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1.
Antonie Van Leeuwenhoek ; 117(1): 83, 2024 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-38806744

RESUMO

An aerobic, Gram-stain-negative, motile rod bacterium, designated as SYSU BS000021T, was isolated from a black soil sample in Harbin, Heilongjiang province, China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate belongs to the genus Methylobacterium, and showed the highest sequence similarity to Methylobacterium segetis KCTC 62267 T (98.51%) and Methylobacterium oxalidis DSM 24028 T (97.79%). Growth occurred at 20-37℃ (optimum, 28 °C), pH 6.0-8.0 (optimum, pH 7.0) and in the presence of 0% (w/v) NaCl. Polar lipids comprised of phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, one unidentified aminolipid and one unidentified polar lipid. The major cellular fatty acids (> 5%) were C18:0 and C18:1 ω7c and/or C18:1 ω6c. The predominant respiratory quinone was Q-10. The genomic G + C content was 68.36% based on the whole genome analysis. The average nucleotide identity (≤ 83.5%) and digital DNA-DNA hybridization (≤ 27.3%) values between strain SYSU BS000021T and other members of the genus Methylobacterium were all lower than the threshold values recommended for distinguishing novel prokaryotic species. Based on the results of phenotypic, chemotaxonomic and phylogenetic analyses, strain SYSU BS000021T represents a novel species of the genus Methylobacterium, for which the name Methylobacterium nigriterrae sp. nov. is proposed. The type strain of the proposed novel species is SYSU BS000021T (= GDMCC 1.3814 T = KCTC 8051 T).


Assuntos
Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano , Ácidos Graxos , Methylobacterium , Filogenia , RNA Ribossômico 16S , Microbiologia do Solo , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , Ácidos Graxos/análise , Ácidos Graxos/química , Methylobacterium/genética , Methylobacterium/classificação , Methylobacterium/isolamento & purificação , China , Hibridização de Ácido Nucleico , Análise de Sequência de DNA , Fosfolipídeos/análise
2.
Microorganisms ; 12(5)2024 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-38792780

RESUMO

The degradation of farmland in China underscores the need for developing and utilizing saline-alkali soil. Soil health relies on microbial activity, which aids in the restoration of the land's ecosystem, and hence it is important to understand microbial diversity. In the present study, two Gram-stain-positive strains HR 1-10T and J-A-003T were isolated from saline-alkali soil. Preliminary analysis suggested that these strains could be a novel species. Therefore, the taxonomic positions of these strains were evaluated using polyphasic analysis. Phylogenetic and 16S rRNA gene sequence analysis indicated that these strains should be assigned to the genus Halalkalibacter. Cell wall contained meso-2,6-diaminopimelic acid. The polar lipids present in both strains were diphosphatidyl-glycerol, phosphatidylglycerol, and an unidentified phospholipid. The major fatty acids (>10%) were anteiso-C15:0, C16:0 and iso-C15:0. Average nucleotide identity and digital DNA#x2013;DNA hybridization values were below the threshold values (95% and 70%, respectively) for species delineation. Based on the above results, the strains represent two novel species of the genus Halalkalibacter, for which the names Halalkalibacter flavus sp. nov., and Halalkalibacter lacteus sp. nov., are proposed. The type strains are HR 1-10T (=GDMCC 1.2946T = MCCC 1K08312T = JCM 36285T), and J-A-003T (=GDMCC 1.2949T = MCCC 1K08417T = JCM 36286T).

3.
Health Secur ; 2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38815144

RESUMO

The purpose of this study was to investigate the feasibility and effectiveness of COVID-19 throat swab samples delivered by medical drones in epidemic prevention and control. This study was carried out in both southern and northern hospital districts of the Affiliated Hospital of Jiangnan University from May to October 2022. The main participants were the Affiliated Hospital of Jiangnan University and Zhejiang Antwork Technology Co., Ltd. We first constructed an urban medical unmanned aerial vehicle (UAV) delivery system and developed a UAV-specific storage box for COVID-19 samples. The UAV system was used to transport COVID-19 throat swab samples from the northern hospital district to the southern hospital district, and the following indexes were obtained: (1) flight time of COVID-19 samples delivered by UAV, (2) real-time temperature of COVID-19 nucleic acid samples during transportation, and (3) the time of distribution of COVID-19 nucleic acid samples by road traffic as measured using the Baidu Maps application, compared with the flight time of UAV. The COVID-19 sample delivery system for urban medical UAV mainly consists of intelligent logistics UAV, low-temperature COVID-19 throat swab sample storage box, unmanned logistics hub, and cloud operation control platform. The flight distance between the northern and southern districts of the Affiliated Hospital of Jiangnan University was 10 km, and the ground distance was 24 km. From May 11 to October 28, 2022, a total of 1,190 UAV flights occurred. The average flight time was 13 minutes, which was 40 to 70 minutes faster than the average road travel time required for manual delivery of COVID-19 throat swab samples. At different time points in the day, UAV delivery efficiency increased by 67.5% to 82%. The use of 5G with the Internet of Things and UAV technology to deliver nucleic acid samples has the characteristics of fast speed, being unaffected by ground traffic conditions, and the ability to ensure the safety of nucleic acid samples in the transportation process, which is worthy of further study.

4.
Burns ; 50(3): 578-584, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38238240

RESUMO

BACKGROUND: The goal of this study is to look into the factors that lead to death in patients with necrotizing soft tissue infections(NSTIs) in the intensive care unit and create a mortality risk model. METHODS: The clinical data of 106 patients with necrotizing soft tissue infections admitted to intensive care unit(ICU) of the First Affiliated Hospital of Wenzhou Medical University between January 2008 and December 2021 were retrospectively analyzed. Univariate analysis and multivariate analysis were performed to evaluate the risk factors impacting patient mortality. The regression coefficient in binary logistic regression analysis was converted into the item score in the model, and then the model score of each patient was calculated. Finally, an ROC curve was constructed to evaluate the efficiency of the model for predicting mortality. Thirteen patients with NSTIs admitted to ICU between January 2022 and November 2022 were used to validate the model. RESULTS: The death group had 44 patients, while the survival group had 62 patients. The overall mortality was 41.5%. Binary logistic regression analysis showed that risk factors for mortality were age≥ 60 years(OR:4.419; 95%CI:1.093-17.862; P = 0.037), creatinine ≥ 132µmol/L(OR:11.166; 95%CI:2.234-55.816; P = 0.003), creatine kinase ≥ 1104 U/L(OR:4.019; 95%CI:1.134-14.250; P = 0.031), prothrombin time ≥ 24.4 s(OR:11.589; 95%CI:2.510-53.506; P = 0.002), and invasive mechanical ventilation (OR:17.404; 95%CI:4.586-66.052; P<0.000). The AUC of the model for predicting mortality was 0.940 (95% CI:0.894-0.986). When the cut-off value for the model was 4 points, the sensitivity was 95.5% and the specificity was 83.9%. CONCLUSION: The death risk model in this study for NSTIs patients in the intensive care unit shows high sensitivity and specificity. Patients with a score of ≥ 4 points have a higher risk of mortality.


Assuntos
Queimaduras , Sepse , Infecções dos Tecidos Moles , Humanos , Pessoa de Meia-Idade , Infecções dos Tecidos Moles/epidemiologia , Estudos Retrospectivos , Prognóstico , Unidades de Terapia Intensiva , Curva ROC
5.
Front Microbiol ; 14: 1095688, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36910216

RESUMO

Rational cropping maintains high soil fertility and a healthy ecosystem. Soil microorganism is the controller of soil fertility. Meanwhile, soil microbial communities also respond to different cropping patterns. The mechanisms by which biotic and abiotic factors were affected by different cropping sequences remain unclear in the major grain-producing regions of northeastern China. To evaluate the effects of different cropping sequences under conventional fertilization practices on soil properties, microbial communities, and crop yield, six types of plant cropping systems were performed, including soybean monoculture, wheat-soybean rotation, wheat-maize-soybean rotation, soybean-maize-maize rotation, maize-soybean-soybean rotation and maize monoculture. Our results showed that compared with the single cropping system, soybean and maize crop rotation in different combinations or sequences can increase soil total organic carbon and nutrients, and promote soybean and maize yield, especially using soybean-maize-maize and maize-soybean-soybean planting system. The 16S rRNA and internal transcribed spacer (ITS) amplicon sequencing showed that different cropping systems had different effects on bacterial and fungal communities. The bacterial and fungal communities of soybean monoculture were less diverse when compared to the other crop rotation planting system. Among the different cropping sequences, the number of observed bacterial species was greater in soybean-maize-maize planting setup and fungal species in maize-soybean-soybean planting setup. Some dominant and functional bacterial and fungal taxa in the rotation soils were observed. Network-based analysis suggests that bacterial phyla Acidobacteria and Actinobacteria while fungal phylum Ascomycota showed a positive correlation with other microbial communities. The phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) result showed the presence of various metabolic pathways. Besides, the soybean-maize-maize significantly increased the proportion of some beneficial microorganisms in the soil and reduced the soil-borne animal and plant pathogens. These results warrant further investigation into the mechanisms driving responses of beneficial microbial communities and their capacity on improving soil fertility during legume cropping. The present study extends our understanding of how different crop rotations effect soil parameters, microbial diversity, and metabolic functions, and reveals the importance of crop rotation sequences. These findings could be used to guide decision-making from the microbial perspective for annual crop planting and soil management approaches.

7.
ACS Chem Biol ; 17(12): 3389-3406, 2022 12 16.
Artigo em Inglês | MEDLINE | ID: mdl-36446024

RESUMO

Ferroptosis is a nonapoptotic form of cell death characterized by iron-dependent lipid peroxidation and has been implicated in multiple pathological conditions. Glutathione peroxidase 4 (GPX4) plays an essential role in inhibiting ferroptosis by eliminating lipid peroxide using glutathione (GSH) as a reductant. In this study, we found Ellman's reagent DTNB and a series of disulfide compounds, including disulfiram (DSF), an FDA-approved drug, which protect cells from erastin-induced ferroptosis. Mechanistically, DTNB or DSF is conjugated to multiple cysteine residues in GPX4 and disrupts GPX4 interaction with HSC70, an adaptor protein for chaperone mediated autophagy, thus preventing GPX4 degradation induced by erastin. In addition, DSF ameliorates concanavalin A induced acute liver injury by suppressing ferroptosis in a mouse model. Our work reveals a novel regulatory mechanism for GPX4 protein stability control. We also discover disulfide compounds as a new class of ferroptosis inhibitors and suggest therapeutic repurposing of DSF in treating ferroptosis-related diseases.


Assuntos
Dissulfetos , Ferroptose , Fosfolipídeo Hidroperóxido Glutationa Peroxidase , Animais , Camundongos , Dissulfetos/farmacologia , Ácido Ditionitrobenzoico , Ferroptose/efeitos dos fármacos , Glutationa/metabolismo , Peroxidação de Lipídeos/fisiologia , Fosfolipídeo Hidroperóxido Glutationa Peroxidase/efeitos dos fármacos , Fosfolipídeo Hidroperóxido Glutationa Peroxidase/metabolismo , Sulfetos , Dissulfiram/farmacologia
9.
Curr Microbiol ; 79(10): 301, 2022 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-36002665

RESUMO

A novel bacterium, designated strain APA_H-1(4)T, was isolated from the saline-alkaline soil, Zhaodong, Heilongjiang Province, China. Phenotypic and chemotaxonomic analyses, and whole-genome sequencing were used to determine the taxonomic position of the strain. Phylogenetic analysis indicated that the isolate belongs to the genus Oceanobacillus, and showed the highest sequence similarity to O. damuensis KCTC 33146T (98.35%, similarity) and 'O. massiliensis' DSM 24644 (98.32%). The average nucleotide identity values between strain APA_H-1(4)T and other members of the genus Oceanobacillus were lower than 82% recommended for distinguishing novel prokaryotic species. The digital DNA-DNA hybridization values of strain APA_H-1(4)T with O. damuensis KCTC 33146T and 'O. massiliensis' DSM 24644 were 13.60 and 17.60%, respectively. Cells of strain APA_H-1(4)T were Gram-staining positive, motile, aerobic, spore-forming rods (0.5-0.7 × 1.8-2.6 µm) with flagella. The growth was found to occur optimally at 37 °C. The whole-cell hydrolysate contained meso-diaminopimelic acid as the diagnostic cell wall diamino acid. The main detected polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, an unidentified phospholipid and an unidentified polar lipid. The predominant respiratory quinone was identified as menaquinone-7 (MK-7). The major cellular fatty acid (>10%) was anteiso-C15:0. The G + C content of the genomic DNA was determined to be 38.4% based on the draft genome sequence. Based on the comparative analysis of polyphasic taxonomic data, strain APA_H-1(4)T represents a novel species of the genus Oceanobacillus, for which the name Oceanobacillus saliphilus sp. nov. is proposed. The type strain is APA_H-1(4)T (=GDMCC 1.2239T = KCTC 43254T).


Assuntos
Bacillaceae , Solo , Álcalis , Técnicas de Tipagem Bacteriana , China , DNA Bacteriano/química , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
10.
Eur Neurol ; 85(3): 224-234, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35100587

RESUMO

INTRODUCTION: We explored microRNA (miRNA) profiles correlated with the penumbra in three different phases of ischaemic stroke, using a permanent middle cerebral artery occlusion (p-MCAO) rat model. MATERIALS AND METHODS: A 2-mm coronal section was cut from the optic chiasma in the caudal direction, and the penumbra was located in the area between a longitudinal line approximately 2 mm from the midline and a transverse diagonal line at the "2-o'clock" position. Total RNA was extracted from tissue specimens and peripheral blood samples, followed by deep sequencing analysis. RESULTS: We identified nine novel miRNA candidates in tissues and evaluated their expression levels using real-time quantitative polymerase chain reaction. In situ hybridization was conducted to assess miRNA localization in the brain. Of these nine candidates, we identified and characterized a novel miRNA, rno-miR-686-3p, which was localized in cell nuclei of the cortex, and associated with the penumbra. rno-miR-686-3p was downregulated at 1 (p = 0.042), 3 (p = 0.032), and 4 h (p = 0.007) post-p-MCAO in the penumbra. A total of 297 potential target genes were predicted. Moreover, functional annotation clustering and pathway enrichment analysis predicted that rno-miR-686-3p participates in transcriptional regulation and the Wnt and cyclic adenosine monophosphate (cAMP) signalling pathways. CONCLUSION: rno-miR-686-3p is a novel miRNA associated with the ischaemic penumbra that is implicated in transcriptional regulation and modulation of the Wnt and cAMP signalling pathways. Furthermore, it may serve as a possible new biomarker with potential value for detecting the existence of the penumbra.


Assuntos
Isquemia Encefálica , MicroRNAs , Acidente Vascular Cerebral , Animais , Biomarcadores , Isquemia Encefálica/genética , Infarto da Artéria Cerebral Média , MicroRNAs/genética , Ratos
11.
Arch Microbiol ; 204(1): 53, 2021 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-34936048

RESUMO

A Gram-positive strain APA H-16(1)T was isolated from a saline-alkali soil sample collected from Heilongjiang Province, China. Cells were rod shaped, non-motile, endospore forming, and aerobic. Growth occurred at 10-45 °C (optimum, 35 °C), pH 7.0-10.5 (optimum, pH 9.5), and could tolerate NaCl up to 15.0% (w/v). Strain showed low 16S rRNA gene sequence similarities with Alteribacter natronophilus (97.8%), Alteribacter aurantiacus (97.7%), and Alteribacter populi (97.1%). The cell wall peptidoglycan was meso-diaminopimelic acid. The predominant menaquinone was MK-7. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, unidentified aminophospholipids, unidentified phospholipid, and unidentified lipid. The major fatty acids were anteiso-C15:0, and iso-C15:0. The genomic G + C content was 45.1%. The average nucleotide identity and digital DNA-DNA hybridization values between strain APA H-16(1)T and the most closely related species were below the cut-off level (95-96%; 70%) for species delineation. Based on phenotypic, phylogenetic, chemotaxonomic, and genome comparison, strain APA H-16(1)T represents a novel species of the genus Alteribacter, for which the name Alteribacter salitolerans sp. nov. is proposed. The type strain is APA H-16(1)T (= KCTC 43228T = CICC 25092T).


Assuntos
Microbiologia do Solo , Solo , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Ácidos Graxos , Fosfolipídeos , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
Front Cell Dev Biol ; 9: 675424, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34336831

RESUMO

A growing number of human diseases have been found to be associated with aberrant DNA methylation, including cancer. Mutations targeting genes encoding DNA methyltransferase (DNMT), TET family of DNA demethylases, and isocitrate dehydrogenase (IDH1, IDH2) that produce TET inhibitory metabolite, 2-hyoxyglutarate (2-HG), are found in more than half of acute myeloid leukemia (AML). To gain new insights into the regulation of DNA de/methylation and consequence of its alteration in cancer development, we searched for genes which are mutated in a manner that is linked with gene mutations involved in DNA de/methylation in multiple cancer types. We found that recurrent CBFB-MYH11 fusions, which result in the expression of fusion protein comprising core-binding factor ß (CBFB) and myosin heavy chain 11 (MYH11) and are found in 6∼8% of AML patients, occur mutually exclusively with DNMT3A mutations. Tumors bearing CBFB-MYH11 fusion show DNA hypomethylation patterns similar to those with loss-of-function mutation of DNMT3A. Expression of CBFB-MYH11 fusion or inhibition of DNMT3A similarly impairs the methylation and expression of target genes of Runt related transcription factor 1 (RUNX1), a functional partner of CBFB. We demonstrate that RUNX1 directly interacts with DNMT3A and that CBFB-MYH11 fusion protein sequesters RUNX1 in the cytoplasm, thereby preventing RUNX1 from interacting with and recruiting DNMT3A to its target genes. Our results identify a novel regulation of DNA methylation and provide a molecular basis how CBFB-MYH11 fusion contributes to leukemogenesis.

13.
Opt Lett ; 46(12): 2815-2818, 2021 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-34129547

RESUMO

In the inverse design of nanophotonic devices, mathematical optimization methods are generally used to perform local optimization in the design region to obtain the physical structure that meets design expectations. These methods usually produce good structures. However, due to the lack of physical considerations, most of the inverse design methods for nanophotonic devices use random initial topology as the initialization for optimization, which will inevitably cause a waste of computing resources. In this Letter, we propose a method based on a time-reversal technique to quickly determine the induced source of the physical structure in the design region and, thus, obtain the initial topological structure of the nanophotonic devices. For a nanophotonic 90°-bend waveguide and 90°-bend power splitter waveguide, numerical examples show that the initial topology obtained by our method not only has good initial performance, but also can be used as a reasonable initialization for inverse design.

14.
Medicine (Baltimore) ; 99(52): e23694, 2020 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-33350749

RESUMO

ABSTRACT: This study aimed to investigate the effects of age on the peak ratio (PR) of ophthalmic artery (OA) Doppler.The initial peak velocity (P1), second peak velocity (P2) and PR of OA were detected by color Doppler ultrasonography in 147 healthy subjects. All of the subjects were divided into 6 groups (G1-G6) according to the age. (G1, 20-29 years; G2, 30-39 years; G3, 40-49 years; G4, 50-59 years; G5, 60-69 years; and G6, 70 years or older). The blood pressure and heart rate were also examined before ultrasonography. The influences of age, blood pressure and heart rate on the P1, P2, and PR were further evaluated.There were significant differences in the P2 and PR among different age groups except for P1. There were no significant differences in the P2 and PR between the first 2 groups, neither among the latter 4 groups. Nevertheless, P2 and PR in the first 2 groups were significantly different from those in the latter 4 groups. In addition, both P2 and PR (not P1) increased significantly with age, systolic and diastolic blood pressure. P1, P2 and PR were not related to heart rate. Both P2 and PR were closely related to the age. PR also had a weak relationship with systolic blood pressure.Both P2 and PR of OA Doppler increase with age. Concern should be raised when P2 and PR are used to evaluate the hemodynamic change of OA.


Assuntos
Artéria Oftálmica/fisiologia , Adulto , Fatores Etários , Idoso , Velocidade do Fluxo Sanguíneo , Feminino , Voluntários Saudáveis , Humanos , Pessoa de Meia-Idade , Artéria Oftálmica/diagnóstico por imagem , Valores de Referência , Fluxo Sanguíneo Regional , Ultrassonografia Doppler em Cores , Adulto Jovem
15.
Med Hypotheses ; 117: 21-27, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30077189

RESUMO

Aberrant expression of miR-146a has been reported to be involved in the progression and metastasis of various types of human cancers; however, its potential role in human neuroblastoma is still poorly understood. The purpose of our study was to investigate the molecular mechanism and possible role of miR-146a in human neuroblastoma. In this study, targeted genes were predicted by bioinformatic analysis and confirmed by dual-Luciferase reporter assay. The expression level of miR-146a in the human neuroblastoma SK-N-SH cell line was detected by quantitative RT-PCR. We used flow cytometric analysis to determine apoptosis and necrosis of SK-N-SH cells after transfection with miR-146a inhibitor, miR-146a mimic, and negative controls. The expression level of target genes was detected by RT-PCR and Western blotting. We identified BCL11A as a target of miR-146a. Thus, miR-146a targets the 3'UTR of BCL11A and inhibits its mRNA and protein expression. Overexpression of miR-146a can inhibit the growth and promote the apoptosis of human neuroblastoma SK-N-SH cells through inhibiting the expression of BCL11A. Furthermore, we found that upregulation of BCL11A by miR-146a inhibitor can promote SK-N-SH cells growth and protect SK-N-SH cells against apoptosis. Our results showed that miR-146a is a potential tumor suppressor gene in human neuroblastoma via directly targeting BCL11A. These findings suggest that miR-146a might be a new candidate target for treatment of human neuroblastoma.


Assuntos
Apoptose , Proteínas de Transporte/genética , MicroRNAs/metabolismo , Neuroblastoma/metabolismo , Proteínas Nucleares/genética , Proteínas de Transporte/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Biologia Computacional , Citometria de Fluxo , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Necrose , Proteínas Nucleares/metabolismo , Plasmídeos/metabolismo , Proteínas Repressoras
16.
Cerebrovasc Dis ; 45(5-6): 204-212, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29627835

RESUMO

BACKGROUND: Early and accurate diagnosis of ischaemic stroke (IS) requires the use of an optimized biomarker. Exosomal microRNAs have the potential to serve as biomarkers owing to their stability and specificity. We investigated the expression levels of plasma-derived exosomal microRNA-21-5p and microRNA-30a-5p in the different phases of IS. METHODS: One hundred forty-three patients with IS and 24 non-stroke controls were enrolled. The patients were divided into the following 5 groups: 1 group for the hyperacute phase IS (HIS, within 6 h); two for the acute phase IS (AIS, including days 1-3 and days 4-7); one for the subacute phase IS (SIS, days 8-14); and one for the recovery phase IS (RIS, days >14). Plasma exosomes were isolated using a QIAGEN exoRNeasy kit and examined by transmission electron -microscopy, nanoparticle tracking, and flow cytometry. The expression levels of miRNA-21-5p and miRNA-30a-5p were detected by quantitative real-time polymerase chain reaction. RESULTS: The plasma exosomal miR-21-5p levels in SIS and RIS were significantly higher than that in controls (p < 0.05 and p < 0.01 respectively). The levels of miR-30a-5p in HIS were significantly higher (p < 0.05) and in AIS (days 1-3) were lower than that in controls (p < 0.05). In AIS (days 1-3), both miRNAs were decreased compared with the HIS group (p = 0.053 and 0.001, respectively). The area under the curve (AUC) of the miR-21-5p was 0.714 for SIS (95% CI 0.570-0.859, p = 0.007), 0.734 for RIS (95% CI 0.596-0.871, p = 0.003); the AUC of the miR-30a-5p was 0.826 for HIS (95% CI 0.665-0.988, p = 0.001), 0.438 for AIS (days 1-3; 95% CI 0.240-0.635, p = 0.516). CONCLUSIONS: The plasma-derived exosomal miR-21-5p and miRNA-30a-5p in combination are promising biomarkers for diagnosing IS and distinguishing among HIS, SIS, and RIS, especially miRNA-30a-5p for the diagnosis of the HIS phase. Our results provide a new reference for clinicians to apply in early-stage diagnosis and identifies the possible value of biomarkers for IS thrombolysis therapy.


Assuntos
Isquemia Encefálica/diagnóstico , Isquemia Encefálica/genética , Exossomos/genética , MicroRNAs/genética , Acidente Vascular Cerebral/diagnóstico , Acidente Vascular Cerebral/genética , Idoso , Isquemia Encefálica/sangue , Estudos de Casos e Controles , Regulação para Baixo , Exossomos/ultraestrutura , Feminino , Marcadores Genéticos , Humanos , Masculino , Microscopia Eletrônica de Transmissão , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Reação em Cadeia da Polimerase em Tempo Real , Acidente Vascular Cerebral/sangue , Fatores de Tempo
17.
Front Aging Neurosci ; 10: 24, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29467645

RESUMO

Background: Differentiation of transient ischaemic attack (TIA) from ischaemic stroke within the thrombolysis time window is difficult. Although TIA may be diagnosed within this window, the latest imaging technologies are complex and costly. Serum markers, which are non-invasive, rapid and economic, are used for diagnosis and prognosis of various diseases. Exosome-derived miRNA markers for TIA are unknown. Methods: We examined focal brain ischaemia produced by occlusion of the middle cerebral artery (MCAo) for 5 min, 10 min, and 2 h in rats. Exosomal miRNAs with consistent trends in cerebrospinal fluid (CSF) and plasma were identified by deep sequencing and quantitative real-time polymerase chain reaction (qRT-PCR). The areas under the curve (AUC) of the receiver operating characteristic (ROC) curve were used to evaluate the diagnostic accuracy of these miRNAs for TIA in rats. Results: Rno-miR-122-5p and rno-miR-300-3p were selected. Plasma exosomal rno-miR-122-5p was significantly downregulated in 10 min ischaemic rats compared with control and 5 min plasma. Plasma exosomal rno-miR-300-3p was significantly upregulated in 5 min ischaemic rats compared with control, 10 min and 2 h rats. Plasma and CSF levels of these miRNAs were correlated. ROC analysis showed high AUC values for rno-miR-122-5p (0.960) and rno-miR-300-3p (0.970) in the 10 and 5 min rats, respectively, compared with controls. Conclusions: Plasma exosomal rno-miR-122-5p and rno-miR-300-3p may be blood-based TIA biomarkers.

18.
Curr Neurovasc Res ; 14(4): 330-337, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28982331

RESUMO

BACKGROUND: MircroRNA (MiRNA) levels are associated with disease pathophysiology and are high in plasma exosomes. Plasma exosomal miRNAs serve as potential therapeutic targets and diagnosis biomarkers in some diseases but few studies have examined them in Ischemic Stroke (IS). Therefore, we explored the potential predictive value of plasma exosomal miR-422a and miR-125b-2-3p in different IS phases (acute and subacute phases). METHODS: Fifty-five IS patients and 25 age and sex matched healthy controls were recruited. Patients were classified into two groups: 27 patients in acute phase (days 1-3) and 28 patients in subacute phase (days 4-14). The plasma exosomal levels of miR-422a and miR-125b-2-3p were examined via quantitative real-time polymerase chain reaction (qRT-PCR). The Areas Under the Curve (AUC) of the Receiver Operating Characteristic (ROC) curve were constructed to evaluate the diagnostic accuracy of these miRNAs in IS. RESULTS: The expression levels of plasma exosomal miR-422a and miR-125b-2-3p were significantly decreased in the subacute phase group (P<0.001, P<0.001, respectively), and the miR-422a levels were increased in the acute phase group (P<0.005) as compared to the controls. Additionally, the expression levels of plasma exosomal miR-422a and miR-125b-2-3p were significantly decreased in the subacute phase group than in the acute phase group (P<0.001, P<0.005, respectively). ROC analysis showed high AUC values for miR-422a and miR-125b-2-3p in the subacute phase group as compared to those in healthy controls: 0.971 and 0.889, respectively, and miR-422a in the acute phase group as compared to healthy controls were 0.769. CONCLUSION: Plasma exosomal miR-422a and miR-125b-2-3p may serve as blood-based biomarkers for monitoring and diagnosing in IS patients, with plasma exosomal miR-422a showing the best diagnostic value. The use of these two plasma exosomal miRNAs in combination may be powerful for determining IS stage.


Assuntos
Isquemia Encefálica/sangue , Exossomos/metabolismo , MicroRNAs/sangue , Acidente Vascular Cerebral/sangue , Idoso , Biomarcadores/sangue , Isquemia Encefálica/diagnóstico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Acidente Vascular Cerebral/diagnóstico
19.
BMC Med Genomics ; 8: 79, 2015 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-26597292

RESUMO

BACKGROUND: Neurogenic neuroprotection is a promising approach for treating patients with ischemic brain lesions. Fastigial nucleus stimulation (FNS) has been shown to reduce the tissue damage resulting from focal cerebral ischemia in the earlier studies. However, the mechanisms of neuroprotection induced by FNS remain unclear. MicroRNAs (miRNAs) are a newly discovered group of non-coding small RNA molecules that negatively regulate target gene expression and involved in the regulation of pathological process. To date, there is a lack of knowledge on the expression of miRNA in response to FNS. Thus, we study the regulation of miRNAs in the rat ischemic brain by the neuroprotection effect of FNS. METHODS: In this study, we used an established focal cerebral ischemia/reperfusion (IR) model in rats. MiRNA expression profile of rat ischemic cortex after 1 h of FNS were investigated using deep sequencing. Microarray was performed to study the expression pattern of miRNAs. Functional annotation on the miRNA was carried out by bioinformatics analysis. RESULTS: Two thousand four hundred ninety three miRNAs were detected and found to be miRNAs or miRNA candidates using deep sequencing technology. We found that the FNS-related miRNAs were differentially expressed according microarray data. Bioinformatics analysis indicated that several differentially expressed miRNAs might be a central node of neuroprotection-associated genetic networks and contribute to neuroprotection induced by FNS. CONCLUSIONS: MiRNA acts as a novel regulator and contributes to FNS-induced neuroprotection. Our study provides a better understanding of neuroprotection induced by FNS.


Assuntos
Núcleos Cerebelares , Biologia Computacional , Estimulação Encefálica Profunda , Sequenciamento de Nucleotídeos em Larga Escala , MicroRNAs/genética , Neuroproteção/genética , Análise de Sequência de RNA , Animais , Isquemia Encefálica/complicações , Masculino , Anotação de Sequência Molecular , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/complicações , Traumatismo por Reperfusão/genética , Traumatismo por Reperfusão/fisiopatologia , Traumatismo por Reperfusão/terapia
20.
Sci Total Environ ; 536: 933-945, 2015 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-26105705

RESUMO

Shanghai is a Chinese megacity in the Yangtze River Delta area, one of the most polluted coastal areas in China. The inhabitants of Shanghai have very high aquatic product consumption rates. A risk-benefit assessment of the co-ingestion of fish nutrients and contaminants has not previously been performed for Shanghai residents. Samples of five farmed fish species (marine and freshwater) with different feeding habits were collected from Shanghai markets in winter and summer. Fatty acids, protein, mercury, cadmium, lead, copper, polychlorinated biphenyls, hexachlorocyclohexanes, and dichlorodiphenyltrichloroethanes were measured in liver, abdominal fat, and dorsal, abdominal, and tail muscles from fish. Tolerable daily intakes and benefit-risk quotients were calculated to allow the benefits and risks of co-ingesting n-3 long-chain polyunsaturated fatty acids and contaminants to be assessed according to the cancer slope factors and reference doses of selected pollutants. All of the contaminant concentrations in the muscle tissues were much lower than the national maximum limits, but the livers generally contained high Hg concentrations, exceeding the regulatory limit. The organic pollutant and n-3 long-chain polyunsaturated fatty acid concentrations correlated with the lipid contents of the fish tissues, and were higher in carnivorous marine fish than in omnivorous and herbivorous freshwater fish. The tolerable daily intakes, risk-benefit quotients, and current daily aquatic product intakes for residents of large Chinese cities indicated that the muscle tissues of most of the fish analyzed can be consumed regularly without significant contaminant-related risks to health. However, attention should be paid to the potential risks posed by dichlorodiphenyltrichloroethane in large yellow croaker and Hg in tilapia. Based on the results of this study, we encourage people to consume equal portions of marine and freshwater fish.


Assuntos
Exposição Ambiental/estatística & dados numéricos , Peixes/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , China , Cidades , Monitoramento Ambiental , Humanos , Nível de Efeito Adverso não Observado , Medição de Risco
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