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The Houttuynia cordata Thunb. belongs to the Saururaceae family and is a well-known medicine and food homologous plant. Herein, the isolation of an α-glucosidase inhibitor from Houttuynia cordata Thunb. and characterization of its in vitro and in vivo hypoglycemic bioactivities are reported. We optimized the extraction conditions and isolated neochlorogenic acid (nCGA), which has α-glucosidase inhibitory activity from Houttuynia cordata Thunb. for the first time. nCGA competed with glucose for the α-glucosidase binding site, with a 50% inhibitory concentration (IC50) of 0.711 mg/mL. In vivo experiments in zebrafish showed that effects of nCGA on blood glucose varied by its concentrations. In particular, 4 mg/L nCGA significantly decreased the blood glucose level and inhibited effects of α-glucosidase in zebrafish. This work provides a theoretical basis for the extraction of hypoglycemic active ingredients from Houttuynia cordata Thunb. and a foundation for the development of natural and effective α-glucosidase inhibitors.
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BACKGROUND: Prior studies have suggested that the chronic inflammatory response has an important role in the pathophysiology of slow coronary flow phenomenon (SCFP). However, data are scarce regarding the role of plasma fibrinogen-to-albumin ratio (PFAR) in patients having SCFP without obstructive coronary artery disease (CAD). In this study, we investigated the relationship between PFAR and the presence of SCFP in patients without obstructive CAD. METHODS: From January 2021 to January 2023, we consecutively recruited 1085 patients without obstructive CAD according to the diagnostic and exclusion criteria. In total, SCFP was diagnosed in 70 patients. A 1:2 age-matched case-control study was then conducted using comparators without SCFP. Ultimately, this study enrolled 70 patients with angiographically normal coronary arteries and SCFP, along with 140 comparators with angiographically normal coronary arteries and normal coronary flow. Plasma fibrinogen and albumin levels were measured, and the PFAR was then calculated for each patient. RESULTS: PFARs were significantly greater in the SCFP group than in the comparators with normal coronary flow (82.8 ± 15.4 vs 73.1 ± 19.5, p < 0.001). PFAR increased with increasing numbers of vessels affected by SCFP. Multivariate logistic regression analysis showed that PFAR was an independent predictor of SCFP (odds ratio: 1.818, p = 0.015). Receiver operating characteristic (ROC) curve analysis indicated that PFAR showed a better predictive value of SCFP than fibrinogen or albumin, although not significantly (p > 0.05). CONCLUSION: PFAR is an independent predictor of SCFP in patients without obstructive CAD. PAFR could improve the predictive value of SFCP than albumin or fibrinogen alone, but not significantly.
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Doença da Artéria Coronariana , Humanos , Estudos de Casos e Controles , Curva ROC , Albuminas , Fibrinogênio , Angiografia CoronáriaRESUMO
The safety valve is the core component of the pressure-relief protection device for pressure-bearing special equipment. When the safety valve leaks, the medium of the pressure vessel will be lost and wasted, which may cause safety accidents. With the aim to solve the problem of accurately locating the multiple leakage sources of safety valves, a localization method combining a uniform circular array acoustic emission detection and an improved multiple signal classification (MUSIC) algorithm is proposed. First, an improved wavelet threshold function denoising method is introduced to extract acoustic emission signals with high SNR, thereby reducing the rank of the covariance matrix, weakening the noise dispersion caused by eigenvalue reconstruction, avoiding signal and noise cross-confusion, and improving positioning accuracy. By introducing a windowed fast Fourier transform (FFT) frequency division processing link to obtain narrowband signal, the premise of using MUSIC positioning algorithm is established. In addition, a forward/backward spatial smoothing algorithm is introduced in the decoherence link to reduce co-channel interference, reduce the rank loss of the signal covariance matrix, and improve the positioning accuracy of the algorithm. The results show that when the working pressure is 0.70 MPa, 0.75 MPa, and 0.80 MPa, the deviation between the azimuth angle and elevation angle positioning results of each leakage source obtained by the improved MUSIC algorithm and the actual angle does not exceed 2°, and the relative error does not exceed 3.5%. Therefore, the improved MUSIC algorithm can accurately locate multiple leakage sources of the safety valve, and as the working pressure of the safety valve increases, the positioning accuracy of the improved MUSIC algorithm also increases accordingly.
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The measured signals of internal leakage detection of the large-diameter pipeline ball valve in natural gas pipeline systems usually contain background noise, which will affect the accuracy of internal leakage detection and sound localization of internal leakage points due to the interference of noise. Aiming at this problem, this paper proposes an NWTD-WP feature extraction algorithm by combining the wavelet packet (WP) algorithm and the improved two-parameter threshold quantization function. The results show that the WP algorithm has a good feature extraction effect on the valve leakage signal, and the improved threshold quantization function can avoid the defects of the traditional soft threshold function and hard threshold function, such as discontinuity and the pseudo-Gibbs phenomenon, when reconstructing the signal. The NWTD-WP algorithm is effective in extracting the features of the measured signals with low signal/noise ratio. The denoise effect is much better than that of the traditional soft and hard threshold quantization functions. It proved that the NWTD-WP algorithm can be used for studying the existing safety valve leakage vibration signals in the laboratory and the internal leakage signals of the scaled-down model of the large-diameter pipeline's ball valve.
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Food safety has long been an area of concern. The selection of stable and efficient model organisms is particularly important for food toxicology studies. Zebrafish (Danio rerio) are small model vertebrates, and 70% of human genes have at least one zebrafish ortholog. Zebrafish have advantages as model organisms due to their short life cycle, strong reproductive ability, easy rearing, and low cost. Zebrafish embryos have the advantage of being sensitive to the breeding environment and thus have been used as biosensors. Zebrafish and their embryos have been widely used for food toxicology assessments. This review provides a systematic and comprehensive summary of food toxicology studies using zebrafish as model organisms. First, we briefly introduce the multidimensional mechanisms and structure-activity relationship studies of food toxicological assessment. Second, we categorize these studies according to eight types of hazards in foods, including mycotoxins, pesticides, antibiotics, heavy metals, endocrine disruptors, food additives, nanoparticles, and other food-related ingredients. Finally, we list the applications of zebrafish in food toxicology studies in line with future research prospects, aiming to provide a valuable reference for researchers in the field of food science.
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Alimentos , Peixe-Zebra , Animais , Humanos , Peixe-Zebra/genética , Tecnologia de AlimentosRESUMO
Dopamine is an essential neurotransmitter and its detection is important for bioanalytical chemistry. Two very different DNA aptamers have been reported for dopamine, one derived from an RNA aptamer (named Apt1) and other obtained via direct aptamer selection (named Apt2). In this study, we used four homogeneous binding assays to compare these two DNA dopamine aptamers. Thiazole orange (TO) fluorescence assay indicated that the Apt2 specifically bound with dopamine with a Kd of 2.37â µM, which was consistent with that from the isothermal titration calorimetry (ITC) assay. However, Apt1 had much less TO fluorescence change and also no signal from ITC. By labeling the two ends of the two aptamers by a fluorophore and a quencher, the aptamer beacons showed binding of dopamine only for Apt2. Finally, the label-free AuNP-based colorimetric assay showed no difference between these two aptamer sequences, and even non-binding random DNA showed the same response, indicating that AuNPs were not a good probe for detecting dopamine. According to the data, Apt1 does not appear to be able to bind dopamine specifically, while Apt2 showed specific binding and could be used for developing related biosensors.
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Aptâmeros de Nucleotídeos/química , Dopamina/análise , Benzotiazóis/química , Calorimetria , Corantes Fluorescentes/química , Quinolinas/químicaRESUMO
Ubiquitin-specific protease 31 (USP31) is a member of deubiquitinase family that is involved in nuclear factor-κB activation and sarcomagenesis. However, little is known about posttranslational modification in the regulation of its activity and cervical cancer cell growth. In our study, we found that the Lys1264 residue of USP31 can be modified with an acetyl group by high-resolution mass spectrometry in HeLa cell line, and site-specific mutagenesis can significantly increase USP31 ubiquitin hydrolase activity and decrease the expression of p65. When being transfected with a plasmid expressing mutated USP31, the number of cancer cells was significantly decreased. We also observed that mutated USP31 could promote apoptosis but not cell cycle by flow cytometer analysis. Overexpression of mutated USP31 could reverse the effect in USP31 knockdown cell line. To further investigate its activity in tumorigenesis, deacetylase sirtuin 1 (Sirt1) was shown to interact with USP31 by co-immunoprecipitation and blocking the function of Sirt1 by knockdown or the inhibitor nicotinamide could increase the acetylation of USP31. When Lys1264 of USP31 mutated, Sirt1 could not remove its acetylation and alter the expression level of p65. Finally, inhibition or knockdown of Sirt1 suppressed USP31 activity in HeLa cell line, leading to cisplatin-induced apoptosis resistance. Therefore, acetylation at Lys1264 suppresses USP31 activity and plays a protective role in cancer cell growth. Our study contributes to understanding the mechanism of USP31 activity regulation and its role in tumorigenesis.
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Proteínas de Neoplasias/metabolismo , Proteases Específicas de Ubiquitina/metabolismo , Neoplasias do Colo do Útero/enzimologia , Acetilação , Feminino , Células HeLa , Humanos , Lisina , Proteínas de Neoplasias/genética , Proteases Específicas de Ubiquitina/genética , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologiaRESUMO
One of the main characteristics of atherosclerosis is vascular calcification, which is linked to adverse cardiovascular events. Increased homocysteine (Hcy), a feature of hyperhomocysteinemia, is correlated with advanced vascular calcification and phenotypic switching of vascular smooth muscle cells (VSMCs). Oxidative stress and high phosphate levels also induce VSMC calcification, suggesting that the Krüppel-like factor 4 (KLF4) signaling pathway may also contribute to vascular calcification. In this study, we investigated this possibility and the role and mechanisms of Hcy in vascular calcification. We found that in atherosclerotic apolipoprotein E-deficient (ApoE-/-) mice, Hcy significantly increases vascular calcification in vivo, as well as VSMC calcification in vitro Of note, the Hcy-induced VSMC calcification was correlated with elevated KLF4 levels. Hcy promoted KLF4 expression in calcified atherosclerotic lesions in vivo and in calcified VSMCs in vitro shRNA-mediated KLF4 knockdown blocked the Hcy-induced up-regulation of runt-related transcription factor 2 (RUNX2) and VSMC calcification. RUNX2 inhibition abolished Hcy-induced VSMC calcification. Using ChIP analysis, we demonstrate that KLF4 interacts with RUNX2, an interaction promoted by Hcy stimulation. Our experiments also revealed that the KLF4 knockdown attenuates Hcy-induced RUNX2 transactivity, indicating that KLF4 is important in modulating RUNX2 transactivity. These findings support a role for Hcy in regulating vascular calcification through a KLF4-RUNX2 interaction and indicate that Hcy-induced, enhanced RUNX2 transactivity increases VSMC calcification. These insights reveal possible opportunities for developing interventions that prevent or manage vascular calcification.
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Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Hiper-Homocisteinemia/metabolismo , Fatores de Transcrição Kruppel-Like/metabolismo , Miócitos de Músculo Liso/citologia , Calcificação Vascular/metabolismo , Animais , Aterosclerose/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Feminino , Homocisteína/farmacologia , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout para ApoE , Músculo Liso Vascular/citologia , Estresse Oxidativo , Fosfatos/sangue , RNA Interferente Pequeno/metabolismo , Calcificação Vascular/genéticaRESUMO
A selection of aptamers specific for di(2-ethylhexyl) phthalate (DEHP) and development of electrochemical impedance spectroscopy (EIS) aptasensor are described in this paper. The aptamers were selected from an immobilized ssDNA library using the systematic evolution of ligands by exponential enrichment (SELEX). The enrichment was monitored using real-time quantitative PCR (Q-PCR), and the aptamers were identified by high-throughput sequencing (HTS), gold nanoparticles (AuNPs) colorimetric assay, and localized surface plasmon resonance (LSPR). The EIS aptasensor was developed to detect DEHP in water samples. After eight rounds of enrichment, HTS, AuNPs colorimetric assay, and LSPR analysis indicated that four aptamers had higher binding activity, and aptamer 31 had the highest affinity (Kd = 2.26 ± 0.06 nM). The EIS aptasensor had a limit of detection (LOD) of 0.103 pg/mL with no cross-reactivity to DEHP analogs and a mean recovery of 76.07% to 141.32% for detection of DEHP in water samples. This aptamer is novel with the highest affinity and sensitivity.
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Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais , DNA de Cadeia Simples/química , Espectroscopia Dielétrica/métodos , Dietilexilftalato/análise , Poluentes Químicos da Água/análise , Colorimetria/métodos , Água Potável/química , Biblioteca Gênica , Ouro/química , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Limite de Detecção , Nanopartículas Metálicas/química , Reação em Cadeia da Polimerase em Tempo Real , Técnica de Seleção de Aptâmeros , Ressonância de Plasmônio de SuperfícieRESUMO
We describe a multiple combined strategy to discover novel aptamers specific for clenbuterol (CBL). An immobilized ssDNA library was used for the selection of specific aptamers using the systematic evolution of ligands by exponential enrichment (SELEX). Progress was monitored using real-time quantitative PCR (Q-PCR), and the enriched library was sequenced by high-throughput sequencing. Candidate aptamers were picked and preliminarily identified using a gold nanoparticles (AuNPs) biosensor. Bioactive aptamers were characterized for affinity, circular dichroism (CD), specificity and sensitivity. The Q-PCR amplification curve increased and the retention rate was about 1% at the eighth round. Use of the AuNPs biosensor and CD analyses determined that six aptamers had binding activity. Affinity analysis showed that aptamer 47 had the highest affinity (Kd = 42.17 ± 8.98 nM) with no cross reactivity to CBL analogs. Indirect competitive enzyme linked aptamer assay (IC-ELAA) based on a 5'-biotin aptamer 47 indicated the limit of detection (LOD) was 0.18 ± 0.02 ng/L (n = 3), and it was used to detect pork samples with a mean recovery of 83.33â»97.03%. This is the first report of a universal strategy including library fixation, Q-PCR monitoring, high-throughput sequencing, and AuNPs biosensor identification to select aptamers specific for small molecules.
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Agonistas Adrenérgicos beta/análise , Aptâmeros de Nucleotídeos/isolamento & purificação , Clembuterol/análise , DNA de Cadeia Simples/metabolismo , Ouro/química , Aptâmeros de Nucleotídeos/química , Aptâmeros de Nucleotídeos/metabolismo , Técnicas Biossensoriais , Dicroísmo Circular , DNA de Cadeia Simples/química , Biblioteca Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Limite de Detecção , Nanopartículas Metálicas/química , Técnica de Seleção de Aptâmeros , Análise de Sequência de DNARESUMO
BACKGROUND: Napping is garnering increased attention as a strategy for adults to sustain alertness and alleviate stress in contemporary society. The nuances of napping habits are emerging as an independent factor influencing the extent of individual benefits. This study aimed to demonstrate the long-term benefits of napping and explore the impact of napping habits on individual alertness, as well as whether this effect was correlated with cortisol levels. METHODS: The study involved 80 healthy adults categorized into two groups based on self-reported napping habits: habitual nappers (n = 49) and non-habitual nappers (n = 31). Karolinska Sleepiness Scale (KSS), psychomotor vigilance task (PVT), and saliva collection were performed every 30 min within 90 min in the absence of napping during the afternoon dip. The measurements were analyzed using repeated measures ANOVA and Pearson correlation analyses. RESULTS: There was an interaction between groups and time in reaction speed and lapse number of PVT and cortisol (all p < 0.05). Post hoc analysis found that habitual nappers maintained higher objective alertness and experienced more significant increases in cortisol over time (all p < 0.05). The cortisol levels at sleepiness time were negatively associated with the slowest 10 % reaction speed of PVT in non-habitual nappers (r = -0.409, p = 0.022). CONCLUSION: Under the premise of mitigating the impacts of acute nap deprivation on sleep homeostasis and rhythm, napping habits emerge as a potential factor influencing the ability of individuals to sustain heightened alertness.
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Hábitos , Hidrocortisona , Desempenho Psicomotor , Saliva , Sono , Humanos , Hidrocortisona/análise , Hidrocortisona/metabolismo , Masculino , Feminino , Sono/fisiologia , Saliva/química , Adulto , Desempenho Psicomotor/fisiologia , Tempo de Reação/fisiologia , Adulto Jovem , Atenção/fisiologia , Vigília/fisiologia , Fatores de Tempo , AutorrelatoRESUMO
Microcystin-LR (MCLR) poses a significant threat to aquatic ecosystems and public health. This study investigated the protective effects of the probiotic Lactobacillus rhamnosus against MCLR-induced developmental toxicity in zebrafish larvae. Zebrafish larvae were exposed to various concentrations of MCLR (0, 0.9, 1.8, and 3.6 mg/L) with or without L. rhamnosus from 72 to 168 h post-fertilization (hpf). Probiotic supplementation significantly improved survival, hatching, and growth rates and reduced malformation rates in MCLR-exposed larvae. L. rhamnosus alleviated MCLR-induced oxidative stress by reducing reactive oxygen species (ROS) levels and enhancing glutathione (GSH) content and catalase (CAT) activity. Probiotics also mitigated MCLR-induced lipid metabolism disorders by regulating key metabolites (triglycerides, cholesterol, bile acids, and free fatty acids) and gene expression (ppara, pparb, srebp1, and nr1h4). Moreover, 16S rRNA sequencing revealed that L. rhamnosus modulated the gut microbiome structure and diversity in MCLR-exposed larvae, promoting beneficial genera like Shewanella and Enterobacter and inhibiting potential pathogens like Vibrio. Significant correlations were found between gut microbiota composition and host antioxidant and lipid metabolism parameters. These findings suggest that L. rhamnosus exerts protective effects against MCLR toxicity in zebrafish larvae by alleviating oxidative stress, regulating lipid metabolism, and modulating the gut microbiome, providing insights into probiotic-based strategies for mitigating MCLR toxicity in aquatic organisms.
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To measure toxins using immunoassays, hazardous toxin standards need to be added for quantification. To solve this problem, we propose to use aptamers as competitors to replace toxin standards. In this work, aptamers specific for ochratoxin A (OTA) nanobodies were selected using a DNA library containing a 36 nucleotide random region. The obtained sequences were highly aligned and the best competitor was identified to be a sequence named apt2-OT based on an indirect competitive enzyme-linked immunosorbent assay (ELISA). The Kd of apt2-OT was measured to be 2.86 µM using local surface plasmon resonance spectroscopy. The optimal apt2-OT was identified to substitute the OTA standard with a concentration needed for 50% inhibition of binding (IC50) of 3.26 µM based on a nontoxic direct competitive ELISA. The equivalence relationship between the aptamer and OTA was established in a flour sample, and a recovery experiment was performed. The detection limit for this method was 0.23 ng/mL, with a linear range from 0.25 to 10.50 ng/mL. The recovery rate was 97.5%-115.5%. This study provides a low-cost, rapid and environmentally friendly alternative to the development of immunoassays for toxins.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Ocratoxinas , Anticorpos de Domínio Único , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Ocratoxinas/toxicidade , Ocratoxinas/análise , Imunoensaio , Limite de DetecçãoRESUMO
Two novel series of RGD-MEKI conjugates derived from a MEK1/2 kinase inhibitor--PD0325901--have been developed for integrin receptor mediated anticancer therapy. The first series, alkoxylamine analog RGD-MEKI conjugates 9a-g showed anti-proliferation activity in melanoma A375 cells by the same mechanism as that of PD0325901. PEGylation increased the IC50 value of 9f three-fold in the A375 assay, and the multi-cRGD peptide cargo significantly improved the receptor specific anti-proliferation activity of 9g in integrin-overexpressing U87 cells. In the second series, RGD-PD0325901 13 exhibited significantly increased antitumor properties compared to the alkoxylamine analogs by both inhibition of the ERK pathway activity and DNA replication of the cancer cells. Furthermore, 13 displayed more potent anti-proliferation activity in the U87 assay than PD0325901 in a dose-dependent manner. All these data demonstrate that RGD-MEKI conjugates with an ester bond linkage enhanced anticancer efficacy with improved targeting capability toward integrin-overexpressing tumor cells.
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Integrinas/metabolismo , MAP Quinase Quinase 1/antagonistas & inibidores , MAP Quinase Quinase 2/antagonistas & inibidores , Inibidores de Proteínas Quinases/farmacologia , Benzamidas/farmacologia , Western Blotting , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Difenilamina/análogos & derivados , Difenilamina/farmacologia , Citometria de Fluxo , Humanos , Integrina alfaVbeta3/metabolismo , OligopeptídeosRESUMO
OBJECTIVE: To analyze the articles and citation published in the Chinese Journal of Internal Medicine from 2008 to 2010, in order to investigate the influence factors of impact factor (IF). METHODS: All articles published in the Chinese Journal of Internal Medicine covered by Chinese Medical Citation Index(CMCI) from 2008 to 2010 were searched and downloaded. Some article related attributions were manual added and their influences to IF were analyzed. RESULTS: A total of 1164 academic papers were published in the journal in 3 years, with 9.95 references per paper. The total citation was 1029 times, with 0.93 time per paper and 0.31 time per page. Among them, 736 articles were not cited, accounting for 63.2%. Original articles, pure clinical articles had good citation output. For fund supporting, the citation of articles without fund was higher than those with fund. The articles on hematology, cardiology and gastroenterology accounted the most, while ICU, emergency and nephrology were the least internal medicine reported specialties. Although there exists citation difference among different subjects, for considering the absolute values, neurology/psychiatry (0.73 time per paper), cardiology (0.65 time per paper) and gastroenterology (0. 54 time per paper) had better citation output, while hematology, basic research and rheumatology had no good performance to IF. CONCLUSION: We should further strengthen acquisition and dissemination of excellent articles, reduce the number of non-cited paper, expand periodical visibility, and provide a quick and convenient way of literature reading.
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Bibliometria , Medicina Interna , Fator de Impacto de Revistas , Publicações Periódicas como Assunto , ChinaRESUMO
OBJECTIVE: To evaluate the academic level of Chinese Journal of Internal Medicine by analyzing its citation status by using bibliometrics method. METHODS: The distribution of articles published in Chinese Journal of Internal Medicine from January, 2005 to December, 2011 indexed by Chinese Science Citation Database (CSCD) was analyzed. RESULTS: A total of 2809 articles were published in Chinese Journal of Internal Medicine between January,2005 and December, 2011. Among them, 832(29.62%) articles were cited for totally 1993 times. There were 14 authors whose total citation number in 2005-2011 was > or = 10 times. Authors of the cited articles came from 25 provinces, autonomous regions and municipalities. The regions from where the articles had the highest citation were Beijing (341 articles), Shanghai (87 articles), Guangdong (64 articles), Jiangsu (45 articles) and Zhejiang (43 articles). The medical institutions with the highest citation were Peking Union Medical College Hospital of Peking Union Medical College and Chinese Academy of Medical Sciences (205 times), Peking University People's Hospital (77 times), and Chinese PLA General Hospital (76 times). CONCLUSIONS: Quite a few high level academic papers had been published in Chinese Journal of Internal Medicine in recent years. Chinese Journal of Internal Medicine plays a good role in enhancing academic exchange.
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Bibliometria , Medicina Interna , Publicações Periódicas como Assunto , ChinaRESUMO
FUS plays a significant role as an RNA-binding protein in several cellular processes, including RNA splicing, DNA repair, and transcriptional regulation. However, the RNA-binding capacity of FUS in atherosclerosis is unclear. We aimed to study the functions of FUS in inflammatory regulation through the role of the splicing factor. We knocked down FUS with siRNA to further study the overall transcriptional level and select alternative splicing (AS) of FUS regulation in human umbilical vein endothelial cells (HUVECs) by RNA sequencing. The results suggested that the knockdown of FUS significantly affected gene expression in HUVECs. In addition, the knockdown of FUS resulted in 200 differentially expressed genes (DEGs) that were highly related to apoptotic process, signal transduction, multicellular organism development, cell adhesion and regulation of transcription, and DNA-templated pathways. Importantly, FUS extensively regulated 2870 AS events with a significant difference. Functional analysis of its modulated AS genes revealed they were highly enriched in cell cycle and cell population proliferation pathways. The qRT-PCR and RNA-seq data showed consistent results. Our findings suggested new knowledge of the mechanisms of FUS associated with atherosclerosis.
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Processamento Alternativo , Aterosclerose , Humanos , Processamento Alternativo/genética , Células Endoteliais/metabolismo , RNA Interferente Pequeno/genética , Proliferação de Células/genética , Aterosclerose/genética , Proteína FUS de Ligação a RNA/genética , Proteína FUS de Ligação a RNA/metabolismoRESUMO
A capture systematic evolution of ligands by exponential enrichment (Capture-SELEX) was described to discover novel aptamers specific for 5-hydroxymethylfurfural (5-HMF), and a biosensor based on molecular beacon was constructed to detect 5-HMF. The ssDNA library was immobilized to streptavidin (SA) resin to select the specific aptamer. The selection progress was monitored using real-time quantitative PCR (Q-PCR), and the enriched library was sequenced by high-throughput sequencing (HTS). Candidate and mutant aptamers were selected and identified by Isothermal Titration Calorimetry (ITC). The FAM-aptamer and BHQ1-cDNA were designed as the quenching biosensor to detect 5-HMF in milk matrix. After the 18th round selection, the Ct value decreased from 9.09 to 8.79, indicating that the library was enriched. The HTS results indicated that the total sequence numbers for 9th, 13th, 16th, and 18th were 417054, 407987, 307666, and 259867, but the number of sequences for the top 300 sequences was gradually increased from 9th to 18th, and the ClustalX2 analysis showed that there were four families with high homology rate. ITC results indicated that the Kd values of H1 and its mutants H1-8, H1-12, H1-14, and H1-21 were 2.5 µM, 1.8 µM, 1.2 µM, 6.5 µM, and 4.7 µM. The linear range of the quenching biosensor was from 0 µM to 75 µM, and it had a similar linear range in the 0.1% milk matrix. This is the first report to select a novel aptamer specific for 5-HMF and develop quenching biosensor for the rapid detection of 5-HMF in milk matrix.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Humanos , Aptâmeros de Nucleotídeos/química , DNA de Cadeia Simples , FuraldeídoRESUMO
The aim of this study was to explore the underlying mechanism of adverse effects caused by tebuconazole (TEB) on the reproduction of aquatic organisms In the present study, in order to explore the effects of TEB on reproduction, four-month-old zebrafish were exposed to TEB (0, DMSO, 0.4 mg/L, 0.8 mg/L, and 1.6 mg/L) for 21 days. After exposure, the accumulations of TEB in gonads were observed and the cumulative egg production was evidently decreased. The decline of fertilization rate in F1 embryos was also observed. Then the changes in sperm motility and histomorphology of gonads were discovered, evaluating that TEB had adverse effects on gonadal development. Additionally, we also found the alternations of social behavior, 17ß-estradiol (E2) level, and testosterone (T) level. Furthermore, the expression levels of genes involved in the hypothalamic-pituitary-gonadal (HPG) axis and social behavior were remarkably altered. Taken together, it could be concluded that TEB affected the egg production and fertilization rate by interfering with gonadal development, sex hormone secretion, and social behavior, which were eventually attributed to the disruption of the expressions of genes associated with the HPG axis and social behavior. This study provides a new perspective to understanding the mechanism of TEB-induced reproductive toxicity.
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Disruptores Endócrinos , Poluentes Químicos da Água , Animais , Masculino , Peixe-Zebra/metabolismo , Motilidade dos Espermatozoides , Gônadas/metabolismo , Reprodução , Comportamento Social , Poluentes Químicos da Água/toxicidade , Disruptores Endócrinos/toxicidadeRESUMO
In the title mol-ecule, C14H26Br2O, the central carbonyl group (C3O) is essentially planar (r.m.s. deviation = 0.0021â Å). The Br atoms lie on the same side of the mol-ecule and are approximately syn, with a Br-Câ¯C-Br torsion angle of -43.52â (13)°. The crystal structure is devoid of any specific inter-molecular inter-actions.