Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 97
Filtrar
1.
Signal Transduct Target Ther ; 6(1): 368, 2021 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-34645784

RESUMO

The long-term immunity and functional recovery after SARS-CoV-2 infection have implications in preventive measures and patient quality of life. Here we analyzed a prospective cohort of 121 recovered COVID-19 patients from Xiangyang, China at 1-year after diagnosis. Among them, chemiluminescence immunoassay-based screening showed 99% (95% CI, 98-100%) seroprevalence 10-12 months after infection, comparing to 0.8% (95% CI, 0.7-0.9%) in the general population. Total anti-receptor-binding domain (RBD) antibodies remained stable since discharge, while anti-RBD IgG and neutralization levels decreased over time. A predictive model estimates 17% (95% CI, 11-24%) and 87% (95% CI, 80-92%) participants were still 50% protected against detectable and severe re-infection of WT SARS-CoV-2, respectively, while neutralization levels against B.1.1.7 and B.1.351 variants were significantly reduced. All non-severe patients showed normal chest CT and 21% reported COVID-19-related symptoms. In contrast, 53% severe patients had abnormal chest CT, decreased pulmonary function or cardiac involvement and 79% were still symptomatic. Our findings suggest long-lasting immune protection after SARS-CoV-2 infection, while also highlight the risk of immune evasive variants and long-term consequences for COVID-19 survivors.


Assuntos
Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , COVID-19/imunologia , Memória Imunológica , Modelos Imunológicos , SARS-CoV-2/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Adulto , COVID-19/diagnóstico por imagem , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Tomografia Computadorizada por Raios X
2.
Nat Commun ; 12(1): 5270, 2021 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-34489413

RESUMO

Following injury, cells in regenerative tissues have the ability to regrow. The mechanisms whereby regenerating cells adapt to injury-induced stress conditions and activate the regenerative program remain to be defined. Here, using the mammalian neonatal heart regeneration model, we show that Nrf1, a stress-responsive transcription factor encoded by the Nuclear Factor Erythroid 2 Like 1 (Nfe2l1) gene, is activated in regenerating cardiomyocytes. Genetic deletion of Nrf1 prevented regenerating cardiomyocytes from activating a transcriptional program required for heart regeneration. Conversely, Nrf1 overexpression protected the adult mouse heart from ischemia/reperfusion (I/R) injury. Nrf1 also protected human induced pluripotent stem cell-derived cardiomyocytes from doxorubicin-induced cardiotoxicity and other cardiotoxins. The protective function of Nrf1 is mediated by a dual stress response mechanism involving activation of the proteasome and redox balance. Our findings reveal that the adaptive stress response mechanism mediated by Nrf1 is required for neonatal heart regeneration and confers cardioprotection in the adult heart.


Assuntos
Coração/fisiologia , Traumatismo por Reperfusão Miocárdica/metabolismo , Fator 1 Relacionado a NF-E2/metabolismo , Animais , Animais Recém-Nascidos , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Doxorrubicina/farmacologia , Feminino , Heme Oxigenase (Desciclizante)/genética , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos , Masculino , Camundongos Knockout , Camundongos Transgênicos , Traumatismo por Reperfusão Miocárdica/patologia , Miócitos Cardíacos/fisiologia , Fator 1 Relacionado a NF-E2/genética , Oxirredução , Proteostase , Ratos Sprague-Dawley , Regeneração
3.
Res Vet Sci ; 140: 198-202, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34525439

RESUMO

Marine Streptomyces S073 was previously shown to have strong anti-Vibrio activity, and its antibacterial mechanism was proposed to be associated with siderophore-mediated iron competition and other antagonistic agents. In this study, anti-Vibrio compounds produced by S073 were isolated by bioassay-guided fractionation using column chromatography and HPLC, and the target compound in the most active fraction was identified as dibutyl phthalate (DBP) by various spectroscopic analyses, including EI-MS, 1H NMR and 13C NMR. The DBP-producing capacity of S073 was 2.39 mg/L in ISP1 culture media. Pure DBP was demonstrated to have strong inhibitory activity on Vibiro parahaemolyticus growth with an MIC of 31.25 mg/L. When standard DBP was supplemented into the S073 fermentation broth in a gradient method, an additive inhibitory effect on V. parahaemolyticus was observed, indicating the important role of DBP in driving anti-Vibrio activity in S073 metabolites pool. A synergistic additive effect between DBP and florfenicol was observed in the Vibrio inhibition. These results indicate that, to achieve Vibrio-inhibition, S073 exerted multifaceted strategies, which included DBP-mediated antagonism and siderophore-governed iron competition. The application potential of S073 as an aquaculture probiotic was evaluated, and the safety risks associated with the endocrine disruptor attributes of DBP were discussed.


Assuntos
Probióticos , Streptomyces , Vibrio , Animais , Aquicultura , Dibutilftalato
5.
Adv Mater ; 33(32): e2101005, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34219279

RESUMO

Coronary artery disease is the "first killer" in the world, while the classical treatment for this disease is to implant stent. An ideal vascular stent should be nontoxic with self-expanding characteristics, quick expanding speed, and appropriate mechanical supporting property. However, no existing vascular stent covers all properties. Herein, a two-way shape-memory cellulose vascular stent, which can realize shape adjustments by mild solutions such as water and alcohol, is constructed. The shape-memory characteristics, mechanical properties, cell toxicity, and biocompatibility, are systemically investigated by ex vivo experiment as well as molecule simulation and theoretical modeling, revealing that the achieved bilayer two-way shape-memory films (BSMFs) can be used as an artificial vascular stent. In particular, this vascular stent made from BSMFs shows superb biocompatibility according to live/dead cell viability assays. Ex vivo experiments reveal that the novel vascular stent can support arteria coronaria sinistra, or the left main coronary artery, at the opening state while the cross-section of the vessel becomes two times larger than that of the initial state after implantation. Thus, it is believed that effective and scalable BSMFs can make meritorious fundamental contributions to biomaterials science and practical applications such as vascular stents.

6.
J Fish Dis ; 44(10): 1647-1655, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34133777

RESUMO

The orange-spotted grouper (Epinephelus coioides) is an important marine farmed fish in China. It is affected by the bacterial pathogen Vibrio alginolyticus, which causes high mortality and substantial economic losses. We studied the transcriptional changes of the IgZ gene in E. coioides following V. alginolyticus stimulation and investigated the distribution of IgZ in different tissues. The highest expression level of IgZ occurred in the head kidney. When fish were stimulated with live and inactivated V. alginolyticus, the expression levels of IgZ in the head kidney, spleen, intestine, gills and blood cells were significantly upregulated. In an in situ hybridization study, IgZ mRNA-positive cells were detected in the head kidney, spleen and gill, but positive signals were not detected in the liver and intestine. IgZ-labelled cells increased in the head kidney, spleen and gills post-infection with V. alginolyticus for 21 days. The present study provides additional evidence that IgZ is involved in mucosal immune responses and helps explain the role of IgZ in E. coioides defence against V. alginolyticus infection.


Assuntos
Bass , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Perfilação da Expressão Gênica/veterinária , Vacinação/veterinária , Vibrioses/veterinária , Vibrio alginolyticus/fisiologia , Animais , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/imunologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/metabolismo , Cadeias Pesadas de Imunoglobulinas/genética , Cadeias Pesadas de Imunoglobulinas/metabolismo , Distribuição Aleatória , Vibrioses/imunologia , Vibrioses/microbiologia
7.
J Biomed Nanotechnol ; 17(6): 1007-1019, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-34167616

RESUMO

In recent years, the emergence of non-toxic but catalytically active inorganic nanoparticles has attracted great attention for cancer treatment, but the therapeutic effect has been affected by the limited reactive oxygen species in tumors. Therefore, the combination of chemotherapy and chemodynamic therapy is regarded as a promising therapeutic strategy. In this paper, we reported the preparation and bioactivity evaluation of poly(lactic acid-co-glycolic acid) (PLGA) grafted-γ-Fe2O3 nanoparticles with dual response of endogenous peroxidase and catalase like activities. Our hypothesis is that PLGAgrafted γ-Fe2O3 nanoparticles could be used as a drug delivery system for the anti-tumor drug doxorubicin to inhibit the growth of lung adenocarcinoma A549 cells; meanwhile, based on its mimic enzyme properties, this kind of nanoparticles could be combined with doxorubicin in the treatment of A549 cells. Our experimental results showed that the PLGAgrafted γ-Fe2O3 nanoparticles could simulate the activity of catalase and decompose hydrogen peroxide into H2O and oxygen in neutral tumor microenvironment, thus reducing the oxidative damage caused by hydrogenperoxide to lung adenocarcinoma A549 cells. In acidic microenvironment, PLGA grafted γ-Fe2O3 nanoparticles could simulate the activity of peroxidase and effectively catalyze the decomposition of hydrogen peroxide to generate highly toxic hydroxyl radicals, which could cause the death of A549 cells. Furthermore, the synergistic effect of peroxidase-like activity of PLGA-grafted γ-Fe2O3 nanoparticles and doxorubicin could accelerate the apoptosisand destruction of A549 cells, thus enhancing the antitumor effect of doxorubicin-loaded PLGA-grafted γ-Fe2O3 nanoparticles. Therefore, this study provides an effective nanoplatform based on dual inorganic biomimetic nanozymes for the treatment of lung cancer.


Assuntos
Adenocarcinoma de Pulmão , Nanopartículas , Células A549 , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Compostos Férricos , Humanos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Microambiente Tumoral
9.
Carbohydr Polym ; 262: 117936, 2021 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-33838813

RESUMO

Inspired by muscle architectures, double network hydrogels with hierarchically aligned structures were fabricated, where cross-linked cellulose nanofiber (CNF)/chitosan hydrogel threads obtained by interfacial polyelectrolyte complexation spinning were collected in alignment as the first network, while isotropic poly(acrylamide-co-acrylic acid) (PAM-AA) served as the second network. After further cross-linking using Fe3+, the hydrogel showed an outstanding mechanical performance, owing to effective energy dissipation of the oriented asymmetric double networks. The average strength and elongation-at-break of PAM-AA/CNF/Fe3+ hydrogel were 11 MPa and 480 % respectively, which the strength was comparative to that of biological tissues. The aligned CNFs in the hydrogels provided probable ion transport channels, contributing to the high ionic conductivity, which was up to 0.022 S/cm when the content of LiCl was 1.5 %. Together with superior biocompatibility, the well-ordered hydrogel showed a promising potential in biological applications, such as artificial soft tissue materials and muscle-like sensors for human motion monitoring.


Assuntos
Materiais Biocompatíveis/química , Celulose/química , Quitosana/química , Hidrogéis/química , Nanofibras/química , Acrilamidas/química , Reagentes para Ligações Cruzadas/química , Condutividade Elétrica , Humanos , Íons/química , Fenômenos Mecânicos , Músculos , Resistência à Tração
10.
Signal Transduct Target Ther ; 6(1): 107, 2021 03 03.
Artigo em Inglês | MEDLINE | ID: mdl-33658482

RESUMO

Synthetic glucocorticoid dexamethasone is the first trial-proven drug that reduces COVID-19 mortality by suppressing immune system. In contrast, interferons are a crucial component of host antiviral immunity and can be directly suppressed by glucocorticoids. To investigate whether therapeutic interferons can compensate glucocorticoids-induced loss of antiviral immunity, we retrospectively analyzed a cohort of 387 PCR-confirmed COVID-19 patients with quasi-random exposure to interferons and conditional exposure to glucocorticoids. Among patients receiving glucocorticoids, early interferon therapy was associated with earlier hospital discharge (adjusted HR 1.68, 95% CI 1.19-2.37) and symptom relief (adjusted HR 1.48, 95% CI 1.06-2.08), while these associations were insignificant among glucocorticoids nonusers. Early interferon therapy was also associated with lower prevalence of prolonged viral shedding (adjusted OR 0.24, 95% CI 0.10-0.57) only among glucocorticoids users. Additionally, these associations were glucocorticoid cumulative dose- and timing-dependent. These findings reveal potential therapeutic synergy between interferons and glucocorticoids in COVID-19 that warrants further investigation.


Assuntos
COVID-19/tratamento farmacológico , Dexametasona/administração & dosagem , Interferons/administração & dosagem , SARS-CoV-2 , Adulto , COVID-19/diagnóstico , COVID-19/mortalidade , Teste de Ácido Nucleico para COVID-19 , Dexametasona/agonistas , Sinergismo Farmacológico , Feminino , Humanos , Interferons/agonistas , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
11.
Proc Natl Acad Sci U S A ; 118(13)2021 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-33758101

RESUMO

Among the large, diverse set of mammalian long noncoding RNAs (lncRNAs), long noncoding primary microRNAs (lnc-pri-miRNAs) are those that host miRNAs. Whether lnc-pri-miRNA loci have important biological function independent of their cognate miRNAs is poorly understood. From a genome-scale lncRNA screen, lnc-pri-miRNA loci were enriched for function in cell proliferation, and in glioblastoma (i.e., GBM) cells with DGCR8 or DROSHA knockdown, lnc-pri-miRNA screen hits still regulated cell growth. To molecularly dissect the function of a lnc-pri-miRNA locus, we studied LOC646329 (also known as MIR29HG), which hosts the miR-29a/b1 cluster. In GBM cells, LOC646329 knockdown reduced miR-29a/b1 levels, and these cells exhibited decreased growth. However, genetic deletion of the miR-29a/b1 cluster (LOC646329-miR29Δ) did not decrease cell growth, while knockdown of LOC646329-miR29Δ transcripts reduced cell proliferation. The miR-29a/b1-independent activity of LOC646329 corresponded to enhancer-like activation of a neighboring oncogene (MKLN1), regulating cell propagation. The LOC646329 locus interacts with the MKLN1 promoter, and antisense oligonucleotide knockdown of the lncRNA disrupts these interactions and reduces the enhancer-like activity. More broadly, analysis of genome-wide data from multiple human cell types showed that lnc-pri-miRNA loci are significantly enriched for DNA looping interactions with gene promoters as well as genomic and epigenetic characteristics of transcriptional enhancers. Functional studies of additional lnc-pri-miRNA loci demonstrated cognate miRNA-independent enhancer-like activity. Together, these data demonstrate that lnc-pri-miRNA loci can regulate cell biology via both miRNA-dependent and miRNA-independent mechanisms.


Assuntos
Proliferação de Células/genética , Loci Gênicos , RNA Longo não Codificante/metabolismo , Apoptose/genética , Técnicas de Silenciamento de Genes , Células HEK293 , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/genética , RNA-Seq
12.
Medicine (Baltimore) ; 100(7): e24727, 2021 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-33607815

RESUMO

PURPOSE: To discuss the value of caudate lobectomy in hilar cholangiocarcinoma (HCCA) treatment. METHODS: A systematic review was performed in PubMed, MEDLINE database, EMBASE, and Cochrane Library for trials comparing combined caudate lobectomy with controls from January 1, 1990 to December 2, 2020. The outcomes were postoperative radical cure information, survival condition, morbidity, and mortality. RESULT: Ten studies were included. No difference was observed in the morbidity (odd ratio (OR) 0.93, 95% confidence interval (CI) 0.65-1.33) and mortality (OR 1.16, 95% CI 0.55-2.42) between the combined caudate lobectomy and control groups. Hepatectomy combined with caudate lobectomy was associated with higher incidence of radical resection (OR 3.88, 95% CI 2.18-6.90) and longer survival (hazard ratio 0.45, 95% CI 0.38-0.55). CONCLUSION: Combining caudate lobectomy can significantly increase the incidence of radical resection of HCCA and the postoperative survival time. The morbidity and mortality were not increased after the operation. Thus, caudate lobectomy should be included when performing partial hepatectomy for HCCA.


Assuntos
Neoplasias dos Ductos Biliares/patologia , Hepatectomia/métodos , Tumor de Klatskin/cirurgia , Estudos de Casos e Controles , Terapia Combinada/métodos , Terapia Combinada/estatística & dados numéricos , Gerenciamento de Dados , Humanos , Mortalidade , Ensaios Clínicos Controlados não Aleatórios como Assunto , Prognóstico , Análise de Sobrevida , Resultado do Tratamento
13.
Environ Sci Pollut Res Int ; 28(13): 16408-16419, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33387322

RESUMO

The novel magnetic biochar (MBC), derived from co-pyrolysis of sewage sludge and biomass loading nanosized iron oxide particles, was used as an environmentally friendly adsorbent. The loading of magnetic particles was in favor of increasing the adsorption capacity and separation from aqueous system for biochar (BC). The physical/chemical characteristics of MBC were revealed by elemental analysis, VSM, SEM-EDS, XRD, FTIR, zeta potential, and batch adsorption-desorption experiments. The nanosized γ-Fe2O3 particles grown on the surface of biochar showed ferromagnetic property. For the remediation of Cu(II) contamination, MBC-5 showed remarkable adsorption capacity of 67.68 mg/g, and presented a wide pH range of 3.0-6.0. The Langmuir isothermal and pseudo-second-order model could describe adsorption process well. The adsorption mechanism of Cu(II) involved physical adsorption, ion exchange, and electrostatic surface complexation on the surface of MBCs. In the desorption experiments, MBC-5 holds the adsorption efficiency of 81.09% after fifth recycle still, which illustrated a remarkable performance of cyclic utilization by the solid waste of sewage sludge and biomass.


Assuntos
Pirólise , Poluentes Químicos da Água , Adsorção , Biomassa , Carvão Vegetal , Cinética , Fenômenos Magnéticos , Esgotos
14.
Lab Invest ; 101(4): 412-422, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33454724

RESUMO

Data processing and learning has become a spearhead for the advancement of medicine, with pathology and laboratory medicine has no exception. The incorporation of scientific research through clinical informatics, including genomics, proteomics, bioinformatics, and biostatistics, into clinical practice unlocks innovative approaches for patient care. Computational pathology is burgeoning subspecialty in pathology that promises a better-integrated solution to whole-slide images, multi-omics data, and clinical informatics. However, computational pathology faces several challenges, including the ability to integrate raw data from different sources, limitation of hardware processing capacity, and a lack of specific training programs, as well as issues on ethics and larger societal acceptable practices that are still solidifying. The establishment of the entire industry of computational pathology requires far-reaching changes of the three essential elements connecting patients and doctors: the local laboratory, the scan center, and the central cloud hub/portal for data processing and retrieval. Computational pathology, unlocked through information integration and advanced digital communication networks, has the potential to improve clinical workflow efficiency, diagnostic quality, and ultimately create personalized diagnosis and treatment plans for patients. This review describes clinical perspectives and discusses the statistical methods, clinical applications, potential obstacles, and future directions of computational pathology.


Assuntos
Inteligência Artificial , Biologia Computacional , Patologia Clínica , Humanos , Informática Médica , Estatística como Assunto
15.
Environ Sci Pollut Res Int ; 28(17): 21396-21410, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33411270

RESUMO

The bio-char was prepared by co-pyrolysis of municipal sewage sludge and biomass with chemical activation. The alkaline activating agents of KOH and K2CO3 were used to develop multilevel pore structure without heavy metal. The proximate analysis, ultimate analysis, SEM, and surface area and porosity analyzer were applied to present the physico-chemical properties and multilevel pore structure of bio-char. After impregnation pretreatment, the KOH provided more functional ingredients and reacted with C to expand pore structure for bio-chars. It was confirmed the specific surface area reached 2122.43 m2/g, and micropore area was 1674.85 m2/g after co-pyrolysis at 800 °C. Through the pretreatment of alkaline activation, the novel evaluation of heavy metal immobilization behavior in bio-char matrix were investigated by BCR sequential extraction and leaching tests. The KOH activation showed prominent immobilization behavior relatively, and the K2CO3 activation had more noticeable effects on leaching behavior. For Cu, Ni, Cr, Cd, Pb, and Zn, after co-pyrolysis at 900 °C, the proportion of unstable fraction decreased significantly, and the residual fractions of heavy metals were above 89.44% according to BCR sequential extraction procedure. Under optimal pyrolysis temperature, the Er value of bio-char reduced to 41.93, and the potential ecological risks decreased from considerable risk to low risk to ensure the further eco-friendly application.


Assuntos
Carvão Vegetal , Metais Pesados , Pirólise , Esgotos
16.
Acta Neurol Belg ; 121(1): 95-106, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32960423

RESUMO

Ischemic stroke is a refractory disease generally caused by cerebral ischemic injury. Remote ischemic preconditioning (RIPC) caused by transient ischemia and reperfusion of the femoral artery exerts a protective effect on ischemic stroke-induced brain injury. This study was designed to investigate the potential molecular mechanism of RIPC-mediated neuroprotection, namely, the biological effects of microRNA-144 on RIPC in mice with ischemic stroke and its effects on PTEN and Akt signaling pathways. Healthy adult C57BL6 mice were selected for the establishment of middle cerebral artery occlusion (MCAO). One hour before the start, remote ischemic preconditioning of limbs was performed in mice. Brain edema and infarct volume were measured. The expressions of microRNA-144, PTEN, and Akt were measured. The results showed that, compared with MCAO group, the RIPC group protected mice from cerebral ischemia-reperfusion injury, systemic accumulation of inflammatory cytokines, and accelerated apoptosis of parenchymal cells. In RIPC group, PTEN expression decreased, and mir-144 and Akt expression increased. The level of phosphorylated PTEN in the transfected microRNA-144 inhibitor group increased and the level of phosphorylated Akt reduced significantly. In conclusion, our results suggest that microRNA-144 may play a protective role in remote ischemic pretreatment by downregulating PTEN and upregulating Akt, suggesting that microRNA-144 via PTEN/Akt pathway may be of therapeutic significance in ischemic stroke.

18.
Cell Rep ; 33(10): 108472, 2020 12 08.
Artigo em Inglês | MEDLINE | ID: mdl-33296652

RESUMO

The adult mammalian heart has limited capacity for regeneration following injury, whereas the neonatal heart can readily regenerate within a short period after birth. Neonatal heart regeneration is orchestrated by multiple cell types intrinsic to the heart, as well as immune cells that infiltrate the heart after injury. To elucidate the transcriptional responses of the different cellular components of the mouse heart following injury, we perform single-cell RNA sequencing on neonatal hearts at various time points following myocardial infarction and couple the results with bulk tissue RNA-sequencing data collected at the same time points. Concomitant single-cell ATAC sequencing exposes underlying dynamics of open chromatin landscapes and regenerative gene regulatory networks of diverse cardiac cell types and reveals extracellular mediators of cardiomyocyte proliferation, angiogenesis, and fibroblast activation. Together, our data provide a transcriptional basis for neonatal heart regeneration at single-cell resolution and suggest strategies for enhancing cardiac function after injury.

19.
STAR Protoc ; 1(2): 100049, 2020 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-33111095

RESUMO

Murine cardiomyocytes undergo proliferation, multinucleation, and polyploidization during the first 3 weeks of postnatal life, resulting in a mixture of diploid and tetraploid cardiomyocytes in the heart. Understanding the molecular differences between diploid and tetraploid cardiomyocytes from these processes has been limited due to lack of unique markers and their heterogenous origins. Here, we apply single-nucleus RNA-sequencing to fluorescence-activated cell sorting-selected diploid and tetraploid cardiomyocytes to characterize their heterogeneity and molecular distinctions. For complete details on the use and execution of this protocol, please refer to Cui et al. (2020).

20.
Mol Biol Rep ; 47(9): 7107-7114, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32880831

RESUMO

To better explore the application potential of heat shock protein Hsp70s in diverse areas including biomonitoring, a further investigation of the details of the regulatory mechanism governing Hsp70 transcription is required. A transcriptional factor ChGATA-4 that displayed affinity to the ChHsp70 promoter of Crassostrea hongkongensis was isolated and identified by DNA affinity purification as well as mass spectrometry analysis. The ChGATA-4 cDNA is 2162 bp in length and the open reading frame encodes a polypeptide containing 482 amino acids with a conserved zinc finger domain. The over-expression of ChGATA-4 significantly inhibited the expression of ChHsp70 promoter in heterologous HEK293T cells. However, the depletion of ChGATA-4 mRNA by RNAi technique resulted in significant increase of ChHsp70 transcription in oyster hemocytes. The RT-PCR results demonstrated that the transcription of both ChHsp70 and ChGATA-4 were induced by heat, Cd, or NP (Nonyl phenol) stress. This suggested a potential correlation between ChHsp70 and ChGATA-4 in the stress-mediated genetic regulatory cascade. This study demonstrated that ChGATA-4 acts in a negative manner in controlling ChHsp70 transcription in C. hongkongensis and promotes to further understand the mechanisms leading Hsp70 transcription.


Assuntos
Crassostrea , Fator de Transcrição GATA4 , Proteínas de Choque Térmico HSP70 , Transcrição Genética , Animais , Crassostrea/genética , Crassostrea/metabolismo , Fator de Transcrição GATA4/genética , Fator de Transcrição GATA4/metabolismo , Proteínas de Choque Térmico HSP70/biossíntese , Proteínas de Choque Térmico HSP70/genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...