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Accumulation of advanced glycation end products (AGEs) causes apoptosis in human nucleus pulposus cells (NPCs), contributing to intervertebral disc degeneration (IVDD). The purpose of this study was to determine the roles of thioredoxin-interacting protein (TXNIP) in the mechanisms underlying AGE-induced apoptosis of NPCs. TXNIP was silenced or overexpressed in HNPCs exposed to AGEs. Glycolysis was assessed using extracellular acidification rate (ECAR), ATP level, GLUT1, and GLUT4 measurements. AGEs, TXNIP, GLUT1, and GLUT4 levels in IVDD patients were measured as well. In NPCs, AGEs reduced cell viability, induced apoptosis, inhibited glycolysis, and increased TXNIP expression. Silencing TXNIP compromised the effects of AGEs on cell viability, apoptosis, and glycolysis in NPCs. Furthermore, TXNIP overexpression resulted in decreased cell viability, increased apoptotic cells, and glycolysis suppression. Furthermore, co-treatment with a glycolysis inhibitor improved TXNIP silencing's suppressive effects on AGE-induced cell injury in NPCs. In IVDD patients with Pfirrmann Grades II-V, increasing trends in AGEs and TXNIP were observed, while decreasing trends in GLUT1 and GLUT4. AGE levels had positive correlations with TXNIP levels. Both AGE and TXNIP levels correlated negatively with GLUT1 and GLUT4. Our study indicates that TXNIP plays a role in mediating AGE-induced cell injury through suppressing glycolysis. The accumulation of AGEs, the upregulation of TXNIP, and the downregulation of GLUT1 and GLUT4 are all linked to the progression of IVDD.
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Degeneração do Disco Intervertebral , Núcleo Pulposo , Humanos , Degeneração do Disco Intervertebral/metabolismo , Núcleo Pulposo/metabolismo , Transportador de Glucose Tipo 1/metabolismo , Apoptose , Produtos Finais de Glicação Avançada/metabolismo , Proteínas de Transporte/metabolismoRESUMO
Preeclampsia (PE) seriously affects pregnant women and fetuses' health and causes maternal near-misses. CD81 has been confirmed to be a novel PE biomarker with great potential. Herein, a hypersensitive dichromatic biosensor based on the plasmonic enzyme-linked immunosorbent assay (plasmonic ELISA) is proposed initially for the application of CD81 in early screening for PE. In this work, a novel chromogenic substrate [(HAuCl4)-(N-methylpyrrolidone)-(Na3C6H5O7)] is designed based on the H2O2 dual catalysis reduction pathway of Au ions. The two reduction pathways of Au ions are controlled by H2O2 which ensures that the synthesis and growth of AuNPs are sensitive to H2O2. The amount of H2O2 correlates with the concentration of CD81 and directs the production of different-sized AuNPs in this sensor. Blue solutions are generated when analytes are present. When analytes are absent, solutions turn red. Therefore, due to different absorption peaks in red and blue, bimodal detection can be performed, and then two detection signals can be generated, one on signal at 550 nm and another off signal at 600 nm. This method exhibits a linear response to the logarithmic CD81 concentrations in the range of 0.1-1000 pg mL-1 with detection limits of 86 fg mL-1 and 152 fg mL-1 at two wavelengths. The false positive rate is low due to the nonspecific coloration caused by serum, which produces a more intense color contrast. The results indicate that the proposed dichromatic sensor could be used as a visual sensing platform for the direct detection of CD81 in biological samples and demonstrate its potential in preeclampsia diagnosis.
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Nanopartículas Metálicas , Pré-Eclâmpsia , Humanos , Feminino , Gravidez , Pré-Eclâmpsia/diagnóstico , Ouro , Peróxido de Hidrogênio , Ensaio de Imunoadsorção Enzimática , Limite de Detecção , Tetraspanina 28RESUMO
Tuberculosis is a major public health issue in Yemen, a country located at the southwestern tip of the Arabian Peninsula, while the situation of tuberculosis had been further exacerbated since the war started in 2015. The objective of this study is to investigate the incidence of tuberculosis in Yemen before the outbreak of COVID-19, from 2006 to 2018. During the 13-year period, 92 482 patients were enrolled in the TB programme records from the 22 governorates. Almost equal number of cases were diagnosed between males and females (a male to female ratio, 1.03:1). A notable rising incidence was observed in all age groups starting from 2011. The sharpest increase occurred in children under age 15, rising by 8.0-fold from 0.5 in the period 2006-2010 to 4.1 in the period 2011-2018. Paediatric TB accounted for 9.6% of all reported cases. In terms of the patient residence, incidence has more than doubled in Sana'a city, Sana'a Gov., Hajjah and Saadah. Concomitant diseases with tuberculosis included diabetes mellitus (14.0%), brucellosis (6.1%), hepatitis (6.0%), rheumatoid arthritis (4.3%), renal disorders (2.5%) and HIV infection (2.5%). Development of interventions to reduce tuberculosis incidence in children and concomitant communicable diseases is urgently needed.
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COVID-19 , Infecções por HIV , Tuberculose , Adolescente , COVID-19/epidemiologia , Criança , Feminino , Humanos , Masculino , Prevalência , Tuberculose/diagnóstico , Tuberculose/epidemiologia , Iêmen/epidemiologiaRESUMO
The target recognition algorithm is one of the core technologies of Zanthoxylum pepper-picking robots. However, most existing detection algorithms cannot effectively detect Zanthoxylum fruit covered by branches, leaves and other fruits in natural scenes. To improve the work efficiency and adaptability of the Zanthoxylum-picking robot in natural environments, and to recognize and detect fruits in complex environments under different lighting conditions, this paper presents a Zanthoxylum-picking-robot target detection method based on improved YOLOv5s. Firstly, an improved CBF module based on the CBH module in the backbone is raised to improve the detection accuracy. Secondly, the Specter module based on CBF is presented to replace the bottleneck CSP module, which improves the speed of detection with a lightweight structure. Finally, the Zanthoxylum fruit algorithm is checked by the improved YOLOv5 framework, and the differences in detection between YOLOv3, YOLOv4 and YOLOv5 are analyzed and evaluated. Through these improvements, the recall rate, recognition accuracy and mAP of the YOLOv5s are 4.19%, 28.7% and 14.8% higher than those of the original YOLOv5s, YOLOv3 and YOLOv4 models, respectively. Furthermore, the model is transferred to the computing platform of the robot with the cutting-edge NVIDIA Jetson TX2 device. Several experiments are implemented on the TX2, yielding an average time of inference of 0.072, with an average GPU load in 30 s of 20.11%. This method can provide technical support for pepper-picking robots to detect multiple pepper fruits in real time.
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Robótica , Zanthoxylum , Algoritmos , Meio Ambiente , Projetos de PesquisaRESUMO
Axially, epitaxially organizing nano-objects of distinct compositions and structures into superlattice nanowires enables full utilization of sunlight, readily engineered band structures, and tunable geometric parameters to fit carrier transport, thus holding great promise for optoelectronics and solar-to-fuel conversion. To maximize their efficiency, the general and high-precision synthesis of colloidal axial superlattice nanowires (ASLNWs) with programmable compositions and structures is the prerequisite; however, it remains challenging. Here, we report an axial encoding methodology toward the ASLNW library with precise control over their compositions, dimensions, crystal phases, interfaces, and periodicity. Using a predesigned, editable nanoparticle framework that offers the synthetic selectivity, we are able to chemically decouple adjacent sub-objects in ASLNWs and thus craft them in a controlled approach, yielding a library of distinct ASLNWs. We integrate therein plasmonic, metallic, or near-infrared-active chalcogenides, which hold great potential in solar energy conversion. Such synthetic capability enables a performance boost in target applications, as we report order-of-magnitude enhanced photocatalytic hydrogen production rates using optimized ASLNWs compared to corresponding solo objects. Furthermore, it is expected that such unique superlattice nanowires could bring out new phenomena.
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The early diagnosis of major diseases, such as malignant tumors, has always been an important field of research. Through screening, early detection of such diseases, and timely and effective treatment can significantly improve the survival rate of patients and reduce medical costs. Therefore, the development of a simple detection method with high sensitivity and strong specificity, and that is low cost is of great significance for the diagnosis and prognosis of the disease. Electrochemical DNA biosensing analysis is a technology based on Watson Crick base complementary pairing, which uses the capture probe of a known sequence to specifically recognize the target DNA and detect its concentration. Because of its advantages of low cost, simple operation, portability, and easy miniaturization, it has been widely researched and has become a cutting-edge topic in the field of biochemical analysis and precision medicine. However, the existing methods for electrochemical DNA biosensing analysis have some shortcomings, such as poor stability and specificity of capture probes, insufficient detection sensitivity, and long detection cycles. In this review, we focus on improving the sensitivity and practicability of electrochemical DNA biosensing analysis methods and summarize a series of research work carried out by using electrically neutral peptide nucleic acid as an immobilized capture probe.
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Técnicas Biossensoriais , Ácidos Nucleicos , Ácidos Nucleicos Peptídicos , DNA , Técnicas Eletroquímicas , Humanos , Hibridização de Ácido NucleicoRESUMO
In this work, we report a new amplification strategy based on electrochemically mediated reversible addition-fragmentation chain transfer (eRAFT) and in situ metalization for electrochemical detection of DNA. First, peptide nucleic acid (PNA) probes were immobilized on the surface of the gold electrode, and when they hybridized with the target DNA, the chain transfer agent (CTA), 4-cyano-4-(phenylcarbonothioylthio)pentanoic acid (CPAD), of RAFT was connected to the PNA/DNA heteroduplex formed by the coordination bonding of Zr4+. Then glycosyloxyethyl methacrylates (GEMA) were assembled on the surface of the electrode by electrochemically mediated surface-initiated reversible addition-fragmentation chain transfer (SI-eRAFT) to form a polymer-containing sugar glucose. Next, the o-hydroxyl groups on the polysaccharide molecular skeleton were oxidized to aldehyde groups by sodium periodate (NaIO4). The aldehyde groups generated then reduce silver ions to silver particles deposited on the electrode surface in situ, and this system was then subjected to differential pulse voltammetry (DPV). Under optimal conditions, the intensity of the stripping current and the logarithm of the target DNA (tDNA) concentration has a good linear relationship in the range of 10 aM to 1 pM (R2 = 0.996), and the detection limit can go down to 5.4 aM (S/N = 3). Moreover, the method is suitable for single-nucleotide polymorphism (SNP) analysis and has strong anti-interference ability for the analysis of target ssDNA in serum samples.
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Técnicas Biossensoriais/métodos , DNA/sangue , Ácidos Nucleicos Peptídicos/química , Complexos de Coordenação/química , DNA/genética , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Eletrodos , Ouro/química , Humanos , Limite de Detecção , Hibridização de Ácido Nucleico , Ácidos Pentanoicos/química , Ácidos Nucleicos Peptídicos/genética , Ácido Periódico/química , Polimorfismo de Nucleotídeo Único , Polissacarídeos/química , Reprodutibilidade dos Testes , Prata/química , Zircônio/químicaRESUMO
A highly sensitive method is described for determination of DNA. It is based on dual signal amplification, viz. (a)DNA-templated metal deposition, and (b) thermally initiated surface-initiated reversible addition-fragmentation chain transfer (SI-RAFT) polymerization. A peptide nucleic acid (PNA) with a terminal thiol group was grasped onto a gold electrode by self-assembly. The modified electrode serves as a probe to selectively capture target DNA (tDNA). In the next step, Zr(IV) ions are bound to the phosphate groups of the tDNA. A chain-transfer agent (CTA) for thermally initiated SI-RAFT polymerization, 4-cyano-4-(phenylcarbonothioylthio)pentanoic acid (CPAD), was immobilized on tDNA by conjugation of the carboxy group to Zr(IV) ions. Subsequently, numerous monomers of glycosyloxyethyl methacrylate (GEMA) were connected to the CPAD by thermally initiated SI-RAFT polymerization with azobisisobutyronitrile (AIBN) serving as the free-radical thermal initiator. Afterwards, hydroxyl groups of the GEMA were oxidized to aldehyde groups reacting with sodium periodate, and silver nanoparticles were further introduced on the surface of electrode via "silver mirror reaction". This results in a large electrochemical signal amplification. Under optimized conditions, the electrochemical signal (best measured at a working potential of 0 V vs. SCE (KCl; 3 M)) increases linearly with the logarithm of tDNA concentration in the 10 to 106 aM concentration range. The detection limit is as low as 5.6 aM (~34 molecules in a 10 µL sample). This is lower by factors between 2 and 1800 times than detection limits of most other ultra-sensitive electrochemical DNA assays. Graphical abstractSchematic representation of a dual signal amplification strategy combining thermally initiated surface-initiated reversible addition-fragmentation chain transfer polymerization (SI-RAFT) and DNA-templated silver nanoparticles for electrochemical determination of DNA.
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Técnicas Biossensoriais , DNA/análise , Técnicas Eletroquímicas , Nanopartículas Metálicas/química , Ácidos Nucleicos Peptídicos/química , Temperatura , Tamanho da Partícula , Polimerização , Prata/química , Propriedades de SuperfícieRESUMO
This systematic review and meta-analysis aim to explore the adjuvant effect of hyperbaric oxygen therapy (HBOT) in patients with venous leg ulcer (VLU) undergoing surgeries and non-surgeries. Literatures were searched from Web of Science, Cochrane Library, Embase, Pubmed, Wan fang, China National Knowledge Infrastructure (CNKI), and VIP from inception to November 15, 2022. The risk ratio (RR) and weighted mean difference (WMD) were used as effect size for categorical variables and continuous variables, respectively, with 95% confidence interval (95%CI). The heterogeneity was assessed using Q-test and quantified as I2. Sensitivity analysis was performed for all outcomes. A total of 11 studies were finally included in this study, with a total of 617 patients (313 in the HBOT group and 304 in the control group). Results showed that HBOT in combination with surgeries was associated with shorter ulcer healing time (WMD: -13.76, 95%CI: -20.42 to -7.10), lower VAS score (WMD: -0.95, 95% CI: -1.83 to -0.07), and smaller ulcer area (WMD: -2.64, 95%CI: -3.86 to -1.42). HBOT in combination with non-surgeries was associated with higher ulcer PAR (WMD: 20.82, 95%CI: 5.86 to 35.79), but no statistical significance was found in the improvement of ulcer area (WMD: 0.79, 95% CI: -1.54 to 3.12). Our results indicating that HBOT had a good adjuvant effect in surgeries to treat VLU, and its effect in non-surgeries needed further studies.
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Oxigenoterapia Hiperbárica , Úlcera Varicosa , Humanos , Úlcera/etiologia , Úlcera/terapia , Oxigenoterapia Hiperbárica/efeitos adversos , Úlcera Varicosa/terapia , Úlcera Varicosa/etiologia , ChinaRESUMO
Quantitative measurement of microRNAs (miRNAs) is extremely important in plenty of biomedical applications especially cancer diagnosis but remains a great challenge. In this work, we developed a logic gate recognition biosensing platform based on the "trinity" molecular recognition mode for quantifying miRNAs with a detection limit of 4.48 aM, along with a linear range from 0.1 nM to 10 aM under optimal experimental conditions. In order to obtain excellent detection performance, we adopted a Zn(TCPP) photocatalytic electron transfer reversible addition-fragmentation chain transfer (PET-RAFT) polymerization signal amplification strategy. The light-induced PET-RAFT has developed green applications of free radical polymerization in the field of biosensors. This is the first report on the preparation of signal amplification biosensors using PET-RAFT for tumor marker detection. With the outstanding detection performance, we can apply the sensor system to the early screening of lung cancer patients.
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Técnicas Biossensoriais , MicroRNAs , Humanos , Polimerização , Técnicas Eletroquímicas , Tomografia por Emissão de Pósitrons , Zinco , Limite de DetecçãoRESUMO
Herein we designed a highly sensitive and selective biosensor for methamphetamine (METH) detection based on aptamer recognition probe and atom transfer radical polymerization (ATRP) signal amplification strategy. In this experiment, METH aptamer and its complementary DNA strand were first attached to the electrode surface. In the presence of METH, the prioritized conjugation between METH and the aptamer will take one strand of DNA from the double-stranded DNA, so that the third segment of azide-modified DNA could be successfully modified onto the electrode surface. Through click chemistry and ATRP polymerization, the monomers with ferrocene were polymerized into a long chain, and the signal was amplified, then high-sensitivity detection of METH can be carried out. The result showed that the sensor could detect METH as low as 17 fM, which is about two orders of magnitude lower than that by traditional METH detection methods. Moreover, when different concentrations of METH were added to serum and urine, the recovery rate of the biosensor was as high as 93%. Therefore, using nucleic acid aptamer as capture probe and ATRP as signal amplification strategy can provide a promising application platform for sensitive detection of low concentration toxicants.
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Técnicas Biossensoriais , Metanfetamina , Técnicas Eletroquímicas , Limite de Detecção , PolimerizaçãoRESUMO
Lower extremity deep venous thrombosis (LEDVT) is a venous reflux disorder caused by abnormal coagulation of blood. LEDVT can obstruct the lumen and LEDVT is the third vascular disease after cerebrovascular diseases and coronary artery diseases. miRNAs are associated with thrombosis, and miR-185 was reported to affect the proliferation and apoptosis of vascular endothelial cells by regulating receptor of advanced glycation end products (RAGE). However, no study has reported the effect of miR-185 on LEDVT. Here, we studied the effects of miR-185 on the PI3K/AKT and MAPK signaling pathways in the LEDVT cells. The results showed that miR-185 promotes cell proliferation through activating the PI3K/AKT and MAPK signaling pathways and then inhibits tissue factor and fibrin expression to reduce thrombosis. In short, our study provides new ideas and a theoretical basis for research on the prevention, diagnosis, and treatment of LEDVT.
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MicroRNAs/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Trombose Venosa/genética , Animais , Proliferação de Células , Células Cultivadas , Cromonas/farmacologia , Modelos Animais de Doenças , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Fibrina/metabolismo , Sistema de Sinalização das MAP Quinases , Masculino , Camundongos , Morfolinas/farmacologia , Ratos , Tromboplastina/metabolismo , Trombose Venosa/etiologia , Trombose Venosa/metabolismoRESUMO
Due to the theoretical ultrahigh energy density of the Li-O2 battery chemistry, it has been hailed as the ultimate battery technology. Yet, practical Li-O2 batteries usually need to be designed in a large-sized pattern to actualize a high specific energy density, and such batteries often cannot be cycled effectively. To understand the inherent reasons, we specially prepared large-sized (13 cm × 13 cm) Li-O2 model batteries with practical energy output (6.9 Ah and 667.4 Wh/kgcell) for investigations. By subregional and postmortem analysis, the cathode interface was found to have severe local inhomogeneity after discharge, which was highly associated with the electrolyte and O2 maldistribution. The quantitative results by X-ray photoelectron spectroscopy (XPS) evidenced that this local inhomogeneity can exacerbate the generation of lithium acetate during charge, where the locally higher ratio of unutilized carbon surface and less Li2O2 after discharge would result in increased lithium acetate formation for a subsequent local overcharge. Moreover, verification experiments proved that the byproduct lithium acetate, which had been of less concern, was recalcitrant and triggered much larger polarization compared with the commonly reported byproduct Li2CO3 during battery operations, further revealing the key limiting factors leading to the poor rechargeability of batteries by its accumulation at a pouch-type cell level.
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OBJECTIVE: To explore relevant mechanisms of miR-139-5p in alleviating the metastasis of non-small cell lung cancer cells (NSCLC) and their resistance against cisplatin. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot (WB) assays were carried out to determine the protein levels of miR-139-5p and YAF2, and cisplatin (DDP)-resistant NSCLC cell strains were established. Subsequently, an MTT assay was employed to evaluate the viability of the cell strains, a Transwell assay to evaluate cell invasion activity, and flow cytometry to analyze cell apoptosis rate. Finally, a Western blot assay was carried out to determine the protein levels of P-PI3K and p-p38. RESULTS: NSCLC tissues showed lower miR-139-5p expression and higher YAF2 expression than paracancerous tissues and human normal lung epithelial cells, and miR-139-5p was related to the prognosis of NSCLC patients. Overexpression of miR-139-5p or knock-down of YAF2 inhibited the proliferation and invasion of NSCLC cells and induced their apoptosis. Additionally, the dual-luciferase reporter assay verified a targeting relationship between miR-139-5p and YAF2. Overexpression of miR-139-5p and knockdown of YAF2 reversed the resistance of A549/DDP cells against DDP, inactivated p38 and Akt proteins, and inhibited the AKT/p38 MAPK signaling pathway. Furthermore, inhibiting the AKT/p38 MAPK signaling pathway with MK2206 resisted the effects of knock-down of miR-139-5p on DDP resistance in NSCLC cells. CONCLUSION: MiR-139-5p targetedly regulates YAF2 and mediates the AKT/p38 MAPK signaling pathway to alleviate the metastasis of NSCLC cells and their resistance against cisplatin, which may be a novel target for improving the therapeutic effect on NSCLC.
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The detection of trace tumor-related serum miRNA biomarkers is in great demand for the early diagnosis of cancer. Herein, for the first time, an electrochemical sensing platform based on atom transfer radical polymerization (ATRP) signal amplification strategy for ultrasensitive determination of the breast and prostate cancer marker miRNA-141 has been developed. The hairpin DNAs were immobilized on the benzoic acid modified electrode to capture the target miRNA-141, the recognition of miRNA-141 released thiol groups on the end of probes, followed by the association of ATRP initiators modified gold nanoparticles with thiol groups, and then triggered the polymerization on electrode surface, causing a great number of ferrocene (Fc) signal molecules grafted on the sensor interface. As a result, the electrochemical signal intensity of signal molecule has been greatly increased. The proposed biosensor has a linear range from 10 pM to 10 aM with a detection limit of 3.23 aM for miRNA-141, opening a new and promising path for ultrasensitive analysis of tumor-related miRNAs.
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Técnicas Biossensoriais , Nanopartículas Metálicas , MicroRNAs , Técnicas Eletroquímicas , Ouro , Humanos , Limite de Detecção , Masculino , PolimerizaçãoRESUMO
In this paper, an ultrasensitive, highly selective and green electrochemical biosensor for quantifying DNA sequences (aM DNA) based on a MnTBAP catalyst for AGET ATRP reaction is proposed. For the first time, a combination of biomimetic catalyzed free radical polymerization and DNA electrochemical biosensing was used as a signal amplification strategy.
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Materiais Biomiméticos/química , Técnicas Biossensoriais , DNA/análise , Técnicas Eletroquímicas , Compostos Organometálicos/química , Catálise , Elétrons , Radicais Livres/química , PolimerizaçãoRESUMO
Metal-N-decorated carbon catalysts are cheap and effective alternatives for replacing the high-priced Pt-based ones in activating the reduction of oxygen for metal-air or fuel cells. The preparation of such heterogeneous catalysts often requires complex synthesis processes, including harsh acid treatment, secondary pyrolysis processes, etching, etc., to make the heteroatoms evenly dispersed in the carbon substrates to obtain enhanced activities. Through combined experimental characterizations, we found that by precise control of the precursors added, a Fe/N uniformly distributed, agglomeration-free Fe/N decorated Super-P carbon material (FNDSP) can be easily obtained by a one-pot synthesis process with distinctly higher pyridinic-N content and elevated catalytic activity. An insight into this phenomenon was carefully demonstrated and also verified in Li-O2 batteries, which delivered a high discharging platform of 2.85 V and can be fully discharged with a capacity of 5811.5 mA h gcarbon+catalyst -1 at the cut-off voltage of 2.5 V by the low-cost Super-P modified catalyst.
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BACKGROUND: Lung cancer is the most prevalent and deadly tumors around the world. Here we aimed to investigate the effect of wogonoside (also called baicalin) on the invasion and migration of lung cancer A549 cells and angiogenesis in xenograft tumors in nude mice. METHODS: A549 cells of lung cancer were treated with different doses of wogonoside. After 24 h, CCK8 was used to detect the survival rate of cells. The non-toxic doses of wogonoside (0, 10, 25, and 50 µM) were selected for subsequent experiments. Transwell and scratch assays were used to detect invasion and migration. The number of microtubule nodules was detected by microtubule formation experiment, and the expressions of VEGF, E-cadherin, N-cadherin, and Vimentin were detected by Western blotting. BALB/c nude mice were subcutaneously injected with lung cancer A549 cells to establish the xenograft model, followed by intraperitoneal injection of 80 mg/kg of wogonoside. After 30 days, tumor volume was measured, and the levels of VEGF and vimentin were detected with immunohistochemistry. The level of CD34 was determined by flow sorting. RESULTS: A549 cell survival decreased in a concentration-dependent manner, with the survival rate significantly reduced when the concentration of wogonoside exceeded 100 µM (P<0.05). A549 cell invasion and the number of microtubule nodules were significantly lower in the wogonoside 20 µM and the wogonoside 50 µM groups (P<0.05) compared with the wogonoside 0 µM group, while the rate of scratch closure and the protein levels of VEGF, N-cadherin, and Vimentin were all significantly reduced (P<0.05), and the expression level of E-cadherin was significantly increased (P<0.05). Compared with the control group, the tumor volumes of wogonoside (80 mg/kg) treated mice were significantly reduced after 30 days (P<0.05), and the levels of VEGF and vimentin positive cells were significantly reduced (P<0.05), as was the level of CD34 (P<0.05). CONCLUSIONS: Wogonoside can inhibit the invasion and migration of lung cancer A549 cells and angiogenesis of xenograft tumors in nude mice.
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SLIT2, a member of neuronal guidance cues, has been reported to regulate inflammation and cancer progression. Periodontitis is an oral inflammatory disease that degenerates periodontal tissue, alveolar bone and tooth. This study aims to explore the expression pattern of SLIT2 in periodontitis and its role in disease progression and bone loss. Gingival tissue of 20 periodontitis patients and 20 healthy-controls was obtained. Ligature-induced periodontitis (LIP) mice-model was developed in Slit2-Tg and wild-type mice. The effect of SLIT2 on inflammation, immune cell infiltration, M1 macrophage polarization, and alveolar bone loss in periodontitis was analyzed extensively. In periodontitis-affected gingival-tissue, SLIT2 expression was 4.4-fold higher compared to healthy-volunteers. LIP enhanced SLIT2 expression in mice periodontitis-affected periodontal tissue (PAPT) and blood circulation of wild-type mice by 4. 6-, and 5.0-fold, respectively. In Slit2-Tg-mice PAPT, SLIT2 expression was 1.8-fold higher compared to wild-type mice. Micro-CT and histomorphometric analysis revealed a 1.3-fold higher cement-enamel-junction to the alveolar-bone-crest (CEJ-ABC) distance and alveolar bone loss in LIP Slit2-Tg-mice compare to LIP wild-type mice. Results from RNA-sequencing, RT-qPCR, and ELISA showed a higher expression of Cxcr2, Il-18, TNFα, IL-6, and IL-1ß in Slit2-Tg-mice PAPT compared to wild-type-mice. Slit2-Tg-mice PAPT showed a higher number of osteoclasts, M1 macrophages, and the upregulation of Robo1 expression. Slit2-Tg-mice PAPT showed upregulation of M1 macrophage marker CD16/32 and osteoclastogenic markers Acp5, Ctsk, and Nfatc1, but osteogenic markers (Alp, Bglap) remained unchanged. Immunohistochemistry unveiled the higher vasculature and infiltration of leucocytes and macrophages in Slit2-Tg-mice PAPT. RNA-sequencing, GO-pathway enrichment analysis, and western blot analysis revealed the activation of the MAPK signaling pathway in Slit2-Tg mice PAPT. In conclusion, SLIT2 overexpression in periodontitis intensifies inflammation, immune cells infiltration, M1 macrophage polarization, osteoclastogenesis, and alveolar bone loss, possibly via activation of MAPK signaling, suggesting the role of SLIT2 on exacerbation of periodontitis and alveolar bone loss.