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The lateral flow immunoassay (LFIA) is a sought-after point-of-care testing platform, yet the insufficient sensitivity of the LFIA limits its application in the detection of tumor biomarkers. Here, a colorimetric signal amplification method, bimetallic nanozyme-mediated in situ-catalyzed reporter deposition (BN-ISCRD), was designed for ultrasensitive cancer diagnosis. The bimetallic nanozyme used, palladium@iridium core-shell nanoparticles (Pd@Ir NPs), had ultrahigh enzyme-like activity, which was further explained by the electron transfer of Pd@Ir NPs and the change in the Gibbs free energy during catalysis through density functional theory calculations. With gastric cancer biomarkers pepsinogen I and pepsinogen II as model targets, this assay could achieve a cutoff value of 10 pg/mL, which was 200-fold lower than that without signal enhancement. The assay was applied to correctly identify 8 positive and 28 negative clinical samples. Overall, this BN-ISCRD-based LFIA showed great merits and potential in the application of ultrasensitive disease diagnosis.
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Nanopartículas Metálicas , Nanopartículas , Neoplasias , Humanos , Imunoensaio/métodos , Biomarcadores Tumorais , Catálise , Neoplasias/diagnóstico , Limite de Detecção , OuroRESUMO
The advantages of rapid amplification of nucleic acid without a template based on terminal deoxyribonucleotidyl transferase (TdT) have been widely used in the field of biosensors. However, the catalytic efficiency of TdT is affected by extension conditions. The sensitivity of TdT- mediated biosensors can be improved only under appropriate conditions. Therefore, in this review, we provide a comprehensive overview of TdT extension characteristics and its applications in biosensors. We focus on the relationship between TdT extension conditions and extension efficiency. Furthermore, the construction strategy of TdT-mediated biosensors according to five different recognition types and their applications in targets are discussed and, finally, several current challenges and prospects in the field are taken into consideration.
Brief introduction to terminal deoxyribonucleotidyl transferase (TdT) characteristics.Provided a systematic and comprehensive summary of TdT extension conditions.Summarized the four effect factors of catalytic efficiency based on extension conditions and enzyme conformation.Sensing strategies of TdT-mediated biosensors for five different recognitions were summarized in detail.The applications of TdT-mediated biosensors in six targets were introduced in detail.
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Semiconductor photocatalytic technology has shown great prospects in converting solar energy into chemical energy to mitigate energy crisis and solve environmental pollution problems. The key issue is the development of high-efficiency photocatalysts. Various strategies in the state-of-the-art advancements, such as heterostructure construction, heteroatom doping, metal/single atom loading, and defect engineering, have been presented for the graphitic carbon nitride (g-C3N4)-based nanocomposite catalysts to design their surface chemical environments and internal electronic structures to make them more suitable for different photocatalytic applications. In this review, nanoarchitecture design, synthesis methods, photochemical properties, potential photocatalytic applications, and related reaction mechanisms of the modified high-efficiency carbon nitride-based photocatalysts were briefly summarized. The superior photocatalytic performance was identified to be associated with the enhanced visible-light response, fast photoinduced electron-hole separation, efficient charge migration, and increased unsaturated active sites. Moreover, the further advance of the visible-light harvesting and solar-to-energy conversions are proposed.
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Trehalose metabolism plays an important role in plant growth and response to abiotic stress. Trehalose-6-phosphate (Tre6P) can help regulate sugar homeostasis and act as an indication signal for intracellular sugar levels. Crop productivity can be greatly increased by altering the metabolic level of endogenous trehalose in plants, which can optimize the source-sink connection. In this study, the upland cotton GhTPP protein family was first homologously compared and 24 GhTPP genes were found. Transcriptome analysis revealed that GhTPP members had obvious tissue expression specificity. Among them, GhTPPA_2 (Gh_A12G223300.1) was predominantly expressed in leaves and bolls. The results of subcellular localization showed that GhTPPA_2 is localized in the chloroplast. Via PlantCare, we analyzed the promoters and found that the expression of GhTPPA_2 may be induced by light, abiotic stress, and hormones such as abscisic acid, ethylene, salicylic acid and jasmonic acid. In addition, GhTPPA_2 was overexpressed (TPPAoe) in tobacco, and we found that the TPPase activity of TPPAoe tobacco increased by 66 %. Soluble sugar content increased by 39 % and starch content increased by 27 %. Whereas, the transgenic tobacco had obvious growth advantages under 100 mM mannitol stress. Transcriptome sequencing results showed that the differential genes between TPPAoe and control were considerably enriched in functions related to photosynthesis, phosphate group metabolism, and carbohydrate metabolism. This study shows that GhTPPA_2 is involved in regulating sugar metabolism, improving soluble sugar accumulation and drought stress tolerance of tobacco, which provides theoretical basis for research on high yield and drought tolerance of crops.
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Resistência à Seca , Açúcares , Trealose/metabolismo , Carboidratos , Fotossíntese/genética , Secas , Estresse Fisiológico/genética , Plantas Geneticamente Modificadas/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
To investigate the effects of nitrogen source supply on microbial protein (MP) production by hydrogen-oxidizing bacteria (HOB) under continuous feed gas provision, a sequencing batch culture comparison (N2 fixation versus ammonium assimilation) was performed. The results confirmed that even under basic cultivation conditions, N2-fixing HOB (NF-HOB) communities showed higher levels of CO2 and N2 fixation (190.45 mg/L Δ CODt and 11.75 mg/L Δ TNbiomass) than previously known, with the highest biomass yield being 0.153 g CDW/g COD-H2. Rich ammonium stimulated MP synthesis and the biomass accumulation of communities (increased by 7.4 ï½ 14.3 times), presumably through the enhancement of H2 and CO2 absorption. The micro mechanism may involve encouraging the enrichment of species like Xanthobacter and Acinetobacter then raising the abundance of nitrogenase and glutamate synthase to facilitate the nitrogen assimilation. This would provide NF-HOB with ideas for optimizing their MP synthesis activity.
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Compostos de Amônio , Fixação de Nitrogênio , Nitrogênio/metabolismo , Compostos de Amônio/metabolismo , Hidrogênio/metabolismo , Dióxido de Carbono/metabolismo , Bactérias/genética , Bactérias/metabolismo , OxirreduçãoRESUMO
The Zingiberis Rhizoma - Jujubae Fructus herb pair (ZJHP) is a classic herb pair in traditional Chinese medicine. The herb pair shows the effect of dispelling cold, harmonizing the middle and improving gastrointestinal function, and is widely used for patients with stomach cold syndrome (SCS), stomachache and anemofrigid cold. The gingerols, shogaols, flavonoids and triterpenic acids are the important bioactive ingredients of ZJHP. However, few pharmacokinetic studies have been investigated in vivo for the above compounds. To comprehend the kinetics of active components and promote their curative application, a fast and sensitive ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS) method was established for simultaneous determination of 12 analytes in normal and SCS rats in this study. The results showed that the pharmacokinetic parameters (Cmax, Tmax, t1/2z, MRT0-t, AUC0-t and AUC0-∞) in SCS model were significantly different from those in normal rats. In addition, the pharmacokinetics of rats given ZJHP were also varied from single herb oral administration, especially in model condition. These results indicated that the in vivo processes of the above analytes changed under pathological conditions and the compatibility of the herb pair could significantly influence the absorption of active components, which might provide an insight and further supports for the clinical application of ZJHP.
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Medicamentos de Ervas Chinesas , Ratos Sprague-Dawley , Espectrometria de Massas em Tandem , Animais , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas em Tandem/métodos , Ratos , Medicamentos de Ervas Chinesas/farmacocinética , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/química , Masculino , Reprodutibilidade dos Testes , Administração Oral , Modelos Lineares , Limite de Detecção , Zingiber officinale/química , Gastropatias/tratamento farmacológico , Gastropatias/veterináriaRESUMO
Transcriptomics of dry age-related macular degeneration (AMD) patients with premature aging revealed the upregulated pathways involved in glycerolipid metabolism, tyrosine metabolism, and pentose and glucuronate interconversion. To investigate natural strategies for modulating these implicated pathways, we examined the impact and underlying mechanism of luteoloside on premature AMD using a stress-induced premature senescence (SIPS)-associated AMD animal model in middle-aged mice that mimicked the dysregulated pathways observed in dry AMD patients with premature aging. Luteoloside supplementation resulted in a significant reduction in serum levels of the pro-inflammatory cytokine IL-1ß and lipofuscin, along with increased serum activity of the antioxidant enzyme superoxide dismutase (SOD) and elevated levels of pigment epithelium-derived factor (PEDF), and preserved retinal thickness and structure in AMD mice. Furthermore, luteoloside supplementation effectively reversed the abnormal serum levels of metabolites, particularly by reducing harmful lysophosphatidylcholine (LysoPC) and increasing beneficial 4-guanidinobutanoic acid. In addition to its impact on metabolites, luteoloside modulated the composition of gut microbiota, promoting the enrichment of beneficial bacterial populations, including Lactobacillus, while reducing the abundance of harmful bacterial populations, including Bacteroides. Overall, our findings highlight the potential of luteoloside supplementation in regulating the dysregulated intestinal microbiota and metabolites in premature AMD, thereby reducing ocular levels of senescence-associated secretory phenotype (SASP) factors through the suppression of the p53-p21-retinoblastoma protein 1 (Rb1) axis.
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Microbioma Gastrointestinal , Degeneração Macular , Metabolômica , Proteína Supressora de Tumor p53 , Animais , Microbioma Gastrointestinal/efeitos dos fármacos , Microbioma Gastrointestinal/fisiologia , Camundongos , Degeneração Macular/tratamento farmacológico , Degeneração Macular/sangue , Masculino , Proteína Supressora de Tumor p53/metabolismo , Metabolômica/métodos , Modelos Animais de Doenças , Humanos , Camundongos Endogâmicos C57BL , Perfilação da Expressão Gênica/métodos , Transcriptoma/efeitos dos fármacosRESUMO
PURPOSE: The microcystic, elongated, and fragmented (MELF) pattern, characterized by myxoid and inflamed stroma, is readily identifiable as a form of myometrial infiltration. This meta-analysis endeavors to assess the prognostic significance of MELF infiltration patterns in patients diagnosed with endometrial cancer. METHODS: A comprehensive literature search, spanning until 11 October 2023, across PubMed, Embase, Cochrane, and Web of Science databases, identified 23 relevant studies involving 5199 patients. Data analysis was performed using Stata 16.0. RESULTS: Analysis indicates that MELF infiltration predicts a higher risk of lymph node metastasis in endometrial cancer patients [hazard ratios (HR)â =â 5.05; 95% confidence interval (CI), 3.62-7.05; Pâ <â 0.05]. Notably, this association remains consistent across various patient demographics, analytical approaches, study designs, and treatment modalities. However, MELF infiltration does not significantly correlate with recurrence (HRâ =â 1.05; 95% CI, 0.73-1.52; Pâ >â 0.05), overall survival (HRâ =â 1.24; 95% CI, 0.91-1.68; Pâ >â 0.05), or disease-free survival (HRâ =â 1.40; 95% CI, 0.85-2.28; Pâ >â 0.05). CONCLUSION: While MELF infiltration heightens the risk of lymph node metastasis in endometrial cancer, its impact on recurrence, overall survival, and disease-free survival remains statistically insignificant.
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Generating diverse ferroelectric ice nanotubes (NTs) efficiently has always been challenging, but matters in nanomaterial synthesis and processing technology. In the present work, we propose a method of growing ice NT forests in a single cooling process. A three-dimensional (3D) graphene structure was selected to behave as a representative container in which a batch of (5, 0) ice NTs was formed simultaneously under the cooling process from molecular dynamics simulation. Other similar 3D graphene structures but with different hole configurations, like uniform triangle or both triangle and pentagon, were also tested, revealing that ice NTs with different tube indices, i.e. both (3, 0) and (5, 0), could also be formed at the same time. Intriguingly, the orientations of the dipole moments of the water molecules of an ice NT formed were independent of each other, making the net ferroelectricity of the whole system weakened or even cancelled. An electric field could help change the orientation of the water molecules of the already obtained ice NTs and even twist the tube to be a spiral (5, 1) one if it was applied during the cooling process, such that the net ferroelectricity was greatly improved. The underlying physical mechanism of all phase transition phenomena, including the improvement of the ferroelectricity under an electric field, were explored in depth from the phase transition curves and structural point of view. The obtained results are of significant application value for improving the preparation efficiency of nano-ferroelectric materials, which are prosperous in nano-devices.
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Sugarcane is a vital crop with significant economic and industrial value. However, the cultivated sugarcane's ultra-complex genome still needs to be resolved due to its high ploidy and extensive recombination between the two subgenomes. Here, we generate a chromosomal-scale, haplotype-resolved genome assembly for a hybrid sugarcane cultivar ZZ1. This assembly contains 10.4 Gb genomic sequences and 68,509 annotated genes with defined alleles in two sub-genomes distributed in 99 original and 15 recombined chromosomes. RNA-seq data analysis shows that sugar accumulation-associated gene families have been primarily expanded from the ZZSO subgenome. However, genes responding to pokkah boeng disease susceptibility have been derived dominantly from the ZZSS subgenome. The region harboring the possible smut resistance genes has expanded significantly. Among them, the expansion of WAK and FLS2 families is proposed to have occurred during the breeding of ZZ1. Our findings provide insights into the complex genome of hybrid sugarcane cultivars and pave the way for future genomics and molecular breeding studies in sugarcane.
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Saccharum , Saccharum/genética , Melhoramento Vegetal , Genômica , Haplótipos/genética , CromossomosRESUMO
(1) Background: leaf structure traits are closely related to leaf photosynthesis, reflecting the ability of trees to obtain external resources in the process of growth. (2) Methods: We studied the morphological, chemical, anatomical, stomatal traits and maximum net photosynthetic rate of six broad-leaf species in northern temperate mixed broad-leaved Korean pine (Pinus koraiensis) forest. (3) Aim: To investigate whether there are differences in leaf structural traits of trees with different shade tolerances and different sizes and the effects of these differences on leaf photosynthetic capacity. (4) Results: the effects of leaf structure traits on leaf photosynthesis were different among trees with different shade tolerances or different sizes. Under the condition of light saturation, the net photosynthetic rate, nitrogen use efficiency, phosphorus use efficiency and stomatal conductance of shade-intolerant trees or small trees were higher than those of shade-tolerant trees or large trees. (5) Conclusions: the shade tolerance of tree species or the size of trees affect the traits of leaf structure and indirectly affect the photosynthetic ability of plants. When constructing the leaf trait-photosynthesis model, the shade tolerance and tree size of tree species should be taken into account.
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Several studies have shown that the dysregulation of the oral microbiota plays a crucial role in human health conditions, such as dental caries, periodontal disease, oral cancer, other oral infectious diseases, cardiovascular diseases, diabetes, bacteremia, and low birth weight. The use of traditional detection methods in conjunction with rapidly advancing molecular techniques in the diagnosis of harmful oral microorganisms has expanded our understanding of the diversity, location, and function of the microbiota associated with health and disease. This review aimed to highlight the latest knowledge in this field, including microbial colonization; the most modern detection methods; and interactions in disease progression. The next decade may achieve the rapid diagnosis and precise treatment of harmful oral microorganisms.
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OBJECTIVE: This study was presented to reveal the most distinct microbial prevalence in recurrent aphthous stomatitis (RAS) lesions compared to healthy controls. DESIGN: The case-control studies were selected in electronic databases until Nov 2022 with key search terms, and the eligible publications were screened and analyzed by independent authors. RESULTS: A total of 14 studies were identified, which included 531 cases of active states of RAS (AS-RAS), 92 cases of passive states of RAS (PS-RAS) and 372 healthy controls. The most sample pattern was the mucosa swab performed in 8 of 14 studies, biopsies in 3 studies, followed by micro-brush, and saliva. A variety of bacteria in higher or lower abundance were observed in RAS lesions. CONCLUSIONS: The etiopathogenesis of RAS may not be ascribed to a single pathogen. A possible explanation is that microbial interactions modify immune response or destroy the epithelial integrity, thus contributing to the development of the condition.
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Microbiota , Estomatite Aftosa , Humanos , Recidiva , Estudos de Casos e ControlesRESUMO
BACKGROUND: Oral leukoplakia (OLK) and oral lichen planus (OLP) are common precancerous lesions of the oral mucosa. The role of circular RNAs (circRNAs) in OLK and OLP is unclear. The aim of this study was to evaluate the circRNA expression profiles of OLK and OLP, and further explore the potential role of circRNAs in the pathogenesis of these two diseases. METHODS: High throughput sequencing technology was performed to detect the differentially expressed circRNA in OLK (n = 6), OLP (n = 6), oral squamous cell carcinoma (n = 6), and normal oral mucosa tissues (n = 6). Expression of selected circRNAs was validated by qRT-PCR, enzyme tolerance assay, and Sanger sequencing. Expanded sample size validation was done in 20 tissue pairs. The biological processes and signal pathways involved in differential circRNA were analyzed by GO and KEGG enrichment. TargetScan and MiRanda were used to predict miRNAs downstream of circRNA and draw competitive endogenous RNA network diagram. RESULTS: Forty-nine circRNAs were significantly altered in OLK and OLP, including 30 upregulated and 19 downregulated circRNAs. The five selected circRNAs were validated by qRT-PCR, Sanger sequencing, and RNase R assay. GO and KEGG analyses indicated that the upregulated circHLA-C may be involved in the biological process of immune function of OLK and OLP. Bioinformatics analysis indicated that circHLA-C may be involved in the progression of OLK and OLP as a ceRNA. In validation with expanded sample size, PCR results showed that circHLA-C expression was significantly upregulated in OLK and OLP. ROC analysis indicated that circHLA-C has potential diagnostic value with good accuracy and specificity. CONCLUSION: Our study revealed that circHLA-C is the most significantly upregulated circRNA co-existing in OLK and OLP, and we preliminarily discuss the role of circHLA-C in the etiopathogenesis and progression of OLK and OLP.
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Carcinoma de Células Escamosas , Líquen Plano Bucal , MicroRNAs , Neoplasias Bucais , Humanos , RNA Circular/genética , RNA Circular/metabolismo , Neoplasias Bucais/genética , Neoplasias Bucais/patologia , Carcinoma de Células Escamosas/patologia , Líquen Plano Bucal/genética , Líquen Plano Bucal/patologia , Leucoplasia Oral/genética , Leucoplasia Oral/patologiaRESUMO
It is known to all that Wnt signaling pathway plays an important role in the early development of tooth. Our previous research found that Wnt signaling pathway played crucial roles in dental development, and mutations in antagonist of Wnt signaling pathway may lead to the formation of supernumerary teeth. However, the expression pattern of Wnt signaling molecules in early development of tooth, especially genes with stage specificity, remains unclear. Hence, we applied RNA-seq analysis to determine the expression levels of wnt signal molecules at five different stages of rat first molar tooth germ. In addition, after literature review we summarized the function of Wnt signaling molecules during tooth development and the relationship between Wnt signaling molecules variation and tooth agenesis. Our research may have implications for exploring the role of Wnt signaling molecules in different stages of tooth development.
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Dente , Via de Sinalização Wnt , Ratos , Animais , Odontogênese/genética , Proteínas Wnt/genética , Proteínas Wnt/metabolismo , Dente Molar/metabolismoRESUMO
The healing of biofilm-infected diabetic wounds characterized by a deteriorative tissue microenvironment represents a substantial clinical challenge. Current treatments remain unsatisfactory due to the limited antibiofilm efficacy caused by weak tissue and biofilm permeability of drugs and the risk of reinfection during the healing process. To address these issues, an integrated therapeutic and preventive nanozyme-based microneedle (denoted as Fe2 C/GOx@MNs) is engineered. The dissolvable tips with enough mechanical strength can deliver and rapidly release Fe2 C nanoparticles (NPs)/glucose oxidase (GOx) in the biofilm active regions, enhancing tissue and biofilm permeability of Fe2 C NPs/GOx, ultimately achieving highly efficient biofilm elimination. Meanwhile, the chitosan backing layer can not only act as an excellent physical barrier between the wound bed and the external environment, but also prevent the bacterial reinvasion during wound healing with its superior antibacterial property. Significantly, the biofilm elimination and reinfection prevention abilities of Fe2 C/GOx@MNs on wound healing are proved on methicillin-resistant Staphylococcus aureus-biofilm-infected diabetic mouse model with full-thickness wound. Together, these results demonstrate the promising clinical application of Fe2 C/GOx@MNs in biofilm-infected wound healing.
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Diabetes Mellitus , Staphylococcus aureus Resistente à Meticilina , Infecção dos Ferimentos , Camundongos , Animais , Reinfecção , Cicatrização , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Biofilmes , Infecção dos Ferimentos/tratamento farmacológico , Infecção dos Ferimentos/prevenção & controleRESUMO
The most commonly used methods for pathogen detection and identification in oral mucosal infectious diseases are DNA or RNA quantitative polymerase chain reaction detection, bacterial or fungal cultures, and immunohistochemical analysis. These traditional methods are time-consuming and can only detect one specific targeted pathogen at a time. An efficient and sensitive method with higher species richness is urgently needed. Metagenomic next-generation sequencing (mNGS) is a new method of pathogen detection with high efficiency and sensitivity. In this case report, mNGS was used to identify the pathogens in oral mucosal tissues of a patient with complex oral mucosal infections and oral leukoplakia. Candida albicans, human gamma herpesvirus 4, and many other pathogens were identified using this method. For complex oral mucosal infections, mNGS is a more efficient and sensitive approach that can replace conventional detection methods.
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Doenças Transmissíveis , Metagenômica , Humanos , Sensibilidade e Especificidade , Metagenômica/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , MucosaRESUMO
Highly pathogenic avian influenza (HPAI) is a fatal zoonotic disease that damages the poultry industry and endangers human lives via exposure to the pathogen. A risk assessment model that precisely predicts high-risk groups and occurrence of HPAI infection is essential for effective biosecurity measures that minimize the socio-economic losses of massive outbreaks. However, the conventional risk prediction approaches have difficulty incorporating the broad range of factors associated with HPAI infections at poultry holdings. Therefore, it is difficult to accommodate the complexity of the dynamic transmission mechanisms and generate risk estimation on a real-time basis. We proposed a continuous risk prediction framework for HPAI occurrences that used machine learning algorithms (MLAs). This integrated environmental, on-farm biosecurity, meteorological, vehicle movement tracking, and HPAI wild bird surveillance data to improve accuracy and timeliness. This framework consisted of (i) the generation of 1788 predictors from six types of data and reconstructed them with an outcome variable into a data mart based on a temporal assumption (i.e. infected period and day-ahead forecasting); (ii) training of the predictors with the temporally rearranged outcome variable that corresponded to HPAI H5N6 infected state at each individual farm on daily basis during the 2016-2017 HPAI epidemic using three different MLAs [Random Forest, Gradient Boosting Machine (GBM), and eXtreme Gradient Boosting]; (iii) predicting the daily risk of HPAI infection during the 2017-2018 HPAI epidemic using the pre-trained MLA models for each farm across the country. The models predicted the high risk to 8-10 out of 19 infected premises during the infected period in advance. The GBM MLAs outperformed the 7-day forecasting of HPAI prediction at individual poultry holdings, with an area under the curve (AUC) of receiver operating characteristic of 0.88. Therefore, this approach enhances the flexibility and timing of interventions against HPAI outbreaks at poultry farms.
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Influenza Aviária , Doenças das Aves Domésticas , Animais , Surtos de Doenças/veterinária , Fazendas , Humanos , Influenza Aviária/epidemiologia , Aprendizado de Máquina , Aves Domésticas , Doenças das Aves Domésticas/epidemiologia , República da Coreia/epidemiologiaRESUMO
OBJECTIVE: To investigate the clinical significance of differentially expressed circRNAs and candidate circRNAs in the transformation of oral leukoplakia (OLK) to oral squamous cell carcinoma (OSCC). METHODS: We performed high-throughput circRNA sequencing in six cases of normal oral mucosal (NOM) tissues, six cases of OLK tissues, and six cases of OSCC tissues. Ten circRNAs with significant differential expression were verified by qRT-PCR. Enzyme tolerance assay and Sanger sequencing were performed on the screened target circRNA hsa_circ_0060927, and a qRT-PCR assay of hsa_circ_0060927 was performed in three tissues (24 cases in each group); this was followed by an ROC analysis. The ceRNA network was predicted using TargetScan and miRanda. MiR-195-5p and TRIM14 were selected as the downstream research objects of hsa_circ_0060927. The sponge mechanism of hsa_circ_0060927 was detected by AGO2 RIP. The interaction between hsa_circ_0060927 and miR-195-5p was verified by RNA pull-down assay and dual luciferase reporter gene assay. The expressions of hsa_circ_0060927, miR-195-5p, and TRIM14 were verified by normal oral epithelial primary cells and cell lines of LEUK1, SCC9, and SCC25. The hsa_circ_0060927 overexpressed plasmid and miR-195-5p mimics were constructed to transfection LEUK1 to detect the changes in cell proliferation, apoptosis, and migration. RESULTS: The results of qRT-PCR validation were consistent with the sequencing results. Hsa_circ_0060927 is a true circRNA with trans-splicing sites. The expression of hsa_circ_0060927 increased in NOM, OLK, and OSCC. Overexpression of hsa_circ_0060927 enhanced the ability of cell proliferation and migration, and decreased cell apoptosis capacity. The prediction of ceRNA network suggested that hsa_circ_0060927 could regulate the target gene TRIM14 through sponging miR-195-5p. AGO2 RIP indicated that hsa_circ_0060927 had a sponge mechanism. RNA pull-down and dual luciferase reporter gene assay suggested that hsa_circ_0060927 interacted with miR-195-5p. Hsa_circ_0060927 was positively correlated with the expression of TRIM14, and could relieve the inhibition of miR-195-5p on TRIM14 to regulate cell proliferation, apoptosis, and migration of LEUK1 cells. CONCLUSION: Hsa_circ_0060927 acted as a potential key ceRNA to sponge downstream miR-195-5p and promote OLK carcinogenesis by upregulating TRIM14. Hsa_circ_0060927 was expected to be a molecular marker for the prevention and treatment of OLK carcinogenesis through the hsa_circ_0060927/miR-195-5p/TRIM14 axis.
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BACKGROUND: This study sought to identify the circular RNAs (circRNAs) differentially expressed in oral lichen planus (OLP) to investigate the possible role of circRNAs in this disease's pathogenesis. METHODS: Six OLP and six normal oral mucosal tissues were used for circRNA detection and sequencing. 10 selected circRNAs were verified by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). A gene ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to predict the functions of circRNAs in OLP. TargetScan and miRanda were applied to predict targeted micro (mi)RNAs and messenger (m)RNAs of circRNAs, and competing endogenous (ce)RNA networks were mapped. RESULTS: One hundred and thirty-five circRNAs were identified differentially expressed in OLP tissues compared to normal control tissues, including 83 upregulated circRNAs, and 52 down-regulated circRNAs. RT-qPCR confirmed that 10 circRNAs were all abnormally expressed in OLP. The GO functional analysis and KEGG pathway analysis showed that differentially expressed circRNAs were involved in 535 GO functional items and 78 signal pathways. A ceRNA network analysis showed that circRNAs might interact with a variety of miRNAs. CONCLUSIONS: This study mapped the expression profile of abnormally expressed circRNAs in OLP tissues for the first time and showed that circRNAs appear to play an important role in the pathogenesis of OLP.