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Super-resolution fluorescence imaging is a crucial method for visualizing the dynamics of the cell membrane involved in various physiological and pathological processes. This requires bright fluorescent dyes with excellent photostability and labeling stability to enable long-term imaging. In this context, we introduce a buffering-strategy-based cyanine dye, SA-Cy5, designed to identify and label carbonic anhydrase IX (CA IX) located in the cell membrane. The unique feature of SA-Cy5 lies in its ability to overcome photobleaching. When the dye on the cell membrane undergoes photobleaching, it is rapidly replaced by an intact probe from the buffer pool outside the cell membrane. This dynamic replacement ensures that the fluorescence intensity on the cell membrane remains stable over time. Under the super-resolution structured illumination microscopy (SIM), the cell membrane can be continuously imaged for 60 min with a time resolution of 20 s. This extended imaging period allows for the observation of substructural dynamics of the cell membrane, including the growth and fusion of filamentous pseudopodia and the fusion of vesicles. Additionally, this buffering strategy introduces a novel approach to address the issue of poor photostability associated with the cyanine dyes.
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Colorantes Fluorescentes , Imagen Óptica , Carbocianinas/química , Colorantes Fluorescentes/química , Membrana CelularRESUMEN
The varied functions of lipid droplets, which encompass the regulation of lipid and energy homeostasis, as well as their association with the occurrence of various metabolic diseases, are intricately linked to their dynamic properties. Super-resolution imaging techniques have emerged to decipher physiological processes and molecular mechanisms on the nanoscale. However, achieving long-term dynamic super-resolution imaging faces challenges due to the need for fluorescent probes with high photostability. This paper introduces LD-CF, a "buffering probe" for imaging lipid droplet dynamics using structured illumination microscopy (SIM). The polarity-sensitive LD-CF eliminates background fluorescence with a "cyan filter" strategy, enabling wash-free imaging of lipid droplets. In the fluorescent "off" state outside droplets, the probes act as a "buffering pool", replacing photobleached probes inside droplets and enabling photostable long-term SIM imaging. With this probe, three modes of lipid droplet fusion were observed, including the discovery of fusion from large to small lipid droplets. Fluorescence intensity tracking also revealed the direction of lipid transport during the lipid droplet fusion.
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Colorantes Fluorescentes , Gotas Lipídicas , Colorantes Fluorescentes/metabolismo , Gotas Lipídicas/metabolismo , Microscopía Fluorescente/métodos , Transporte Biológico , LípidosRESUMEN
BACKGROUND: The aim was to investigate the value of blood Septin9, SRSF1, and PAX8 gene methylation detection techniques in early screening of colorectal cancer (CRC). METHODS: A prospective cohort study enrolled 3,000 participants undergoing routine physical examination at Shizong County People's Hospital Health Management Center from December 2021 through November 2022, including 1,512 males and 1,488 females, ranging in age from 20 to 90 years, with a median age of 49 years. Fresh blood samples were collected and tested for Septin9, SRSF1, and PAX8 gene methylation. Positive or negative results were reported. Colonoscopy was recommended for positive results and telephone follow-up for negative results. A chi-squared test analyzed the positive rate of initial screening, colonoscopy compliance, and the detection rate of colorectal lesions. Finally, combined with the follow-up data, the screening effect of Septin9, SRSF1, and PAX8 methylation detection on CRC was evaluated. RESULTS: Among 3,000 cases, 215 cases were preliminarily positive, with a positive rate of 7.1% (215/3,000). The positive rate of Septin9 gene methylation was the highest (6%, 180/3000), followed by SRSF1 (4.1%, 124/3000) and PAX8 (3.6%, 108/3000). The sensitivity of combined detection of Septin9, SRSF1, and PAX8 methylation in the diagnosis of CRC was higher than that of the three alone, and the specificity, positive predictive value, and negative predictive value of combined detection were higher than that of the single detection of blood Septin9, SRSF1, and PAX8 DNA methylation. In addition, the positive rate of initial screening increased with age (χ2 = 32.135, p < 0.001). A total of 150 cases underwent further colonoscopy, and the colonoscopy compliance rate was 69.8% (150/215). Among 150 cases who completed colonoscopy, 5 cases of CRC (3.4%), 25 cases of advanced adenoma (16.0%), 78 cases of non-advanced adenoma (52.0%), and 24 cases of non-adenomatous polyps (22.7%) were detected. The positive predictive value of Septin9, SRSF1, and PAX8 methylation was 94% (141/150) for all colorectal lesions, and 70.0% (105/150) for colorectal cancer and precancerous lesions. CONCLUSIONS: Blood Septin9, SRSF1, and PAX8 gene methylation detection, combined with colonoscopy, can effectively detect colorectal cancer and precancerous lesions. This strategy may be an effective way to carry out large-scale colorectal cancer screening in the general risk population. Combined detection of the three genes can improve the detection rate of colorectal cancer, but Septin9 methylation is the most sensitive, which can be used for screening and efficacy evaluation of CRC.
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Adenoma , Neoplasias Colorrectales , Metilación de ADN , Lesiones Precancerosas , Septinas , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Adenoma/diagnóstico , Colonoscopía , Neoplasias Colorrectales/diagnóstico , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Proteínas del Citoesqueleto/genética , Detección Precoz del Cáncer/métodos , Tamizaje Masivo/métodos , Factor de Transcripción PAX8/genética , Examen Físico , Lesiones Precancerosas/genética , Estudios Prospectivos , Factores de Empalme Serina-Arginina/genética , Septinas/genéticaRESUMEN
The interfacial barrier of charge transfer from semiconductors to cocatalysts means that the photogenerated charges cannot be fully utilized, especially for the challenging water oxidation reaction. Using cobalt cubane molecules (Co4 O4 ) as water oxidation cocatalysts, we rationally assembled partially oxidized graphene (pGO), acting as a charge-transfer mediator, on the hole-accumulating {-101} facets of lead chromate (PbCrO4 ) crystal. The assembled pGO enables preferable immobilization of Co4 O4 molecules on the {-101} facets of the PbCrO4 crystal, which is favorable for the photogenerated holes transferring from PbCrO4 to Co4 O4 molecules. The surface charge-transfer efficiency of PbCrO4 was boosted by selective assembly of pGO between PbCrO4 and Co4 O4 molecules. An apparent quantum efficiency for photocatalytic water oxidation on the Co4 O4 /pGO/PbCrO4 photocatalyst exceeded 10 % at 500â nm. This strategy of rationally assembling charge-transfer mediator provides a feasible method for acceleration of charge transfer and utilization in semiconductor photocatalysis.
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Relative permeability of polymer flooding plays a very important role in oil field development. This paper aimed to measure and calculate the relative permeability curves of polymer flooding more accurately. First, viscosity variation law of polymer in porous media was studied. Rock particles of different diameters and cementing agent were used to make artificial cores and hydrophobically associating polymer solutions were prepared for experiments. Polymer solutions were injected into the cores filled with crude oil and irreducible water. In the process of polymer flooding, produced fluid was collected at different water saturations and locations of the core. Polymer solutions were separated and their viscosities were measured. With the experimental data, the viscosity variation rule of polymer transporting in porous media was explored. The result indicates that the viscosity retention rate of polymer solutions transporting in porous media has power function relationship with the water saturation and the dimensionless distance from the core inlet. Finally, the relative permeability curves of polymer flooding were measured by unsteady state method and the viscosity variation rule was applied to the calculation of the relative permeability curves.
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Polímeros , Agua , Permeabilidad , Porosidad , ViscosidadRESUMEN
A crucial issue in artificial photosynthesis is how to modulate the behaviors of photogenerated charges of semiconductor photocatalysts. Here, using lead chromate (PbCrO4 ) as an example, we conducted the morphology tailoring from parallelepiped (p-PbCrO4 ) to truncated decahedron (t-PbCrO4 ) and elongated rhombic (r-PbCrO4 ), resulting in exposed anisotropic facets. The spatial separation of photogenerated charges closely correlates to the anisotropic facets of crystals, which can only be realized for t-PbCrO4 and r-PbCrO4 . The charge-separation efficiencies exhibit a quasilinear relation with the surface photovoltage difference between anisotropic facets. The r-PbCrO4 gives an apparent quantum efficiency of 6.5 % at 500â nm for photocatalytic water oxidation using Fe3+ ions as electron acceptors. Moreover, the oxidation reverse reaction from Fe2+ to Fe3+ ions was completely blocked with â¼100 % of Fe3+ conversion achieved on the anisotropic PbCrO4 crystals.
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The visualization of self-assembled structure and dynamics at the molecular level has become a powerful method to understand structure-function relationships of self-assembly. Herein, we in situ real-time imaged the dynamic process of benzyl-naphthalimide dyes at the nanoscale and inspected their internal structure with minimum 2.8â nm localization accuracy through single-molecule localization microscopy (SMLM) imaging. We monitored the growth process of three different assemblies in situ, which possessed highly heterogeneous dynamics with different shapes and growth rates. Furthermore, diverse growth rates were also found at different sites in the same assembly. These results highlight the application of super-resolution microscopy techniques for real-time visualization of internal assembled structure and dynamics in situ.
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Colorantes Fluorescentes , Imagen Individual de Molécula , Colorantes Fluorescentes/química , Microscopía , Imagen Individual de Molécula/métodosRESUMEN
The feasibility of an advanced oxidation process based upon sodium persulfate (SPS) activated simultaneously by heat (50 °C) and nanoscale zero-valent iron (nZVI) on benzene removal was investigated. The experimental results strongly showed the synergistic effect of thermal and nZVI activation to SPS and benzene removal was enhanced with the increase of SPS/nZVI/benzene molar ratio. Specifically, 94% of benzene could be removed in 1 hr at 50 °C at the SPS/nZVI/benzene molar ratio of 10/5/1. The radical scavenger tests and electron paramagnetic resonance (EPR) analysis confirmed that SO4â¢- was the predominant species contributing to benzene degradation. Further, the effects of the solution matrix on benzene elimination were investigated. The results indicated that benzene destruction in the thermally activated SPS/nZVI system performed better under acidic conditions, and the high concentration of both Cl- and HCO3 - had adverse effects on benzene elimination. The test for the performance of benzene degradation in the actual groundwater demonstrated that benzene could be degraded entirely at SPS/nZVI/benzene molar ratio of 40/40/1 at 50 °C, indicating that the synergistic catalysis of thermal and nZVI activation to SPS is exploitable and the thermally activated SPS/nZVI system can be applicable to the remediation of benzene contaminated groundwater.
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Agua Subterránea , Contaminantes Químicos del Agua , Benceno , Hierro , Oxidación-Reducción , Contaminantes Químicos del Agua/análisisRESUMEN
This study investigated the reductive initiation for the depletion of highly oxidized/perhalogenated pollutants, specifically the degradation of carbon tetrachloride (CT) was induced by adding methanol (MeOH) into a ferrous ion (Fe(II)) activated calcium peroxide (CaO2) system. The results indicated that CT could be completely degraded within 20 min at CaO2/Fe(II)/MeOH/CT molar ratio of 30/40/10/1 in this system. Scavenging tests suggested that both superoxide radical anion (O2 â¢-) and carbon dioxide radical anion (CO2 â¢-) were predominant reactive species responsible for CT destruction. Hydroxymethyl radicals (â¢CH2OH), an intermediate in the transformation of MeOH, could also initiate CT degradation by reducing C-Cl bond. GC/MS analysis identified CHCl3, C2Cl4, and C2Cl6 as the intermediates accompanied by CT destruction, and a reduction mechanism for CT degradation was proposed accordingly. In addition, the impact of solution matrix and initial solution pH were evaluated, and the results showed that Cl-, NO3 -, and HCO3 - had adverse effects on CT degradation. Moreover, the alkaline condition was unfavorable to CT depletion. In conclusion, the results obtained in the actual groundwater tests encouragingly demonstrated that the CaO2/Fe(II)/MeOH process is a highly promising technique for the remediation of CT-contaminated groundwater.
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In this study, nCaO2 was synthesized successfully and applied in the Fe(II)-based catalytic environments in investigating trichloroethylene (TCE) removal performance. nCaO2 with the particle sizes in the range of 50-200 nm was prepared, and it performed better for TCE removal when compared to the conventional CaO2. Further experimental results showed that 70.4% of TCE could be removed in 180 min at the nCaO2/Fe(II)/TCE molar ratio of 1/2/1, while this data was elevated to 86.1% in the presence of citric acid (CA) at the nCaO2/Fe(II)/CA/TCE molar ratio of 1/2/2/1 in the same test period. Probe compound tests, specifically designed for free radicals confirmation, demonstrated the presence of HO⢠and O2 -â¢. Moreover, scavenging tests indicated that HO⢠was the major radical responsible for TCE degradation but O2 -⢠promoted the removal of TCE in both nCaO2/Fe(II) and nCaO2/Fe(II)-CA system. In addition, the effects of initial solution pH and anions (Cl-, HCO3 -) were also evaluated. The performance of TCE degradation in actual groundwater demonstrated that both nCaO2/Fe(II) and nCaO2/Fe(II)-CA systems can be applicable for TCE removal in ISCO practice and the nCaO2/Fe(II)-CA system is much promising technique. These fundamental data strongly confirmed the feasibility and potential of nCaO2 based technique in the remediation of TCE contaminated groundwater.
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Silk sericin has great potential as a biomaterial for biomedical applications due to its good hydrophilicity, reactivity, and biodegradability. To develop multifunctional sericin materials for potential antibacterial application, a one-step synthesis method for preparing silver nanoparticles (AgNPs) modified on polydopamine-coated sericin/polyvinyl alcohol (PVA) composite films was developed. Polydopamine (PDA) acted as both metal ion chelating and reducing agent to synthesize AgNPs in situ on the sericin/PVA composite film. Scanning electron microscopy and energy dispersive spectroscopy analysis revealed that polydopamine could effectively facilitate the high-density growth of AgNPs as a 3-D matrix. X-ray diffractometry studies suggested the synthesized AgNPs formed good face-centered cubic crystalline structures. Contact angle measurement and mechanical test indicated AgNPs modified PDA-sericin/PVA composite film had good hydrophilicity and mechanical property. The bacterial growth curve and inhibition zone assays showed the AgNPs modified PDA-sericin/PVA composite film had long-term antibacterial activities. This work develops a new method for the preparation of AgNPs modified PDA-sericin/PVA film with good hydrophilicity, mechanical performance and antibacterial activities for the potential antimicrobial application in biomedicine.
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Antibacterianos/síntesis química , Indoles/química , Nanopartículas del Metal/química , Polímeros/química , Alcohol Polivinílico/química , Sericinas/química , Plata/química , Adsorción , Antibacterianos/farmacología , Vendajes , Materiales Biocompatibles Revestidos/química , Pruebas Antimicrobianas de Difusión por Disco , Escherichia coli/efectos de los fármacos , Escherichia coli/crecimiento & desarrollo , Nanocompuestos/química , Tamaño de la Partícula , Espectroscopía Infrarroja por Transformada de Fourier , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Resistencia a la Tracción , HumectabilidadRESUMEN
Supercapacitors (SCs) are energy storage devices which have high power density and long cycle life. Conventional SCs have two-dimensional planar structures. As a new family of SCs, fiber SCs utilize one-dimensional cylindrically shaped fibers as electrodes. They have attracted significant interest since 2011 and have shown great application potential either as micro-scale devices to complement or even replace micro-batteries in miniaturized electronics and microelectromechanical systems or as macro-scale devices for wearable electronics or smart textiles. This tutorial review provides an essential introduction to this new field. We first introduce the basics of performance evaluation for fiber SCs as a foundation to understand different research approaches and the diverse performance metrics reported in the literature. Next, we summarize the current state-of-the-art progress in structure design and electrode fabrication of fiber SCs. This is followed by a discussion on the integration of multiple fiber SCs and the combination with other energy harvesting or storage devices. Last, we present our perspectives on the future development of fiber SCs and highlight key technical challenges with the hope of stimulating further research progress.
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Nickel hydroxide (Ni(OH)2) is a promising pseudocapacitive material to increase the energy storage capacity of supercapacitors. Ni(OH)2 has three common crystalline structures: amorphous (amor-), α-, and ß-Ni(OH)2. There is a lack of good understanding on their pros and cons as supercapacitor electrodes. In this work, we synthesized three nanocomposites with thin layers (10-15 nm) of amor-, α-, and ß-Ni(OH)2 deposited on conductive multi-walled carbon nanotubes (MWCNTs). The mass loading of Ni(OH)2 is analogous in these nanocomposites, ranging from 49.1-52.2 wt% with a comparable narrow-pore size distribution centered around 4-5 nm. They were fabricated into supercapacitor electrodes at a mass loading of 6 mg cm(-2) with a thickness of â¼250 µm, similar to the electrodes used in commercial supercapacitors. Our results show that MWCNT/amor-Ni(OH)2 has the highest specific capacitance (1495 or 2984 F g(-1), based on the mass of total active materials or Ni(OH)2 only at the scan rate of 5 mV s(-1) in 1 M KOH electrolyte). It also has the best rate capability among the three nanocomposites. Better performances can be attributed to its disordered structure, which increases its effective surface area and reduces diffusion resistance for redox reactions. However, superior performances gradually deteriorate to the same level as that of MWCNT/ß-Ni(OH)2 over 3000 charge/discharge cycles, because amor- and α-Ni(OH)2 transform slowly to more ordered ß-Ni(OH)2. Our results highlight that the electrochemical performances of MWCNT/Ni(OH)2 nanocomposites depend on the crystallinity of Ni(OH)2, and the performances of electrodes change upon the crystalline structure transformation of Ni(OH)2 under repeated redox reactions. Future research should focus on improving the structure stability of amor-Ni(OH)2.
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BACKGROUND: Multi-category response models are very important complements to binary logistic models in medical decision-making. Decomposing model construction by aggregating computation developed at different sites is necessary when data cannot be moved outside institutions due to privacy or other concerns. Such decomposition makes it possible to conduct grid computing to protect the privacy of individual observations. METHODS: This paper proposes two grid multi-category response models for ordinal and multinomial logistic regressions. Grid computation to test model assumptions is also developed for these two types of models. In addition, we present grid methods for goodness-of-fit assessment and for classification performance evaluation. RESULTS: Simulation results show that the grid models produce the same results as those obtained from corresponding centralized models, demonstrating that it is possible to build models using multi-center data without losing accuracy or transmitting observation-level data. Two real data sets are used to evaluate the performance of our proposed grid models. CONCLUSIONS: The grid fitting method offers a practical solution for resolving privacy and other issues caused by pooling all data in a central site. The proposed method is applicable for various likelihood estimation problems, including other generalized linear models.
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Toma de Decisiones Clínicas , Confidencialidad , Difusión de la Información , Modelos Logísticos , Modelos Estadísticos , HumanosRESUMEN
UNLABELLED: WebGLORE is a free web service that enables privacy-preserving construction of a global logistic regression model from distributed datasets that are sensitive. It only transfers aggregated local statistics (from participants) through Hypertext Transfer Protocol Secure to a trusted server, where the global model is synthesized. WebGLORE seamlessly integrates AJAX, JAVA Applet/Servlet and PHP technologies to provide an easy-to-use web service for biomedical researchers to break down policy barriers during information exchange. AVAILABILITY AND IMPLEMENTATION: http://dbmi-engine.ucsd.edu/webglore3/. WebGLORE can be used under the terms of GNU general public license as published by the Free Software Foundation.
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Investigación Biomédica , Confidencialidad , Difusión de la Información , Modelos Logísticos , Programas Informáticos , Área Bajo la Curva , Redes de Comunicación de Computadores , Conducta Cooperativa , Humanos , Internet , Máquina de Vectores de SoporteRESUMEN
We achieve a successful transition of Co4O4 molecules from a homogeneous to a heterogeneous system by modifying the functional groups at their termini. The resulting cocatalyst, denoted as Co4O4-poly, not only preserved the catalytic sites of Co4O4 molecules but also exhibited outstanding performance in catalyzing water oxidation.
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Regulating the interfacial charge transfer behavior between cocatalysts and semiconductors remains a critical challenge for attaining efficient photoelectrochemical water oxidation reactions. Herein, using bismuth vanadate (BiVO4 ) photoanode as a model, it introduces an Au binding bridge as holes transfer channels onto the surfaces of BiVO4 , and the cyano-functionalized cobalt cubane (Co4 O4 ) molecules are preferentially immobilized on the Au bridge due to the strong adsorption of cyano groups with Au nanoparticles. This orchestrated arrangement facilitates the seamless transfer of photogenerated holes from BiVO4 to Co4 O4 molecules, forming an orderly charge transfer pathway connecting the light-absorbing layer to reactive sites. An exciting photocurrent density of 5.06 mA cm-2 at 1.23 V versus the reversible hydrogen electrode (3.4 times that of BiVO4 ) is obtained by the Co4 O4 @Au(A)/BiVO4 photoanode, where the surface charge recombination is almost completely suppressed accompanied by a surface charge transfer efficiency over 95%. This work represents a promising strategy for accelerating interfacial charge transfer and achieving efficient photoelectrochemical water oxidation reaction.
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In the realm of cell research, membraneless organelles have become a subject of increasing interest. However, their ever-changing and amorphous morphological characteristics have long presented a formidable challenge when it comes to studying their structure and function. In this paper, a fluorescent probe Nu-AN is reported, which exhibits the remarkable capability to selectively bind to and visualize the nucleolus morphology, the largest membraneless organelle within the nucleus. Nu-AN demonstrates a significant enhancement in fluorescence upon its selective binding to nucleolar RNA, due to the inhibited twisted intramolecular charge-transfer (TICT) and reduced hydrogen bonding with water. What sets Nu-AN apart is its neutral charge and weak interaction with nucleolus RNA, enabling it to label the nucleolus selectively and reversibly. This not only reduces interference but also permits the replacement of photobleached probes with fresh ones outside the nucleolus, thereby preserving imaging photostability. By closely monitoring morphology-specific changes in the nucleolus with this buffering fluorogenic probe, screenings for agents are conducted that induce nucleolar stress within living cells.
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Nucléolo Celular , ARN , Nucléolo Celular/metabolismo , ARN/metabolismoRESUMEN
The early lesions of Hashimoto's thyroiditis are inconspicuous, and the ultrasonic features of these early lesions are indistinguishable from other thyroid diseases. This paper proposes a Hashimoto Thyroiditis ultrasound image classification model HT-RCM which consists of a Residual Full Convolution Transformer (Res-FCT) model and a Residual Channel Attention Module (Res-CAM). To collect the low-order information caused by hypoechoic signals accurately, the residual connection is injected between FCTs to form Res-FCT which helps HT-RCM superimpose the low-order input information and high-order output information together. Res-FCT can make HT-RCM focus more on hypoechoic information while avoiding gradient dispersion. The initial feature map is inserted into Res-FCT again through a down-sampling component, which further helps HT-RCM exact multi-level original semantic information in the ultrasound image. Res-CAM is constructed by implementing a residual connection between a channel attention module and a convolution layer. Res-CAM can effectively increase the weights of the lesion channels while suppressing the weights of the noise channels, which makes HT-RCM focus more on the lesion regions. The experimental results on our collected dataset show that HT-RCM outperforms the mainstream models and obtains state-of-the-art performance in HT ultrasound image classification.
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Enfermedad de Hashimoto , Humanos , Enfermedad de Hashimoto/diagnóstico por imagen , Enfermedad de Hashimoto/patología , UltrasonografíaRESUMEN
Silk is a natural protein fiber that is predominantly comprised of fibroin and sericin. In addition, it contains seroins, protease inhibitors, enzymes, and other proteins. We found an ecdysone oxidase BmGMC2, notably, which is specifically and highly expressed only in the silk glands of silkworms (Bombyx mori L.). It is also one of the main components of non-cocoon silk, however, its precise function remains unclear. In this study, we examined the spatiotemporal expression pattern of this protein and obtained a homozygous mutant strain (K-GMC2) using the CRISPR-Cas9 system. Compared to the wild-type strain (WT), the silk production and main silk proteins significantly decreased in the larval stage, and the adhesive strength of native silk proteins decreased in the final instar. Proteomic data indicated the abundance of ribosomal proteins decreased significantly in K-GMC2, differentially expressed proteins (DEPs) were enriched in pathways related to neurodegenerative diseases and genetic information processing, indicating that knockout may lead to a certain degree of cell stress, affecting the synthesis of silk proteins. This study investigated the expression pattern and gene function of ecdysone oxidase BmGMC2 in silk and silk glands, laying the groundwork for understanding the role of enzymes in the production of silk fibers.