality of acute and chronic neurological insults, and contribute to long-term sequelae. Preserving or rescuing BBB integrity and homeostasis therefore represents a translational research area of high therapeutic potential. METHODS: Induction of general and localized BBB disruption in mice was carried out using systemic administration of LPS and focal photothrombotic ischemic insult, respectively, in the presence and absence of the monoacylglycerol lipase (MAGL) inhibitor, CPD-4645. The effects of CPD-4645 treatment were assessed by gene expression analysis performed on neurovascular-enriched brain fractions, cytokine and inflammatory mediator measurement, and functional assessment of BBB permeability. The mechanism of action of CPD-4645 was studied pharmacologically using inverse agonists/antagonists of the cannabinoid receptors CB1 and CB2. RESULTS: Here, we demonstrate that the neurovasculature exhibits a unique transcriptional signature following inflammatory insults, and pharmacological inhibition of MAGL using a newly characterized inhibitor rescues the transcriptional profile of brain vasculature and restores its functional homeostasis. This pronounced effect of MAGL inhibition on blood-brain barrier permeability is evident following both systemic inflammatory and localized ischemic insults. Mechanistically, the protective effects of the MAGL inhibitor are partially mediated by cannabinoid receptor signaling in the ischemic brain insult. CONCLUSIONS: Our results support considering MAGL inhibitors as potential therapeutics for BBB dysfunction and cerebral edema associated with inflammatory brain insults.


Assuntos
Ácidos Araquidônicos/antagonistas & inibidores , Ácidos Araquidônicos/metabolismo , Barreira Hematoencefálica/metabolismo , Lesões Encefálicas/tratamento farmacológico , Lesões Encefálicas/metabolismo , Permeabilidade Capilar/fisiologia , Endocanabinoides/antagonistas & inibidores , Endocanabinoides/metabolismo , Glicerídeos/antagonistas & inibidores , Glicerídeos/metabolismo , Animais , Barreira Hematoencefálica/efeitos dos fármacos , Lesões Encefálicas/induzido quimicamente , Permeabilidade Capilar/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/uso terapêutico , Hidrólise/efeitos dos fármacos , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Monoacilglicerol Lipases/antagonistas & inibidores , Monoacilglicerol Lipases/metabolismo
11.
J Biol Chem ; 291(39): 20692-706, 2016 09 23.
Artigo em Inglês | MEDLINE | ID: mdl-27451391

RESUMO

Cell migration is orchestrated by dynamic interactions of microtubules with the plasma membrane cortex. How these interactions facilitate these dynamic processes is still being actively investigated. TIP150 is a newly characterized microtubule plus end tracking protein essential for mitosis and entosis (Ward, T., Wang, M., Liu, X., Wang, Z., Xia, P., Chu, Y., Wang, X., Liu, L., Jiang, K., Yu, H., Yan, M., Wang, J., Hill, D. L., Huang, Y., Zhu, T., and Yao, X. (2013) Regulation of a dynamic interaction between two microtubule-binding proteins, EB1 and TIP150, by the mitotic p300/CBP-associated factor (PCAF) orchestrates kinetochore microtubule plasticity and chromosome stability during mitosis. J. Biol. Chem. 288, 15771-15785; Xia, P., Zhou, J., Song, X., Wu, B., Liu, X., Li, D., Zhang, S., Wang, Z., Yu, H., Ward, T., Zhang, J., Li, Y., Wang, X., Chen, Y., Guo, Z., and Yao, X. (2014) Aurora A orchestrates entosis by regulating a dynamic MCAK-TIP150 interaction. J. Mol. Cell Biol. 6, 240-254). Here we show that TIP150 links dynamic microtubules to steer cell migration by interacting with cortactin. Mechanistically, TIP150 binds to cortactin via its C-terminal tail. Interestingly, the C-terminal TIP150 proline-rich region (CT150) binds to the Src homology 3 domain of cortactin specifically, and such an interaction is negatively regulated by EGF-elicited tyrosine phosphorylation of cortactin. Importantly, suppression of TIP150 or overexpression of phospho-mimicking cortactin inhibits polarized cell migration. In addition, CT150 disrupts the biochemical interaction between TIP150 and cortactin in vitro, and perturbation of the TIP150-cortactin interaction in vivo using a membrane-permeable TAT-CT150 peptide results in an inhibition of directional cell migration. We reason that a dynamic TIP150-cortactin interaction orchestrates directional cell migration via coupling dynamic microtubule plus ends to the cortical cytoskeleton.


Assuntos
Movimento Celular/fisiologia , Cortactina/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Microtúbulos/metabolismo , Cortactina/genética , Células HEK293 , Humanos , Proteínas Associadas aos Microtúbulos/genética , Microtúbulos/genética , Ligação Proteica , Domínios de Homologia de src
12.
BMC Genomics ; 17: 39, 2016 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-26747388

RESUMO

BACKGROUND: RNA sequencing (RNA-seq), a next-generation sequencing technique for transcriptome profiling, is being increasingly used, in part driven by the decreasing cost of sequencing. Nevertheless, the analysis of the massive amounts of data generated by large-scale RNA-seq remains a challenge. Multiple algorithms pertinent to basic analyses have been developed, and there is an increasing need to automate the use of these tools so as to obtain results in an efficient and user friendly manner. Increased automation and improved visualization of the results will help make the results and findings of the analyses readily available to experimental scientists. RESULTS: By combing the best open source tools developed for RNA-seq data analyses and the most advanced web 2.0 technologies, we have implemented QuickRNASeq, a pipeline for large-scale RNA-seq data analyses and visualization. The QuickRNASeq workflow consists of three main steps. In Step #1, each individual sample is processed, including mapping RNA-seq reads to a reference genome, counting the numbers of mapped reads, quality control of the aligned reads, and SNP (single nucleotide polymorphism) calling. Step #1 is computationally intensive, and can be processed in parallel. In Step #2, the results from individual samples are merged, and an integrated and interactive project report is generated. All analyses results in the report are accessible via a single HTML entry webpage. Step #3 is the data interpretation and presentation step. The rich visualization features implemented here allow end users to interactively explore the results of RNA-seq data analyses, and to gain more insights into RNA-seq datasets. In addition, we used a real world dataset to demonstrate the simplicity and efficiency of QuickRNASeq in RNA-seq data analyses and interactive visualizations. The seamless integration of automated capabilites with interactive visualizations in QuickRNASeq is not available in other published RNA-seq pipelines. CONCLUSION: The high degree of automation and interactivity in QuickRNASeq leads to a substantial reduction in the time and effort required prior to further downstream analyses and interpretation of the analyses findings. QuickRNASeq advances primary RNA-seq data analyses to the next level of automation, and is mature for public release and adoption.


Assuntos
Sequenciamento de Nucleotídeos em Larga Escala/métodos , Análise de Sequência de RNA/métodos , Software , Transcriptoma/genética , Algoritmos , Sequência de Bases , RNA/genética
13.
J Neurosci ; 34(1): 66-78, 2014 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-24381269

RESUMO

Developing axons can locally synthesize proteins, with roles in axon growth, guidance, and regeneration, but the mechanisms that regulate axonal mRNA translation are not well understood. MicroRNAs (miRNAs) are important regulators of translation but have still been little characterized in developing axons. Here we study mouse dorsal root ganglion (DRG) axons and show that their extension is impaired by conditional deficiency of the miRNA-processing enzyme Dicer in vitro and in vivo. A screen for axonal localization identifies a specific set of miRNAs preferentially enriched within the developing axon. High axonal expression and preferential localization were observed for miR-132, a miRNA previously known for roles in dendrites and dysregulation in major neurologic diseases. miR-132 knockdown reduced extension of cultured DRG axons, whereas overexpression increased extension. Mechanistically, miR-132 regulated the mRNA for the Ras GTPase activator Rasa1, a novel target in neuronal function. Moreover, miR-132 regulation of Rasa1 translation was seen in severed axons, demonstrating miRNA function locally within the axon. miR-132 expression in DRGs peaked in the period of maximum axon growth in vivo, consistent with its effect on axon growth, and suggesting a role as a developmental timer. Together, these findings identify miR-132 as a positive regulator of developing axon extension, acting through repression of Rasa1 mRNA, in a mechanism that operates locally within the axon.


Assuntos
Axônios/fisiologia , Gânglios Espinais/crescimento & desenvolvimento , MicroRNAs/fisiologia , RNA Mensageiro/fisiologia , Proteína p120 Ativadora de GTPase/fisiologia , Animais , Axotomia , Células Cultivadas , Feminino , Masculino , Camundongos , Camundongos da Linhagem 129 , Camundongos Transgênicos
14.
BMC Genomics ; 16: 675, 2015 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-26334759

RESUMO

BACKGROUND: While RNA-sequencing (RNA-seq) is becoming a powerful technology in transcriptome profiling, one significant shortcoming of the first-generation RNA-seq protocol is that it does not retain the strand specificity of origin for each transcript. Without strand information it is difficult and sometimes impossible to accurately quantify gene expression levels for genes with overlapping genomic loci that are transcribed from opposite strands. It has recently become possible to retain the strand information by modifying the RNA-seq protocol, known as strand-specific or stranded RNA-seq. Here, we evaluated the advantages of stranded RNA-seq in transcriptome profiling of whole blood RNA samples compared with non-stranded RNA-seq, and investigated the influence of gene overlaps on gene expression profiling results based on practical RNA-seq datasets and also from a theoretical perspective. RESULTS: Our results demonstrated a substantial impact of stranded RNA-seq on transcriptome profiling and gene expression measurements. As many as 1751 genes in Gencode Release 19 were identified to be differentially expressed when comparing stranded and non-stranded RNA-seq whole blood samples. Antisense and pseudogenes were significantly enriched in differential expression analyses. Because stranded RNA-seq retains strand information of a read, we can resolve read ambiguity in overlapping genes transcribed from opposite strands, which provides a more accurate quantification of gene expression levels compared with traditional non-stranded RNA-seq. In the human genome, it is not uncommon to find genomic loci where both strands encode distinct genes. Among the over 57,800 annotated genes in Gencode release 19, there are an estimated 19 % (about 11,000) of overlapping genes transcribed from the opposite strands. Based on our whole blood mRNA-seq datasets, the fraction of overlapping nucleotide bases on the same and opposite strands were estimated at 2.94 % and 3.1 %, respectively. The corresponding theoretical estimations are 3 % and 3.6 %, well in agreement with our own findings. CONCLUSIONS: Stranded RNA-seq provides a more accurate estimate of transcript expression compared with non-stranded RNA-seq, and is therefore the recommended RNA-seq approach for future mRNA-seq studies.


Assuntos
Perfilação da Expressão Gênica , Genes , Análise de Sequência de RNA/métodos , Moléculas de Adesão Celular/genética , Humanos , Interleucinas/genética , Masculino , Reprodutibilidade dos Testes
15.
Biomedicines ; 12(7)2024 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-39061952

RESUMO

Neuropathic pain (NP) results from lesions or diseases affecting the peripheral or central somatosensory system. However, there are currently no drugs that are particularly effective in treating this condition. SKI306X is a blend of purified extracts of three oriental herbs (Clematis mandshurica, Trichosanthes kirilowii, and Prunella vulgaris) commonly used to treat osteoarthritis for their chondroprotective effects. Chronic postischemic pain (CPIP) and spinal nerve ligation (SNL) models were created by binding the upper left ankle of mice with an O-ring for 3 h and ligating the L5 spinal nerve, respectively. Mice with allodynia were injected intraperitoneally with 0.9% normal saline (NS group) or different doses (25, 50, or 100 mg/kg) of SKI306X (SKI groups). We assessed allodynia using von Frey filaments before injection and 30, 60, 90, 120, 180, and 240 min and 24 h after injection to confirm the antiallodynic effect of SKI306X. We also measured glial fibrillary acidic protein (GFAP) levels in the spinal cord and dorsal root ganglia to confirm the change of SKI306X administration. Both models exhibited significant mechanical allodynia. The intraperitoneal injection of SKI306X significantly increased the paw withdrawal threshold in a dose-dependent manner, as the paw withdrawal threshold was significantly increased after SKI306X administration compared with at baseline or after NS administration. GFAP levels in the SKI group decreased significantly (p < 0.05). Intraperitoneal administration of SKI306X dose-dependently attenuated mechanical allodynia and decreased GFAP levels, suggesting that GFAP is involved in the antiallodynic effect of SKI306X in mice with CPIP and SNL-induced NP.

16.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 44(2): 295-9, 2013 Mar.
Artigo em Zh | MEDLINE | ID: mdl-23745276

RESUMO

OBJECTIVE: To investigate the methodology and influential factors of real-time elastography (RTE) in liver examination. METHODS: Forty normal volunteers received the examination of liver with RTE. All strain images were analyzed by the Strain Histogram Measurement and the liver fibrosis index (LFI) values were recorded. Two-tailed t-test was used to evaluate the significance of the potential influence factors of RTE, including inter-lobar variations, respiratory phase, different sections and gender. A paired two-tailed t-test and Bland-Altman test were used in the analysis of the inter- and intra-observer consistency. RESULTS: There were significant differences between the LFI values in the left lobe and those in the right lobe (2.52 +/- 0.47 vs. 1.58 +/- 0.41), also between right intercostal and right sub-costal approach (1.58 +/- 0.41 vs. 1. 59 +/- 0.45). There were no significant differences either between the LFI values of end-expiration and those of end-inspiration (2.61 +/- 0.54 vs. 1.58 +/- 0.41) or between male and female (1.57 +/- 0.37 vs. 1.60 +/- 0.46). RTE showed goodness of fit between the inter- and intra-observer consistency. CONCLUSION: Liver stiffness measurement performed by RTE at end-inspiration in the right lobe with inter-costal approach may reveal liver elasticity more accurately.


Assuntos
Técnicas de Imagem por Elasticidade/métodos , Elasticidade , Fígado/diagnóstico por imagem , Adulto , Área Sob a Curva , Feminino , Voluntários Saudáveis , Humanos , Masculino , Respiração , Adulto Jovem
17.
World Neurosurg ; 180: e506-e513, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37774790

RESUMO

PURPOSE: To determine the reliability of dynamic magnetic resonance imaging (MRI) perfusion parameters for the evaluation of blood supply to spinal metastatic tumors. METHODS: A total of 36 patients with spinal metastasis who underwent dynamic contrast-enhanced magnetic resonance spinal perfusion imaging at Tianjin Hospital from December 2018 to December 2020 were reviewed. Subsequently, the patients underwent corresponding preoperative examination using digital subtraction angiography of the spine at the hospital and were divided into 2 groups accordingly. Differences in dynamic MRI perfusion parameters between the 2 groups were analyzed. RESULTS: There were statistically significant differences between the 2 groups in the quantitative dynamic contrast-enhanced MRI perfusion parameters vascular permeability and plasma volume, as well as semi-quantitative peak enhancement and blood flow ratio parameters. CONCLUSIONS: Dynamic MRI perfusion may distinguish spinal metastatic lesions with rich blood supply from those with poor blood supply and may help clinicians identify patients that can benefit from invasive spinal angiography and preoperative embolization. This technique may also provide guidance on decision taking for surgery basing on dynamic MRI perfusion parameters.


Assuntos
Meios de Contraste , Neoplasias , Humanos , Reprodutibilidade dos Testes , Imageamento por Ressonância Magnética/métodos , Angiografia Digital/métodos , Perfusão
18.
J Org Chem ; 77(2): 1148-53, 2012 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-22191387

RESUMO

The construction of a new family of endo-functionalized multiferrocenyl hexagons with various sizes via coordination-driven self-assembly is described. The structures of these novel metallacycles, containing several ferrocenyl moieties at their interior surface, are characterized by multinuclear NMR ((31)P and (1)H) spectroscopy, cold-spray ionization mass spectrometry (CSI-TOF-MS), elemental analysis, and molecular modeling. Insight into the structural and electrochemical properties of these endo-functionalized multiferrocenyl hexagons was obtained through cyclic voltammetry investigation.


Assuntos
Complexos de Coordenação/química , Compostos de Ferro/química , Complexos de Coordenação/síntese química , Cristalografia por Raios X , Eletroquímica , Compostos Ferrosos/química , Compostos de Ferro/síntese química , Espectroscopia de Ressonância Magnética , Metalocenos , Estrutura Molecular , Espectrometria de Massas por Ionização por Electrospray
19.
J Org Chem ; 77(7): 3426-32, 2012 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-22428775

RESUMO

A new family of 60° dendritic di-Pt(II) acceptor tectons have been successfully designed and synthesized, from which a series of novel "three-component" triangular metallodendrimers were prepared via [3 + 3] coordination-driven self-assembly. The structures of newly designed triangular metallodendrimers are characterized by multinuclear NMR ((1)H and (31)P), (1)H DOSY NMR, mass spectrometry (CSI-TOF-MS), and elemental analysis. The shape and size of all supramolecular dendritic triangles were investigated with PM6 semiempirical molecular orbital methods.


Assuntos
Dendrímeros/química , Dendrímeros/síntese química , Compostos Organoplatínicos/química , Compostos Organoplatínicos/síntese química , Cristalografia por Raios X , Espectroscopia de Ressonância Magnética , Modelos Moleculares
20.
J Chem Phys ; 136(11): 114701, 2012 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-22443784

RESUMO

The adsorption and reaction behaviors of HF on the α-Al(2)O(3)(0001) surface are systematically investigated using density functional theory method. By increasing the number of HF molecules in a p(2 × 1) α-Al(2)O(3)(0001) slab, we find that HF is chemically dissociated at low coverage; while both physical and dissociative adsorption occurs at a 3/2 monolayer (ML) coverage. At the same coverage (1.0 ML), diverse configurations of the dissociated HF are obtained in the p(2 × 1) model; while only one is observed in the p(1 × 1) slab due to its smaller surface area compared with the former one. Preliminary fluorination reaction study suggests that the total energy of two dissociated HF in the p(2 × 1) slab increases by 1.00 and 0.72 eV for the formation and desorption of water intermediate, respectively. The coadsorption behaviors of HF and H(2)O indicate that the pre-adsorbed water is unfavorable for the fluorination of Al(2)O(3), which is well consistent with the experimental results. The calculated density of states show that the peak of σ(H-F) disappears, while the peaks of σ(H-O) and σ(Al-F) are observed at -8.4 and -5 to -3 eV for the dissociated HF. Charge density difference analysis indicates that the dissociated F atom attracts electrons, while no obvious changes on electrons are observed for the surface Al atoms.

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