RESUMO
Dwarfism is an important agronomic trait in fruit breeding programs. However, the germplasm resources required to generate dwarf pear (Pyrus spp.) varieties are limited. Moreover, the mechanisms underlying dwarfism remain unclear. In this study, "Yunnan" quince (Cydonia oblonga Mill.) had a dwarfing effect on "Zaosu" pear. Additionally, the dwarfism-related NAC transcription factor gene PbNAC71 was isolated from pear trees comprising "Zaosu" (scion) grafted onto "Yunnan" quince (rootstock). Transgenic Nicotiana benthamiana and pear OHF-333 (Pyrus communis) plants overexpressing PbNAC71 exhibited dwarfism, with a substantially smaller xylem and vessel area relative to the wild-type controls. Yeast one-hybrid, dual-luciferase, chromatin immunoprecipitation-qPCR, and electrophoretic mobility shift assays indicated that PbNAC71 downregulates PbWalls are thin 1 expression by binding to NAC-binding elements in its promoter. Yeast two-hybrid assays showed that PbNAC71 interacts with the E3 ubiquitin ligase PbRING finger protein 217 (PbRNF217). Furthermore, PbRNF217 promotes the ubiquitin-mediated degradation of PbNAC71 by the 26S proteasome, thereby regulating plant height as well as xylem and vessel development. Our findings reveal a mechanism underlying pear dwarfism and expand our understanding of the molecular basis of dwarfism in woody plants.
Assuntos
Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Plantas Geneticamente Modificadas , Pyrus , Fatores de Transcrição , Xilema , Xilema/metabolismo , Xilema/genética , Pyrus/genética , Pyrus/metabolismo , Pyrus/crescimento & desenvolvimento , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Nicotiana/genética , Nicotiana/metabolismo , Nicotiana/crescimento & desenvolvimento , Regiões Promotoras Genéticas/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Complexo de Endopeptidases do Proteassoma/genéticaRESUMO
Fourier ptychographic microscopy (FPM) is used to achieve high resolution and a large field of view. However, traditional FPM image reconstruction methods often yield poor image quality when encountering out-of-focus issues during reconstruction. Therefore, this study proposes a defocus-distance regression network based on convolutional neural networks. In an experimental validation, the root-mean-square error calculated from 1000 sets of predicted and true values was approximately 6.2 µm. The experimental results suggest that the proposed method has good generalization, maintains high accuracy in predicting defocus distances even for different biological samples, and extends the imaging depth-of-field of the FPM system by a factor of more than 3.
RESUMO
The regulation of fruit development is a complex process and a core issue in the fruit tree industry. To investigate the role of PbGIF1 in pear fruit development, we identified a transcription factor PbbHLH137 that regulates pear (Pyrus bretschneideri) fruit development by screening a yeast library constructed from fruit cDNA. Yeast two-hybrid (Y2H), bimolecular fluorescence complementation (BiFC), and split luciferase complementation (split-LUC) assays were performed to confirm the PbbHLH137-PbGIF1 interaction. By tracing the complete fruit development process, we found that PbbHLH137 expression was closely related to fruit size and highly involved at the late pear fruit development stage. Transgenic experiments showed that heterologous expression of PbbHLH137 or PbGIF1 promoted fruit enlargement. PbbHLH137 promoted mainly the expansion of fruit cell volume, whereas PbGIF1 mainly increased the number of cells. Further LUC experiments demonstrated that PbGIF1 promoted the transcriptional activation ability of PbbHLH137. Our work identified PbbHLH137 as a transcription factor that regulates fruit development, and showed that PbGIF1 played an ongoing role during fruit development, making it a candidate gene for genetic improvement of pear fruit development.
Assuntos
Frutas , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Pyrus , Fatores de Transcrição , Pyrus/genética , Pyrus/crescimento & desenvolvimento , Pyrus/metabolismo , Frutas/crescimento & desenvolvimento , Frutas/genética , Frutas/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Plantas Geneticamente Modificadas , Técnicas do Sistema de Duplo-HíbridoRESUMO
Redclaw crayfish (Cherax quadricarinatus) is a large, tropical freshwater crustacean species with considerable potential of commercial production. In recent years, infection with DIV1 in redclaw crayfish is being reported in aquaculture industries, causing high mortality and huge economic losses. However, many characteristics of this virus, including pathogenesis, transmission mechanism, and host immunity, remain largely unknown.MicroRNAs are known to play important roles in numerous biological processes, and many microRNAs are reported to be involved in the regulation of immune responses. In this study, nine-small RNA libraries were constructed using hemocytes of redclaw crayfish to characterize the differentially expressed miRNAs (DE-miRNAs) at 24 and 48 h postinfection (hpi). A total of 14 and 22 DE-miRNAs were identified in response to DIV1 infection at 24 and 48 hpi, respectively. Further, functional annotation of the predicted host target genes using GO and KEGG pathway enrichment analyses indicated that relevant biological processes and signal pathways underwent miRNA-mediated regulation after DIV1 infection. Our results enhanced the understanding of the mechanisms of miRNA-mediated regulation of immune responses under DIV1 infection in crustaceans.
RESUMO
Nucleic acid sequence-based amplification (NASBA) provides a fast and convenient approach for nucleic acid amplification under isothermal conditions, and its combination with an immunoassay-based lateral flow dipstick (LFD) could produce a higher detection efficiency for M. rosenbergii nodavirus isolated from China (MrNV-chin). In this study, two specific primers and a labelled probe of the capsid protein gene of MrNV-chin were constructed. The process of this assay mainly included a single-step amplification at a temperature of 41 â for 90 min, and hybridization with an FITC-labeled probe for 5 min, with the hybridization been required for visual identification during LFD assay. The test results indicated that, the NASBA-LFD assay showed sensitivity for 1.0 fg M. rosenbergii total RNA with MrNV-chin infection, which was 104 times that of the present RT-PCR approach for the detection of MrNV. In addition, no products were created for shrimps with infection of other kinds of either DNA or RNA virus, which indicated that the NASBA-LFD was specific for MrNV. Therefore, the combination of NASBA and LFD is a new alternative detection method for MrNV which is rapid, accurate, sensitive and specific without expensive equipment and specialised personnel. Early detection of this infectious disease among aquatic organisms will help implement efficient therapeutic strategy to prevent its spread, enhance animal health and limit loss of aquatic breeds in the event of an outbreak.
Assuntos
Nodaviridae , Palaemonidae , Vírus de RNA , Animais , Replicação de Sequência Autossustentável , Nodaviridae/genética , Vírus de RNA/genética , Técnicas de Amplificação de Ácido Nucleico/métodosRESUMO
BACKGROUND: Parthenocarpy results in traits attractive to both consumers and breeders, and it overcomes the obstacle of self-incompatibility in the fruit set of horticultural crops, including pear (Pyrus bretshneider). However, there is limited knowledge regarding the genetic and molecular mechanisms that regulate parthenogenesis. RESULTS: Here, in a transcriptional comparison between pollination-dependent fruit and GA4-induced parthenocarpy, PbCYP78A6 was identified and proposed as a candidate gene involved in parthenocarpy. PbCYP78A6 is similar to Arabidopsis thaliana CYP78A6 and highly expressed in pear hypanthia. The increased PbCYP78A6 expression, as assessed by RT-qPCR, was induced by pollination and GA4 exposure. The ectopic overexpression of PbCYP78A6 contributed to parthenocarpic fruit production in tomato. The PbCYP78A6 expression coincided with fertilized and parthenocarpic fruitlets development and the expression of fruit development-related genes as assessed by cytological observations and RT-qPCR, respectively. PbCYP78A6 RNA interference and overexpression in pear calli revealed that the gene is an upstream regulator of specific fruit development-related genes in pear. CONCLUSIONS: Our findings indicate that PbCYP78A6 plays a critical role in fruit formation and provide insights into controlling parthenocarpy.
Assuntos
Ciclo Celular , Sistema Enzimático do Citocromo P-450/genética , Genes de Plantas/genética , Partenogênese , Proteínas de Plantas/genética , Pyrus/metabolismo , Ciclo Celular/genética , Ciclo Celular/fisiologia , Sistema Enzimático do Citocromo P-450/fisiologia , Perfilação da Expressão Gênica , Genes de Plantas/fisiologia , Partenogênese/genética , Partenogênese/fisiologia , Filogenia , Proteínas de Plantas/fisiologia , Polinização , Pyrus/genética , Pyrus/crescimento & desenvolvimento , Pyrus/fisiologiaRESUMO
ABSTRACT: This study assessed the efficacy and safety of tirofiban in combination with dual-antiplatelet therapy (DAPT) in progressive ischemic stroke. One hundred and four patients equally divided into a tirofiban group or DAPT group were enrolled from June 2018 to December 2019. Efficacy outcomes included National Institutes of Health Stroke Scale score for 14 days, and modified Rankin scale (mRs) scores as excellent (mRs 0-1) or favorable (mRs 0-2) measured 90 days after stroke. At 14 days, the tirofiban group had a lower National Institutes of Health Stroke Scale score compared with the DAPT group (F = 14.959, P = 0.000). The mRS scores of the 2 groups at 90 days after treatment were significantly different from those before treatment. At 90 days, excellent favorable functional outcome (mRS ≤ 2) was achieved in 33 of 52 (63.43%) patients in the tirofiban group compared with 25 of 52 (48.08%) patients in the DAPT group. The incidence of bleeding was 5.77% in the tirofiban group, compared with 0% in DAPT group. Intravenous (IV) tirofiban alone or combined with DAPT was shown to be safe and effectively improved clinical outcome in progressive ischemic stroke patients. IV tirofiban was shown to be superior to the DAPT regimen.
Assuntos
Aspirina/administração & dosagem , Clopidogrel/administração & dosagem , Terapia Antiplaquetária Dupla , AVC Isquêmico/tratamento farmacológico , Inibidores da Agregação Plaquetária/administração & dosagem , Tirofibana/administração & dosagem , Idoso , Aspirina/efeitos adversos , Clopidogrel/efeitos adversos , Avaliação da Deficiência , Terapia Antiplaquetária Dupla/efeitos adversos , Feminino , Estado Funcional , Hemorragia/induzido quimicamente , Humanos , Infusões Intravenosas , AVC Isquêmico/diagnóstico , AVC Isquêmico/fisiopatologia , Masculino , Pessoa de Meia-Idade , Inibidores da Agregação Plaquetária/efeitos adversos , Recuperação de Função Fisiológica , Estudos Retrospectivos , Fatores de Tempo , Tirofibana/efeitos adversos , Resultado do TratamentoRESUMO
SYJ15 is a highly pathogenic Gram-positive Bacillus sp. with top bud spore newly isolated from dying soft shell turtle. 16SrDNA sequencing showed that it is highly homologous to B. cereus, B. thuringiensis and B. anthracis. Biochemical examinations showed that it belongs to B. cereus. To further study the new pathogen, we conducted whole-genome sequencing based on single-molecular sequencing technology from PacBio. Genome assembly analysis showed that the strain has a 5,296,886 bp chromosome, a 218,649 bp plasmid and a 5221 bp plasmid with GC content of 35.51%, 31.91% and 29.75%, respectively. The genome contains 5736 coding sequences and 6 CRISPR systems located in the chromosome as well as 11 genomic islands in the chromosome and the large plasmid. Genome function analyses were annotated by nr database, SwissProt, KEGG, COG, GO, PHI, VFDB, ARDB, Secretory_Protein and T3SS. In addition, 13 gene clusters of secondary metabolism were predicted by antiSMASH. Comparison of SYJ15 with B. subtilis, B. anthracis, B. cereus and B. thuringiensis identified 1031 core genes of the five strains and 816 genes specific to SYJ15. In addition, SYJ15 had the most common core genes with B. thuringiensis, and the least with B. subtilis. Phylogenetic analysis demonstrated that SYJ15 is between B. thuringiensis and B. cereus, suggesting that SYJ15 belongs to Bacillus cereus group. We designed a specific primer pair to distinguish SYJ15 from B. pumilus, B. licheniformis, B.subtilis, B. thuringiensis and B. cereus. In conclusion, information of SYJ15 genome will help to enhance our understanding of pathogenesis of SYJ15 and find effective treatment.
Assuntos
Bacillus cereus/classificação , Bacillus cereus/genética , Genoma Bacteriano/genética , Filogenia , Tartarugas/microbiologia , Animais , Bacillus/classificação , Bacillus/genética , Plasmídeos/genética , Análise de Sequência , Sequenciamento Completo do GenomaRESUMO
We developed a convenient technique to detect Herpesviral haematopoietic necrosis attributed to cyprinid herpes virus 2 (CyHV-2), a serious disease of Crucian carp and goldfish related to high mortality. In the present study, we employed a lateral flow dipstick (LAMP-LFD) to present a loop-mediated isothermal amplification assay. The specificity was ascertained via other six viruses, and the sensitivity was compared using PCR method, which are the reaction conditions changes for the method improved. The results revealed that CyHV-2 performance was observable at 64 °C in a separated tube within 60 min, when the samples hybridized using an FITC-labeled probe. As the LAMP-LFD method's specificity was high, with its sensitivity identical to that of traditional PCR, the overall DNA collected revealed the lowest detection limit of 0.18 pg/µl from goldfish diseased by CyHV-2. In summary, the development of LAMP-LFD's method does not require expensive instruments, and it can be regarded as a fast, simple, and reliable method for CyHV-2 detection.
Assuntos
Herpesviridae/genética , Herpesviridae/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Reologia/métodos , Animais , Cyprinidae/virologia , Doenças dos Peixes/virologia , Sensibilidade e EspecificidadeRESUMO
Trionyx sinensis Hemorrhagic Syndrome Virus (TSHSV) is the firstly discovered aquatic arterivirus inducing high mortality of Trionyx sinensis. So far, the lack of genomic resources has hindered further research on revealing the immunological characteristics of T. sinensis in response to TSHSV. In the present study, we performed a transcriptome analysis from the lungs of T. sinensis challenged by TSHSV using Illumina-based RNA-Seq. The validity of transcriptomic data was confirmed with the gradual increase of TSHSV RNA copies detected in lung. A total of 103079339 clean reads were generated, and 58374764 unique mapped reads were analyzed. Assembly of the sequence data allowed identifying 16383 unigenes consisting of 36 significant differentially expressed genes (DEGs). These DEGs were categorized into 30 GO-enriched bioprocesses and 9â¯KEGG pathways. The combinational analysis of GO-enriched bioprocesses and KEGG pathways demonstrated that TSHSV modulated several immune genes of T. sinensis related to various biological processes, including virus recognition (RIG-I/MDA-5), immune initiation (IFIT-1 and IFIT-5), endocytosis (CUBN, ENPP2 and LRP2) and steroid metabolism (FCNIL and STAR). In summary, the finding of this study revealed several immune pathways and candidated genes involved in the immune response of T. sinensis against TSHSV-infection. These results will provide helpful information to investigate molecular mechanism of T. sinensis in response to TSHSV.
Assuntos
Arteriviridae/fisiologia , Pulmão/metabolismo , Infecções por Vírus de RNA/veterinária , Transcriptoma , Tartarugas , Animais , Perfilação da Expressão Gênica/veterinária , Pulmão/virologia , Infecções por Vírus de RNA/metabolismo , Infecções por Vírus de RNA/virologia , RNA-Seq/veterinária , Proteínas de Répteis/análiseRESUMO
Parthenocarpy, the productions of seedless fruit without pollination or fertilization, is a potentially desirable trait in many commercially grown fruits, especially in pear, which is self-incompatible. Phytohormones play important roles in fruit set, a process crucial for parthenocarpy. In this study, 2,4-dichlorophenoxyacetic acid (2,4-D), an artificially synthesized plant growth regulator with functions similar to auxin, was found to induce parthenocarpy in pear. Histological observations revealed that 2,4-D promoted cell division and expansion, which increased cortex thickness, but the effect was weakened by paclobutrazol (PAC), a gibberellin (GA) biosynthesis inhibitor. Phenotypic differences in pear may therefore be due to different GA contents. Hormone testing indicated that 2,4-D mainly induced the production of bioactive GA4 , rather than GA3. Three key oxidase genes function in the GA biosynthetic pathway: GA20ox, GA3ox and GA2ox. In a pear group treated with only 2,4-D, PbGA20ox2-like and PbGA3ox-1 were significantly upregulated. When treated with 2,4-D supplemented with PAC, however, expression levels of these genes were significantly downregulated. Additionally, PbGA2ox1-like and PbGA2ox2-like expression levels were significantly downregulated in pear treated with either 2,4-D only or 2,4-D supplemented with PAC. We thus hypothesize that 2,4-D can induce parthenocarpy by enhancing GA4 biosynthesis.
Assuntos
Ácido 2,4-Diclorofenoxiacético/farmacologia , Giberelinas/metabolismo , Pyrus/efeitos dos fármacos , Pyrus/fisiologia , Frutas/efeitos dos fármacos , Frutas/fisiologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Triazóis/farmacologiaRESUMO
Trionyx sinensis hemorrhagic syndrome virus (TSHSV) is a newly discovered lethal arterivirus that causes serious disease in Trionyx sinensis in China. In this study, the complete genome sequence of TSHSV was determined by RACE cloning, and the functions of the predicted proteins were predicted. The complete genome of TSHSV was found to be 17,875 bp in length, and a 3'-end poly(A) tail was detected. Eight TSHSV hypothetical proteins (TSHSV-HPs) were predicted by gene model identification. TSHSV-HP2, 3 and 4 were associated with replicase activity, since papain-like protease (PLPs), serine-type endopeptidase, P-loop-containing nucleoside triphosphate hydrolase, and EndoU-like endoribonuclease motifs were detected. Phylogenetic analysis showed that TSHSV clusters with an arterivirus from a Chinese broad-headed pond turtle.
Assuntos
Infecções por Arterivirus/veterinária , Arterivirus/classificação , Arterivirus/isolamento & purificação , Filogenia , Tartarugas/virologia , Animais , Arterivirus/genética , Infecções por Arterivirus/virologia , China , Genoma Viral , RNA Mensageiro , Análise de Sequência de DNA , Proteínas Virais/genéticaRESUMO
In this letter, we propose an indoor visible light positioning technique using a Modified Momentum Back-Propagation (MMBP) algorithm based on received signal strength (RSS) with sparse training data set. Unlike other neural network algorithms that require a large number of training data points to locate accurately, we have realized high-precision positioning for 100 test points with only 20 training points in a 1.8 m × 1.8 m × 2.1 m localization area. In order to verify the adaptability of the MMBP algorithm, we experimentally demonstrate two different training data acquisition methods adopting either even or arbitrary training sets. In addition, we also demonstrate the positioning accuracy of the traditional RSS algorithm. Experimental results show that the average localization accuracy optimized by our proposed algorithm is only 1.88 cm for the arbitrary set and 1.99 cm for the even set, while the average positioning error of the traditional RSS algorithm reaches 14.34 cm. Comparison indicates that the positioning accuracy of our proposed algorithm is 7.6 times higher. Results also show that the performance of our system is higher than some previous reports based on RSS and RSS fingerprint databases using complex machine learning algorithms trained by a large amount of training points.
RESUMO
The large yellow croaker Larimichthys crocea (L. crocea) is one of the most economically important marine fish in China and East Asian countries. It also exhibits peculiar behavioral and physiological characteristics, especially sensitive to various environmental stresses, such as hypoxia and air exposure. These traits may render L. crocea a good model for investigating the response mechanisms to environmental stress. To understand the molecular and genetic mechanisms underlying the adaptation and response of L. crocea to environmental stress, we sequenced and assembled the genome of L. crocea using a bacterial artificial chromosome and whole-genome shotgun hierarchical strategy. The final genome assembly was 679 Mb, with a contig N50 of 63.11 kb and a scaffold N50 of 1.03 Mb, containing 25,401 protein-coding genes. Gene families underlying adaptive behaviours, such as vision-related crystallins, olfactory receptors, and auditory sense-related genes, were significantly expanded in the genome of L. crocea relative to those of other vertebrates. Transcriptome analyses of the hypoxia-exposed L. crocea brain revealed new aspects of neuro-endocrine-immune/metabolism regulatory networks that may help the fish to avoid cerebral inflammatory injury and maintain energy balance under hypoxia. Proteomics data demonstrate that skin mucus of the air-exposed L. crocea had a complex composition, with an unexpectedly high number of proteins (3,209), suggesting its multiple protective mechanisms involved in antioxidant functions, oxygen transport, immune defence, and osmotic and ionic regulation. Our results reveal the molecular and genetic basis of fish adaptation and response to hypoxia and air exposure. The data generated by this study will provide valuable resources for the genetic improvement of stress resistance and yield potential in L. crocea.
Assuntos
Adaptação Fisiológica , Proteínas de Peixes/genética , Genoma , Pressão Osmótica , Estresse Oxidativo , Perciformes/genética , Animais , Proteínas de Peixes/metabolismo , Perciformes/metabolismo , TranscriptomaRESUMO
Real-time sensing of reactive oxygen species (ROS) and timely scavenging of excessive ROS in physiological environments are critically important in the diagnosis and prevention of ROS-related diseases. To solve the mismatch problem between conventional rigid ROS biosensors and biological tissues in terms of both modulus and composition, here, we present a flexible ferrocene-based hydrogel biosensor designed for oxidative stress detection and antioxidation treatment. The hydrogel was fabricated through a supramolecular assembly of ferrocene-grafted polyethylenimine (PEI-Fc), sodium alginate (SA), and polyvinyl alcohol (PVA). Multiple non-covalent interactions, including electrostatic interactions between PEI-Fc and SA, hydrophobic interactions and π-π stacking among ferrocene groups, and the PVA crystalline domain, synergistically improve the mechanical properties of the PVA/SA/PEI-Fc hydrogel. The flexible PVA/SA/PEI-Fc hydrogel biosensor exhibited a broad detection range for hydrogen peroxide (H2O2), from 0 to 120 µM, using the differential pulse voltammetry method. Furthermore, the hydrogel demonstrated effective ROS scavenging and oxygen generation performance, desirable biocompatibility, and satisfactory antibacterial activity, making it suitable for biological interfaces. In vitro studies revealed that the PVA/SA/PEI-Fc hydrogel could monitor H2O2 concentration in the proximity of inflammatory cells, and effectively scavenge ROS to protect cells from oxidative stress damage. This all-in-one multifunctional hydrogel, integrating both sensing and treatment functions, holds great promise for clinical applications in the diagnosis and management of ROS-related diseases.
Assuntos
Técnicas Biossensoriais , Compostos Ferrosos , Hidrogéis , Hidrogéis/química , Antioxidantes , Peróxido de Hidrogênio , Metalocenos , Espécies Reativas de Oxigênio , Técnicas Biossensoriais/métodos , Estresse Oxidativo , Alginatos/químicaRESUMO
OBJECTIVE: To investigate the expression patterns and clinical significance of specific lymphocyte subsets in the peripheral blood of patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. METHODS: Between December 2022 and February 2023, a cohort of 165 patients from the First Affiliated Hospital of Guangzhou University of Chinese Medicine were analyzed. The participants represented various stages of coronavirus infection severity: mild, moderate, severe, and critical. Additionally, 40 healthy individuals constituted the control group. The FC 500MPL flow cytometer and associated reagents for flow cytometry. RESULTS: Compared with the healthy control group, activated B lymphocytes witnessed a pronounced increase (p < .05). A significant decrease was observed in the levels of Breg, Cytotoxic T cells or Suppressor T-cell (Tc/s), late-activated T, late-activated Th, and late-activated Tc/s lymphocytes (p < .05). Th, initial Th, initial Tc/s, total Treg, natural Treg, induced Treg, early activated T, and early activated Th lymphocyte levels showed no significant difference (p > .05). As the disease progressed, there was an uptick in midterm activated T lymphocytes (p < .05), while Breg, T, Tc/s, senescent Tc/s, and total senescent T levels dwindled (p < .05). Noteworthy patterns emerged across different groups for B1, T-lymphocytes, Tc/s, B2, CD8+ Treg cells, and other subsets, highlighting variance in immune responses relative to disease severity. When juxtaposed, no significant difference was found in the expression levels of lymphocyte subsets between patients who died and those deemed critically ill (p > .05). CONCLUSION: Subsets of Treg and B-cells could act as yardsticks for the trajectory of SARS-CoV-2 infection and might have potential in forecasting patient trajectories. A comprehensive evaluation of lymphocyte subsets, especially in real-time, holds the key to discerning the clinical severity in those with COVID-19. This becomes instrumental in monitoring treatment outcomes, tracking disease evolution, and formulating prognostications. Moreover, the results provide a deeper understanding of the cellular immune defense mechanisms against the novel coronavirus infection.
Assuntos
COVID-19 , Subpopulações de Linfócitos , SARS-CoV-2 , Índice de Gravidade de Doença , Humanos , COVID-19/imunologia , COVID-19/sangue , Masculino , Feminino , Pessoa de Meia-Idade , SARS-CoV-2/imunologia , Adulto , Prognóstico , Subpopulações de Linfócitos/imunologia , Subpopulações de Linfócitos/metabolismo , Idoso , Linfócitos B/imunologia , Linfócitos T Reguladores/imunologia , Citometria de Fluxo , Ativação Linfocitária/imunologiaRESUMO
Matrix metalloproteinase 9 (MMP-9) plays an important role in wound healing. However, overexpression of MMP-9 leads to the degradation of the newly formed extracellular matrix, which delays wound healing, ultimately leading to chronic wounds. Therefore, timely monitoring of the MMP-9 activity using simple, cost-effective methods is important to prevent the formation of chronic wounds. In this work, ferrocene-modified MMP-9 cleavage peptide (Fc-MG) modified carboxymethyl chitosan hydrogels were prepared as electrochemical biosensors. In the presence of MMP-9, the peptide chain is sheared, and the electrochemically active ferrocene segment is released. Therefore, analyzing the electrochemical activity of hydrogels using differential pulse voltammetry (DPV) can be used to determine MMP-9 activity. The results showed that the DPV peaks were correlated with the MMP-9 concentration in phosphate-buffered saline (PBS, pH 7.4) and Dulbecco's modified Eagle's medium (DMEM). Specifically, the corresponding coefficient of determination (R2) were 0.918 and 0.993. The limit of detections were 73.08 ng/mL and 131.71 ng/mL, respectively. Compared with the enzyme-linked immunosorbent assay, the hydrogel biosensor determined the concentration of MMP-9 in solution with simpler steps. This study demonstrates a novel strategy based on Fc-MG-modified hydrogels to monitor MMP-9 activity in cell secretion samples and shows the potential application in chronic wounds.
Assuntos
Técnicas Biossensoriais , Quitosana , Técnicas Eletroquímicas , Compostos Ferrosos , Hidrogéis , Metaloproteinase 9 da Matriz , Metalocenos , Quitosana/química , Quitosana/análogos & derivados , Metaloproteinase 9 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/análise , Metalocenos/química , Hidrogéis/química , Compostos Ferrosos/química , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , HumanosRESUMO
Quantitative phase microscopy (QPM) is indispensable in biomedical research due to its advantages in unlabeled transparent sample thickness quantification and obtaining refractive index information. Fourier ptychographic microscopy (FPM) is among the most promising QPM methods, incorporating multi-angle illumination and iterative phase recovery for high-resolution quantitative phase imaging (QPI) of large cell populations over a wide field of-view (FOV) in a single pass. However, FPM is limited by data redundancy and sequential acquisition strategies, resulting in low imaging efficiency, which in turn limits its real-time application in in vitro label-free imaging. Here, we report a fast QPM based on Fourier ptychography (FQP-FPM), which uses an optimized annular downsampling and parallel acquisition strategy to minimize the amount of data required in the front end and reduce the iteration time of the back-end algorithm (3.3% and 4.4% of conventional FPM, respectively). Theoretical and data redundancy analyses show that FQP-FPM can realize high-throughput quantitative phase reconstruction at thrice the resolution of the coherent diffraction limit by acquiring only ten raw images, providing a precondition for in vitro label-free real-time imaging. The FQP-FPM application was validated for various in vitro label-free live-cell imaging. Cell morphology and subcellular phenomena in different periods were observed with a synthetic aperture of 0.75â NA at a 10× FOV, demonstrating its advantages and application potential for fast high-throughput QPI.
RESUMO
Chemical reorganization and interfacial engineering in hybrid nanomaterials are promising strategies for enhancing electrocatalytic performance. Herein, MoO3@zeolitic imidazolate framework-67 (ZIF-67) heterogeneous nanoribbons are designed through coordination assembly. By following heat treatment, a Mo/Mo2C@CoO@NC heterostructure with nitrogen-doped carbon-encapsulated CoO hexagons (CoO@NC) anchored on the Mo/Mo2C jag matrix was fabricated. Notably, through controllable experimental optimization, the as-prepared Mo/Mo2C@CoO@NC heterostructure exhibits numerous active centers (e.g. Mo, Mo2C, CoO, and NC), fully exposed active sites (numerous pores and jagged structures), and abundant heterointerfaces (Mo/Mo2C, Mo2C/CoO@NC, Mo2C/amorphous, and CoO@NC/amorphous), and exhibits good conductivity (localized single-crystal behavior, graphitized carbon). As a result, the as-developed Mo/Mo2C@CoO@NC heterostructures inherit impressive oxygen evolution reaction (OER) performance with an overpotential of only 215 mV at 10 mA cm-2. Furthermore, Mo/Mo2C@CoO@NC heterostructures exhibit excellent stability with a current density retention of 98.4% after 20 h chronoamperometry. This work provides deep insights into chemical reconstructions and tuning heterointerfaces to efficiently enhance the OER activity of heterostructure-based electrocatalysts.
RESUMO
The Chinese soft-shelled turtle (Pelodiscus sinensis) is an important freshwater aquaculture animal in China. The Wnt gene family plays important regulatory roles in the development and growth of mammals. However, the precise function of these family genes has not been well understood in the sex differentiation of Chinese soft-shelled turtles. Here, we cloned a member of the Wnt family, Wnt2, which obtained a 1077 bp open reading frame that encoded a 358-aa protein. The putative amino acid sequences of proteins are exceeded 80% identical to other turtles. The expression level of Wnt2 peaked at the 14th stage both in female and male embryos during the early gonadal differentiation period of Chinese soft-shelled turtles, which occurred before gonadal differentiation. Wnt2 mRNA was expressed at higher levels in the brains and gonads of mature P. sinensis females compared with those in mature males. Wnt agonists significantly affected the expression level of Wnt2 during the gonadal differentiation period. After Wnt agonists (1.0 µg/µL, 2.5 µg/µL, 5.0 µg/µL) treatment, the expression level of the Wnt2 generally appeared to have an inverted-V trend over time in female embryonic gonads. The results suggested that Wnt2 may participate in the regulation of gonad development in P. sinensis during the early embryonic stages. These results could provide a theoretical basis for the reproduction process of the Chinese soft-shelled turtle.