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1.
Mar Pollut Bull ; 202: 116396, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38657493

RESUMO

The Ledong Sea Area is located on the southwest side of Hainan Island. In recent years, with the development of industrialization and urbanization, the problem of heavy metals in marine sediments has gradually become a global problem, and research on this topic is of great significance for nearshore environmental protection and coastal management. This paper analysed the heavy metal content of 97 surface sediments in the Ledong Sea, indicating unpolluted to moderately polluted and low to moderate risk. Cu, Zn, Hg, Pb, Cr, and Cd are highly correlated, with similar origins, and originate from rivers carrying industrial wastewater, domestic sewage, and weathered material from the parent rocks, which are subsequently redistributed under the action of ocean dynamics. The distribution of Hg is mainly influenced by feed and biological metabolites during the farming process. As originates from rivers carrying large amounts of agricultural pesticide and fertilizer residues.

2.
Anal Chim Acta ; 1302: 342474, 2024 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-38580403

RESUMO

BACKGROUND: N6-methyladenosine (m6A) is a common modification in RNA, crucial for various cellular functions and associated with human diseases. Quantification of m6A at single-base resolution is of great significance for exploring its biological roles and related disease research. However, existing analysis techniques, such as polymerase chain reaction (PCR) or loop-mediated isothermal amplification (LAMP), face challenges like the requirement for thermal cycling or intricate primer design. Therefore, it is urgent to establish a simple, non-thermal cycling and highly sensitive assay for m6A. RESULTS: Leveraging the inhibitory effect of m6A on primer elongation and uncomplicated feature of the isothermal exponential amplification reaction (IEXPAR), we have developed an extension-based IEXPAR (E-IEXPAR). This approach requires just a single extension primer and one template, simplifying the design process in comparison to the more complex primer requirements of the LAMP methods. The reactions are conducted at constant temperatures, therby elimiating the use of thermal cycling that needed in PCR methods. By combining IEXPAR with an extension reaction, E-IEXPAR can identify m6A in RNA concentrations as low as 4 fM. We have also introduced a new analytical model to process E-IEXPAR results, which can aid to minimize the impact of unmodified adenine (A) on m6A measurements, enabling accurate m6A quantification in small mixed samples and cellular RNA specimens. SIGNIFICANCE AND NOVELTY: E-IEXPAR streamlines m6A detection by eliminating the need for intricate primer design and thermal cycling, which are common in current analytical methods. Its utilization of an extension reaction for the initial identification of m6A, coupled with a novel calculation model tailored to E-IEXPAR outcomes, ensures accurate m6A selectivity in mixed samples. As a result, E-IEXPAR offers a reliable, straightforward, and potentially economical approach for specifically assaying m6A in both biological function studies and clinical research.


Assuntos
Adenosina/análogos & derivados , Técnicas de Amplificação de Ácido Nucleico , RNA , Humanos , Primers do DNA/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Temperatura , Sensibilidade e Especificidade
3.
Arch Microbiol ; 206(4): 174, 2024 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-38493436

RESUMO

The present study focuses on investigating 60 strains of yeast isolated from the natural fermentation broth of Vitis labruscana Baily × Vitis vinifera L. These strains underwent screening using lysine culture medium and esculin culture medium, resulting in the identification of 27 local non-Saccharomyces yeast strains exhibiting high ß-glucosidase production. Subsequent analysis of their fermentation characteristics led to the selection of four superior strains (Z-6, Z-11, Z-25, and Z-58) with excellent ß-glucosidase production and fermentation performance. Notably, these selected strains displayed a dark coloration on esculin medium and exhibited robust gas production during Duchenne tubules' fermentation test. Furthermore, all four non-Saccharomyces yeast strains demonstrated normal growth under specific conditions including SO2 mass concentration ranging from 0.1 to 0.3 g/L, temperature between 25 and 30 °C, glucose mass concentration ranging from 200 to 400 g/L, and ethanol concentration at approximately 4%. Molecular biology identification confirmed that all selected strains belonged to Pichia kudriavzevii species which holds great potential for wine production.


Assuntos
Vitis , Vinho , Saccharomyces cerevisiae/metabolismo , Fermentação , beta-Glucosidase/metabolismo , Esculina/análise , Leveduras/metabolismo , Vinho/análise , Pichia/metabolismo
4.
Thorac Cancer ; 15(10): 764-777, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38400818

RESUMO

BACKGROUND: Non-small cell lung cancer (NSCLC) is a type of lung cancer that occurs in the cells of the respiratory tract, and its development is influenced by the regulation of circular RNAs (circRNAs). However, the role of circRNA carboxypeptidase A4 (circCPA4) in the progression of NSCLC and the underlying mechanism remain relatively clear. METHODS: The study utilized both real-time quantitative polymerase chain reaction (RT-qPCR) and western blot techniques to evaluate the levels of circCPA4, microRNA-145-5p (miR-145-5p), alanine, serine, or cysteine-preferring transporter 2 (ASCT2). To assess cell proliferation, cell counting kit-8 (CCK8) and 5-ethynyl-2'-deoxyuridine (EdU) assays were performed. Apoptosis was determined using flow cytometry, while cell migration and invasive capacity were evaluated through transwell and wound-healing assays. Intracellular levels of glutamine, glutamate, and α-KG were measured using specific kits. The relationship between miR-145-5p and circCPA4 or ASCT2 was confirmed using dual-luciferase reporter assay and RNA immunoprecipitation assay. RESULTS: CircCPA4 and ASCT2 RNA levels were elevated, while miR-145-5p was downregulated in both NSCLC tissues and cells. Depletion of circCPA4 significantly inhibited NSCLC cell proliferation, migration, invasion, and intracellular levels of glutamine, glutamate, and α-KG, and promoted apoptosis. Moreover, circCPA4 knockdown delayed tumor growth in vivo. Furthermore, circCPA4 was found to bind to miR-145-5p, thereby regulating the progression of NSCLC in vitro. ASCT2 was also identified as a downstream target of miR-145-5p, and its upregulation rescued the effects of miR-145-5p overexpression on NSCLC cell processes. CONCLUSION: CircCPA4 knockdown inhibited tumor property of NSCLC cells by modulating the miR-145-5p/ASCT2 axis.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , MicroRNAs , Humanos , Alanina , Carcinoma Pulmonar de Células não Pequenas/genética , Proliferação de Células , Cisteína , Glutamatos , Glutamina , Neoplasias Pulmonares/genética , MicroRNAs/genética , Serina
5.
Mol Brain ; 17(1): 12, 2024 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-38409127

RESUMO

BACKGROUND: Autism spectrum disorder (ASD) encompasses a diverse range of neurodevelopmental disorders, but the precise underlying pathogenesis remains elusive. This study aim to explore the potential mechanism of TREM2 in regulating microglia function in ASD. MATERIALS AND METHODS: The offspring rat model of ASD was established through prenatal exposure to valproic acid (VPA), and the behavioral symptoms of the ASD model were observed. On postnatal day (PND) 7 and PND 28, the effects of prenatally exposure to VPA on synaptic development and microglia phenotype of offspring rats were observed. Primary microglia were cultured in vitro. Lentivirus and adenovirus were utilized to interfere with TREM2 and overexpress TREM2. RESULTS: Prenatally VPA exposure induced offspring rats to show typical ASD core symptoms, which led to abnormal expression of synapse-related proteins in the prefrontal cortex of offspring rats, changed the phenotype of microglia in offspring rats, promoted the polarization of microglia to pro-inflammatory type, and increased inflammatory response. The experimental results in vitro showed that overexpression of TREM2 could increase the expression of Gephyrin, decrease the content of CD86 protein and increase the content of CD206 protein. In addition, after the expression of TREM2 was interfered, the content of p-P38 MAPK protein increased and the content of p-ELK-1 protein decreased. CONCLUSION: The protective influence of TREM2 on the VPA-induced ASD model is attributed to its inhibition of the P38 MAPK pathway, this protective effect may be achieved by promoting the polarization of microglia to anti-inflammatory phenotype and improving the neuronal synaptic development.


Assuntos
Transtorno do Espectro Autista , Animais , Feminino , Gravidez , Ratos , Transtorno do Espectro Autista/metabolismo , Modelos Animais de Doenças , Microglia/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Transdução de Sinais , Ácido Valproico/farmacologia
6.
Protein Expr Purif ; 217: 106432, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38232795

RESUMO

Natural ginsenoside needs to be converted into rare ginsenoside before it can be readily absorbed into the bloodstream for action. In this study, an α-l-arabinofuranosidase (α-l-AFase) gene Bsafs2 was cloned from Bacillus subtilis (B. subtilis). Bsafs2 was ligated to the expression vector pET28a(+), and the expression vector was constructed and transformed into Escherichia coli (E. coli) BL21 heterologous recombinant expression to obtain α-l-AFase. α-l-AFase can hydrolyze at the C20 site of Ginsenoside Rc to obtain rare ginsenoside Rd. Studies on the enzymatic property showed that α-l-AFase had good tolerance to ethanol, glucose, and l-arabinose. The optimum temperature of α-l-AFase was 40 °C and pH = 5.5. Kinetic parameters Km of α-l-AFase for pNPαAraf and Ginsenoside Rc were 1.93 and 8.9 mmol/L, the Vmax were 26 and 154 µmol/min/mg, the Kcat were 24.14 and 1.48 S-1, respectively. This study provides the enzyme source for the biotransformation of Ginsenoside Rc.


Assuntos
Ginsenosídeos , Ginsenosídeos/metabolismo , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Clonagem Molecular , Proteínas Recombinantes/química , Escherichia coli/metabolismo , Glicosídeo Hidrolases/química
7.
J Colloid Interface Sci ; 660: 458-468, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38246049

RESUMO

The high activity barriers of Li2S nucleation and deposition limit the redox reaction kinetics of lithium polysulfides (LiPSs), meanwhile, the significant shuttle effect of LiPSs hampers the advancement of Li-S batteries (LSBs). In this work, a NiSe2/CoSe2-rGO (NiSe2/CoSe2-G) sulfur host with bifunctional catalytic activity was prepared through a hard template method. Electrochemical experiment results confirm that the combination of NiSe2 and CoSe2 not only facilitates the bidirectional catalytic function during charge and discharge processes, but also increases the active sites toward LiPSs adsorption. Simultaneously, the highly conductive rGO network enhances the electronic conductivity of NiSe2/CoSe2-G/S and provides convenience for loading NiSe2/CoSe2 catalysts. Benefitting from the exceptional catalytic-adsorption capability of NiSe2/CoSe2 and the presence of rGO, the NiSe2/CoSe2-G/S electrode exhibits excellent electrochemical properties. At 1C, it demonstrates a low capacity attenuation of 0.087 % per cycle during 500 cycles. The electrode can maintain a discharge capacity of 927 mAh/g at a sulfur loading of 3.3 mg cm-2. The bidirectional catalytic activity of NiSe2/CoSe2-G offers a prospective approach to expedite the redox reactions of active S, meanwhile, this work also offers an ideal approach for designing efficient S hosts for LSBs.

8.
Int J Biometeorol ; 68(1): 133-141, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37950095

RESUMO

Dengue is one of the world's most rapidly spreading mosquito-borne viral diseases. As it is found mostly in urban and semi-urban areas, urbanization and associated human activities that affect the environment and larval habitats could become risk factors (e.g., lane width, conditions of street ditches) for the spread of dengue. However, there are currently no systematic studies of micro-scale urbanization-based risk factors for the spread of dengue epidemics. We describe the study area, two micro-scale environmental risk factors associated with urbanization, and meteorological data. Since the observations involve spatial and temporal correlations, we also use some statistical methods for the analysis of spatial and spatial-temporal data for the relationship between urbanization and dengue. In this study, we analyzed data from Kaohsiung, a densely populated city in southern Taiwan, and found a positive correlation between environmental risk factors associated with urbanization (ditches positive for mosquito larvae and closely packed streets termed "dengue lanes") and clustering effects in dengue cases. The statistical analysis also revealed that the occurrence of positive ditches was significantly associated with that of dengue lanes in the study area. The relationship between climate variables and positive ditches was also analyzed in this paper, indicating a relationship between dengue and both rainfall and temperature, with temperature having a greater effect. Overall, this work is immediately relevant and applicable for policymakers in government, who will need to reduce these favorable habitats for vector-born disease spreaders and implement regulations for new urban constructions to thus reduce dengue spread in future outbreaks.


Assuntos
Dengue , Epidemias , Animais , Humanos , Urbanização , Dengue/epidemiologia , Cidades/epidemiologia , Fatores de Risco , Larva
9.
J Allergy Clin Immunol Glob ; 2(4): 100163, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37781663

RESUMO

Background: Reinforcement of the immune-regulatory pathway is a feasible strategy for prevention and therapy of allergic asthma. The short-chain fatty acids (SCFAs) acetate, propionate, and butyrate are pleiotropic microbial fermentation products known to induce regulatory T (Treg) cells and exert an immune-regulatory effect. The cellular mechanism underlying SCFA immune regulation in asthma is not fully understood. Objective: We investigated the role of myeloid-derived suppressor cells (MDSCs) and Treg cells, the immune-regulatory cells of innate and adaptive origin, respectively, in SCFA-elicited protection against allergic airway inflammation. Methods: BALB/c mice were given SCFA-containing drinking water before being rendered asthmatic in response to ovalbumen. When indicated, mice were given a GR1-depleting antibody to investigate the function of MDSCs in allergic inflammation of the airways. MDSCs were sorted to examine their immunosuppressive function and interaction with T cells. Results: The mice receiving SCFAs developed less severe asthma that was accompanied by expansion of PMN-MDSCs and Treg cells. Mice depleted of PMN-MDSCs exhibited aggravated asthma, and the protective effect of SCFAs was abrogated after PMN-MDSC depletion. SCFAs were able to directly induce T-cell differentiation toward Treg cells. Additionally, we found that PMN-MDSCs enhanced Treg cell expansion in a cell contact-dependent manner. Whilst membrane-bound TGF-ß has been shown to induce Treg cell differentiation, we found that MDSCs upregulated surface expression of TGF-ß after coculture with T-cells and that MDSC-induced Treg cell differentiation was partially inhibited by TGF-ß blockage. Conclusions: Although previous studies revealed Treg cells as the effector mechanism of SCFA immune regulation, we found that SCFAs ameliorate allergic airway inflammation by relaying immune regulation, with sequential induction of PMN-MDSCs and Treg cells.

10.
BMC Pediatr ; 23(1): 480, 2023 09 21.
Artigo em Inglês | MEDLINE | ID: mdl-37735378

RESUMO

BACKGROUND: Contiguous gene gain syndrome including entire ZEB2 may be a novel syndrome. In the past, there were no easily distinct and recognizable features as a guide for precise clinical and genetic diagnosis of the syndrome. CASE PRESENTATION: We report a novel case with the syndrome with a novel de novo 22.16 Mb duplication at 2q21.2-q24.1. The syndrome is characterized by multiple anomalies including the same typical craniofacial phenotype that is entirely different from Mowat-Wilson syndrome (MWS), and other quite similar features of MWS consisting of development delay, congenital heart disease, abdominal abnormalities, urogenital abnormalities, behavioral problems and so on, in which the distinctive craniofacial features can be more easily recognized. CONCLUSIONS: Contiguous gene gain syndrome including entire ZEB2 characterized with similar multiple congenital anomalies of MWS and the distinctive craniofacial features is mainly caused by large 2q22 repeats including ZEB2 leading to dominant singe ZEB2 gene gain mutation, which is recommended to be named "Liu-Liang-Chung" syndrome. We diagnose this novel syndrome to distinguish it from MWS. Some variable additional features in the syndrome including remarkable growth and development retardation and protruding ears were recognized for the first time.


Assuntos
Anormalidades Múltiplas , Doença de Hirschsprung , Humanos , Anormalidades Múltiplas/genética , Mutação , Doença de Hirschsprung/diagnóstico , Doença de Hirschsprung/genética , Fenótipo , Homeobox 2 de Ligação a E-box com Dedos de Zinco/genética
11.
Biosens Bioelectron ; 238: 115578, 2023 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-37573644

RESUMO

Quantification of microRNAs (miRNAs) at the single-molecule level is of great significance for clinical diagnostics and biomedical research. The challenges lie in the limits to transforming single-molecule measurements into quantitative signals. To address these limits, here, we report a new approach called a Single Microbead-based Space-confined Digital Quantification (SMSDQ) to measure individual miRNA molecules by counting gold nanoparticles (AuNPs) with localized surface plasmon resonance (LSPR) light-scattering imaging. One miRNA target hybridizes with the alkynyl-modified capture DNA probe immobilized on a microbead (60 µm) and the azide-modified report DNA probe anchored on AuNP (50 nm), respectively. Through the click reaction between the alkynyl and azide group, a single microbead can covalently link the AuNPs in the confined space within the view of the microscope. By digitally counting the light-scattering spots of AuNPs, we demonstrated the proposed approach with single-molecule detection sensitivity and high specificity of single-base discrimination. Taking the advantages of ultrahigh sensitivity, specificity, and the digital detection manner, the approach is suitable for evaluating cell heterogeneity and small variations of miRNA expression and has been successfully applied to direct quantification of miRNAs in one-tenth single-cell lysates and serum samples without RNA-isolated and nucleic acid amplification steps.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , MicroRNAs , MicroRNAs/genética , Ouro , Azidas , Microesferas , Técnicas Biossensoriais/métodos , Limite de Detecção
12.
Chem Commun (Camb) ; 59(72): 10769-10772, 2023 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-37592916

RESUMO

We developed a method for quantifying N6-methyladenosine at one-nucleotide resolution based on double blocking gap-filling-ligation and cascade isothermal amplification. This proposed method can detect as low as 1 fM target RNA, achieving selectivity up to approximately 100-fold between m6A and A, and has been successfully applied to the analysis of m6A at specific sites in cell samples.


Assuntos
Adenosina , RNA , Adenosina/análise , RNA/análise
13.
Hum Gene Ther ; 34(13-14): 629-638, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37279283

RESUMO

The cytosine base editor (CBE) has shown promise as a gene editing tool for gene therapy, as it can convert cytidine to thymidine. Adeno-associated virus (AAV) has been widely used for in vivo gene therapy, but its limited 4.7 kb packing capacity presents challenges in delivering CBE by a single AAV. To address this, one feasible solution is to split CBE into two sections for dual-AAV delivery. In this study, we utilized BE3 as an example and constructed 22 potential split-BE3 pairs with the combination of 11 splitting sites and two split-inteins (Npu and Rma). These split-BE3 pairs were initially screened in the green fluorescent protein (GFP) reporter system, with six split-BE3 pairs selected for further evaluation. The subsequent screening of split-BE3 pairs was performed at two endogenous sites in 293T and HeLa cells, revealing that the split-BE3-Rma674, split-BE3-Rma713, and split-BE3-Rma1005 displayed effective C-to-T conversion after transfection. The effectiveness of dual-AAV split-BE3 was further validated in culture cells and adult mouse eyes. Of note, the split-BE3-Rma674 demonstrated the most efficient C-to-T editing after AAV infection, with a maximal editing efficiency of 23.29% ± 10.98% in the mouse retinal pigment epithelium cells in vivo. Overall, our study presents a novel split-BE3 system with effective C-to-T conversion, which could be applied to CBE-based in vivo gene therapy.


Assuntos
Citosina , Dependovirus , Humanos , Animais , Camundongos , Dependovirus/genética , Dependovirus/metabolismo , Citosina/metabolismo , Células HeLa , Edição de Genes , Transfecção , Sistemas CRISPR-Cas
14.
PLoS Negl Trop Dis ; 17(6): e0011346, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37289665

RESUMO

Dengue fever is a vector-borne disease that has become a serious global public health problem over the past decade. An essential aspect of controlling and preventing mosquito-borne diseases is reduction of mosquito density. Through the process of urbanization, sewers (ditches) have become easy breeding sources of vector mosquitoes. In this study, we, for the first time, used unmanned ground vehicle systems (UGVs) to enter ditches in urban areas to observe vector mosquito ecology. We found traces of vector mosquitoes in ~20.7% of inspected ditches, suggesting that these constitute viable breeding sources of vector mosquitoes in urban areas. We also analyzed the average gravitrap catch of five administrative districts in Kaohsiung city from May to August 2018. The gravitrap indices of Nanzi and Fengshan districts were above the expected average (3.26), indicating that the vector mosquitoes density in these areas is high. Using the UGVs to detect positive ditches within the five districts followed by insecticide application generally yielded good control results. Further improving the high-resolution digital camera and spraying system of the UGVs may be able to effectively and instantly monitor vector mosquitoes and implement spraying controls. This approach may be suitable to solve the complex and difficult task of detecting mosquito breeding sources in urban ditches.


Assuntos
Aedes , Culicidae , Dengue , Animais , Mosquitos Vetores , Ecologia , Cidades , Urbanização , Controle de Mosquitos/métodos , Dengue/epidemiologia
15.
Cancer Med ; 12(12): 13811-13820, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-37132165

RESUMO

BACKGROUND: This study aims to provide a theoretical basis for the prevention of oral cancer by analyzing the epidemiological trends of oral cancer. MATERIALS AND METHODS: The data on oral cancer from 1990 to 2019 were extracted from the Global Burden of Disease 2019 database. The incidence, mortality, disability-adjusted life years (DALYs), and age-standardized rate as well as attributable risk factors of oral cancer were used for the analysis. Estimated annual percentage change (EAPC) was calculated to describe the changes in age-standardized incidence rate (ASIR), age-standardized mortality rate (ASMR), and age-standardized DALYs rate (ASDR). RESULTS: The global ASIR of oral cancer showed an increasing trend from 1990 to 2019. ASIR in high SDI regions showed a decreasing trend during the studied period, with high SDI regions having the lowest ASMR in 2019. In 2019, the highest ASIR, ASMR, and ASDR were detected in South Asia. At the national level, Pakistan had the highest ASMR and ASDR in 2019. The increasing disease burden was observed in younger populations aged below 45 during the studied period. Smoking and alcohol use still exerted profound impacts on the oral cancer burden, with South Asia having the greatest increase in the percentage of deaths due to oral cancer attributable to chewing tobacco from 1990 to 2019. CONCLUSION: In conclusion, there is a large variability in the temporal and spatial burden of oral cancer, and it is essential for priority countries to take targeted intervention policies and measures to reduce the disease burden of oral cancer. In addition, the oral cancer burden caused by attributable risk factors should also receive close attention.


Assuntos
Neoplasias Bucais , Humanos , Idoso , Neoplasias Bucais/epidemiologia , Neoplasias Bucais/etiologia , Fatores de Risco , Consumo de Bebidas Alcoólicas/efeitos adversos , Consumo de Bebidas Alcoólicas/epidemiologia , Fumar , Ásia Meridional , Saúde Global , Incidência
16.
Materials (Basel) ; 16(6)2023 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-36984078

RESUMO

The machining process of aluminum alloy usually produces built-up edge and tool sticking problems due to their low hardness and large plastic deformation, which may further affect the machined surface quality and tool life. This paper aims to investigate the influence of different cutting fluids on the machined surface quality and tool life during the milling process of 7050 aluminum alloy. A novel cutting fluid (QC-2803) was considered in the study, which is synthesized by addition of alkyl alcohol amide and chlorinated polyolefin, and the traditional cutting fluid (CCF-10) was used as the control group. The physical and chemical properties of two cutting fluids were characterized. The milling process of 7050 aluminum alloy was carried out under two different cutting fluid conditions. The machined surface morphology, cutting force and tool wear morphology were observed during the process. Results show that the surface tension of the novel cutting fluid is significantly lower than that of the traditional cutting fluid, which makes it easier to produce a lubricating film between the aluminum alloy and tool, and further benefits the machined surface quality and tool life. As a result, the surface roughness and cutting force are reduced by ~20.0% and ~42.9%, respectively, and the tool life is increased by 25.6% in the case of the novel cutting fluid (QC-2803). The results in this paper revealed the important laws of cutting fluid with metal surface quality, cutting performance and tool wear, which helps to control the machined surface quality and tool life by the selection of cutting fluid during metal milling.

17.
Anal Chem ; 95(13): 5729-5737, 2023 04 04.
Artigo em Inglês | MEDLINE | ID: mdl-36944919

RESUMO

The target-dependent endonuclease activity (also known as the trans-cleavage activity) of CRISPR-Cas systems has stimulated great interest in the development of nascent sensing strategies for nucleic acid diagnostics. Despite many attempts, the majority of the sensitive CRISPR-Cas diagnostics strategies mainly rely on nucleic acid preamplification, which generally needs complex probes/primers designs, multiple experimental steps, and a longer testing time, as well as introducing the risk of false-positive results. In this work, we propose the CRISPR-Cas-Driven Single Micromotor (Cas-DSM), which can directly detect the nucleic acid targets at a single-molecule level with high specificity. We have demonstrated that the Cas-DSM is a reliable and practical method for the quantitative detection of DNA/RNA in various complex clinical samples as well as in individual cells without any preamplification processes. Due to the excellent features of the CRISPR/Cas system, including constant temperature, simple design, high specificity, and flexible programmability, the Cas-DSM could serve as a simple and universal platform for nucleic acid detection. More importantly, this work will provide a breakthrough for the development of next-generation amplification-free CRISPR/Cas sensing toolboxes.


Assuntos
Técnicas Biossensoriais , Ácidos Nucleicos , Sistemas CRISPR-Cas/genética , RNA , Biomarcadores , Primers do DNA
18.
J Biomed Sci ; 30(1): 12, 2023 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-36803804

RESUMO

BACKGROUND: Zika virus (ZIKV) infection is clinically known to induce testicular swelling, termed orchitis, and potentially impact male sterility, but the underlying mechanisms remain unclear. Previous reports suggested that C-type lectins play important roles in mediating virus-induced inflammatory reactions and pathogenesis. We thus investigated whether C-type lectins modulate ZIKV-induced testicular damage. METHODS: C-type lectin domain family 5 member A (CLEC5A) knockout mice were generated in a STAT1-deficient immunocompromised background (denoted clec5a-/-stat1-/-) to enable testing of the role played by CLEC5A after ZIKV infection in a mosquito-to-mouse disease model. Following ZIKV infection, mice were subjected to an array of analyses to evaluate testicular damage, including ZIKV infectivity and neutrophil infiltration estimation via quantitative RT-PCR or histology and immunohistochemistry, inflammatory cytokine and testosterone detection, and spermatozoon counting. Furthermore, DNAX-activating proteins for 12 kDa (DAP12) knockout mice (dap12-/-stat1-/-) were generated and used to evaluate ZIKV infectivity, inflammation, and spermatozoa function in order to investigate the potential mechanisms engaged by CLEC5A. RESULTS: Compared to experiments conducted in ZIKV-infected stat1-/- mice, infected clec5a-/-stat1-/- mice showed reductions in testicular ZIKV titer, local inflammation and apoptosis in testis and epididymis, neutrophil invasion, and sperm count and motility. CLEC5A, a myeloid pattern recognition receptor, therefore appears involved in the pathogenesis of ZIKV-induced orchitis and oligospermia. Furthermore, DAP12 expression was found to be decreased in the testis and epididymis tissues of clec5a-/-stat1-/- mice. As for CLEC5A deficient mice, ZIKV-infected DAP12-deficient mice also showed reductions in testicular ZIKV titer and local inflammation, as well as improved spermatozoa function, as compared to controls. CLEC5A-associated DAP12 signaling appears to in part regulate ZIKV-induced testicular damage. CONCLUSIONS: Our analyses reveal a critical role for CLEC5A in ZIKV-induced proinflammatory responses, as CLEC5A enables leukocytes to infiltrate past the blood-testis barrier and induce testicular and epididymal tissue damage. CLEC5A is thus a potential therapeutic target for the prevention of injuries to male reproductive organs in ZIKV patients.


Assuntos
Orquite , Infecção por Zika virus , Zika virus , Humanos , Masculino , Camundongos , Animais , Sêmen/metabolismo , Camundongos Knockout , Inflamação/genética , Lectinas Tipo C/genética , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo
19.
Front Bioeng Biotechnol ; 11: 1100968, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36741759

RESUMO

An essential aspect of controlling and preventing mosquito-borne diseases is to reduce mosquitoes that carry viruses. We designed a smart mosquito trap system to reduce the density of mosquito vectors and the spread of mosquito-borne diseases. This smart trap uses computer vision technology and deep learning networks to identify features of live Aedes aegypti and Culex quinquefasciatus in real-time. A unique mechanical design based on the rotation concept is also proposed and implemented to capture specific living mosquitoes into the corresponding chambers successfully. Moreover, this system is equipped with sensors to detect environmental data, such as CO2 concentration, temperature, and humidity. We successfully demonstrated the implementation of such a tool and paired it with a reliable capture mechanism for live mosquitos without destroying important morphological features. The neural network achieved 91.57% accuracy with test set images. When the trap prototype was applied in a tent, the accuracy rate in distinguishing live Ae. aegypti was 92%, with a capture rate reaching 44%. When the prototype was placed into a BG trap to produce a smart mosquito trap, it achieved a 97% recognition rate and a 67% catch rate when placed in the tent. In a simulated living room, the recognition and capture rates were 90% and 49%, respectively. This smart trap correctly differentiated between Cx. quinquefasciatus and Ae. aegypti mosquitoes, and may also help control mosquito-borne diseases and predict their possible outbreak.

20.
J Colloid Interface Sci ; 629(Pt B): 279-287, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36155923

RESUMO

With the increasing requirement for high capacity energy storage systems, a large amount of recent work has focused on the development of zinc-iodine batteries (ZIBs) on account of high energy density, fast redox kinetics, and excellent reversibility. Nevertheless, low electron conductivity, the shuttle effect, and highly soluble iodine species (I2, I-, and I3-) have impeded their widespread application. In this study, metal organic framework-5 (MOF-5)-derived mesoporous carbon (MPC) loaded iodine (MPC/I2) cathode and the single-sided ketjen black modified cotton fiber (KB@CF) separator are designed to solve the problems mentioned above. That is, the double fixation strategy using MPC and KB@CF separators for iodine species suppresses the shuttle effect. Therefore, the ZIBs constructed with the MPC/I2 cathode and the KB@CF separator can exhibit excellent electrochemical performance. At the current density of 0.1 A g-1, a high discharge specific capacity of 137 mAh g-1 is still available after 300 cycles. Meanwhile, it exhibits a low capacity decay rate at long cycling (0.030% per cycle over 2000 cycles).

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