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1.
Am J Hum Genet ; 2024 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-39226898

RESUMO

Variants that alter gene splicing are estimated to comprise up to a third of all disease-causing variants, yet they are hard to predict from DNA sequencing data alone. To overcome this, many groups are incorporating RNA-based analyses, which are resource intensive, particularly for diagnostic laboratories. There are thousands of functionally validated variants that induce mis-splicing; however, this information is not consolidated, and they are under-represented in ClinVar, which presents a barrier to variant interpretation and can result in duplication of validation efforts. To address this issue, we developed SpliceVarDB, an online database consolidating over 50,000 variants assayed for their effects on splicing in over 8,000 human genes. We evaluated over 500 published data sources and established a spliceogenicity scale to standardize, harmonize, and consolidate variant validation data generated by a range of experimental protocols. According to the strength of their supporting evidence, variants were classified as "splice-altering" (∼25%), "not splice-altering" (∼25%), and "low-frequency splice-altering" (∼50%), which correspond to weak or indeterminate evidence of spliceogenicity. Importantly, 55% of the splice-altering variants in SpliceVarDB are outside the canonical splice sites (5.6% are deep intronic). These variants can support the variant curation diagnostic pathway and can be used to provide the high-quality data necessary to develop more accurate in silico splicing predictors. The variants are accessible through an online platform, SpliceVarDB, with additional features for visualization, variant information, in silico predictions, and validation metrics. SpliceVarDB is a very large collection of splice-altering variants and is available at https://splicevardb.org.

2.
J Am Chem Soc ; 146(21): 14422-14426, 2024 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-38709624

RESUMO

Here we report a concise and divergent synthesis of scabrolide A and havellockate, representative members of polycyclic marine natural product furano(nor)cembranoids. The synthesis features a highly efficient exo-exo-endo radical cascade. Through the generation of two rings, three C-C bonds, and three contiguous stereocenters in one step, this remarkable transformation not only assembles the bowl-shaped, common 6-5-5 fused ring system from simple building blocks but also precisely installs the functionalities at desired positions and sets the stage for further divergent preparation of both target molecules. Further studies reveal that the robust and unusual 6-endo radical addition in the cascade is likely facilitated by the rigidity of the substrate.

3.
BMC Plant Biol ; 24(1): 390, 2024 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-38730367

RESUMO

Granulation of juice sacs is a physiological disorder, which affects pomelo fruit quality. Here, the transcriptome and ubiquitinome of the granulated juice sacs were analyzed in Guanxi pomelo. We found that lignin accumulation in the granulated juice sacs was regulated at transcription and protein modification levels. In transcriptome data, we found that the genes in lignin biosynthesis pathway and antioxidant enzyme system of the granulated juice sacs were significantly upregulated. However, in ubiquitinome data, we found that ubiquitinated antioxidant enzymes increased in abundance but the enzyme activities decreased after the modification, which gave rise to reactive oxygen species (ROS) contents in granulated juice sacs. This finding suggests that ubiquitination level of the antioxidant enzymes is negatively correlated with the enzyme activities. Increased H2O2 is considered to be a signaling molecule to activate the key gene expressions in lignin biosynthesis pathway, which leads to the lignification in granulated juice sacs of pomelo. This regulatory mechanism in juice sac granulation of pomelo was further confirmed through the verification experiment using tissue culture by adding H2O2 or dimethylthiourea (DMTU). Our findings suggest that scavenging H2O2 and other ROS are important for reducing lignin accumulation, alleviating juice sac granulation and improving pomelo fruit quality.


Assuntos
Citrus , Lignina , Lignina/metabolismo , Citrus/metabolismo , Citrus/genética , Sucos de Frutas e Vegetais/análise , Espécies Reativas de Oxigênio/metabolismo , Transcriptoma , Peróxido de Hidrogênio/metabolismo , Regulação da Expressão Gênica de Plantas , Frutas/metabolismo , Frutas/genética , Antioxidantes/metabolismo
4.
Artigo em Inglês | MEDLINE | ID: mdl-39225840

RESUMO

PURPOSE: Identification of mature sperm at microdissection testicular sperm extraction (mTESE) is a crucial step of sperm retrieval to help patients with non-obstructive azoospermia (NOA) proceed to intracytoplasmic sperm injection. Touch print smear (TPS) cytology allows immediate interpretation and prompt sperm identification intraoperatively. In this study, we leverage machine learning (ML) to facilitate TPS reading and conquer the learning curve for new operators. MATERIALS AND METHODS: One hundred seventy-six microscopic TPS images from the testicular specimen of patients with azoospermia at Taipei Veterans General Hospital were retrospectively collected, including categories of Sertoli cell, primary spermatocytes, round spermatids, elongated spermatids, immature sperm, and mature sperm. Among them, 118 images were assigned as the training set and 29 images as the validation set. RetinaNet (Lin et al. in IEEE Trans Pattern Anal Mach Intell. 42:318-327, 2020), a one-stage detection framework, was adopted for cell detection. The performance was evaluated at the cell level with average precision (AP) and recall, and the precision-recall (PR) curve was displayed among an independent testing set that contains 29 images that aim to assess the model. RESULTS: The training set consisted of 4772 annotated cells, including 1782 Sertoli cells, 314 primary spermatocytes, 443 round spermatids, 279 elongated spermatids, 504 immature sperm, and 1450 mature sperm. This study demonstrated the performance of each category and the overall AP and recall on the validation set, which were 80.47% and 96.69%. The overall AP and recall were 79.48% and 93.63% on the testing set, while increased to 85.29% and 93.80% once the post-meiotic cells were merged into one category. CONCLUSIONS: This study proposed an innovative approach that leveraged ML methods to facilitate the diagnosis of spermatogenesis at mTESE for patients with NOA. With the assistance of ML techniques, surgeons could determine the stages of spermatogenesis and provide timely histopathological diagnosis for infertile males.

5.
Int J Mol Sci ; 25(17)2024 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-39273633

RESUMO

The maize Snf2 gene family plays a crucial role in chromatin remodeling and response to environmental stresses. In this study, we identified and analyzed 35 members of the maize Snf2 gene family (ZmCHR1 to ZmCHR35) using the Ensembl Plants database. Each protein contained conserved SNF2-N and Helicase-C domains. Phylogenetic analysis revealed six groups among the Snf2 proteins, with an uneven distribution across subfamilies. Physicochemical analysis indicated that the Snf2 proteins are hydrophilic, with varied amino acid lengths, isoelectric points, and molecular weights, and are predominantly localized in the nucleus. Chromosomal mapping showed that these genes are distributed across all ten maize chromosomes. Gene structure analysis revealed diverse exon-intron arrangements, while motif analysis identified 20 conserved motifs. Collinearity analysis highlighted gene duplication events, suggesting purifying selection. Cis-regulatory element analysis suggested involvement in abiotic and biotic stress responses. Expression analysis indicated tissue-specific expression patterns and differential expression under various stress conditions. Specifically, qRT-PCR validation under drought stress showed that certain Snf2 genes were upregulated at 12 h and downregulated at 24 h, revealing potential roles in drought tolerance. These findings provide a foundation for further exploration of the functional roles of the maize Snf2 gene family in development and stress responses.


Assuntos
Regulação da Expressão Gênica de Plantas , Família Multigênica , Filogenia , Proteínas de Plantas , Estresse Fisiológico , Zea mays , Zea mays/genética , Zea mays/metabolismo , Estresse Fisiológico/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Secas , Cromossomos de Plantas/genética , Mapeamento Cromossômico , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
6.
Anal Chem ; 95(39): 14665-14674, 2023 10 03.
Artigo em Inglês | MEDLINE | ID: mdl-37679861

RESUMO

A sandwich immunoassay theoretically exhibits higher sensitivity and specificity compared to a competitive counterpart; however, it is extremely difficult to obtain a pair of antibodies that can bind to a small molecule simultaneously, which is always thought to be a single epitope. In the present study, abamectin (ABM) was selected to prove the effect of hapten design and antibody recognition properties on the development of a sandwich immunoassay for small molecules. First, the epitopes of ABM were roughly located, and epitope distances were determined. Then, two haptens were designed by introducing spacer arms at the C4″-OH and C5-OH of ABM, respectively, aiming to provide the longest epitope distances. A total of seven rabbit polyclonal antibodies (pAbs) and 21 mouse monoclonal antibodies (mAbs) with various recognition properties were obtained. Extensive combinatorial associations of antibody pairs for simultaneously binding to ABM were performed, and only two mAb-mAb pairs were observed to achieve a sandwich immunoassay for ABM with a total success rate of 0.27%. The best mAb pair for sandwich immunoassay was confirmed by surface plasmon resonance, used to develop a sandwich immunoassay, and then evaluated by cross-reactivities and molecular docking with structurally similar analogues and abamectin. Altogether, the study provided a theoretical foundation as well as practical experience and demonstrated the importance of careful hapten design and extensive antibody screening to successfully establish the sandwich immunoassay for small molecules.


Assuntos
Anticorpos Monoclonais , Haptenos , Animais , Camundongos , Coelhos , Simulação de Acoplamento Molecular , Anticorpos Monoclonais/química , Imunoensaio , Epitopos , Ensaio de Imunoadsorção Enzimática
7.
Anal Chem ; 94(51): 17843-17852, 2022 12 27.
Artigo em Inglês | MEDLINE | ID: mdl-36519948

RESUMO

The pursuit of the limit between dimensionalities is a scientific goal with high applicability. Sandwich immunoassay, usually based on two antibodies binding two epitopes, is one of the most popular mainstay tools in both academic and industrial fields. Herein, we determined and evaluated the minimum distance of two epitopes in sandwich immunoassays for small molecules. Briefly, nine model analytes comprising two hapten epitopes, that is, melamine (MEL) and p-nitroaniline (NIA), were designed by increasing the linear chain linkers brick by brick. Two groups of monoclonal antibodies (mAbs) were produced with different recognition properties toward MEL and NIA using 12 new haptens with different spacer arms. The results indicated that two epitopes of the analyte with a distance of only 2.4 Å could be simultaneously bound by two mAbs, which is the known limit of epitope distance in sandwich immunoassays thus far. We further found that an epitope distance of below 8.8 Å for the analyte generally induces noticeable steric hindrance of antibodies, preventing a sandwich immunoassay with high probability. These observations were investigated and evaluated by molecular docking, molecular dynamics, and surface plasmon resonance and using model and real analytes. Altogether, we determined the minimum distance of two epitopes and explored the molecular mechanism of the antibody-analyte-antibody ternary complex in sandwich immunoassays, providing a theoretical basis for hapten design, antibody discovery and development, and sandwich immunoassay establishment for small molecules.


Assuntos
Anticorpos Monoclonais , Haptenos , Epitopos , Simulação de Acoplamento Molecular , Imunoensaio/métodos , Anticorpos Monoclonais/química
8.
Sensors (Basel) ; 22(9)2022 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-35591170

RESUMO

In recent decades, zebrafish have become an increasingly popular laboratory organism in several fields of research due to their ease of reproduction and rapid maturation. In particular, shoaling behavior has attracted the attention of many researchers. This article presents a fully printed robotic model used to sense and stimulate shoaling behavior in zebrafish (Danio rerio). Specifically, we exposed laboratory-fabricated replicated materials to critical acid/base/salt environments and evaluated the mechanical, optical, and surface properties after a three-month immersion period. Focusing on weatherability, these test samples maintained high tensile strength (~45 MPa) and relatively similar transmission (>85%T in the visible region), as determined by UV−vis/FTIR spectroscopy. Three-dimensional (3D) printing technology allowed printing of models with different sizes and appearances. We describe the sense of zebrafish responses to replicas of different sizes and reveal that replicas approximating the true zebrafish size (3 cm) are more attractive than larger replicas (5 cm). This observation suggests that larger replicas appear as predators to the zebrafish and cause fleeing behavior. In this study, we determined the weatherability of a high-transparency resin and used it to fabricate a fully printed driving device to induce shoaling by zebrafish. Finally, we demonstrate a weathering-resistant (for three months) 3D-printed decoy model with potential utility for future studies of outdoor shoaling behavior, and the result has the potential to replace the traditional metal frame devices used in outdoor experiments.


Assuntos
Procedimentos Cirúrgicos Robóticos , Peixe-Zebra , Animais , Comportamento Animal/fisiologia , Impressão Tridimensional , Comportamento Social
9.
Nano Lett ; 20(4): 2857-2864, 2020 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-32163291

RESUMO

Two-dimensional spiral plasmonic structures have emerged as a versatile approach to generate near-field vortex fields with tunable topological charges. We demonstrate here a far-field approach to observe the chiral second-harmonic generation (SHG) at designated visible wavelengths from a single plasmonic vortex metalens. This metalens comprises an Archimedean spiral slit fabricated on atomically flat aluminum epitaxial film, which allows for precise tuning of plasmonic resonances and subsequent transfer of two-dimensional materials on top of the spiral slit. The nonlinear optical measurements show a giant SHG circular dichroism. Furthermore, we have achieved an enhanced chiral SHG conversion efficiency (about an order of magnitude greater than the bare aluminum lens) from monolayer tungsten disulfide (WS2)/aluminum metalens, which is designed at the C-exciton resonance of WS2. Since the C-exciton is not a valley exciton, the enhanced chiral SHG in this hybrid system originates from the plasmonic vortex field-enhanced SHG under the optical spin-orbit interaction.

10.
Int J Mol Sci ; 22(20)2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-34681776

RESUMO

Anthocyanins are natural pigments with antioxidant effects that exist in various fruits and vegetables. The accumulation of anthocyanins is induced by environmental signals and regulated by transcription factors in plants. Numerous evidence has indicated that among the environmental factors, light is one of the most signal regulatory factors involved in the anthocyanin biosynthesis pathway. However, the signal transduction of light and molecular regulation of anthocyanin synthesis remains to be explored. Here, we focus on the research progress of signal transduction factors for positive and negative regulation in light-dependent and light-independent anthocyanin biosynthesis. In particular, we will discuss light-induced regulatory pathways and related specific regulators of anthocyanin biosynthesis in plants. In addition, an integrated regulatory network of anthocyanin biosynthesis controlled by transcription factors is discussed based on the significant progress.


Assuntos
Antocianinas/biossíntese , Luz , Redes e Vias Metabólicas/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Redes e Vias Metabólicas/genética , Desenvolvimento Vegetal/genética , Desenvolvimento Vegetal/efeitos da radiação , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/efeitos da radiação , Plantas/genética , Plantas/metabolismo , Plantas/efeitos da radiação
11.
Molecules ; 26(18)2021 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-34577037

RESUMO

Theoretically, sandwich immunoassay is more sensitive and has a wider working range than that of competitive format. However, it has been thought that small molecules cannot be detected by the sandwich format due to their limited size. In the present study, we proposed a novel strategy for achieving sandwich immunoassay of ß-lactams with low molecular weights. Firstly, five ß-lactam antibiotics were selected to bind with penicillin binding protein (PBP)2x* to form complexes. Then, monoclonal and polyclonal antibodies against PBP2x*-ß-lactams complexes were produced by animal immunization. Subsequently, the optimal pairing antibodies were utilized to establish sandwich immunoassay for detection of 18 PBP2x*-ß-lactam complexes. Among them, ceftriaxone could be detected at as low as 1.65 ng/mL with working range of 1-1000 ng/mL in milk. To reveal the detection mechanism, computational chemistry and molecular recognition study were carried out. The results showed that ß-lactams with a large size and complex structures maybe conducive to induce conformational changes of PBP2x*, and then exhibit greater possibility of being detected by sandwich immunoassay after combination with PBP2x*. This study provides insights for subsequent investigations of anti-metatype antibody screening and sandwich immunoassay establishment for small-molecule detection.


Assuntos
Proteínas de Ligação às Penicilinas , Streptococcus pneumoniae , beta-Lactamas , Antibacterianos , Testes de Sensibilidade Microbiana
12.
Biochem Biophys Res Commun ; 527(4): 1057-1063, 2020 07 05.
Artigo em Inglês | MEDLINE | ID: mdl-32439176

RESUMO

Salidroside possesses excellent anti-tumor activity in many types of malignant tumor. In present study, we focused on the effects of salidroside on hepatocellular carcinoma (HCC). The viability of human HCC cells was assayed using MTT. Apoptosis in the cells and tissues samples were detected by Annexin V/PI or TUNEL staining assays. The levels of apoptosis and endoplasmic reticulum (ER) stress related proteins were measured by western blotting analysis. We found salidroside significantly suppressed cell viability and promoted apoptosis in HCC cells. Salidroside could activate intrinsic and extrinsic apoptotic pathways, by increasing activities of caspase-3, caspase-8 and caspase-9, up-regulating levels of Bax, Cytochrome c and decreasing level of Bcl-2 in HepG2 cells. Moreover, it was found salidroside induced ER stress and increased expression of p-PERK, eIF2a, p-eIF2a, ATF-6 and CHOP in HepG2 cells. Interestingly, knockdown of CHOP impaired salidroside induced inhibitory effects on HepG2 cells, suggesting the important role of ER stress in cytotoxic effect of salidroside. Finally, we have confirmed salidroside induced ER stress and inhibited development of HepG2 in an xenograft mouse model. In conclusion, our data suggest salidroside inhibits viability and induces apoptosis of HCC both in vitro and vivo, and this effect is partially mediated by activation of ER stress.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/tratamento farmacológico , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Glucosídeos/farmacologia , Neoplasias Hepáticas/tratamento farmacológico , Fenóis/farmacologia , Animais , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/uso terapêutico , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Glucosídeos/química , Glucosídeos/uso terapêutico , Células Hep G2 , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Fenóis/química , Fenóis/uso terapêutico , Rhodiola/química
13.
Mol Cell ; 45(6): 777-89, 2012 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-22364742

RESUMO

Abnormal inflammatory signaling activation occurs commonly in cancer cells. However, how it is initiated and maintained and its roles in early stages of tumorigensis are largely unknown. Here, we report that the monocyte-derived MCP-1-induced transformation of immortal breast epithelial cells is triggered by transient activation of MEK/ERK and IKK/NF-κB pathways and maintained by constitutive activation of a feed-forward inflammatory signaling circuit composed of miR-200c, p65, JNK2, HSF1, and IL6. Suppression of miR-200c by IL6 constitutively activates p65/RelA and JNK2, and the latter phosphorylates and activates HSF1. In turn, HSF1 triggers demethylation of the IL6 promoter that facilitates the binding of p65 and c-Jun, which together drive constitutive IL6 transcription. Importantly, this signaling circuit is manifest in human cancer cells and in a mouse model of ErbB2-driven breast cancer, where IL6 loss significantly impairs tumorigenesis. Therefore, targeting this signaling circuit represents an effective therapeutic avenue for breast cancer prevention and treatment.


Assuntos
Transformação Celular Neoplásica/genética , Inflamação/metabolismo , Interleucina-6/metabolismo , MicroRNAs/metabolismo , Animais , Mama/citologia , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Receptor alfa de Estrogênio/metabolismo , Feminino , Fatores de Transcrição de Choque Térmico , Humanos , Inflamação/genética , Interleucina-6/genética , Proteínas Quinases JNK Ativadas por Mitógeno/genética , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Camundongos , Camundongos Transgênicos , Proteína Quinase 9 Ativada por Mitógeno/genética , Proteína Quinase 9 Ativada por Mitógeno/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , Regiões Promotoras Genéticas , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/genética , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
14.
J Comput Assist Tomogr ; 44(2): 255-261, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32195805

RESUMO

OBJECTIVE: We aimed to determine the effects of recombinant human growth hormone (rhGH) replacement on cognitive function in subjects with poststroke cognitive impairment using resting-state functional magnetic resonance imaging. METHODS: We included 60 patients with a first-ever stroke for 3 months and a diagnosis of cognitive impairment who were randomized 1:1 to receive either rhGH subcutaneously or placebo injection for 6 months. All subjects were required to receive the same rehabilitative therapy program. Both groups were subjected to pretreatment and posttreatment neuropsychological assessment using the Montreal Cognitive Assessment, serum neurotrophic factors, biomarkers of glucose and lipid metabolism, and functional magnetic resonance imaging during 6 months of the study period. The pattern of brain activity was determined by examining the functional connectivity and amplitude of low-frequency fluctuations (ALFF) of blood oxygen level dependent signal. RESULTS: Forty-three (82.7%) completed the study. Treatment with rhGH reduced levels of triglycerides and low-density lipoprotein cholesterol but did not significantly altered plasma concentrations of glucose and glycated hemoglobin. We found a significant increase in serum insulin-like growth factor 1 levels (32.6%; P < 0.001) in the rhGH-treated group compared with that in the controls. After 6 months of rhGH treatment, mean Montreal Cognitive Assessment score improved from 16.31 (5.32) to 21.19 (6.54) (P < 0.001). The rhGH group showed significant increased area of activation with increased ALFF values in the regions of the frontal lobe, putamen, temporal lobe, and thalamus (P < 0.05), relative to the baseline conditions. The correlation analysis revealed that the ALFF and functional connectivity of default mode network was positively correlated with the ΔMoCA score and ΔIGF-1 levels; that is, the more the scale score increased, the higher the functional connection strength. No undesirable adverse effects were observed. CONCLUSIONS: The rhGH replacement has a significant impact on global and domain cognitive functions in poststroke cognitive impairment.


Assuntos
Disfunção Cognitiva/complicações , Disfunção Cognitiva/tratamento farmacológico , Hormônio do Crescimento Humano/uso terapêutico , Imageamento por Ressonância Magnética/métodos , Acidente Vascular Cerebral/complicações , Acidente Vascular Cerebral/tratamento farmacológico , Encéfalo/diagnóstico por imagem , Encéfalo/efeitos dos fármacos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Testes Neuropsicológicos , Resultado do Tratamento
15.
Mediators Inflamm ; 2020: 3019378, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32104147

RESUMO

CD19+CD24hiCD38hi B cells are immature transitional B cells that, in normal individuals, exert suppressive effects by IL-10 production but are quantitatively altered and/or functionally impaired in individuals with various autoimmune diseases. Primary biliary cholangitis (PBC), an autoimmune disease, clinically presents as chronic cholestasis and nonsuppurative destructive cholangitis. A role for CD19+CD24hiCD38hi B cells in PBC is unknown. This study investigated the frequency and functional variation of circulating CD19+CD24hiCD38hi B cells in PBC patients. Flow cytometry was employed to quantify the percentage of CD19+CD24hiCD38hi B cells in peripheral blood samples. Correlations between CD19+CD24hiCD38hi B cells and routine laboratory parameters were assessed. Levels of IL-10, TNF-α, IL-6 and IL-12, and Tim-1 in CD19+CD24hiCD38hi B cells from PBC patients were analyzed. The effect of CD19+CD24hiCD38hi B cells on CD4+T cell differentiation was evaluated. The percentage of CD19+CD24hiCD38hi B cells in PBC patients was significantly higher than in healthy controls and was positively correlated with liver cholestasis. After activation by anti-B cell receptor and CpG, the production of IL-10 was decreased and the production of IL-6 and IL-12 was increased in CD19+CD24hiCD38hi B cells from PBC patients. Moreover, Tim-1 levels were significantly downregulated in CD19+CD24hiCD38hi B cells from PBC patients. Coculture showed that PBC-derived CD19+CD24hiCD38hi B cells were less capable of CD4+T cell inhibition, but promoted Th1 cell differentiation. In conclusion, PBC patients have expanded percentages, but impaired CD19+CD24hiCD38hi B cells, which correlate with disease damage. In PBC patients, this B cell subset has a skewed proinflammatory cytokine profile and a decreased capacity to suppress immune function, which may contribute to the pathogenesis of PBC.


Assuntos
ADP-Ribosil Ciclase 1/metabolismo , Antígenos CD19/metabolismo , Linfócitos B/metabolismo , Linfócitos B/patologia , Antígeno CD24/metabolismo , Cirrose Hepática Biliar/imunologia , Cirrose Hepática Biliar/patologia , Idoso , Diferenciação Celular/fisiologia , Feminino , Citometria de Fluxo , Receptor Celular 1 do Vírus da Hepatite A/metabolismo , Humanos , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Interleucina-6/metabolismo , Cirrose Hepática Biliar/metabolismo , Masculino , Pessoa de Meia-Idade , Células-Tronco de Sangue Periférico/imunologia , Células-Tronco de Sangue Periférico/metabolismo , Células-Tronco de Sangue Periférico/patologia , Fator de Necrose Tumoral alfa/metabolismo
16.
J Virol ; 90(4): 1694-704, 2016 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-26581994

RESUMO

UNLABELLED: Vaccine manufacturing costs prevent a significant portion of the world's population from accessing protection from vaccine-preventable diseases. To enhance vaccine production at reduced costs, a genome-wide RNA interference (RNAi) screen was performed to identify gene knockdown events that enhanced poliovirus replication. Primary screen hits were validated in a Vero vaccine manufacturing cell line using attenuated and wild-type poliovirus strains. Multiple single and dual gene silencing events increased poliovirus titers >20-fold and >50-fold, respectively. Host gene knockdown events did not affect virus antigenicity, and clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9-mediated knockout of the top candidates dramatically improved viral vaccine strain production. Interestingly, silencing of several genes that enhanced poliovirus replication also enhanced replication of enterovirus 71, a clinically relevant virus to which vaccines are being targeted. The discovery that host gene modulation can markedly increase virus vaccine production dramatically alters mammalian cell-based vaccine manufacturing possibilities and should facilitate polio eradication using the inactivated poliovirus vaccine. IMPORTANCE: Using a genome-wide RNAi screen, a collection of host virus resistance genes was identified that, upon silencing, increased poliovirus and enterovirus 71 production by from 10-fold to >50-fold in a Vero vaccine manufacturing cell line. This report provides novel insights into enterovirus-host interactions and describes an approach to developing the next generation of vaccine manufacturing through engineered vaccine cell lines. The results show that specific gene silencing and knockout events can enhance viral titers of both attenuated (Sabin strain) and wild-type polioviruses, a finding that should greatly facilitate global implementation of inactivated polio vaccine as well as further reduce costs for live-attenuated oral polio vaccines. This work describes a platform-enabling technology applicable to most vaccine-preventable diseases.


Assuntos
Poliomielite/prevenção & controle , Vacinas contra Poliovirus/isolamento & purificação , Poliovirus/isolamento & purificação , Poliovirus/fisiologia , Tecnologia Farmacêutica/métodos , Replicação Viral , Animais , Vacinas Atenuadas/isolamento & purificação , Células Vero , Cultura de Vírus/métodos
17.
J Gen Virol ; 97(3): 659-668, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26683768

RESUMO

Vaccination with live-attenuated polio vaccine has been the primary reason for the drastic reduction of poliomyelitis worldwide. However, reversion of this attenuated poliovirus vaccine occasionally results in the emergence of vaccine-derived polioviruses that may cause poliomyelitis. Thus, the development of anti-poliovirus agents remains a priority for control and eradication of the disease. MicroRNAs (miRNAs) have been shown to regulate viral infection through targeting the viral genome or reducing host factors required for virus replication. However, the roles of miRNAs in poliovirus (PV) replication have not been fully elucidated. In this study, a library of 1200 miRNA mimics was used to identify miRNAs that govern PV replication. High-throughput screening revealed 29 miRNAs with antiviral properties against Sabin-2, which is one of the oral polio vaccine strains. In particular, miR-555 was found to have the most potent antiviral activity against three different oral polio attenuated vaccine strains tested. The results show that miR-555 reduced the level of heterogeneous nuclear ribonucleoprotein C1/C2 (hnRNP C) required for PV replication in the infected cells, which in turn resulted in reduction of PV positive-strand RNA synthesis and production of infectious progeny. These findings provide the first evidence for the role of miR-555 in PV replication and reveal that miR-555 could contribute to the development of antiviral therapeutic strategies against PV.


Assuntos
MicroRNAs/imunologia , Poliomielite/imunologia , Poliovirus/fisiologia , Replicação Viral , Regulação Viral da Expressão Gênica , Ribonucleoproteínas Nucleares Heterogêneas/genética , Ribonucleoproteínas Nucleares Heterogêneas/imunologia , Interações Hospedeiro-Patógeno , Humanos , MicroRNAs/genética , Poliomielite/genética , Poliomielite/virologia , Poliovirus/genética , RNA Viral/genética , RNA Viral/metabolismo
18.
ISA Trans ; 2024 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-39278813

RESUMO

To reduce the cross-regulation and improve the dynamic response performance of a single-inductor double-output Boost converter, an adaptive backstepping sliding mode control (ABSMC) strategy is proposed in this paper. The nonlinear mathematical model of the converter is established, an output function satisfying the exact feedback linearization (EFL) is constructed based on the inverse system theory, and the linearization and decoupling of the model are implemented. Meanwhile, the problem of EFL heavily relying on an exact model is solved by combining backstepping control with sliding mode control. Furthermore, an adaptive reaching law is proposed to adjust the gain of the switching function, and the chattering phenomenon of sliding mode control is reduced. The stability of the system is proven according to the Lyapunov stability theorem. Finally, compared with the existing control methods, both the simulation results and experimental results verify the effectiveness and superiority of the proposed ABSMC strategy.

19.
PLoS One ; 19(6): e0304691, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38833435

RESUMO

With the rapid development of intelligent connected vehicles, there is an increasing demand for hardware facilities and onboard systems of driver assistance systems. Currently, most vehicles are constrained by the hardware resources of onboard systems, which mainly process single-task and single-sensor data. This poses a significant challenge in achieving complex panoramic driving perception technology. While the panoramic driving perception algorithm YOLOP has achieved outstanding performance in multi-task processing, it suffers from poor adaptability of feature map pooling operations and loss of details during downsampling. To address these issues, this paper proposes a panoramic driving perception fusion algorithm based on multi-task learning. The model training involves the introduction of different loss functions and a series of processing steps for lidar point cloud data. Subsequently, the perception information from lidar and vision sensors is fused to achieve synchronized processing of multi-task and multi-sensor data, thereby effectively improving the performance and reliability of the panoramic driving perception system. To evaluate the performance of the proposed algorithm in multi-task processing, the BDD100K dataset is used. The results demonstrate that, compared to the YOLOP model, the multi-task learning network performs better in lane detection, drivable area detection, and vehicle detection tasks. Specifically, the lane detection accuracy improves by 11.6%, the mean Intersection over Union (mIoU) for drivable area detection increases by 2.1%, and the mean Average Precision at 50% IoU (mAP50) for vehicle detection improves by 3.7%.


Assuntos
Algoritmos , Condução de Veículo , Humanos , Condução de Veículo/psicologia , Análise e Desempenho de Tarefas
20.
PLoS One ; 19(2): e0296782, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38306357

RESUMO

Based Correctly handling the creativity of employees who have not been adopted is not only conducive to continuously stimulating employees' creativity and improving individual innovation performance, but also conducive to making the best use of organizational resources. This study integrates conservation of resource theory (COR) and social information processing theory to explore the influence of bootleg innovation behavior in organizations on individual innovation performance, as well as the mediating role of cognitive flexibility and the moderating role of leadership emotional intelligence. A three-stage time-lagged research design is used to obtain a valid sample of 327 employees from China. The PROCESS macro for SPSS was applied to test the hypothesized relationships. Findings demonstrated that bootleg innovation is positively related to individual innovation performance; cognitive flexibility mediates the relationship between bootleg innovation and individual innovation performance. Moreover, leadership emotional intelligence moderates the relationship between bootleg innovation and individual innovation performance and between bootleg innovation and cognitive flexibility and between cognitive flexibility and individual innovation performance respectively. The conclusion of the study not only provides a theoretical basis for individuals and leaders to deal with employees' creative abortion, but also provides a new thinking mode for how to maximize the effectiveness of unaccepted ideas and promote individual innovation performance.


Assuntos
Aborto Induzido , Liderança , Feminino , Gravidez , Humanos , Cognição , China , Inteligência Emocional
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