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The presence of adherens junctions and the associated protein ß-catenin are requirements for the development of glucose-stimulated insulin secretion (GSIS) in ß-cells. Evidence indicates that modulation of ß-catenin function in response to changes in glucose levels can modulate the levels of insulin secretion from ß-cells but the role of ß-catenin phosphorylation in this process has not been established. We find that a Ser552Ala version of ß-catenin attenuates glucose-stimulated insulin secretion indicating a functional role for Ser552 phosphorylation of ß-catenin in insulin secretion. This is associated with alterations F/G actin ratio but not the transcriptional activity of ß-catenin. Both glucose and GLP-1 stimulated phosphorylation of the serine 552 residue on ß-catenin. We investigated the possibility that an EPAC-PAK1 pathway might be involved in this phosphorylation event. We find that reduction in PAK1 levels using siRNA attenuates both glucose and GLP-1 stimulated phosphorylation of ß-catenin Ser552 and the effects of these on insulin secretion in ß-cell models. Furthermore, both the EPAC inhibitor ESI-09 and the PAK1 inhibitor IPA3 do the same in both ß-cell models and mouse islets. Together this identifies phosphorylation of ß-catenin at Ser552 as part of a cell signalling mechanism linking nutrient and hormonal regulation of ß-catenin to modulation of insulin secretory capacity of ß-cells and indicates this phosphorylation event is regulated downstream of EPAC and PAK1 in ß-cells.
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Fatores de Troca do Nucleotídeo Guanina/genética , Células Secretoras de Insulina/metabolismo , Insulina/genética , Ilhotas Pancreáticas/metabolismo , beta Catenina/genética , Quinases Ativadas por p21/genética , Actinas/genética , Actinas/metabolismo , Junções Aderentes/efeitos dos fármacos , Junções Aderentes/metabolismo , Animais , Linhagem Celular Transformada , Dissulfetos/farmacologia , Regulação da Expressão Gênica , Peptídeo 1 Semelhante ao Glucagon/farmacologia , Glucose/metabolismo , Glucose/farmacologia , Fatores de Troca do Nucleotídeo Guanina/antagonistas & inibidores , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Hidrazonas/farmacologia , Insulina/metabolismo , Células Secretoras de Insulina/citologia , Células Secretoras de Insulina/efeitos dos fármacos , Ilhotas Pancreáticas/citologia , Ilhotas Pancreáticas/efeitos dos fármacos , Isoxazóis/farmacologia , Masculino , Camundongos , Naftóis/farmacologia , Fosforilação , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Ratos , Transdução de Sinais , Técnicas de Cultura de Tecidos , beta Catenina/metabolismo , Quinases Ativadas por p21/antagonistas & inibidores , Quinases Ativadas por p21/metabolismoRESUMO
Electroencephalogram (EEG) analysis has been widely used in the diagnosis of stroke diseases for its low cost and noninvasive characteristics. In order to classify the EEG signals of stroke patients with cerebral infarction and cerebral hemorrhage, this paper proposes a novel EEG stroke signal classification method. This method has two highlights. The first is that a multi-feature fusion method is given by combining wavelet packet energy, fuzzy entropy and hierarchical theory. The second highlight is that a suitable classification model based on ensemble classifier is constructed for perfectly classification stroke signals. Entropy is an accessible way to measure information and uncertainty of time series. Many entropy-based methods have been developed these years. By comparing with the performances of permutation entropy, sample entropy, approximate entropy in measuring the characteristic of stroke patient's EEG signals, it can be found that fuzzy entropy has best performance in characterization stroke EEG signal. By combining hierarchical theory, wavelet packet energy and fuzzy entropy, a multi-feature fusion method is proposed. The method first calculates wavelet packet energy of EEG stroke signal, then extracts hierarchical fuzzy entropy feature by combining hierarchical theory and fuzzy entropy. The experimental results show that, compared with the fuzzy entropy feature, the classification accuracy based on the fusion feature of wavelet packet energy and hierarchical fuzzy entropy is much higher than benchmark methods. It means that the proposed multi-feature fusion method based on stroke EEG signal is an efficient measure in classifying ischemic and hemorrhagic stroke. Support vector machine (SVM), decision tree and random forest are further used as the stroke signal classification models for classifying ischemic stroke and hemorrhagic stroke. Experimental results show that, based on the proposed multi-feature fusion method, the ensemble method of random forest can get the best classification performance in accuracy among three models.
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Algoritmos , Eletroencefalografia/métodos , Acidente Vascular Cerebral/classificação , Acidente Vascular Cerebral/diagnóstico por imagem , Lógica Fuzzy , Humanos , Reconhecimento Automatizado de Padrão , Processamento de Sinais Assistido por Computador , Acidente Vascular Cerebral/fisiopatologia , Análise de OndaletasRESUMO
Autism is a severe neurodevelopmental disorder with a large population prevalence, characterized by abnormal reciprocal social interactions, communication deficits, and repetitive behaviors with restricted interests. The BTBR T(+)Itpr3(tf) (BTBR) mice have emerged as strong candidates to serve as models of a range of autism-relevant behaviors. Increasing evidences suggest that interleukin (IL)-6, one of the most important neuroimmune factors, was involved in the pathophysiology of autism. It is of great importance to further investigate whether therapeutic interventions in autism can be achieved through the manipulation of IL-6. Our previous studies showed that IL-6 elevation in the brain could mediate autistic-like behaviors, possibly through the imbalances of neural circuitry and impairments of synaptic plasticity. In this study, we evaluate whether inhibiting IL-6 signaling in the brain is sufficient to modulate the autism-like behaviors on the BTBR mice. The results showed that chronic infusion of an analog of the endogenous IL-6 trans-signaling blocker sgp130Fc protein increased the sociability in BTBR mice. Furthermore, no change was observed in the number of excitatory synapse, level of synaptic proteins, density of dentitic spine and postsynaptic density in BTBR cortices after inhibiting IL-6 trans-signaling. However, inhibition of IL-6 trans-signaling increased the evoked glutamate release in synaptoneurosomes from the cerebral cortex of BTBR mice. Our findings suggest that inhibition of excessive production of IL-6 may have selective therapeutic efficacy in treating abnormal social behaviors in autism.
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Transtorno Autístico/metabolismo , Comportamento Animal , Córtex Cerebral/metabolismo , Interleucina-6/metabolismo , Plasticidade Neuronal , Animais , Transtorno Autístico/tratamento farmacológico , Transtorno Autístico/genética , Transtorno Autístico/patologia , Córtex Cerebral/patologia , Receptor gp130 de Citocina/uso terapêutico , Modelos Animais de Doenças , Humanos , Fragmentos Fc das Imunoglobulinas/farmacologia , Interleucina-6/antagonistas & inibidores , Interleucina-6/genética , Camundongos , Camundongos Transgênicos , Densidade Pós-Sináptica/genética , Densidade Pós-Sináptica/metabolismo , Densidade Pós-Sináptica/patologia , Proteínas Recombinantes/farmacologia , Transdução de SinaisRESUMO
BACKGROUND: Cerebralcare Granule (CG) is a polyherbal Chinese medicine that has been shown to have neuroprotective effects in experimental models of stroke. We compared the efficacy and safety of CG with aspirin in patients with acute stroke. METHODS: For this open-label, controlled trial, we recruited patients with angiographically confirmed strokes and US National Institutes of Health Stroke Scale (NIHSS) scores of 4-22 within 2 weeks of symptom onset; recruitment was performed at 55 sites in China. Patients received CG or aspirin. The primary efficacy end-point was neurological function. Analyses were done by intention to treat. Patients were measured for NIHSS, Montreal Cognitive Assessment, and Mini-Mental State Examination scores and Barthel index at baseline and at 4, 8, and 12 weeks after treatment. RESULTS: Between January 2013 and January 2014, we treated 1963 patients with CG and 1288 patients with aspirin. Baseline NIHSS, Mini-Mental State Examination, and Montreal Cognitive Assessment scores were comparable between the two groups. Patients in the CG group had a greater improvement than the aspirin group in terms of NIHSS (P < 0.01) and Barthel index at 4, 8, and 12 weeks. At 12 weeks, patients in the CG group had a greater improvement than the aspirin group in terms of Mini-Mental State Examination (P < 0.01) and Montreal Cognitive Assessment (P < 0.05). Adverse reactions were similar between the two groups. CONCLUSIONS: This large-scale, controlled trial indicated that CG may be a useful treatment in the management of post-stroke patients.
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Aspirina/uso terapêutico , Cognição/efeitos dos fármacos , Medicamentos de Ervas Chinesas/administração & dosagem , Fármacos Neuroprotetores/uso terapêutico , Acidente Vascular Cerebral/tratamento farmacológico , Idoso , China , Medicamentos de Ervas Chinesas/efeitos adversos , Feminino , Humanos , Acontecimentos que Mudam a Vida , Masculino , Pessoa de Meia-Idade , Fármacos Neuroprotetores/efeitos adversos , Testes Neuropsicológicos , Inibidores da Agregação Plaquetária/uso terapêutico , Estudos Prospectivos , Resultado do TratamentoRESUMO
Autism spectrum disorder (ASD) is a developmental disorder characterized by impairments in social and communication abilities, as well as by restricted and repetitive behaviors. The BTBR T + Itpr3 tf (BTBR) mice have emerged as a well characterized and widely used mouse model of a range of ASD-like phenotype, showing deficiencies in social behaviors and unusual ultrasonic vocalizations as well as increased repetitive self-grooming. However, the inherited neurobiological changes that lead to ASD-like behaviors in these mice are incompletely known and still under active investigation. The aim of this study was to further evaluate the structure and neurotransmitter release of the glutamatergic synapse in BTBR mice. C57BL/6J (B6) mice were used as a control strain because of their high level of sociability. The important results showed that the evoked glutamate release in the cerebral cortex of BTBR mice was significantly lower than in B6 mice. And the level of vesicle docking-related protein Syntaxin-1A was reduced in BTBR mice. However, no significant changes were observed in the number of glutamatergic synapse, level of synaptic proteins, density of dendritic spine and postsynaptic density between BTBR mice and B6 mice. Overall, our results suggest that abnormal vesicular glutamate activity may underlie the ASD relevant pathology in the BTBR mice.
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Transtorno Autístico/metabolismo , Comportamento Animal/fisiologia , Espinhas Dendríticas/metabolismo , Comportamento Social , Transmissão Sináptica/fisiologia , Animais , Transtorno Autístico/fisiopatologia , Modelos Animais de Doenças , Ácido Glutâmico/metabolismo , CamundongosRESUMO
Chinese scorpion Buthus martensii Karsch (BmK) venom is a rich source of neurotoxins which bind to various ion channels with high affinity and specificity and thus widely used as compounds to modulate channel gating or channel currents. To promote the insecticidal effects of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), the gene encoding an excitatory insect toxin, BmK IT, was inserted into the genome of AcMNPV to construct a recombinant baculovirus, AcMNPV-BmK IT. Spodopter frugiperda 9 (Sf9) cells were infected with AcMNPV and AcMNPV-BmK IT respectively for 24 h. Results from the MTT assay, TUNEL assay, analysis of the expression level of apoptosis-related proteins (c-Myc, cleaved-Caspase3, Bcl-2 and Bax) of Sf9 cells, the transcription level of key genes (38K, C42, P78, F) of AcMNPV, and viral propagation assay demonstrated that AcMNPV-mediated expression of BmK IT promoted the apoptosis of Sf9 cells and replication of AcMNPV. The results laid a foundation for further structural and functional analysis of BmK IT.
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Apoptose , Nucleopoliedrovírus/fisiologia , Venenos de Escorpião/biossíntese , Animais , Linhagem Celular , Nucleopoliedrovírus/metabolismo , Células Sf9/efeitos dos fármacos , Replicação ViralRESUMO
OBJECTIVE: To explore the prevalence, diagnosis and treatment of restless leg syndrome (RLS) among military elders. METHODS: A cross-sectional study was conducted among retired military elders in Shanxi Province from 2010 to 2011. According to age, they were divided into 4 groups to compare the prevalence of RLS and evaluate the severity of symptoms as "mild, moderate and severe". And the compositions of symptom severity were compared among different age groups. Also the rate of correct diagnosis of RLS was examined. Univariate analyses were conducted with SPSS 13. 0. RESULTS: There were 559 RLS patients in 3 472 elder subjects. The prevalence of RLS was 16. 1% and 15. 8% in male elders. The prevalence of RLS in male elders increased significantly with advancing age (P <0. 05). No significant difference existed in the compositions of symptom severity among different age groups of RLS patients (P > 0. 05). Only 3. 22% RLS patients were previously diagnosed and none received a proper therapy. CONCLUSION: The prevalence of RLS among military elders is higher than that of their foreign counterparts. The prevalence of RLS in male elders increases significantly with advancing age. Symptom severity of RLS has no obvious relationship with advancing age. Since the rate of correct diagnosis and therapy is rather low for RLS elders in China, we should pay greater attention to the knowledge of RLS.
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Militares , Síndrome das Pernas Inquietas , Idoso , China , Estudos Transversais , Humanos , Masculino , PrevalênciaRESUMO
The gene encoding isocitrate dehydrogenase (IDH) is somatically mutated predominantly in secondary glioblastoma multiforme. Mutations of IDH1 and IDH2 lead to simultaneous loss and gain of activities in the production of α-ketoglutarate and 2-hydroxyglutarate, respectively. Lithium chloride was recently proved efficient in inhibiting glioma cell migration. The mechanism of lithium chloride on C6 glioma cells harboring IDH2 mutation has not been studied. Here, we found lithium chloride induced inhibitive effects on cell proliferation of both C6 glioma cells with and without IDH2 mutation, although IDH2 mutation increased the stability of HIF-1α. GSK-3ß could be phosphorylated at Ser9 and its activity was inhibited when C6 glioma cells were treated by lithium chloride. The degree of phosphorylation in IDH2(R172G) treatment group was lower than that as compared to the control and IDH2 treatment groups. At the same time, the accumulation of ß-catenin in C6 cell nucleus was decreased. Moreover, although the ß-catenin and HIF-1α increased the secretion of metalloproteinase-2,-9 in C6 glioma cells harboring IDH2 mutation, the migration potential of lithium chloride-treated C6 glioma cells harboring the IDH2 and its mutant was uniform. These results indicated lithium chloride could decrease the proliferation and migration potential of C6 glioma cells harboring IDH2 mutation.
Assuntos
Glioma/genética , Quinase 3 da Glicogênio Sintase/genética , Isocitrato Desidrogenase/genética , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Glioma/patologia , Glutaratos/metabolismo , Glicogênio Sintase Quinase 3 beta , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Isocitrato Desidrogenase/metabolismo , Ácidos Cetoglutáricos/metabolismo , Cloreto de Lítio/administração & dosagem , Mutação , beta Catenina/metabolismoRESUMO
BACKGROUND AND PURPOSE: Stroke may cause extensive after-effects such as motor function impairments and disorder of consciousness (DoC). Detecting these after-effects of stroke and monitoring their changes are challenging jobs currently undertaken via traditional clinical examinations. These behavioural examinations often take a great deal of manpower and time, thus consuming significant resources. Computer-aided examinations of the electroencephalogram (EEG) microstates derived from bedside EEG monitoring may provide an alternative way to assist medical practitioners in a quick assessment of the after-effects of stroke. METHODS: In this study, we designed a framework to extract microstate maps and calculate their statistical parameters to input to classifiers to identify DoC in ischemic stroke patients automatically. As the dataset is imbalanced with the minority of patients being DoC, an ensemble of support vector machines (EOSVM) is designed to solve the problem that classifiers always tend to be the majority classes in the classification on an imbalanced dataset. RESULTS: The experimental results show EOSVM get better performance (with accuracy and F1-Score both higher than 89%), improving sensitivity the most, from lower than 60% (SVM and AdaBoost) to higher than 80%. This highlighted the usefulness of the EOSVM-aided DoC detection based on microstates parameters. CONCLUSION: Therefore, the classifier EOSVM classification based on features of EEG microstates is helpful to medical practitioners in DoC detection with saved resources that would otherwise be consumed in traditional clinic checks.
Assuntos
AVC Isquêmico , Acidente Vascular Cerebral , Humanos , Eletroencefalografia/métodos , Acidente Vascular Cerebral/diagnóstico , Máquina de Vetores de SuporteRESUMO
As the global population ages, ischemic stroke has risen to become the second leading cause of disability and mortality worldwide, placing an immense burden on both society and families. Although treatments such as intravenous thrombolysis and endovascular interventions can substantially improve the outcomes for patients with acute ischemic stroke, only a small percentage of individuals benefit from these therapies. To advance our understanding of the disease and to discover more effective treatments, researchers are continuously developing and refining animal models. Among these, the middle cerebral artery occlusion (MCAO) model stands out as the most commonly used model in cerebrovascular disease research. The filament used in this model is crucial for its development. This protocol outlines a method for creating filaments with consistent diameters and varying lengths of silicone coating. The MCAO model produced using this method in C57 mice has demonstrated high success and consistency, offering a valuable tool for tailored investigations into ischemic cerebrovascular diseases.
Assuntos
Modelos Animais de Doenças , Infarto da Artéria Cerebral Média , Camundongos Endogâmicos C57BL , Silicones , Animais , Camundongos , Silicones/química , MasculinoRESUMO
Introduction: Ischemic stroke patients commonly experience disorder of consciousness (DOC), leading to poorer discharge outcomes and higher mortality risks. Therefore, the identification of applicable electrophysiological biomarkers is crucial for the rapid diagnosis and evaluation of post-stroke disorder of consciousness (PS-DOC), while providing supportive evidence for cerebral neurology. Methods: In our study, we conduct microstate analysis on resting-state electroencephalography (EEG) of 28 post-stroke patients with awake consciousness and 28 patients with PS-DOC, calculating the temporal features of microstates. Furthermore, we extract the Lempel-Ziv complexity of microstate sequences and the delta/alpha power ratio of EEG on spectral. Statistical analysis is performed to examine the distinctions in features between the two groups, followed by inputting the distinctive features into a support vector machine for the classification of PS-DOC. Results: Both groups obtain four optimal topographies of EEG microstates, but notable distinctions are observed in microstate C. Within the PS-DOC group, there is a significant increase in the mean duration and coverage of microstates B and C, whereas microstate D displays a contrasting trend. Additionally, noteworthy variations are found in the delta/alpha ratio and Lempel-Ziv complexity between the two groups. The integration of the delta/alpha ratio with microstates' temporal and Lempel-Ziv complexity features demonstrates the highest performance in the classifier (Accuracy = 91.07%). Discussion: Our results suggest that EEG microstates can provide insights into the abnormal brain network dynamics in DOC patients post-stroke. Integrating the temporal and Lempel-Ziv complexity microstate features with spectral features offers a deeper understanding of the neuro mechanisms underlying brain damage in patients with DOC, holding promise as effective electrophysiological biomarkers for diagnosing PS-DOC.
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OBJECTIVE: EGR3 is implicated in angiogenesis in rats with cerebral ischemia/reperfusion injury (CIRI). This research aimed to explore the effect and in vivo and ex vivo mechanisms of EGR3 in CIRI. METHODS: CIRI rat models were established via middle cerebral artery occlusion. Cell models were established via oxygen-glucose deprivation/reoxygenation (OGD/R). Brain injury was assessed by neurological scoring, HE, and TTC staining. Inflammatory factors and oxidative stress markers were measured using corresponding kits. Mitochondrial membrane potential and mitochondrial respiration were examined by flow cytometry and respirometry. EGR3-miR-146 network was predicted on TransmiR v2.0 database. Target genes of miR-146 were screened on Starbase, Targetscan, and miRDB databases. miR-146 expression was determined by RT-qPCR. Levels of EGR3 and SORT1 were determined by Western blot. Binding relationships among EGR3, miR-146, and SORT1 were validated by dual-luciferase assay. EGR3, miR-146, and SORT1 levels were altered by injection or cell transfection to observe their functions. RESULTS: EGR3 was poorly-expressed in CIRI rats and OGD/R-induced neurons. EGR3 overexpression reduced inflammatory factor levels and attenuated oxidative stress and mitochondrial injury in CIRI rats and OGD/R-induced neurons. EGR3 bound to miR-146b promoter region. EGR3 promoted pri-miR-146a/146b processing and stimulated miR-146 transcription. miR-146 overexpression ameliorated oxidative stress and mitochondrial injury and miR-146 downregulation abolished the effect of EGR3 overexpression in vitro. miR-146 targeted SORT1. SORT1 overexpression invalidated the protective function of miR-146 overexpression on oxidative stress and mitochondrial injury in vitro. CONCLUSION: EGR3 protected against CIRI by mitigating oxidative stress and mitochondrial injury via the miR-146/SORT1 axis.
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Taxillus chinensis (DC.) Danser, a parasitic plant of the Loranthaceae family, grows by attacking other plants. It has a long history of being used in Chinese medicine to treat multiple chronic diseases. We previously observed that T. chinensis seeds are sensitive to cold. In this study, we performed transcriptome sequencing for T. chinensis seeds treated by cold (0°C) for 0 h, 12 h, 24 h, and 36 h. TRINITY assembled 257,870 transcripts from 223,512 genes. The GC content and N50 were calculated as 42.29% and 1,368, respectively. Then, we identified 42,183 CDSs and 35,268 likely proteins in the assembled transcriptome, which contained 1,622 signal peptides and 6,795 transmembrane domains. Next, we identified 17,217 genes (FPKM > 5) and 2,333 differentially expressed genes (DEGs) in T. chinensis seeds under cold stress. The MAPK pathway, as an early cold response, was significantly enriched by the DEGs in the T. chinensis seeds after 24 h of cold treatment. Known cold-responsive genes encoding abscisic acid-associated, aquaporin, C-repeat binding factor (CBF), cold-regulated protein, heat shock protein, protein kinase, ribosomal protein, transcription factor (TF), zinc finger protein, and ubiquitin were deregulated in the T. chinensis seeds under cold stress. Notably, the upregulation of CBF gene might be the consequences of the downregulation of MYB and GATA TFs. Additionally, we identified that genes encoding CDC20, YLS9, EXORDIUM, and AUX1 and wound-responsive family protein might be related to novel mechanisms of T. chinensis seeds exposed to cold. This study is first to report the differential transcriptional induction in T. chinensis seeds under cold stress. It will improve our understanding of parasitic plants in response to cold and provide a valuable resource for future studies.
Assuntos
Loranthaceae , Temperatura Baixa , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Sementes/genética , Transcriptoma/genéticaRESUMO
Background: There has been a gradual trend towards younger ageing of acute cerebral infarction in recent years. Atherosclerotic plaque rupture followed by dislodgement of emboli and resulting arterial embolism is an important mechanism for the development of acute cerebral infarction. Traditional independent risk factors for cerebral infarction have received attention from clinicians, but the risk factors for large artery atherosclerotic cerebral infarction are still unclear. Various blood biomarkers have an important role in the early diagnosis of large artery atherosclerotic cerebral infarction. Objective: To assess the diagnostic predictive value of a group of biomarkers for large artery atherosclerotic cerebral infarction. Methods: Lipoprotein-associated phospholipase A2 (LP-PLA2), trypsin-like protein (TPS), serum amyloid A (SAA), and supersensitive C-reactive protein (hs-CRP) levels were measured in the case group (30 cases) and control group (54 cases), respectively. Results: The differences in the general data between the two groups were not statistically significant (P > 0.05). Logistic regression and ROC curve analysis showed that Lp-PLA2, TPS, and SAA were positively associated with the diagnosis of large atherosclerotic cerebral infarction (P < 0.05). The area under the ROC curve of the multivariate model for the biomarker group reached 0.995. Conclusion: Biomarkers are closely associated with the occurrence of large atherosclerotic cerebral infarction and can be used as clinical adjuncts for diagnosis and assessment of prognosis.
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Taxillus chinensis is an important medicinal and parasitic plant that attacks other plants for living. The development of haustorium is a critical process, imperative for successful parasitic invasion. To reveal the mechanisms underlying haustorium development, we performed an iTRAQ-based proteomics analysis which led to the identification of several differentially abundant proteins (DAPs) in fresh seeds (CK), baby (FB), and adult haustoria (FD). A total of 563 and 785 DAPs were identified and quantified in the early and later developmental stages, respectively. Pathway enrichment analysis revealed that the DAPs are mainly associated with metabolic pathways, ribosome, phenylpropanoid biosynthesis, and photosynthesis. In addition, DAPs associated with the phytohormone signaling pathway changed markedly. Furthermore, we evaluated the content of various phytohormones during different stages of haustoria development. These results indicated that phytohormones are very important for haustorium development. qRT-PCR results validated that the mRNA expression levels were consistent with the expression of proteins, suggesting that our results are reliable. This is the first report on haustoria proteomes in the parasitic plant, Taxillus chinensis, to the best of our knowledge. Our findings will enhance our understanding of the molecular mechanism of haustoria development.
Assuntos
Loranthaceae , Proteômica , Regulação da Expressão Gênica de Plantas , Reguladores de Crescimento de Plantas , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Sementes/metabolismoRESUMO
The hemiparasitic Taxillus chinensis (DC.) Danser is a root-parasitizing medicinal plant with photosynthetic ability, which is lost in other parasitic plants. However, the cultivation and medical application of the species are limited by the recalcitrant seeds of the species, and even though the molecular mechanisms underlying this recalcitrance have been investigated using transcriptomic and proteomic methods, genome resources for T. chinensis have yet to be reported. Accordingly, the aim of the present study was to use nanopore, short-read, and high-throughput chromosome conformation capture sequencing to construct a chromosome-level assembly of the T. chinensis genome. The final genome assembly was 521.90â Mb in length, and 496.43â Mb (95.12%) could be grouped into nine chromosomes with contig and scaffold N50 values of 3.80 and 56.90â Mb, respectively. In addition, a total of 33,894 protein-coding genes were predicted, and gene family clustering identified 11 photosystem-related gene families, thereby indicating photosynthetic ability, which is a characteristic of hemiparasitic plants. This chromosome-level genome assembly of T. chinensis provides a valuable genomic resource for elucidating the genetic basis underlying the recalcitrant characteristics of T. chinensis seeds and the evolution of photosynthesis loss in parasitic plants.
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Loranthaceae , Cromossomos , Genoma , Loranthaceae/genética , Filogenia , ProteômicaRESUMO
OBJECTIVE: This study is aimed to investigate the role of the long noncoding RNA SOX2 overlapping transcript (SOX2OT) in cerebral ischemia-reperfusion injury (CIRI) and the underlying regulatory mechanisms. METHODS: The oxygen-glucose deprivation/reoxygenation (OGD/R)-treated PC12 cells and middle cerebral artery occlusion/reperfusion (MCAO/R)-treated rats were established to simulate CIRI condition in vitro and in vivo. Quantitative real-time polymerase chain reaction was performed to detect the expression of SOX2OT, microRNA-135a-5p (miR-135a-5p), and nuclear receptor subfamily 3 group C member 2 (NR3C2). The cell viability and apoptosis were analyzed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assays. The levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), and reactive oxygen species (ROS) or interleukin (IL)-1ß and IL-6 were used to evaluate the oxidative stress or inflammation. Dual-luciferase reporter assay was conducted to validate the interactions among SOX2OT, miR-135a-5p, and NR3C2. Additionally, neurological deficit scores (NDS), infarct volume, and brain edema were used to assess brain impairments in vivo. RESULTS: The expression of SOX2OT and NR3C2 was increased, while miR-135a-5p was decreased in OGD/R-treated PC12 cells. SOX2OT silencing repressed the levels of LDH, MDA, ROS, IL-1ß, IL-6, reduced the numbers of TUNEL positive cells, and elevated viability and SOD level in OGD/R-treated PC12 cells. Besides, SOX2OT targeted miR-135a-5p, and miR-135a-5p targeted NR3C2. Both miR-135a-5p downregulation and NR3C2 upregulation reversed the suppressive effects of SOX2OT knockdown on oxidative stress, apoptosis, and inflammation of OGD/R-treated PC12 cells. Furthermore, injection of sh-SOX2OT reduced the NDS, cerebral infarct, and cerebral edema in MCAO/R-treated rats. CONCLUSIONS: Silencing of SOX2OT attenuated CIRI via regulation of the miR-135a-5p/NR3C2 axis, which may provide a novel therapeutic target for CIRI.
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Isquemia Encefálica/metabolismo , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Receptores de Mineralocorticoides/metabolismo , Traumatismo por Reperfusão/metabolismo , Animais , Apoptose/fisiologia , Encéfalo/metabolismo , Isquemia Encefálica/genética , Sobrevivência Celular/fisiologia , Citocinas/metabolismo , Regulação da Expressão Gênica , Malondialdeído/metabolismo , MicroRNAs/genética , Neurônios/metabolismo , Células PC12 , RNA Longo não Codificante/genética , Ratos , Espécies Reativas de Oxigênio/metabolismo , Receptores de Mineralocorticoides/genética , Traumatismo por Reperfusão/genéticaRESUMO
OBJECTIVE: MicroRNA (miR)-532-5p has been reported to protect against ischemic stroke (IS), while the underlying mechanism of miR-532-5p targeting BTB and CNC homology 1 (BACH1) in IS remains unknown. Thus, we aim to detect the role of miR-532-5p in IS via targeting BACH1. METHODS: Blood samples were collected from IS patients and healthy controls. Rat middle cerebral artery occlusion (MCAO) models were established and intracerebrally injected with altered miR-532-5p or BACH1 plasmid vectors to reveal their roles in neurological function, brain tissue pathology and inflammation in MCAO. Expression of miR-532-5p and BACH1 in patients' blood samples and rat brain tissues was assessed, and the targeting relationship between miR-532-5p and BACH1 was confirmed. RESULTS: MiR-532-5p was downregulated and BACH1 was upregulated in IS. BACH1 was targeted by miR-532-5p. Restored miR-532-5p or inhibited BACH1 improved neurological function and inhibited inflammation and apoptosis in MCAO rats. On the contrary, miR-532-5p reduction or BACH1 overexpression had totally opposite effects on MCAO rats. The protective role of miR-532-5p for MCAO rats was reversed by upregulated BACH1. CONCLUSION: MiR-532-5p upregulation protects against neurological deficits after IS through inhibition of BACH1.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/genética , Encéfalo/fisiopatologia , AVC Isquêmico/imunologia , MicroRNAs/metabolismo , Proteínas Repressoras/genética , Idoso , Animais , Fatores de Transcrição de Zíper de Leucina Básica/antagonistas & inibidores , Encéfalo/imunologia , Encéfalo/patologia , Estudos de Casos e Controles , Modelos Animais de Doenças , Regulação para Baixo , Feminino , Voluntários Saudáveis , Humanos , AVC Isquêmico/complicações , AVC Isquêmico/genética , AVC Isquêmico/fisiopatologia , Masculino , Pessoa de Meia-Idade , Ratos , Proteínas Repressoras/antagonistas & inibidores , Regulação para CimaRESUMO
INTRODUCTION: This aim of this study was to delineate current clinical scenarios of painful diabetic peripheral neuropathy (PDN) and associated anxiety and depression among patients in Mainland China, and to report current therapy and clinical practices. METHODS: A total of 1547 participants were enrolled in the study between 14 June 2018 and 11 November 2019. Recruitment was conducted using a multilevel sampling method. Participants' demographics, medical histories, glucose parameters, Douleur Neuropathique 4 Questionnaire (DN4) scores, visual analogue scale (VAS) pain scores, Patient Health Questionnaire 9 (PHQ-9) scores, Generalised Anxiety Disorder 7 (GAD-7) scores and therapies were recorded. RESULTS: The male-to-female ratio was 1.09:1 (807:740), and the mean age at onset was 61.28 ± 11.23 years. The mean DN4 score (± standard deviation) was 4.91 ± 1.88. The frequencies of DN4 sub-item phenotypes were: numbness, 81%; tingling, 68.71%; pins and needles, 62.90%; burning, 53.59%; hypoaesthesia to touch, 50.16%; electronic shocks, 43.31%; hypoaesthesia to pinprick, 37.94%; brushing, 37.82%; painful cold, 29.61%; and itching, 25.86%. Age, diabetic duration, depression history, PHQ-9 score and GAD-7 score were identified as risk factors for VAS pain score. Peripheral artery disease (PAD) was a protective factor for VAS pain score. For all participants currently diagnosed with PDN and for those previously diagnosed PDN, fasting blood glucose (FBG) was a risk factor for VAS; there was no association between FBG and VAS pain score for PDN diagnosed within 3 months prior to recruitment. Utilisation rate of opium therapies among enrolled participants was 0.71% , contradiction of first-line guideline recommendation for pain relief accounted for 9.43% (33/350) and contradiction of second-line guideline recommendation for opium dosage form was 0.57% (2/350). CONCLUSION: Moderate to severe neuropathic pain in PDN was identified in 73.11% of participants. Age, diabetic duration, depression history, PHQ-9 score, GAD-7 score and FBG were risk factors for VAS pain scores. PAD was protective factor. The majority of pain relief therapies prescribed were in accordance with guidelines. TRIAL REGISTRATION: ClinicalTrials.gov identifier, NCT03520608, retrospectively registered, 2018-05-11.
RESUMO
OBJECTIVES: To investigate the clinical feature of Parkinson's disease (PD) with restless leg syndrome (RLS) and the pathogenesis of RLS. METHODS: We conducted a cross-sectional and control study. The case group concluded 31 PD with RLS patients, meanwhile 39 PD patients were selected as the control group. Clinical history, clinical manifestations, complications and laboratory examinations were compared respectively between the two groups. RESULTS: All the RLS symptoms did not appear in RLS patients until the PD symptoms came out. Significant differences were found in complications such as swallow disturbance, constipation and illusion, when we compared the two PD groups (P < 0.05). Compared with the PD or healthy group, the level of serum ferritin and the H-reflex latency of tibial nerve were significantly decreased in PD with RLS group (P < 0.05). CONCLUSIONS: Secondary RLS is a complication of PD. Deficiency of iron and decreased inhibition function of spinal cord may lead to the occurrence of RLS in PD patients. When their motor symptoms are serious and complications are more common, PD patients are more possible to have RLS symptoms.