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1.
Plant Physiol ; 2024 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-39374536

RESUMO

Apple replant disease (ARD) is a worldwide problem that threatens the industry. However, the genetic mechanism underlying plant disease resistance against ARD remains unclear. In this study, a negative regulatory microRNA in Malus domestica, mdm-miR397b \, and its direct target MdLAC7b (Laccase) was selected for examination based on our previous small RNA and degradome sequencing results. Overexpressing the mdm-miR397b-MdLAC7b module altered the lignin deposition and JA contents in apple roots, which also led to increased resistance to Fusarium solani. Additionally, Y1H library screening using mdm-miR397b promoter recombinants identified a transcription factor, MdERF61, that represses mdm-miR397b transcriptional activity by directly binding to two GCC-boxes in the mdm-miR397b promoter. In summary, our results suggest that the MdERF61-mdm-miR397b-MdLAC7b module plays a crucial role in apple resistance to F. solani and offers insights for enhancing plant resistance to soilborne diseases in apple.

2.
Hum Brain Mapp ; 45(8): e26750, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38853710

RESUMO

The triple-network model has been widely applied in neuropsychiatric disorders including autism spectrum disorder (ASD). However, the mechanism of causal regulations within the triple-network and their relations with symptoms of ASD remains unclear. 81 male ASD and 80 well matched typically developing control (TDC) were included in this study, recruited from Autism Brain Image Data Exchange-I datasets. Spatial reference-based independent component analysis was used to identify the anterior and posterior part of default-mode network (aDMN and pDMN), salience network (SN), and bilateral executive-control network (ECN) from resting-state functional magnetic resonance imaging data. Spectral dynamic causal model and parametric empirical Bayes with Bayesian model reduction/average were adopted to explore the effective connectivity (EC) within triple-network and the relationship between EC and autism diagnostic observation schedule (ADOS) scores. After adjusting for age and site effect, ASD and TDC groups both showed inhibition patterns. Compared with TDC, ASD group showed weaker self-inhibition in aDMN and pDMN, stronger inhibition in pDMN→aDMN, weaker inhibition in aDMN→LECN, pDMN→SN, LECN→SN, and LECN→RECN. Furthermore, negative relationships between ADOS scores and pDMN self-inhibition strength, as well as with the EC of pDMN→aDMN were observed in ASD group. The present study reveals imbalanced effective connections within triple-networks in ASD children. More attentions should be focused at the pDMN, which modulates the core symptoms of ASD and may serve as an important region for ASD diagnosis and the target region for ASD treatments.


Assuntos
Transtorno do Espectro Autista , Rede de Modo Padrão , Imageamento por Ressonância Magnética , Humanos , Transtorno do Espectro Autista/diagnóstico por imagem , Transtorno do Espectro Autista/fisiopatologia , Masculino , Criança , Rede de Modo Padrão/diagnóstico por imagem , Rede de Modo Padrão/fisiopatologia , Conectoma , Rede Nervosa/diagnóstico por imagem , Rede Nervosa/fisiopatologia , Função Executiva/fisiologia , Adolescente , Teorema de Bayes
3.
Mol Breed ; 44(10): 69, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39359407

RESUMO

The PHOSPHATE STARVATION RESPONSE REGULATOR (PHR) plays a crucial regulatory role in plants during the process of responding to phosphate starvation. In this study, we combined reverse genetics and biotechnology to investigate the function of ZmPHR1 and ZmPHR2, including proteins containing the Myb_DNA_banding and Myb_CC-LHEQLE structural domains, in maize seedlings. Phylogenetic analysis revealed that ZmPHR1 and ZmPHR2 have high homology with AtPHR1 and OsPHR2, and share the characteristic features of nuclear localisation and transcriptional self-activation. Real-time quantitative PCR analysis showed that low phosphate (Pi) stress significantly induced the expression of ZmPHR1 and ZmPHR2 in maize seedling stage, and candidate gene association analysis further revealed the close association of these two genes with root traits under Pi stress conditions. Transgenic plants overexpressing ZmPHR1 and ZmPHR2 in Arabidopsis show a significant increase in lateral root number, fresh weight and total phosphorus accumulation under low-Pi stress. Besides, CHIP-PCR experiments identified target genes involved in hormone regulation, metal ion transport and homeostasis, phosphatase encoding, and photosynthesis, providing new insights into the biological functions of ZmPHR1 and ZmPHR2. Furthermore, our study showed that ZmPHR1 interacts with six SPX domain-only proteins (ZmSPXs) in maize, while ZmPHR2 interacts with five of these proteins. ZmPHR1 and ZmPHR2 expression was repressed in low Pi conditions, but was up-regulated in ZmSPX1 knockout material, according to our study of transgenic seedlings overexpressing ZmSPX1 in maize. We identified downstream target genes involved in the phosphorus signaling pathway, which are mainly involved in plant-pathogen interactions, ascorbic acid and arabinose metabolism, and ABC transporter proteins, by RNA-seq analysis of transgenic seedlings grown under low Pi stress for 7 days. Collectively, these results provide important clues to elucidate the role and functional significance of ZmPHR1 and ZmPHR2 under low Pi stress and also provide insights into understand the molecular mechanism of phosphorus homeostasis in maize. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-024-01508-2.

4.
Eur Arch Psychiatry Clin Neurosci ; 274(2): 433-443, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-37400684

RESUMO

BACKGROUND: Dopamine receptor D2 (DRD2) TaqIA polymorphism has an influence on addiction treatment response and prognosis by mediating brain dopaminergic system efficacy. Insula is crucial for conscious urges to take drugs and maintain drug use. However, it remains unclear about the contribution of DRD2 TaqIA polymorphism to the regulation of insular on addiction behavioral and its relation with the therapeutic effect of methadone maintenance treatment (MMT). METHODS: 57 male former heroin dependents receiving stable MMT and 49 matched male healthy controls (HC) were enrolled. Salivary genotyping for DRD2 TaqA1 and A2 alleles, brain resting-state functional MRI scan and a 24-month follow-up for collecting illegal-drug-use information was conducted and followed by clustering of functional connectivity (FC) patterns of HC insula, insula subregion parcellation of MMT patients, comparing the whole brain FC maps between the A1 carriers and non-carriers and analyzing the correlation between the genotype-related FC of insula sub-regions with the retention time in MMT patients by Cox regression. RESULTS: Two insula subregions were identified: the anterior insula (AI) and the posterior insula (PI) subregion. The A1 carriers had a reduced FC between the left AI and the right dorsolateral prefrontal cortex (dlPFC) relative to no carriers. And this reduced FC was a poor prognostic factor for the retention time in MMT patients. CONCLUSION: DRD2 TaqIA polymorphism affects the retention time in heroin-dependent individuals under MMT by mediating the functional connectivity strength between left AI and right dlPFC, and the two brain regions are promising therapeutic targets for individualized treatment.


Assuntos
Dependência de Heroína , Heroína , Humanos , Masculino , Heroína/uso terapêutico , Córtex Pré-Frontal Dorsolateral , Polimorfismo Genético/genética , Dependência de Heroína/diagnóstico por imagem , Dependência de Heroína/tratamento farmacológico , Dependência de Heroína/genética , Metadona/uso terapêutico , Imageamento por Ressonância Magnética , Receptores de Dopamina D2/genética
5.
Plant J ; 111(3): 768-784, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35648423

RESUMO

Two factors are proposed to account for the unusual features of organellar genomes: the disruptions of organelle-targeted DNA replication, repair, and recombination (DNA-RRR) systems in the nuclear genome and repetitive elements in organellar genomes. Little is known about how these factors affect organellar genome evolution. The deep-branching vascular plant family Selaginellaceae is known to have a deficient DNA-RRR system and convergently evolved organellar genomes. However, we found that the plastid genome (plastome) of Selaginella sinensis has extremely accelerated substitution rates, a low GC content, pervasive repeat elements, a dynamic network structure, and it lacks direct or inverted repeats. Unexpectedly, its organelle DNA-RRR system is short of a plastid-targeted Recombinase A1 (RecA1) and a mitochondrion-targeted RecA3, in line with other explored Selaginella species. The plastome contains a large collection of short- and medium-sized repeats. Given the absence of RecA1 surveillance, we propose that these repeats trigger illegitimate recombination, accelerated mutation rates, and structural instability. The correlations between repeat quantity and architectural complexity in the Selaginella plastomes support these conclusions. We, therefore, hypothesize that the interplay of the deficient DNA-RRR system and the high repeat content has led to the extraordinary divergence of the S. sinensis plastome. Our study not only sheds new light on the mechanism of plastome divergence by emphasizing the power of cytonuclear integration, but it also reconciles the longstanding contradiction on the effects of DNA-RRR system disruption on genome structure evolution.


Assuntos
Genomas de Plastídeos , Selaginellaceae , DNA , Evolução Molecular , Genomas de Plastídeos/genética , Filogenia , Selaginellaceae/genética
6.
Apoptosis ; 28(3-4): 627-638, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36719469

RESUMO

Recent evidence have indicated that ferroptosis, a novel iron-dependent form of non-apoptotic cell death, plays a critical role in human cancers. Besides, emerging literatures have revealed the ovel function of N6-methyladenosine (m6A) in bladder cancer physiological. However, the underlying mechanism of m6A on bladder cancer is still unclear. Here, present work revealed that m6A methyltransferase ('writer') WTAP up-regulated in bladder cancer tissue and cells, indicating the poor prognosis of bladder cancer patients. Functionally, gain/loss-of-functional experiments illustrated that WTAP promoted the viability of bladder cancer cells and inhibited the erastin-induced ferroptosis. Mechanistically, there was a remarkable m6A modification site on 3'-UTR of endogenous antioxidant factor NRF2 RNA and WTAP could install its methylation. Moreover, m6A reader YTHDF1 recognized the m6A site on NRF2 mRNA and enhanced its mRNA stability. Therefore, these findings demonstrated potential therapeutic strategyies for bladder cancer via m6A-dependent manner.


Assuntos
Ferroptose , Neoplasias da Bexiga Urinária , Humanos , Regiões 3' não Traduzidas , Apoptose , Proteínas de Ciclo Celular , Ferroptose/genética , Fator 2 Relacionado a NF-E2/genética , Fatores de Processamento de RNA , Neoplasias da Bexiga Urinária/genética
7.
Psychol Med ; 53(6): 2216-2228, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-34702384

RESUMO

BACKGROUND: Based on hubs of neural circuits associated with addiction and their degree centrality (DC), this study aimed to construct the addiction-related brain networks for patients diagnosed with heroin dependence undertaking stable methadone maintenance treatment (MMT) and further prospectively identify the ones at high risk for relapse with cluster analysis. METHODS: Sixty-two male MMT patients and 30 matched healthy controls (HC) underwent brain resting-state functional MRI data acquisition. The patients received 26-month follow-up for the monthly illegal-drug-use information. Ten addiction-related hubs were chosen to construct a user-defined network for the patients. Then the networks were discriminated with K-means-clustering-algorithm into different groups and followed by comparative analysis to the groups and HC. Regression analysis was used to investigate the brain regions significantly contributed to relapse. RESULTS: Sixty MMT patients were classified into two groups according to their brain-network patterns calculated by the best clustering-number-K. The two groups had no difference in the demographic, psychological indicators and clinical information except relapse rate and total heroin consumption. The group with high-relapse had a wider range of DC changes in the cortical-striatal-thalamic circuit relative to HC and a reduced DC in the mesocorticolimbic circuit relative to the low-relapse group. DC activity in NAc, vACC, hippocampus and amygdala were closely related with relapse. CONCLUSION: MMT patients can be identified and classified into two subgroups with significantly different relapse rates by defining distinct brain-network patterns even if we are blind to their relapse outcomes in advance. This may provide a new strategy to optimize MMT.


Assuntos
Dependência de Heroína , Heroína , Humanos , Masculino , Heroína/uso terapêutico , Metadona/uso terapêutico , Encéfalo , Dependência de Heroína/psicologia , Recidiva , Tratamento de Substituição de Opiáceos
8.
BMC Psychiatry ; 23(1): 379, 2023 05 30.
Artigo em Inglês | MEDLINE | ID: mdl-37254082

RESUMO

BACKGROUND: During the COVID-19 outbreak, most Chinese college students were home-quarantined to prevent the spread of the virus. COVID-19-associated impact has been shown to be a risk factor for the development of post-traumatic symptoms disorder (PTSD). However, little is known about the psychological processes that mediate this association. This study investigated the association between COVID-19-associated impact and PTSD and examined whether past stressful events, psychological resilience, and social support have mediating effects on this association. METHODS: The 12,397 valid responses from 31cities in China via an online survey assessed PTSD symptoms, past stressful events, psychological resilience, social support and social-demographic variables. AMOS was used to test the hypotheses of mediating effects. RESULTS: On the 39th day of the declared COVID-19 epidemic in China, 6.75% of the surveyed sample showed PTSD symptoms. A positive mediating effect of past stressful events was found between COVID-19-associated impact and PTSD, whereas psychological resilience and social support had negative mediating effects. The fit indices for the path model were found to be significant (ß = 0.28, p < 0.001), COVID-19-associated impact indirectly affects the risk of PTSD through mediating pathways (past stressful events → psychological resilience → social support) on PTSD. CONCLUSIONS: Attention should be paid to the effects of past stressful events of Chinese college students who were home-quarantined during the COVID-19 epidemic, and strategies should also be implemented to improve social support and develop psychological resilience. TRIAL REGISTRATION: The study was approved by the ethics committee of the Southwest Minzu University.


Assuntos
COVID-19 , Resiliência Psicológica , Transtornos de Estresse Pós-Traumáticos , Humanos , COVID-19/epidemiologia , Transtornos de Estresse Pós-Traumáticos/epidemiologia , Transtornos de Estresse Pós-Traumáticos/etiologia , Transtornos de Estresse Pós-Traumáticos/psicologia , Pandemias , Apoio Social , Estudantes/psicologia
9.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 40(7): 815-820, 2023 Jul 10.
Artigo em Zh | MEDLINE | ID: mdl-37368382

RESUMO

OBJECTIVE: To analyze the clinical significance of combined newborn hearing and deafness gene screening in Yuncheng area of Shanxi Province. METHODS: Results of audiological examinations, including transient evoked otoacoustic emission and automatic discriminative auditory brainstem evoked potentials, for 6 723 newborns born in Yuncheng area from January 1, 2021 to December 31, 2021, were retrospectively analyzed. Those who failed one of the tests were considered to have failed the examination. A deafness-related gene testing kit was used to detect 15 hot spot variants of common deafness-associated genes in China including GJB2, SLC26A4, GJB3, and mtDNA12S rRNA. Neonates who had passed the audiological examinations and those who had not were compared using a chi-square test. RESULTS: Among the 6 723 neonates, 363 (5.40%) were found to carry variants. These have included 166 cases (2.47%) with GJB2 gene variants, 136 cases (2.03%) with SLC26A4 gene variants, 26 cases (0.39%) with mitochondrial 12S rRNA gene variants, and 33 cases (0.49%) with GJB3 gene variants. Among the 6 723 neonates, 267 had failed initial hearing screening, among which 244 had accepted a re-examination, for which 14 cases (5.73%) had failed again. This has yielded an approximate prevalence of hearing disorder of 0.21% (14/6 723). Among 230 newborns who had passed the re-examination, 10 (4.34%) were found to have carried a variant. By contrast, 4 out of the 14 neonates (28.57%) who had failed the re-examination had carried a variant, and there was a significant difference between the two groups (P < 0.05). CONCLUSION: Genetic screening can provide an effective supplement to newborn hearing screening, and the combined screening can provide a best model for the prevention of hearing loss, which can enable early detection of deafness risks, targeted prevention measures, and genetic counseling to provide accurate prognosis for the newborns.


Assuntos
Conexinas , Surdez , Recém-Nascido , Humanos , Conexinas/genética , Estudos Retrospectivos , Surdez/diagnóstico , Surdez/genética , Conexina 26/genética , Triagem Neonatal/métodos , Mutação , Testes Genéticos/métodos , China/epidemiologia , Audição , Análise Mutacional de DNA
10.
Plant J ; 104(6): 1657-1672, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33073395

RESUMO

Plastids and mitochondria are endosymbiotic organelles that store genetic information. The genomes of these organelles generally exhibit contrasting patterns regarding genome architecture and genetic content. However, they have similar genetic features in Selaginellaceae, and little is known about what causes parallel evolution. Here, we document the multipartite plastid genomes (plastomes) and the highly divergent mitochondrial genomes (mitogenomes) from spikemoss obtained by combining short- and long-reads. The 188-kb multipartite plastome has three ribosomal operon copies in the master genomic conformation, creating the alternative subgenomic conformation composed of 110- and 78-kb subgenomes. The long-read data indicated that the two different genomic conformations were present in almost equal proportions in the plastomes of Selaginella nipponica. The mitogenome of S. nipponica was assembled into 27 contigs with a total size of 110 kb. All contigs contained directly arranged repeats at both ends, which introduced multiple conformations. Our results showed that plastomes and mitogenomes share high tRNA losses, GC-biased nucleotides, elevated substitution rates and complicated organization. The exploration of nuclear-encoded organelle DNA replication, recombination and repair proteins indicated that, several single-targeted proteins, particularly plastid-targeted recombinase A1, have been lost in Selaginellaceae; conversely, the dual-targeted proteins remain intact. According to the reported function of recombinase A1, we propose that the plastomes of spikemoss often fail to pair homologous sequences during recombination, and the dual-targeted proteins play a key role in the convergent genetic features of plastomes and mitogenomes. Our results provide a distinctive evolutionary pattern of the organelle genomes in Selaginellaceae and evidence of their convergent evolution.


Assuntos
Genoma de Planta/genética , Genomas de Plastídeos/genética , Selaginellaceae/genética , Evolução Molecular , Rearranjo Gênico/genética , Genes de Plantas/genética , Genoma Mitocondrial/genética , Huperzia/genética , Organelas/genética , Recombinação Genética/genética
11.
Nat Methods ; 15(1): 73-80, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29176592

RESUMO

Stable, high-resolution intravital imaging of the lung has become possible through the utilization of vacuum-stabilized imaging windows. However, this technique is extremely invasive and limited to only hours in duration. Here we describe a minimally invasive, permanently implantable window for high-resolution intravital imaging of the murine lung that allows the mouse to survive surgery, recover from anesthesia, and breathe independently. Compared to vacuum-stabilized windows, this window produces the same high-quality images without vacuum-induced artifacts; it is also less invasive, which allows imaging of the same lung tissue over a period of weeks. We further adapt the technique of microcartography for reliable relocalization of the same cells longitudinally. Using commonly employed experimental, as well as more clinically relevant, spontaneous metastasis models, we visualize all stages of metastatic seeding, including: tumor cell arrival; extravasation; growth and progression to micrometastases; as well as tumor microenvironment of metastasis function, the hallmark of hematogenous dissemination of tumor cells.


Assuntos
Modelos Animais de Doenças , Processamento de Imagem Assistida por Computador/métodos , Microscopia Intravital/métodos , Neoplasias Pulmonares/secundário , Neoplasias Mamárias Experimentais/patologia , Animais , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Células Tumorais Cultivadas
12.
Neurochem Res ; 46(12): 3365-3374, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34514556

RESUMO

Parkinson's disease (PD) is an age-related neurodegenerative disease caused by the loss of dopaminergic neurons in the substantia nigra. LncRNA MIAT has been shown to be critical in Alzheimer's disease, but its role and mechanism in PD are still unknown. Differentiated PC12 cells were treated with 1-methyl-4-phenylpyridinium (MPP+) to establish in vitro cell injury model of PD. MTT, Annexin V-PI double staining test and Western blot were used to detect cell viability and apoptosis. Reactive oxygen species (ROS), superoxide dismutase (SOD) and phospholipid hydroperoxide glutathione peroxidase (GSH-PX) kits were used to evaluate oxidative stress in cells. These results showed that LncRNA MIAT was down-regulated in MPP+-induced PC12 cells. Overexpression of LncRNA MIAT remarkably increased cell viability, inhibited cell apoptosis and oxidative stress in MPP+-treated cells. In addition, we proved that miR-132 is a target of LncRNA MIAT. Overexpression of miR-132 could reverse the positive effect of LncRNA MIAT overexpression on MPP+-induced cell oxidative stress injury. SIRT1 is a target of miR-132 and silencing of SIRT1 attunated the positive effect of LncRNA MIAT overexpression on oxidative stress injury in MPP+-induced PC12 cells. In conclusion, this study indicated that LncRNA MIAT suppressed MPP+-induced oxidative stress injury by regulating miR-132/SIRT1 axis in PC12 cells.


Assuntos
1-Metil-4-fenilpiridínio/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , MicroRNAs/genética , Doenças Neurodegenerativas/tratamento farmacológico , Doença de Parkinson/tratamento farmacológico , RNA Longo não Codificante/administração & dosagem , Sirtuína 1/metabolismo , Animais , Apoptose , Proliferação de Células , Herbicidas/toxicidade , Doenças Neurodegenerativas/metabolismo , Doenças Neurodegenerativas/patologia , Células PC12 , Doença de Parkinson/etiologia , Doença de Parkinson/metabolismo , Doença de Parkinson/patologia , Substâncias Protetoras/farmacologia , RNA Longo não Codificante/genética , Ratos , Sirtuína 1/genética
13.
Anal Bioanal Chem ; 413(3): 901-909, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33184760

RESUMO

We developed a fluorescent aptamer/graphene oxide (GO)-based biosensor to detect sulfamethazine (SMZ) residues in animal-derived foods. The SMZ-bound aptamers were identified and screened with an improved GO-SELEX technique using non-immobilizing ssDNA library. After seven rounds of selection, six SMZ aptamers were sequenced and analyzed for secondary structure, and their affinity and specificity were assessed by binding assays. The truncated aptamer (SMZ1S: 5'-CGTTAGACG-3') with a unique stem-loop structure showed the highest affinity (Kd = 24.6 nM) to SMZ and was used to develop a GO-based fluorescent aptasensor. The binding mechanism between SMZ1S and SMZ was further analyzed by molecular docking. Under optimal conditions, the fluorescent aptasensor showed low detection limits (0.35 ng/mL) and a wide dynamic linear range (from 2 to 100 ng/mL). The aptasensor was also validated against real samples spiked with SMZ, which showed a fluorescence recovery from 93.9 to 108.8% and a coefficient of variation of < 12.7%. Taken together, these results suggest that this novel aptasensor can be used to sensitively, selectively, and accurately detect SMZ residues in foods. Schematic illustration of fluorescent aptasensor based on aptamer/graphene oxide complex detection of of SMZ.


Assuntos
Anti-Infecciosos/análise , Aptâmeros de Nucleotídeos/análise , Contaminação de Alimentos/análise , Grafite/química , Sulfametazina/análise , Animais , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/instrumentação , DNA de Cadeia Simples/química , Fluorescência , Transferência Ressonante de Energia de Fluorescência , Limite de Detecção , Simulação de Acoplamento Molecular , Estrutura Molecular , Reação em Cadeia da Polimerase/métodos , Técnica de Seleção de Aptâmeros/métodos
14.
Molecules ; 26(14)2021 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-34299518

RESUMO

To monitor the illegal used of furaltadone, a highly sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and fluorescence-linked immunosorbent assay (FLISA) based on a monoclonal antibody (mAb) were developed for the detection of 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ), the major metabolite of furaltadone in animal tissues. The highly specific mAb, which was very sensitive to a nitrophenyl derivative of AMOZ (2-NP-AMOZ) with IC50 values of 0.11 and 0.09 ng/mL for ic-ELISA and FLISA, respectively, was selected for the development of immunoassays. For both the ic-ELISA and FLISA for AMOZ-spiked experiments, acceptable recovery rates of 81.1-105.3% and coefficients of variation of 4.7-9.8% were obtained. In addition, results from both ic-ELISA and FLISA methods for spiked samples' data showed excellent correlation coefficients ranging from 0.9652 to 0.9927. Meanwhile, the proposed ic-ELISA and FLISA for thirty spiked samples were confirmed by standard LC-MS/MS with high correlation coefficients of 0.9911 and 0.9921, respectively. These results suggest that the developed ic-ELISA and FLISA are valid and cost-effective tools for high-throughput monitoring methods for AMOZ residues in animal tissues.


Assuntos
Anticorpos Monoclonais/imunologia , Morfolinos/análise , Morfolinos/imunologia , Animais , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Fluorimunoensaio/métodos , Contaminação de Alimentos/análise , Imunoadsorventes/química , Camundongos Endogâmicos BALB C , Modelos Moleculares
15.
Opt Express ; 28(20): 30234-30247, 2020 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-33114907

RESUMO

Though three-dimensional (3D) fluorescence microscopy has been an essential tool for modern life science research, the light scattering by biological specimens fundamentally prevents its more widespread applications in live imaging. We hereby report a deep-learning approach, termed ScatNet, that enables reversion of 3D fluorescence microscopy from high-resolution targets to low-quality, light-scattered measurements, thereby allowing restoration for a blurred and light-scattered 3D image of deep tissue. Our approach can computationally extend the imaging depth for current 3D fluorescence microscopes, without the addition of complicated optics. Combining ScatNet approach with cutting-edge light-sheet fluorescence microscopy (LSFM), we demonstrate the image restoration of cell nuclei in the deep layer of live Drosophilamelanogaster embryos at single-cell resolution. Applying our approach to two-photon excitation microscopy, we could improve the signal-to-noise ratio (SNR) and resolution of neurons in mouse brain beyond the photon ballistic region.


Assuntos
Encéfalo/diagnóstico por imagem , Embrião não Mamífero/diagnóstico por imagem , Imageamento Tridimensional/métodos , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Microscopia de Fluorescência/métodos , Animais , Aprendizado Profundo , Drosophila melanogaster , Processamento de Imagem Assistida por Computador , Camundongos , Neuroimagem/métodos , Neurônios/citologia , Razão Sinal-Ruído
16.
Exp Mol Pathol ; 114: 104434, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32240615

RESUMO

The over-activation of Toll-like receptors (TLRs) is a typical immune response to injury. Previous work has suggested that controlling the over-activation of TLR4-MyD88-NF-κB may represent a new therapeutic option for diabetic kidney disease (DKD). 1,25(OH)2D3 has also been shown to exert a protective effect on DKD, although the mechanism involved has yet to be elucidated. The aim of this study was to investigate whether 1,25(OH)2D3 protects against DKD by down-regulating the innate immune TLR-NF-κB pathway. NRK-52E cells were cultured under normal or high-glucose conditions. We then used siRNA to knock down TLR4 expression under high-glucose conditions. NRK-52E cells cultured under high-glucose conditions, and streptozotocin (STZ)-induced diabetic rats, were treated with different doses of 1,25(OH)2D3 and used as in vitro and in vivo models, respectively. Renal biochemical indicators were then measured to evaluate the influence of 1,25(OH)2D3 treatment on DKD in diabetic rats. Histological analysis was also performed to determine the extent of infiltration by inflammatory cells and tubulointerstitial fibrosis. Using RT-qPCR, western blotting, immunohistochemistry and immunofluorescence, we determined the expression levels of TLR4, MyD88, NF-κB p65, MCP-1 and α-SMA to investigate whether 1,25(OH)2D3 could reduce the development of tubulointerstitial fibrosis. Knocking down TLR4 abolished the tubulointerstitial fibrosis caused by high-glucose conditions. High doses of 1,25(OH)2D3 consistently reduced the expression of TLR4-MyD88-NF-κB in NRK-52E cells. Moreover, high doses of 1,25(OH)2D3 had an obvious protective effect on kidney injury and inhibited the infiltration of inflammatory cells and tubulointerstitial fibrosis in diabetic rats. In conclusion, high doses of 1,25(OH)2D3 protected against tubulointerstitial fibrosis both in vitro and in vivo by downregulating the expression of TLR4-MyD88-NF-κB.


Assuntos
Nefropatias Diabéticas/tratamento farmacológico , Fator 88 de Diferenciação Mieloide/genética , Esteroide Hidroxilases/farmacologia , Receptor 4 Toll-Like/genética , Fator de Transcrição RelA/genética , Actinas/genética , Animais , Quimiocina CCL2/genética , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Nefropatias Diabéticas/genética , Nefropatias Diabéticas/patologia , Modelos Animais de Doenças , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , NF-kappa B/genética , Ratos , Transdução de Sinais/efeitos dos fármacos
17.
Exp Cell Res ; 370(2): 273-282, 2018 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-29953877

RESUMO

Nonmuscle myosin-IIA (NMHC-IIA) heavy chain phosphorylation has gained recognition as an important feature of myosin-II regulation. In previous work, we showed that phosphorylation on S1943 promotes myosin-IIA filament disassembly in vitro and enhances EGF-stimulated lamellipod extension of breast tumor cells. However, the contribution of NMHC-IIA S1943 phosphorylation to the modulation of invasive cellular behavior and metastasis has not been examined. Stable expression of phosphomimetic (S1943E) or non-phosphorylatable (S1943A) NMHC-IIA in breast cancer cells revealed that S1943 phosphorylation enhances invadopodia function, and is critical for matrix degradation in vitro and experimental metastasis in vivo. These studies demonstrate a novel link between NMHC-IIA S1943 phosphorylation, the regulation of extracellular matrix degradation and tumor cell invasion and metastasis.


Assuntos
Proteínas do Citoesqueleto/metabolismo , Metástase Neoplásica/patologia , Miosina não Muscular Tipo IIA/metabolismo , Podossomos/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/fisiologia , Humanos , Cadeias Pesadas de Miosina/metabolismo , Fosforilação , Podossomos/genética
18.
Cell Physiol Biochem ; 45(1): 119-130, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29339630

RESUMO

BACKGROUND/AIMS: A network meta-analysis is used to compare the efficacy of ropinirole, rasagiline, rotigotine, entacapone, apomorphine, pramipexole, sumanirole, bromocriptine, piribedil and levodopa, with placebo as a control, for non-motor symptoms in Parkinson's disease (PD). METHODS: PubMed, Embase and the Cochrane Library were searched from their establishment dates up to January 2017 for randomized controlled trials (RCTs) investigating the efficacy of the above ten drugs on the non-motor symptoms of PD. A network meta-analysis combined the evidence from direct comparisons and indirect comparisons and evaluated the pooled weighted mean difference (WMD) values and surfaces under the cumulative ranking curves (SUCRA). The network meta-analysis included 21 RCTs. RESULTS: The analysis results indicated that, using the United Parkinson's Disease Rating Scale (UPDRS) III, the efficacies of placebo, ropinirole, rasagiline, rotigotine, entacapone, pramipexole, sumanirole and levodopa in treating PD were lower than that of apomorphine (WMD = -10.90, 95% CI = -16.12∼-5.48; WMD = -11.85, 95% CI = -17.31∼-6.16; WMD = -11.15, 95% CI = -16.64∼-5.04; WMD = -11.70, 95% CI = -16.98∼-5.60; WMD = -11.04, 95% CI = -16.97∼-5.34; WMD = -13.27, 95% CI = -19.22∼-7.40; WMD = -10.25, 95% CI = -15.66∼-4.32; and WMD = -11.60, 95% CI = -17.89∼-5.57, respectively). Treatment with ropinirole, rasagiline, rotigotine, entacapone, pramipexole, sumanirole, bromocriptine, piribedil or levodopa, with placebo as a control, on PD exhibited no significant differences on PD symptoms when the UPDRS II was used for evaluation. Moreover, using the UPDRS III, the SUCRA values indicated that a pomorphine had the best efficacy on the non-motor symptoms of PD (99.0%). Using the UPDRS II, the SUCRA values for ropinirole, rasagiline, rotigotine, entacapone, pramipexole, sumanirole, bromocriptine, piribedil and levodopa treatments, with placebo as a control, indicated that bromocriptine showed the best efficacy on the non-motor symptoms of PD (75.6%). CONCLUSION: Among ropinirole, rasagiline, rotigotine, entacapone, apomorphine, pramipexole, sumanirole, bromocriptine, piribedil and levodopa, with placebo as a control, apomorphine may be the most efficacious drug for therapy in treating the non-motor symptoms of PD.


Assuntos
Agonistas de Dopamina/uso terapêutico , Doença de Parkinson/tratamento farmacológico , Apomorfina/uso terapêutico , Teorema de Bayes , Bases de Dados Factuais , Humanos , Doença de Parkinson/patologia , Índice de Gravidade de Doença , Resultado do Tratamento
19.
J Psychiatry Neurosci ; 43(1): 48-57, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29252165

RESUMO

BACKGROUND: It is unknown whether impaired coupling among 3 core large-scale brain networks (salience [SN], default mode [DMN] and executive control networks [ECN]) is associated with relapse behaviour in treated heroin-dependent patients. METHODS: We conducted a prospective resting-state functional MRI study comparing the functional connectivity strength among healthy controls and heroin-dependent men who had either relapsed or were in early remission. Men were considered to be either relapsed or in early remission based on urine drug screens during a 3-month follow-up period. We also examined how the coupling of large-scale networks correlated with relapse behaviour among heroin-dependent men. RESULTS: We included 20 controls and 50 heroin-dependent men (26 relapsed and 24 early remission) in our analyses. The relapsed men showed greater connectivity than the early remission and control groups between the dorsal anterior cingulate cortex (key node of the SN) and the dorsomedial prefrontal cortex (included in the DMN). The relapsed men and controls showed lower connectivity than the early remission group between the left dorsolateral prefrontal cortex (key node of the left ECN) and the dorsomedial prefrontal cortex. The percentage of positive urine drug screens positively correlated with the coupling between the dorsal anterior cingulate cortex and dorsomedial prefrontal cortex, but negatively correlated with the coupling between the left dorsolateral prefrontal cortex and dorsomedial prefrontal cortex. LIMITATIONS: We examined deficits in only 3 core networks leading to relapse behaviour. Other networks may also contribute to relapse. CONCLUSION: Greater coupling between the SN and DMN and lower coupling between the left ECN and DMN is associated with relapse behaviour. These findings may shed light on the development of new treatments for heroin addiction.


Assuntos
Dependência de Heroína/fisiopatologia , Dependência de Heroína/psicologia , Vias Neurais/fisiopatologia , Adulto , Estudos de Casos e Controles , Neuroimagem Funcional , Humanos , Imageamento por Ressonância Magnética , Masculino , Recidiva , Adulto Jovem
20.
Methods ; 128: 65-77, 2017 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-28911733

RESUMO

Pathologists rely on microscopy to diagnose disease states in tissues and organs. They utilize both high-resolution, high-magnification images to interpret the staining and morphology of individual cells, as well as low-magnification overviews to give context and location to these cells. Intravital imaging is a powerful technique for studying cells and tissues in their native, live environment and can yield sub-cellular resolution images similar to those used by pathologists. However, technical limitations prevent the straightforward acquisition of low-magnification images during intravital imaging, and they are hence not typically captured. The serial acquisition, mosaicking, and stitching together of many high-resolution, high-magnification fields of view is a technique that overcomes these limitations in fixed and ex vivo tissues. The technique however, has not to date been widely applied to intravital imaging as movements caused by the living animal induce image distortions that are difficult to compensate for computationally. To address this, we have developed techniques for the stabilization of numerous tissues, including extremely compliant tissues, that have traditionally been extremely difficult to image. We present a novel combination of these stabilization techniques with mosaicked and stitched intravital imaging, resulting in a process we call Large-Volume High-Resolution Intravital Imaging (LVHR-IVI). The techniques we present are validated and make large volume intravital imaging accessible to any lab with a multiphoton microscope.


Assuntos
Corantes Fluorescentes , Microscopia Intravital/métodos , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Análise de Célula Única/métodos , Imagem com Lapso de Tempo/métodos , Animais , Movimento Celular/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Técnicas de Janela Pericárdica
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