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Lung cancer (LC) is the most prevalent cancer type, with a high mortality rate worldwide. The current treatment options for LC have not been particularly successful in improving patient outcomes. Yifei Sanjie (YFSJ), a well-applicated traditional Chinese medicine formula, is widely used to treat pulmonary diseases, especially LC, yet little is known about its molecular mechanisms. This study was conducted to explore the molecular mechanism by which YFSJ ameliorated LC progression. The A549, NCI-H1975, and Calu-3 cells were treated with the YFSJ formula and observed for colony number, apoptosis, migration, and invasion properties recorded via corresponding assays. The PRMT6-YBX1-CDC25A axis was tested and verified through luciferase reporter, RNA immunoprecipitation, and chromatin immunoprecipitation assays and rescue experiments. Our results demonstrated that YFSJ ameliorated LC cell malignant behaviors by increasing apoptosis and suppressing proliferation, migration, and invasion processes. We also noticed that the xenograft mouse model treated with YFSJ significantly reduced tumor growth compared with the control untreated group in vivo. Mechanistically, it was found that YFSJ suppressed the expression of PRMT6, YBX1, and CDC25A, while the knockdown of these proteins significantly inhibited colony growth, migration, and invasion, and boosted apoptosis in LC cells. In summary, our results suggest that YFSJ alleviates LC progression via the PRMT6-YBX1-CDC25A axis, confirming its efficacy in clinical use. The findings of our study provide a new regulatory network for LC growth and metastasis, which could shed new insights into pulmonary medical research.
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Neoplasias Pulmonares , MicroRNAs , Humanos , Animais , Camundongos , Neoplasias Pulmonares/patologia , Proliferação de Células/genética , Movimento Celular/genética , Pulmão/patologia , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Proteína 1 de Ligação a Y-Box/genética , Proteína 1 de Ligação a Y-Box/metabolismo , Proteínas Nucleares/genética , Proteína-Arginina N-Metiltransferases/genética , Proteína-Arginina N-Metiltransferases/metabolismo , Proteína-Arginina N-Metiltransferases/uso terapêutico , Fosfatases cdc25/genética , Fosfatases cdc25/metabolismoRESUMO
1,3,6-Trigalloylglucose is a natural compound that can be extracted from the aqueous extracts of ripe fruit of Terminalia chebula Retz, commonly known as "Haritaki". The potential anti-Helicobacter pylori (HP) activity of this compound has not been extensively studied or confirmed in scientific research. This compound was isolated using a semi-preparative liquid chromatography (LC) system and identified through Ultra-high-performance liquid chromatography-MS/MS (UPLC-MS/MS) and Nuclear Magnetic Resonance (NMR). Its role was evaluated using Minimum inhibitory concentration (MIC) assay and minimum bactericidal concentration (MBC) assay, scanning electron microscope (SEM), inhibiting kinetics curves, urea fast test, Cell Counting Kit-8 (CCK-8) assay, Western blot, and Griess Reagent System. Results showed that this compound effectively inhibits the growth of HP strain ATCC 700392, damages the HP structure, and suppresses the Cytotoxin-associated gene A (Cag A) protein, a crucial factor in HP infection. Importantly, it exhibits selective antimicrobial activity without impacting normal epithelial cells GES-1. In vitro studies have revealed that 1,3,6-Trigalloylglucose acts as an anti-adhesive agent, disrupting the adhesion of HP to host cells, a critical step in HP infection. These findings underscore the potential of 1,3,6-Trigalloylglucose as a targeted therapeutic agent against HP infections.
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Helicobacter pylori , Terminalia , Extratos Vegetais/química , Terminalia/química , Cromatografia Líquida , Espectrometria de Massas em Tandem , ÁguaRESUMO
Copper ions play vital roles in regulating life processes and being closely involved in several diseases such as cancer. Although detection methods based on fluorescent sensors or other strategies have been developed, it still remains a challenge to simultaneously realize the convenience, specificity, and accuracy in intracellular copper ion analysis. Herein, we propose an aptamer-functionalized DNA fluorescent sensor (AFDS) for accurate and specific detection of Cu(II) both in vitro and in cells by engineering the linkage of two DNA aptamers, namely, Lettuce aptamer and AS1411 aptamer, to achieve the manner of recognition response. Taking advantage of the functions of each aptamer, the tumor cell recognition capability and the high-contrast detection performance are simultaneously equipped in the AFDS. Furthermore, the AFDS shows high specificity and selectivity in Cu(II) response to avoid interference from common metal ions, chelators, and reactants by being associated with the irreversible interaction between nucleobases and Cu(II), which can destroy the topological structures and switch off the fluorescence of the AFDS. It also enables a sensitive in vitro detection of Cu(II) with a detection limit as lower as 0.1 µM and a wide detection linear range from 0.1 to 300 µM. The feasibility and superiority of the AFDS provide an opportunity to reveal both concentration-dependent and time-dependent intracellular Cu(II) responses in living cells. Therefore, the AFDS has achieved the novel detection performance of Cu(II) to exhibit great potential in exploring copper-related biological and pathological research.
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Cobre , Metais , Cobre/química , DNA , Íons , Corantes Fluorescentes/químicaRESUMO
Metal cylindrical shaft parts are critical components in industrial manufacturing that require high standards for roundness error and surface roughness. When using the self-developed multi-beam angle sensor (MBAS) to detect metal cylindrical shaft parts, the distorted multi-spots degrade the measurement accuracy due to the nonlinear distortion caused by the metal material's reflective properties and surface roughness. In this study, we propose a spot coordinate prediction network (SCPNet), which is a deep-learning neural network designed to predict spot coordinates, in combination with Hough circle detection for localization. The singular value decomposition (SVD) model is employed to eliminate the tilt error to achieve high-precision, three-dimensional (3D) surface reconstruction of metal cylindrical shaft parts. The experimental results demonstrate that SCPNet can effectively correct distorted multi-spots, with an average error of the spot center of 0.0612 pixels for ten points. The proposed method was employed to measure metal cylindrical shaft parts with radii of 10 mm, 20 mm, 35 mm, and 50 mm, with resulting standard deviation (STD) values of 0.0022â µm, 0.0026â µm, 0.0028â µm, and 0.0036â µm, respectively.
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To solve limited efficiency and reliability issues caused by current manual quality control processes in optical lens (OL) production environments, we propose an automatic micro vision-based inspection system named MVIS used to capture the surface defect images and make the OL dataset and predictive inference. Because of low resolution and recognition, OL defects are weak, due to their ambiguous morphology and micro size, making a poor detection effect for the existing method. A deep-learning algorithm for a weak micro-defect detector named ISE-YOLO is proposed, making the best for deep layers, utilizing the ISE attention mechanism module in the neck, and introducing a novel class loss function to extract richer semantics from convolution layers and learning more information. Experimental results on the OL dataset show that ISE-YOLO demonstrates a better performance, with the mean average precision, recall, and F1 score increasing by 3.62%, 6.12% and 3.07% respectively, compared to the YOLOv5. In addition, compared with YOLOv7, which is the latest version of YOLO serials, the mean average precision of ISE-YOLO is improved by 2.58%, the weight size is decreased by more than 30% and the speed is increased by 16%.
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Chronic disease following Nocardia seriolae infection in a wide range of aquatic animals has been reported in many Asian countries and recently in America and Mexico. This study aimed to investigate the epidemiological relationship among N. seriolae isolates in Taiwan by investigating their genotype and enzymatic activities. A total of 66 strains isolated from 14 known and four unknown host fish from five sites in Taiwan were characterized using five combined methods. High genotypic diversity was recognized among the isolates with 10 pulsotypes being identified from the pulsed-field gel electrophoresis method and 21 reptypes from the repetitive extragenic palindromic amplification method; however, no natural plasmids were detected in this bacterial population. Pulsotypes A8 and RI analysed by PFGE and repPCR, respectively, were found to be predominant within five sites in Taiwan over 17 years of isolation. Enzymatically, the majority of isolates displayed high leucine arylamidase, ß-glucosidase and α-glucosidase activities but were negative for lipase, α-galactosidase, ß-glucuronidase, N-acetyl-glucosaminidase, α-mannosidase and α-fucosidase activities. We identified a strong association between genotype and enzymatic activity since the majority of pulsotypes displayed the same type of enzymatic profile. This study provides comprehensive and potential epidemiological data, which will aid the fish farming activities and prevention method development.
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Doenças dos Peixes , Nocardiose , Nocardia , Animais , Taiwan/epidemiologia , Doenças dos Peixes/epidemiologia , Doenças dos Peixes/microbiologia , Nocardia/genética , Nocardiose/epidemiologia , Nocardiose/veterinária , Nocardiose/microbiologia , Genótipo , Peixes/microbiologiaRESUMO
Large curvature aspheric optical elements are widely used in visual system. But its morphological detection is very difficult because its accuracy requirement is very high. When we use the self-developed multi-beam angle sensor (MBAS) to detect large curvature aspheric optical elements, the accuracy will be reduced due to spot distortion. Therefore, we propose a scheme combining distorted spot correction neural network (DSCNet) and gaussian fitting method to improve the detection accuracy of distorted spot center. We develop a spot discrimination method to determine spot region in multi-spot images. The spot discrimination threshold is obtained by the quantitative distribution of pixels in the connected domain. We design a DSCNet, which corrects the distorted spot to Gaussian spot, to extract the central information of distorted spot images by multiple pooling. The experimental results demonstrate that the DSCNet can effectively correct the distorted spot, and the spot center can be extracted to sub-pixel level, which improves the measurement accuracy of the MBAS. The standard deviations of plano-convex lenses with curvature radii of 500â mm, 700â mm and 1000â mm measured with the proposed method are respectively 0.0112 um, 0.0086 um and 0.0074 um.
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Lactococcus garvieae is the etiological agent of Lactococcosis, an evolving disease affecting many fish species and causing significant economic losses worldwide. Assessing pathogen relatedness and bacterial population structure is critical for determining the epidemiology of L. garvieae infections and in establishing effective pathogen management methods. The previously published morphological and genetic studies point to a clonal population structure, as seen in other fish bacteria. In the present study, the pulsed-field gel electrophoresis (PFGE) method was utilized to define a population of 41 Taiwanese isolates from outbreaks with comparisons to four well-characterized non-Taiwanese isolates previously published. Two restriction enzymes (ApaI and SmaI) were utilized individually for PFGE analysis (cut-off value = 90.0%), revealing genetic heterogeneity across L. garvieae isolates, with ApaI and SmaI yielding 12 and seven distinct PFGE band patterns, respectively. The phylogenic analysis using internal transcribed spacer region clustered all L. garvieae isolates in the same clad. Furthermore, the electron microscopic results confirmed the absence of capsular gene cluster (CGC) in previously characterized Taiwanese vaccine strain (S3) from grey mullet. Overall, our findings emphasize the importance of analysing the morphological and genetic diversity in L. garvieae being correlated for proper taxonomic classification in vaccine strain selection and epidemiological studies.
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Doenças dos Peixes , Animais , Eletroforese em Gel de Campo Pulsado/veterinária , Doenças dos Peixes/microbiologia , Peixes , Genótipo , Lactococcus/genética , Microscopia EletrônicaRESUMO
Edwardsiella spp. is a gram-negative, facultatively anaerobic, intracellular bacteria threatening the aquaculture industry worldwide. Noticeably, E. tarda is now genotypically classified into three distinct groups (E. tarda, E. piscicida and E. anguillarum), but morphologically, it is unclear due to varying degrees of virulence in different fish hosts. Hence, to reclassify E. tarda, we investigated differences in genotypes, phenotypes and pathogenicity. We collected Edwardsiella isolates from five different counties of Taiwan between 2017 and 2021. At first, gyrB gene was amplified for a phylogenetic tree from 40 isolates from different fish and one reference isolate, BCRC10670, from the human. Thirty-nine strains clustered into E. anguillarum, 1 strain into E. piscicida and 1 strain into E. tarda from human strain. Second, all isolates were characterized using various phenotypic (API 20E biochemical profiles) and genotypic (pulsed-field gel electrophoresis [PFGE], and virulence-related gene detection). SpeI digestion revealed 10 pulsotypes and I-CeuI into 7 pulsotypes. Virulent genes (citC, gadB, katB, mukF and fimA) confirmed in 35, 31, 28, 37 and 38 isolates, respectively. Finally, in vivo challenge test in milkfish (Chanos chanos) indicated the highest mortality from E. anguillarum. Overall, results revealed unique features with Edwardsiella spp. genotypes and pathogenicity, which are relevant to the host and provide useful insights for future vaccine development.
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Edwardsiella , Infecções por Enterobacteriaceae , Doenças dos Peixes , Animais , Edwardsiella/genética , Edwardsiella tarda/genética , Infecções por Enterobacteriaceae/microbiologia , Infecções por Enterobacteriaceae/veterinária , Doenças dos Peixes/microbiologia , Peixes/microbiologia , Humanos , Fenótipo , Filogenia , TaiwanRESUMO
We propose a compressive Hadamard computational ghost imaging (CGI) method to restore clear images of objects in the underwater environment. We construct an underwater CGI system model and develop a total variation regularization prior-based compressed-sensing algorithm for the CGI image reconstruction. We design a wavelet enhancement algorithm to further denoise and enhance the quality of the CGI image. We build an experimental setup and implement a series of experiments. The effectiveness and advantages of the proposed method are experimentally investigated. The results show that the proposed method can achieve clear imaging for underwater objects with a sub-Nyquist sampling ratio. The proposed method is helpful for improving the image quality of the underwater CGI.
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We designed this retrospective study with aims to investigate the incidence and risk factors associated with surgical site infection (SSI) following posterior lumbar interbody fusion (PLIF) and instrumentation in patients with lumbar degenerative disease. Eligible patients treated between January 2016 and June 2019 were included. Electronic medical records were inquired for data extraction and collection. Patients with SSI and without SSI were compared using the univariate analyses, and the association between variables and risk of SSI was investigated using multivariate logistics regression analyses. Among 1269 patients, 43 were found to have SSI, indicating a rate of 3.4%. Microbiological culture tests showed 88.4% patients had a positive result. Four SSIs were caused by mixed bacterial, and the remaining 34 by single bacteria. Multiple drug-resistant strains were detected in 25 (65.8%) SSIs, with meticillin-resistant coagulase-negative staphylococcus (MRCNS) predominating (12, 48.0%). ASA III and above (odd ratio (OR), 1.67; 95% confidence interval (CI), 1.11 to 3.07), preoperative stay (OR, 1.13; 95% CI, 1.04 to 1.23), heart disease (OR, 2.88; 95% CI, 1.24 to 6.71), diabetes mellitus (OR, 3.28; 95% CI, 1.66 to 6.47) and renal insufficiency (OR, 4.23; 95% CI, 1.26 to 10.21), prolonged prophylactic antibiotics use (OR, 4.43; 95% CI, 2.30 to 8.54), and the reduced lymphocyte count (OR, 2.11; 95% CI, 1.03 to 4.33) were identified as independent risk factors associated with SSI. These factors, although most not modifiable, should be kept in mind, optimised for surgical conditions, or readily adjusted in the future postoperative management of antibiotics, to reduce postoperative SSIs.
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Infecção da Ferida Cirúrgica , Humanos , Incidência , Modelos Logísticos , Estudos Retrospectivos , Fatores de Risco , Infecção da Ferida Cirúrgica/epidemiologia , Infecção da Ferida Cirúrgica/etiologiaRESUMO
A new method for spectroscopic interferometry using rotating diffraction grating was developed for industrial measurements. Two diffraction gratings increase the spectroscopic resolution, and the effective measuring range can be extended considerably. Instead of calibrating the wavelength, we used the Fabry-Perot Etalon (standard) to calibrate the system and determine the absolute position. The rotation diffraction gratings may also be used as a spectroscopic element over extensive ranges for low-cost and high-speed measurement. Our experiments indicate a length range of approximately 4.00 mm with repeatability of 0.17µm (0.0167%) for the narrow range and 3.84 µm (0.0955%) for the wide range.
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Spinal cannabinoid receptor 1 (CB1R) and purinergic P2X receptors (P2XR) play a critical role in the process of pathological pain. Both CB1R and P2XR are expressed in spinal dorsal horn (DH) neurons. It is not clear whether CB1 receptor activation modulates the function of P2X receptor channels within dorsal horn. For this reason, we observed the effect of CP55940 (cannabinoid receptor agonist) on ATP-induced Ca2+ mobilization in cultured rat DH neurons. The changes of intracellular calcium concentration ([Ca2+]i) were detected with confocal laser scanning microscopy using fluo-4/AM as a calcium fluorescent indicator. 100 µM ATP caused [Ca2+]i increase in cultured DH neurons. ATP-evoked [Ca2+]i increase in DH neurons was blocked by chelating extracellular Ca2+ and P2 purinoceptor antagonist PPADS. At the same time, ATP-γ-S (a non-hydrolyzable ATP analogue) mimicked the ATP action, while P2Y receptor agonist ADP failed to evoke [Ca2+]i increase in cultured DH neurons. These data suggest that ATP-induced [Ca2+]i elevation in cultured DH neurons is mediated by P2X receptor. Subsequently, we noticed that, in cultured rat DH neurons, ATP-induced Ca2+ mobilization was inhibited after pretreated with CP55940 with a concentration-dependent manner, which implies that the opening of P2X receptor channels are down-regulated by activation of cannabinoid receptor. The inhibitory effect of CP55940 on ATP-induced Ca2+ response was mimicked by ACEA (CB1R agonist), but was not influenced by AM1241 (CB2R agonist). Moreover, the inhibitory effect of CP55940 on ATP-induced Ca2+ mobilization was blocked by AM251 (CB1 receptor antagonist), but was not influenced by AM630 (CB2 receptor antagonist). In addition, we also observed that forskolin (an activator of adenylate cyclase) and 8-Br-cAMP (a cell-permeable cAMP analog) reversed the inhibitory effect of CP55940, respectively. In a summary, our observations raise a possibility that CB1R rather than CB2R can downregulate the opening of P2X receptor channels in DH neurons. The reduction of cAMP/PKA signaling is a key element in the inhibitory effect of CB1R on P2X-channel-induced Ca2+ mobilization.
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Cálcio/metabolismo , Células do Corno Posterior/metabolismo , Receptor CB1 de Canabinoide/metabolismo , Receptores Purinérgicos P2X/efeitos dos fármacos , Receptores Purinérgicos P2/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Piperidinas/farmacologia , Pirazóis/farmacologia , Ratos , Receptores Purinérgicos P2X/metabolismo , Medula Espinal/metabolismoRESUMO
Hepatitis B virus (HBV) acts as a severe public health threat, causing chronic liver diseases. Although the quantified evaluation of HBV infection can be obtained by estimating the capacity of the HBV DNA genome, it still lacks an effective and robust detection method without using enzymes or chemical labeling. Herein, we have designed a binary split fluorescent DNA aptasensor (bsFDA) by rationally splitting the lettuce aptamer into two functional DNA short chains and utilizing the HBV DNA segment complementary sequences (HDs). In this strategy, the bsFDA has been investigated to specifically recognize the HDs, forming a triplex DNA with the lettuce aptamer structure. Meanwhile, the turn-on fluorescence of bsFDA is obtained upon formation of a fluorescent complex between DFHO and the triplex DNA structure, allowing the enzyme-free, label-free, fast-responsive, and reliable fluorescence readout for detecting HDs and the potential HDs mutants. Moreover, bsFDA has been applied for spiked HDs analysis in different real matrixes, including human serum and cell lysate. The satisfactory recovery rates and reproducibility of the bsFDA reveal its potential detection efficacy for HDs analysis in biological samples. Overall, bsFDA holds great potential in developing functionalized aptasensors and realizing viral genome analysis in biological research.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , DNA Viral , Vírus da Hepatite B , Lactuca , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Vírus da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , DNA Viral/análise , Humanos , Lactuca/virologia , Lactuca/química , Corantes Fluorescentes/química , Espectrometria de Fluorescência/métodos , Limite de Detecção , Hepatite B/diagnóstico , Hepatite B/sangue , Reprodutibilidade dos TestesRESUMO
BACKGROUND: The aqueous extract of the dried buds of Syzygium aromaticum (SAAE) have the potential to alleviate Helicobacter pylori infection, but the specific molecular mechanism has not been fully elucidated. PURPOSE: This study aimed to investigate the underlying mechanisms of SAAE on H. pylori pathogenicity. METHODS: The inhibitory kinetics and anti-H. pylori adhesive capacity assays were conducted to examine the effects of SAAE on the growth and adhesive capability of H. pylori. The H. pylori outer membrane vesicles (OMVs) were purified from the culture supernatant through high-speed centrifugation, filtration, and two rounds of ultracentrifugation. Their characteristics and protein composition were then identified using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and qualitative proteomics study. Subsequently, the effect of SAAE on the pathogenicity of H. pylori OMVs was investigated using the Griess reagent assay, enzyme-linked immunosorbent assay (ELISA), quantitative proteomics study, TEM, and western blotting assay. RESULTS: SAAE exhibited inhibitory effects on H. pylori growth and adhesion. The isolated H. pylori OMVs showed particle size of 27-242 nm and Zeta potential of -9.67 ± 0.53 mV. A total of 599 proteins were identified in the OMVs. Proteomics study indicated that the differential expressed proteins induced by OMVs with or without SAAE commonly enriched in P53 and autophagy pathways. Besides, SAAE counteracted the increased production of pro-inflammatory cytokines and attenuated the induction of cell autophagy caused by H. pylori OMVs. Furthermore, SAAE normalized the abnormal regulation of downstream targets (AIFM2 and IGFBP3) in the P53 signaling pathway caused by H. pylori OMVs. CONCLUSION: SAAE can inhibit the growth and adhesion of H. pylori, reduce the inflammation and autophagy induced by H. pylori OMVs, and combated the abnormal regulation of P53 signaling pathway caused by H. pylori OMVs. These findings may help elucidate the mechanisms through which SAAE reduces the pathogenicity of H. pylori.
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Helicobacter pylori , Extratos Vegetais , Syzygium , Helicobacter pylori/efeitos dos fármacos , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Syzygium/química , Humanos , Aderência Bacteriana/efeitos dos fármacos , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Proteômica , Proteína Supressora de Tumor p53/metabolismo , Antibacterianos/farmacologia , Autofagia/efeitos dos fármacosRESUMO
OBJECTIVE: Metastases in the advanced stages of colorectal cancer (CRC) present a major challenge to its treatment. Epithelial-Mesenchymal Transition (EMT) plays a crucial role in enhancing the metastasis and invasion ability of cancer cells. However, the progress of E2F transcription factor 1 (E2F1) and Regulator of chromatin condensation 1 (RCCD1) in CRC on EMT has not been studied. METHODS: The CRC differential expression data from The Cancer Genome Atlas database were analyzed by Gene Set Enrichment Analysis to verify the difference in expression of E2F1 and RCCD1 in cancerous and para-cancerous tissues.DNA-pull down and dual luciferase experiments confirmed that E2F1 regulates RCCD1. Western-blot and q-PCR experiments confirmed that E2F1 regulates RCCD1 and participates in the EMT-related progress of CRC.EDU, Wound healing and Transwell experiments verified the effects of regulation of E2F1 and RCCD1 on the proliferation, migration and invasion of CRC cells. RESULTS: E2F1 and RCCD1 are highly expressed in cancer tissues and cancer cells. E2F1 binds to the upstream promoter of RCCD1 to regulate RCCD1 and affect the expression of EMT-related targets in CRC cells. It also affects the proliferation, migration and invasion of CRC cells. CONCLUSIONS: E2F1 regulates the involvement of RCCD1 in CRC EMT and affects the proliferation, migration and invasion ability of CRC cells.
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Neoplasias Colorretais , Fator de Transcrição E2F1 , Transição Epitelial-Mesenquimal , Regulação Neoplásica da Expressão Gênica , Humanos , Transição Epitelial-Mesenquimal/genética , Neoplasias Colorretais/patologia , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Fator de Transcrição E2F1/metabolismo , Fator de Transcrição E2F1/genética , Movimento Celular/genética , Proteínas de Ciclo Celular/metabolismo , Proteínas de Ciclo Celular/genética , Proliferação de Células/genética , Invasividade Neoplásica/genética , Progressão da Doença , Linhagem Celular TumoralRESUMO
Background: Helicobacter pylori (H. pylori) is thought to primarily colonize the human stomach and lead to various gastrointestinal disorders, such as gastritis and gastric cancer. Currently, main eradication treatment is triple or quadruple therapy centered on antibiotics. Due to antibiotic resistance, the eradication rate of H. pylori is decreasing gradually. Therefore, searching for anti-H. pylori drugs from herbal sources has become a strategy for the treatment. Our team proposed a Hezi Qingyou Formula (HZQYF), composed of Chebulae Fructus, Ficus hirta Vahl and Cloves, and studied its anti-H. pylori activity and mechanism. Methods: Chemical components of HZQYF were studied using UHPLC-MS/MS and HPLC. Broth microdilution method and agar dilution method were used to evaluate HZQYF's antibacterial activity. The effects of HZQYF on expression of adhesion genes (alpA, alpB, babA), urease genes (ureE, ureF), and flagellar genes (flaA, flaB) were explored using Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) technology. Effects on morphology and permeability of the extracellular membrane were studied using scanning electron microscopy (SEM) and N-phenylnaphthalen-1-amine (NPN) uptake. Effect on urease activity was studied using a urease kinetics analysis in vitro. Immunofluorescence staining method was used to examine the effect on adhesion. Western blot was used to examine the effect on cagA protein. Results: Minimum inhibitory concentration (MIC) values of the formula against H. pylori clinical strains and standard strains were 80-160 µg/mL, and minimum bactericidal concentration (MBC) values were 160-320 µg/mL. The formula could down-regulate the expression of adhesion genes (alpA, alpB, babA), urease genes (ureE, ureF) and flagellar genes (flaA, flaB), change the morphology of H. pylori, increase its extracellular membrane permeability, and decrease its urease activity. Conclusion: Present studies confirmed that HZQYF had promising in vitro anti-H. pylori activities and demonstrated its possible mechanism of action by down-regulating the bacterial adhesion, urease, and flagellar gene expression, which provided scientific bases for further clinical investigations.
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ETHNOPHARMACOLOGICAL RELEVANCE: Phellodendron chinense C.K.Schneid(P. chinense Schneid) is known in TCM as Huang Bo, is traditionally used to support gastrointestinal function and alleviate stomach-related ailments, including gastric ulcer bleeding and symptoms of gastroesophageal reflux disease. Helicobacter pylori (H. pylori) is classified by the WHO as a Group 1 carcinogen. However, the specific activity and mechanism of action of P. chinense Schneid against H. pylori infection remain unclear. It has been noted that Huangjiu processing may alter the bitter and cold properties of P. chinense Schneid, but its effect on antimicrobial activity requires further investigation. Additionally, it remains uncertain whether berberine is the sole antimicrobial active component of P. chinense Schneid. AIM OF STUDY: This study aims to elucidate the anti-H. pylori infection activity of P. chinense Schneid, along with its mechanism of action and key antimicrobial active components. MATERIALS AND METHODS: Phytochemical analysis was carried out by UPLC-MS/MS. HPLC was employed to quantify the berberine content of the extracts. Antimicrobial activity was assessed using the micro broth dilution method. Morphology was observed using SEM. The impact on urease activity was analyzed through in vitro urease enzyme kinetics. RT-qPCR was employed to detect the expression of virulence genes, including adhesin, flagellum, urease, and cytotoxin-related genes. The adhesion effect was evaluated by immunofluorescence staining and agar culture. RESULTS: P. chinense Schneid exhibited strong antimicrobial activity against both antibiotic-sensitive and resistant H. pylori strains, with MIC ranging from 40 to 160 µg/mL. Combination with amoxicillin, metronidazole, levofloxacin, and clarithromycin did not result in antagonistic effects. P. chinense Schneid induced alterations in bacterial morphology and structure, downregulated the expression of various virulence genes, and inhibited urease enzyme activity. In co-infection systems, P. chinense Schneid significantly attenuated H. pylori adhesion and urease relative content, thereby mitigating cellular damage caused by infection. Huangjiu processing enhanced the anti-H. pylori activity of P. chinense Schneid. Besides berberine, P. chinense Schneid contained seven other components with anti-H. pylori activity, with palmatine exhibiting the strongest activity, followed by jatrorrhizine. CONCLUSIONS: This study sheds light on the potential therapeutic mechanisms of P. chinense Schneid against H. pylori infection, demonstrating its capacity to disrupt bacterial structure, inhibit urease activity, suppress virulence gene transcription, inhibit adhesion, and protect host cells. The anti-H. pylori activity of P. chinense Schneid was potentiated by Huangjiu processing, and additional components beyond berberine were identified as possessing strong anti-H. pylori activity. Notably, jatrorrhizine, a core component of P. chinense Schneid, exhibited significant anti-H. pylori activity, marking a groundbreaking discovery.
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Antibacterianos , Berberina , Helicobacter pylori , Phellodendron , Extratos Vegetais , Helicobacter pylori/efeitos dos fármacos , Phellodendron/química , Antibacterianos/farmacologia , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Berberina/farmacologia , Testes de Sensibilidade Microbiana , Urease/metabolismo , Humanos , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologiaRESUMO
Heavy metal pollution poses a great threat to the ecological environment and human health. In particular, copper ions (Cu2+) play a vital role in regulating fundamental life behavior, and the homeostasis of Cu2+ is closely related to many physiological processes. The excessive accumulation of Cu2+ in the human body through food and drinking water will cause severe diseases. However, current conventional Cu2+ detection methods for evaluating the content of Cu2+ are unable to meet the complete requirements of practical Cu2+ analysis in the practical aquatic environment. In this work, we successfully constructed a novel fluorescent DNA aptasensor, which originated from the binding reaction between the improved DNA fluorescent light-up aptamer termed S2T3AT-GC and a small fluorescent molecule termed DFHBI-1T (S2T3AT-GC/DFHBI-1T) to realize fast and anti-interference response for Cu2+via the competitive interaction between Cu2+ and S2T3AT-GC (Cu2+/S2T3AT-GC) destroying the contained G-quadruplex structure of S2T3AT-GC. Moreover, it enables the sensitive detection of Cu2+ with a detection limit of 0.3 µM and a wide detection linear range from 0.3 to 300 µM. Moreover, with the verification of high stability in real industrial sewage samples, this aptasensor exhibits excellent detection performance for Cu2+ analysis in real water samples. Therefore, the proposed aptasensor exhibits great potential in exploring Cu2+-related environmental and ecological research.
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Cobre , Esgotos , Humanos , Cobre/análise , Cobre/química , DNA , Íons , Corantes Fluorescentes/químicaRESUMO
To automatically measure the surface profile of a cylindrical workpiece, a high-precision multi-beam optical method is proposed in this paper. First, some successive images for the cylindrical workpiece's surface are acquired by a multi-beam angle sensor under different light directions. Then, the light directions are estimated based on the feature regions in the images to calculate surface normal vectors. Finally, according to the relationship of the surface normal vector and the vertical section of the workpiece's surface, a depth map is reconstructed to achieve the curvature surface, which can be employed to measure the curvature radius of the cylindrical workpiece's surface. Experimental results indicate that the proposed measurement method can achieve good measurement precision with a mean error of the curvature radius of a workpiece's surface of 0.89% at a reasonable speed of 10.226 s, which is superior to some existing methods.