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1.
Org Biomol Chem ; 21(40): 8170-8175, 2023 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-37782212

RESUMO

A new strategy for the preparation of 3-halobenzo[b]thiophenes via a photo-driven halocyclization/demethylation of 2-alkynylthioanisoles with simple alkyl halides was developed. The reaction can proceed smoothly at room temperature under visible-light irradiation without any external photocatalyst, and the protocol has a range of advantages, including simplicity and mildness of the reaction conditions, good functional-group tolerance, and excellent yields of the products.

2.
Phytother Res ; 37(5): 1864-1882, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36740450

RESUMO

Shenlian (SL) extract has been proven to be effective in the prevention and treatment of atherosclerosis and myocardial ischemia. However, the function and molecular mechanisms of SL on coronary artery no-reflow have not been fully elucidated. This study was designed to investigate the contribution of SL extract in repressing excessive mitochondrial autophagy to protect the mitochondrial function and prevent coronary artery no-reflow. The improvement of SL on coronary artery no-reflow was observed in vivo experiments and the molecular mechanisms were further explored through vitro experiments. First, a coronary artery no-reflow rat model was built by ligating the left anterior descending coronary artery for 2 hr of ischemia, followed by 24 hr of reperfusion. Thioflavin S (6%, 1 ml/kg) was injected into the inferior vena cava to mark the no-reflow area. Transmission electron microscopy was performed to observe the cellular structure, mitochondrial structure, and mitochondrial autophagy of the endothelial cells. Immunofluorescence was used to observe the microvascular barrier function and microvascular inflammation. Cardiac microvascular endothelial cells (CMECs) were isolated from rats. The CMECs were deprived of oxygen-glucose deprivation (OGD) for 2 hr and reoxygenated for 4 hr to mimic the Myocardial ischemia-reperfusion (MI/R) injury-induced coronary artery no-reflow in vitro. Mitochondrial membrane potential was assessed using JC-1 dye. Intracellular adenosine triphosphate (ATP) levels were determined using an ATP assay kit. The cell total reactive oxygen species (ROS) levels and cell apoptosis rate were analyzed by flow cytometry. Colocalization of mitochondria and lysosomes indirectly indicated mitophagy. The representative ultrastructural morphologies of the autophagosomes and autolysosomes were also observed under transmission electron microscopy. The mitochondrial autophagy-related proteins (LC3II/I, P62, PINK, and Parkin) were analyzed using Western blot analysis. In vivo, results showed that, compared with the model group, SL could reduce the no-reflow area from 37.04 ± 9.67% to 18.31 ± 4.01% (1.08 g·kg-1 SL), 13.79 ± 4.77% (2.16 g·kg-1 SL), and 12.67 ± 2.47% (4.32 g·kg-1 SL). The extract also significantly increased the left ventricular ejection fraction (EF) and left ventricular fractional shortening (FS) (p < 0.05 or p < 0.01). The fluorescence intensities of VE-cadherin, which is a junctional protein that preserves the microvascular barrier function, decreased to ~74.05% of the baseline levels in the no-reflow rats and increased to 89.87%(1.08 g·kg-1 SL), 82.23% (2.16 g·kg-1 SL), and 89.69% (4.32 g·kg-1 SL) of the baseline levels by SL treatment. SL administration repressed the neutrophil migration into the myocardium. The oxygen-glucose deprivation/reoxygenation (OGD/R) model was induced in vitro to mimic microvascular ischemia-reperfusion injury. The impaired mitochondrial function after OGD/R injury led to decreased ATP production, calcium overload, the excessive opening of the Mitochondrial Permeability Transition Pore, decreased mitochondrial membrane potential, and reduced ROS scavenging ability (p < 0.05 or p < 0.01). The normal autophagosomes (double-membrane vacuoles with autophagic content) in the sham group were rarely found. The large morphology and autophagosomes were frequently observed in the model group. By contrast, SL inhibited the excessive activation of mitochondrial autophagy. The mitochondrial autophagy regulated by the PINK/Parkin pathway was excessively activated. However, administration of SL prevented the activation of the PINK/Parkin pathway and inhibited excessive mitochondrial autophagy to regulate mitochondrial dysfunction. Results also demonstrated that mitochondrial dysfunction stimulated endothelial cell barrier dysfunction, but Evans blue transmission was significantly decreased and transmembrane resistance was increased significantly by SL treatment (p < 0.05 or p < 0.01). Carbonylcyanide-3-chlorophenylhydrazone (CCCP) could activate the PINK/Parkin pathway. CCCP reversed the regulation of SL on mitochondrial autophagy and mitochondrial function. SL could alleviate coronary artery no-reflow by protecting the microvasculature by regulating mitochondrial function. The underlying mechanism was related to decreased mitochondrial autophagy by the PINK/Parkin pathway.


Assuntos
Vasos Coronários , Traumatismo por Reperfusão Miocárdica , Ratos , Animais , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Vasos Coronários/metabolismo , Células Endoteliais/metabolismo , Carbonil Cianeto m-Clorofenil Hidrazona/metabolismo , Carbonil Cianeto m-Clorofenil Hidrazona/farmacologia , Volume Sistólico , Função Ventricular Esquerda , Autofagia , Mitocôndrias , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/farmacologia , Oxigênio/metabolismo , Trifosfato de Adenosina/metabolismo , Glucose/metabolismo
3.
Cell Mol Biol (Noisy-le-grand) ; 68(4): 202-207, 2022 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-35988275

RESUMO

COVID-19 vaccines have become an important hope for slowing down or stopping the pandemic. As the population ages, older adults will comprise a greater proportion of the vaccinated population. We aimed to assess influencing factors of COVID-19 vaccine hesitancy in older adults. For this aim, We conducted a cross-sectional study on a questionnaire survey of the elderly over 65 years living in the community of Haikou City from August 1st to September 30th, 2021. Univariate and multivariate Logistic regression analyses were performed to identify factors related to vaccine hesitancy. We analyzed completed questionnaires from 225 respondents (42.2% women, mean age 73.4±6.2 years). There were 99 people in the vaccine hesitation group and 126 people in the vaccine acceptance group, the incidence of vaccine hesitation in the elderly population is about44%(99/225). The incidence of frailty in the vaccine hesitation group was much higher than that in the vaccine trust group (62.63 vs. 30.95%, P<0.001). The risk factors of vaccine hesitancy in the elderly aged 70-75years and over 75 years were 2.987 times and 3.587 times higher than that in the population aged 65-70 years (OR=2.987,95%CI: 1.424-6.265, P<0.001; OR=3.587,95% CI:1.804-7.131, P<0.001). Frailty is also an independent risk factor of vaccine hesitancy in the elderly population (OR=2.624,95%CI: 1.447-4.757, P<0.001). Then the vaccination rate of the COVID-19 vaccine is far from reaching the requirements of herd immunity, and more flexible and comprehensive efforts are needed to increase the vaccination willingness of the frail elderly.


Assuntos
COVID-19 , Fragilidade , Vacinas , Idoso , COVID-19/epidemiologia , COVID-19/prevenção & controle , Vacinas contra COVID-19/uso terapêutico , Estudos Transversais , Feminino , Fragilidade/epidemiologia , Humanos , Masculino , Hesitação Vacinal
4.
Environ Sci Technol ; 55(24): 16770-16782, 2021 12 21.
Artigo em Inglês | MEDLINE | ID: mdl-34855387

RESUMO

A complex dataset with 140 sampling events was generated using triple quadrupole gas chromatography-mass spectrometer to track the occurrence of 95 odorants in raw and finished water from 98 drinking water treatment plants in 31 cities across China. Data analysis identified more than 70 odorants with concentrations ranging from not detected to thousands of ng/L. In raw water, Pearson correlation analysis determined that thioethers, non-oxygen benzene-containing compounds, and pyrazines were classes of chemicals that co-occurred, and geosmin and p(m)-cresol, as well as cyclohexanone and benzaldehyde, also co-occurred, indicating similar natural or industrial sources. Based on classification and regression tree analysis, total dissolved organic carbon and geographical location were identified as major factors affecting the occurrence of thioethers. Indoles, phenols, and thioethers were well-removed through conventional and advanced treatment processes, while some aldehydes could be generated. For other odorants, higher removal was achieved by ozonation-biological activated carbon (39.3%) compared to the conventional treatment process (14.5%). To our knowledge, this is the first study to systematically identify the major odorants in raw water and determine suitable treatment strategies to control their occurrence by applying data analytics and statistical methods to the complex dataset. These provide informative reference for odor control and water quality management in drinking water industry.


Assuntos
Água Potável , Poluentes Químicos da Água , Purificação da Água , Ciência de Dados , Matéria Orgânica Dissolvida , Odorantes/análise , Poluentes Químicos da Água/análise
5.
Zhongguo Zhong Yao Za Zhi ; 46(13): 3402-3409, 2021 Jul.
Artigo em Zh | MEDLINE | ID: mdl-34396761

RESUMO

This study aimed to investigate the effect and the possible mechanism of Shenlian( SL) extract on tumor necrosis factor-α( TNF-α)-induced ECV304 injury. After the establishment of TNF-α-induced ECV304 cells injure model,MTT assay was used to detect cell viability and the level of reactive oxygen species( ROS) was measured by flow cytometry. The contents of superoxide dismutase( SOD),malondialdehyde( MDA),nitric oxide( NO),endothelin-1( ET-1) and interleukin-1ß( IL-1ß) in the supernatant were detected by biochemical method and enzyme linked immunosorbent assay( ELISA). The expression levels of apoptosis-related proteins B-lymphoma-2 gene( Bcl-2),Bcl-2 associated X protein( Bax),caspase-3,caspase-9 and nuclear factor E2 associated factor2( Nrf2)/Kelch like epichlorohydrin associated protein-1( Keap1) signaling pathway related proteins Nrf2,Keap1,quinone oxidoreductase( NQO1) and heme oxygenase 1( HO-1) were detected by Western blot. The results showed that 50 µg·L-1 TNF-α significantly damaged ECV304 cells,induced the impairment of cell viability( P<0. 01),the increase of ROS production,the decrease of SOD activity,and the increase of MDA,NO,ET-1 and IL-1ß( P<0. 01),meanwhile,it caused the up-regulation of Keap1,caspase-9 and Bax protein expression,and down-regulation of NQO1 and Bcl-2 protein expression( P<0. 05) compared with the control group.Compared with the model group,SL extract reduced the damage of ECV304 cells induced by TNF-α,improved cell viability,reduced ROS production,increased SOD activity and decreased MDA,NO,ET-1,IL-1ß content( P<0. 01 or P<0. 05). In addition,SL extract also down-regulated the protein expression levels of Keap1,caspase-3,caspase-9 and Bax,and increased the protein expressions of Nrf2,NQO1,HO-1 and Bcl-2( P<0. 01 or P<0. 05). The above results indicate that SL extract can provide protective effect on ECV304 cells injury induced by TNF-α,alleviate oxidative stress injury,inflammation and apoptosis,and its mechanism may be related to regulating Nrf2/Keap1 signaling pathway.


Assuntos
Fator 2 Relacionado a NF-E2 , Fator de Necrose Tumoral alfa , Apoptose , Proteína 1 Associada a ECH Semelhante a Kelch/genética , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo , Extratos Vegetais , Transdução de Sinais , Fator de Necrose Tumoral alfa/genética
6.
Int J Syst Evol Microbiol ; 70(10): 5271-5279, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32833615

RESUMO

A psychrotolerant actinobacterium, designated strain J5903T, was isolated from an alkaline soil sample from the rhizosphere of Suaeda salsa collected in desertification land surrounding Jiuliancheng Nur in Hebei Province, PR China. Cells of the isolate were Gram-stain-positive, aerobic, non-motile and non-spore-forming cocci. Strain J5903T grew optimally at 20‒25 °C, at pH 7.0‒7.5 and with <1 % (w/v) NaCl. The cell-wall peptidoglycan type was B2γ with d-2,4-diaminobutyric acid and l-2,4-diaminobutyric acid as diagnostic amino acids. The muramyl residue was acetyl type. The menaquinones were MK-11, MK-12, MK-10 and MK-13. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and one unidentified glycolipid. The major whole-cell fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. The genomic DNA G+C content was 69.1 mol%. It shared the highest average nucleotide identity and digital DNA-DNA hybridization values with Planctomonas deserti 13S1-3T. Phylogenies based on genome sequence showed that strain J5903T and P. deserti 13S1-3T formed a robust cluster with high bootstrap support. Strain J5903T shared typical chemotaxonomic characteristics with P. deserti 13S1-3T. Combining the polyphasic taxonomic evidence, strain J5903T represents a novel species of the genus Planctomonas, for which the name Planctomonas psychrotolerans sp. nov. is proposed. The type strain is J5903T (=DSM 101894T=CGMCC 1.15523T).


Assuntos
Actinobacteria/classificação , Chenopodiaceae/microbiologia , Filogenia , Rizosfera , Microbiologia do Solo , Actinobacteria/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Glicolipídeos/química , Hibridização de Ácido Nucleico , Peptidoglicano/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/química
7.
Int J Syst Evol Microbiol ; 70(7): 4179-4185, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32558636

RESUMO

A novel moderately halophilic, filamentous actinobacterium, designated as XMNu-373T, was isolated from a saline-alkaline soil sample collected from the Mongolia Plateau, Dongwu County, Inner Mongolia Autonomous Region, PR China. The isolate grew optimally at 28‒37 °C, pH 7.0‒8.0 and with 2-5 % (w/v) NaCl. The substrate mycelia fragmented into rod-like elements, and the white aerial mycelia formed spore chains at maturity. The predominant menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, three unidentified phosphoglycolipids, an unidentified aminophospholipid, two phosphatidylinositol mannosides, four unidentified phospholipids, phosphatidylglycerol and two unidentified lipids. The major cellular fatty acids were iso-C16 : 0, anteiso-C17 : 0 and anteiso-C15 : 0. The genomic DNA G+C content was 66.2 mol%. It shared high 16S rRNA gene sequence similarities to Phytoactinopolyspora halotolerans YIM 96448T (96.1 %) and Phytoactinopolyspora endophytica EGI 60009T (96.0 %). Phylogenetic trees based on 16S rRNA gene sequences revealed that strain XMNu-373T resided in the clade of family Jiangellaceae, and it formed a monophyletic branch distinct from four other recognized type species in the subclade of the genus Phytoactinopolyspora. On the basis of polyphasic taxonomic evidence, strain XMNu-373T represents a novel species of the genus Phytoactinopolyspora, for which the name Phytoactinopolyspora mesophila sp. nov. is proposed. The type strain is XMNu-373T (=JCM 33740T=CGMCC 4.7654T).


Assuntos
Actinobacteria/classificação , Álcalis , Filogenia , Salinidade , Microbiologia do Solo , Actinobacteria/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Solo/química , Vitamina K 2/análogos & derivados , Vitamina K 2/química
8.
Int Heart J ; 61(4): 822-830, 2020 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-32684596

RESUMO

This study aims to analyze the expression level and correlation of miR-182-5p and its target gene PAPPA in coronary atherosclerosis (CAD).Real time PCR, ELISA, and Western blotting methods were used to detect the expression levels. Dual-luciferase reporter gene assays were used to analyze the interaction between the 3'-UTR of PAPPA and miR-182-5p.The expression level of miR-182-5p in CAD was significantly lower than that in normal population, while the content of serum PAPPA was significantly increased, and the expression level of miR-182-5p was negatively correlated with the PAPPA content. The expression level of miR-182-5p decreased, while the expression level of PAPPA increased significantly in the ox-LDL treated HA-VSMC cells. Researchers found that PAPPA could promote the activation of IGF signaling pathway in HA-VSMC cells treated by ox-LDL, further activate NF-kB, PI3K/AKT and ERK signaling pathway, and promote cell proliferation. However, miR-182-5p could inhibit the expression of PAPPA, block the activation of IGF signal pathway, and inhibit the proliferation of HA-VSMC cells induced by ox-LDL. miR-182-5p had a targeted action site in the 3'-UTR of PAPPA by bioinformatics prediction. The analysis of luciferase reporter gene further confirmed that miR-182-5p could target the 3'-UTR of PAPPA to inhibit its expression.miR-182-5p demonstrated a protective effect on atherosclerosis and may be a potential therapeutic target for atherosclerosis.


Assuntos
Doença da Artéria Coronariana/sangue , MicroRNAs/sangue , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Proteína Plasmática A Associada à Gravidez/metabolismo , Células Cultivadas , Humanos , Sistema de Sinalização das MAP Quinases
9.
J Environ Sci (China) ; 79: 100-110, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30784435

RESUMO

A wide range of compounds with various structural features can cause taste and odor (T&O) problems in drinking water. It would be desirable to determine all of these compounds using a simple analytical method. In this paper, a sensitive method combining liquid-liquid extraction (LLE) with gas chromatography-triple quadrupole tandem mass spectrometry (GC-MS/MS) was established to simultaneously analyze 51 odor-causing compounds in drinking water, including organic sulfides, aldehydes, benzenes, phenols, ethers, esters, ketones, nitrogenous heterocyclic compounds, 2-methylisoborneol and geosmin. Three deuterated analogs of target analytes, dimethyl disulfide-d6, benzaldehyde-d6 and o-cresol-3,4,5,6-d4, were used to correct the variations in recovery, and five isotope-labeled internal standards (4-chlorotoluene-d4, 1, 4-dichlorobenzene-d4, naphthalene-d8, acenaphthene-d10, phenanthrene-d10 respectively) were used prior to analysis to correct the variations arising from instrument fluctuations and injection errors. The calibration curves of the target compounds showed good linearity (R2 > 0.99, level = 7), and method detection limits (MDLs) below 1/10 of the odor threshold concentrations were achieved for most of the odorants (0.10-20.55 ng/L). The average recoveries of most of the analytes in tap water samples were between 70% and 120%, and the method was reproducible (RSD < 20%, n = 7). Additionally, concentrations of odor-causing compounds in water samples collected from three drinking water treatment plants (DWTPs) were analyzed by this method. According to the results, dimethyl trisulfide, dimethyl disulfide and indole were considered to be the key odorants responsible for the swampy/septic odor. 2-Methylisoborneol and geosmin were detected as the main odor-causing compounds for musty/earthy odor in DWTP B.


Assuntos
Água Potável/análise , Odorantes/análise , Poluentes Químicos da Água/análise , Monitoramento Ambiental , Cromatografia Gasosa-Espectrometria de Massas , Espectrometria de Massas em Tandem
10.
J Cell Biochem ; 118(10): 3150-3157, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-28004426

RESUMO

The aim of this study was to investigate the possibility of APC/CCdh1 as a potential therapeutic target in the radiosensitivity of nasopharyngeal carcinoma (NPC) cell CNE-1, and explain the role of APC subunits after silence of Cdh1 combined with radiotherapy. Transfection with Cdh1 shRNA significantly increased the radiosensitivity of CNE-1 cells and the radiation enhancement ratio (RER) of sh-Cdh1 cells was 1.76. Knockdown of Cdh1 in CNE-1 cells increased irradiation induced apoptosis and G2/M phase cell cycle arrest. The levels of CDC20 and CylinB1 increased and the levels of Ku70 and APC3 decreased after irradiation. APC/CCdh1 is involved in regulation of radiosensitivity in human NPC CNE-1 cells. Our study may provide a promising therapeutic strategy for NPC by targeting Cdh1. J. Cell. Biochem. 118: 3150-3157, 2017. © 2016 Wiley Periodicals, Inc.


Assuntos
Subunidade Apc3 do Ciclossomo-Complexo Promotor de Anáfase/metabolismo , Apoptose , Caderinas/metabolismo , Carcinoma/metabolismo , Neoplasias Nasofaríngeas/metabolismo , Tolerância a Radiação , Antígenos CD , Subunidade Apc3 do Ciclossomo-Complexo Promotor de Anáfase/genética , Caderinas/genética , Carcinoma/genética , Carcinoma/radioterapia , Linhagem Celular Tumoral , Técnicas de Silenciamento de Genes , Humanos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/radioterapia
11.
Pacing Clin Electrophysiol ; 40(7): 803-807, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28585685

RESUMO

In patients without associated myocardial diseases, characterized by left bundle branch block and inferior axis morphologies, repetitive idiopathic right ventricular tachycardias and ventricular premature contractions typically arise from right ventricular outflow tract (RVOT). Accumulated evidences have shown that radiofrequency catheter ablation is a useful treatment for patients with RVOT ventricular arrhythmias (VAs). Interestingly, several medical centers have shown that pulmonary artery (PA) is a potential novel site for catheter ablation in RVOT-like VAs, particularly in patients where termination of RVOT VAs at the usual site fails. In this review, we comprehensively demonstrated that RVOT VAs were successfully terminated at the site of PA, analyzed the characteristics of surface electrocardiogram and endocardial potentials, and explored the underlying mechanisms for these cases.


Assuntos
Ablação por Cateter , Artéria Pulmonar/fisiopatologia , Artéria Pulmonar/cirurgia , Taquicardia Ventricular/cirurgia , Complexos Ventriculares Prematuros/cirurgia , Técnicas Eletrofisiológicas Cardíacas , Humanos , Taquicardia Ventricular/fisiopatologia , Complexos Ventriculares Prematuros/fisiopatologia
12.
Korean J Parasitol ; 55(1): 77-79, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28285510

RESUMO

The infection by Limnodrilus hoffmeisteri Claparède, 1862 (Oligochaeta: Tubificinae) in humans is relatively uncommon. The present report is to describe an incidental human infection with oligochaetes in the nasal cavity of a Chinese man, a 25-year-old man residing in Zhangjiakou city, Hebei province, China presenting with nose bleed, severe itching, continuous sneezing, and rhinorrhea. A lot of oligochaete worms were found in the nasal discharge of the patient. The detected worms were identified as Limnodrilus hoffmeisteri (Annelida: Oligochaeta) based on morphological and molecular characteristics. This incidental L. hoffmeisteri nasal infection is the first case in China and indicates that oligochaete worms can be encountered in humans.


Assuntos
Cavidade Nasal/parasitologia , Oligoquetos/crescimento & desenvolvimento , Doenças Parasitárias/diagnóstico , Doenças Parasitárias/patologia , Adulto , Animais , China , Análise por Conglomerados , DNA de Helmintos/química , DNA de Helmintos/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Humanos , Masculino , Microscopia , Oligoquetos/anatomia & histologia , Oligoquetos/classificação , Oligoquetos/genética , Doenças Parasitárias/parasitologia , Filogenia , RNA Ribossômico 18S/genética , Análise de Sequência de DNA
13.
Artigo em Zh | MEDLINE | ID: mdl-26541029

RESUMO

OBJECTIVE: To construct a recombinant eukaryotic expression plasmid containing ROP18-ROP12 (encoding rhoptry protein 18 and 12) complex gene of Toxoplasma gondii, and examine its expression in eukaryotic cells. METHODS: Recombinant plasmids pVAX1-ROP18 and pVAX1-ROP12 were digested by restriction enzymes BamH I and Xba I . ROP12 gene was cloned into pVAX1-ROP18 to construct the eukaryotic expression plasmid pVAX1-ROP18- ROP12. After colony PCR, enzyme digestion and sequencing, the correct recombinant plasmid pVAX1-ROP18-ROP12 was transfected into HeLa cells. Along with it were groups of empty plasmid, pVAX1-ROP18 and pVAX1-ROP12. Total RNA was extracted from HeLa cells and reverse-transcribed to cDNA. RT-PCR was performed to evaluate mRNA expression of the housekeeping gene ß-actin and ROP18-ROP12 complex gene. Immunofluorescence assay and Western blotting were performed to determine the protein levels of ROP18-ROP12 fusion protein. RESULTS: Colony PCR in recombinant plasmid pVAX1-ROP18-ROP12 showed a specific band at about 2 373 bp, consistent with expectation. The extracted recombinant plasmids were confirmed by Hind III, BamH I and Xba I digestion. Sequencing results showed that the sequence of pVAX1-ROP18-ROP12 was 100% identical to that of T. gondii RH strain ROP18 gene (Accession No. AM075204.1) and 99% identical to that of T. gondii RH strain ROP12 gene (Accession No. DQ096559.1). Further, RT-PCR showed amplification products at 613 bp for ß-actin in all the groups, while only the pVAX1-ROP18-ROP12 transfection group showed amplification products for the ROP18-ROP12 complex at 2,373 bp. In addition, the indirect immunofluorescence assay showed yellow-green fluorescence in HeLa cells transfected with pVAX1-ROP18-ROP12, but not in control cells. Western blotting showed that the ROP18-ROP12 fusion protein was expressed in HeLa cells transfected with recombinant plasmid pVAX1-ROP18-ROP12. CONCLUSIONS: The recombinant eukaryotic plasmid pVAX1-ROP18-ROP-2 is constructed and can be expressed in eukaryotic system.


Assuntos
Toxoplasma , Actinas , Western Blotting , Vetores Genéticos , Células HeLa , Humanos , Plasmídeos , Reação em Cadeia da Polimerase , Proteínas Recombinantes , Transfecção
14.
Oral Oncol ; 153: 106838, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38729038

RESUMO

Actinomycetes are predominantly pathogenic bacteria that lack aerial hyphae and do not form spores. They are generally anaerobic or facultative anaerobic Gram-positive bacteria, belonging to the prokaryotic group. Actinomycetes are widely distributed in nature, similar to other bacteria, and are mostly saprophytic, with a few being parasitic. They are named as such due to their colony's radial form. The symptoms and signs of actinomycosis are atypical and rarely manifest in the nasopharynx. Consequently, it can be challenging to distinguish actinomycosis from nasopharyngeal carcinoma, making diagnosis difficult. Histopathology is usually relied upon for diagnosis, although culture may pose challenges. Fortunately, actinomycetes are highly sensitive to penicillin. Therefore, timely treatment with high doses of penicillin is crucial for successful recovery. In this case study, we present the details of a 53-year-old female patient with no history of mucosal damage or tooth decay but with a previous partial thyroidectomy. The patient experienced recurring neck pain accompanied by progressive limitation of neck movement. Nasopharyngoscopy revealed the presence of a smooth-surfaced mass. Subsequent biopsy, clinical imaging, microbiological analysis, and histological findings confirmed the diagnosis of actinomycosis.Following a comprehensive treatment plan involving a combination of penicillin and doxycycline for a duration of two months, the disease was successfully eradicated.


Assuntos
Actinomicose , Humanos , Actinomicose/diagnóstico , Feminino , Pessoa de Meia-Idade , Antibacterianos/uso terapêutico , Diagnóstico Diferencial
15.
Eur J Pharm Sci ; 198: 106789, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38710335

RESUMO

BACKGROUND: Cytoplasmic epidermal growth factor receptor (EGFR) is overexpressed in both nasopharyngeal carcinoma (NPC) and triple-negative breast cancer (TNBC), while clinical outcome and prognosis vary greatly among patients treated with gefitinib, and all patients eventually develop resistance to this agent. Therefore, we propose a new concept for synthesizing multitarget compounds and reveal new therapeutic strategies for NPC and TNBC expressing EGFR. METHODS: Compound H was synthesized in our previous study. Molecular docking, and cell thermal shift assays (CETSAs) and drug affinity responsive target stability(DARTS) were used to confirm the binding of compound H to EGFR and GLUT1. Methylthiazolyldiphenyl-tetrazolium bromide(MTT), annexin V-PE assays, mitochondrial membrane potential (MMP) assays, and animal models were used to evaluate the inhibitory effect of compound H on TNBC cell lines. Energy metabolism tests, Western blotting, and immunofluorescence staining were performed to evaluate the synergistic effects on EGFR- and glucose transporter type 1(GLUT1)-mediated energy metabolism. RESULTS: Compound H can simultaneously act on the EGFR tyrosine kinase ATP-binding site and inhibit GLUT1-mediated energy metabolism, resulting in reductions in ATP, MMP, intra-cellular lactic acid, and EGFR nuclear transfer. The anti-tumor activity of compound H is significantly superior to the combination of GLUT1 inhibitor BAY876 and EGFR inhibitor gefitinib. Compound H has remarkable anti-proliferative effects on TNBC MDA-MB231 cells, and importantly, no obvious toxicity effects of compound H were found in vivo. CONCLUSIONS: Synergistic effects of inhibition of EGFR- and GLUT1-mediated energy metabolism by compound H may present a new strategy for the treatment of TNBC and NPC.


Assuntos
Antineoplásicos , Receptores ErbB , Transportador de Glucose Tipo 1 , Carcinoma Nasofaríngeo , Neoplasias de Mama Triplo Negativas , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/metabolismo , Transportador de Glucose Tipo 1/antagonistas & inibidores , Transportador de Glucose Tipo 1/metabolismo , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/metabolismo , Humanos , Animais , Linhagem Celular Tumoral , Feminino , Carcinoma Nasofaríngeo/tratamento farmacológico , Carcinoma Nasofaríngeo/metabolismo , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Simulação de Acoplamento Molecular , Neoplasias Nasofaríngeas/tratamento farmacológico , Neoplasias Nasofaríngeas/metabolismo , Camundongos Nus , Camundongos Endogâmicos BALB C , Gefitinibe/farmacologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos
16.
Environ Sci Ecotechnol ; 21: 100392, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38434492

RESUMO

Activated carbon is employed for the adsorption of organic micropollutants (OMPs) from water, typically present in concentrations ranging from ng L-1 to µg L-1. However, the efficacy of OMP removal is considerably deteriorated due to competitive adsorption from background dissolved organic matter (DOM), present at substantially higher concentrations in mg L-1. Interpreting the characteristics of competitive DOM is crucial in predicting OMP adsorption efficiencies across diverse natural waters. Molecular weight (MW), aromaticity, and polarity influence DOM competitiveness. Although the aromaticity-related metrics, such as UV254, of low MW DOM were proposed to correlate with DOM competitiveness, the method suffers from limitations in understanding the interplay of polarity and aromaticity in determining DOM competitiveness. Here, we elucidate the intricate influence of aromaticity and polarity in low MW DOM competition, spanning from a fraction level to a compound level, by employing direct sample injection liquid chromatography coupled with ultrahigh-resolution Fourier-transform ion cyclotron resonance mass spectrometry. Anion exchange resin pre-treatment eliminated 93% of UV254-active DOM, predominantly aromatic and polar DOM, and only minimally alleviated DOM competition. Molecular characterization revealed that nonpolar molecular formulas (constituting 26% PAC-adsorbable DOM) with medium aromaticity contributed more to the DOM competitiveness. Isomer-level analysis indicated that the competitiveness of highly aromatic LMW DOM compounds was strongly counterbalanced by increased polarity. Strong aromaticity-derived π-π interaction cannot facilitate the competitive adsorption of hydrophilic DOM compounds. Our results underscore the constraints of depending solely on aromaticity-based approaches as the exclusive interpretive measure for DOM competitiveness. In a broader context, this study demonstrates an effect-oriented DOM analysis, elucidating counterbalancing interactions of DOM molecular properties from fraction to compound level.

17.
Chemosphere ; 350: 141043, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38154675

RESUMO

Fishy odor, as an offensive and unpleasant odor, could occur in drinking water source with poor nutrition, it is usually considered to be associated with odor-producing microalgae. However, the specific relations among fishy odor, fishy odorants and responsible microalgae were not elucidated comprehensively. In this paper, the odor-causing compounds generated from six microalgae with fishy odor characteristic, isolated in drinking water source Tongyu River, were identified simultaneously. The sensory evaluation result indicated that Tongyu River was principally related to fishy odor (odor intensity 6.5-7.6). Correspondingly, seven, nine, seven, six, seven and seven olfactory detection peaks (ODP) were screened by combining data of GC/O/MS and GC/GC/TOFMS in Cyclotella, Cryptomonas ovate, Melosira, Dinobryon sp., Synedra and Ochromonas sp., which were isolated in Tongyu River and cultured in laboratory. Totally twenty odor-causing compounds, including hexanal, 2-hexenal, 3-hexen-1-ol, heptanal, 1-octen-3-one, 2,4-heptadienal, 2-tetradecanone, 3,5-octadien-2-one, octanal, 1-octen-3-ol, 2-octenal, nonanal, 2,4-octadienal, 2-nonenal, decanal, 2,6-nonadienal, 2-decenal, undecanal, 2,4-decadienal and dodecanal, were screened and identified in all isolated microalgae. Additionally, fishy odor intensity for all identified microalgae increased obviously as microalgae cell number increased and microalgae cell ruptured in cultivation cycles through pearson and spearman correlation analysis. For the first time, twenty odor-causing compounds associating with fishy odor were recognized from six isolated microalgae, which would provide more scientific basis and theoretical support for preventing and treating fishy odor episode of drinking water source.


Assuntos
Diatomáceas , Água Potável , Microalgas , Compostos Orgânicos Voláteis , Odorantes/análise , Água Potável/análise , Olfato , Cromatografia Gasosa-Espectrometria de Massas , Compostos Orgânicos Voláteis/análise
18.
Sci Total Environ ; 921: 171155, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38387591

RESUMO

The occurrence and distribution of 1,4-dioxane was investigated in 280 source and finished drinking water samples from 31 Chinese cities, based on which its ecological and health risks were systematically evaluated. The findings demonstrated that 1,4-dioxane was detected in about 80.0 % samples with values ranging from n.d. to 7757 ng/L in source water and n.d. to 2918 ng/L in drinking water. 1,4-Dioxane showed limited removal efficiency using conventional coagulation-sedimentation-filtration processes (14 % ± 48 %), and a removal efficiency of 35 % ± 44 % using ozonation-biological activated carbon advanced treatment processes. Relatively higher concentrations, detection frequency and environmental risk were observed in Taihu Lake, Yellow River, Yangtze River, Zhujiang River, and Huaihe River mainly in the eastern and southern regions, where there are considerable industrial activities and comparatively high population densities. The widespread presence as by-products during manufacturing consumer products e.g., ethoxylated surfactants, suggested municipal wastewater discharges were the dominant source for the ubiquitous occurrence of 1,4-dioxane, while industrial activities, e.g. resin manufacturing, also contribute considerably to the elevated concentrations of 1,4-dioxane. The estimated risk quotients were in the range of <1.5 × 10-4 for ecological risk, <5.0 × 10-3 by oral exposure and < 5.0 × 10-2 by inhalation exposure for health risk, illustrating limited ecological harm to water environment or chronic toxicity to human health. For carcinogenic risk, 1,4-Dioxane presented a mean risk of 1.8 × 10-6 by oral exposure, which slightly surpassed the recommended acceptable levels of U.S. EPA (<10-6), and risk from inhalation exposure could be negligible. The pervasiveness in drinking water, low removal efficiencies during water treatment processes, and suspected health impacts, highlighted the necessity to set related water quality standards of 1,4-dioxane in order to improve water environment in China.


Assuntos
Dioxanos , Água Potável , Poluentes Químicos da Água , Humanos , Poluentes Químicos da Água/análise , Qualidade da Água , China , Rios , Monitoramento Ambiental
19.
Artigo em Zh | MEDLINE | ID: mdl-24818410

RESUMO

Total RNA was extracted from tachyzoites of RH strain of Toxoplasma gondii. The open reading frame of ROP11 gene was amplified by using a pair of specific primers designed according to the coding sequence of ROP11 gene (Accession No. DQ077905). The RT-PCR product was digested by restriction enzyme EcoR I and Not I, and then ligated into a pGEX-6P-2 vector. The recombinant plasmid was transferred into E. coli XL-Blue. The positive clones was selected by colony PCR, and confirmed by the double restriction enzyme digestion and sequencing. The RT-PCR product was 1,548 bp. The recombinant plasmid was confirmed by colony PCR and double restriction enzyme digestion. Sequencing results showed that the obtained ROP11 gene was 1 548 bp (Accession No. KC456639). There was a high sequence consistency (99%) between the obtained ROP11 gene sequence and the Toxoplasma ROP11 gene from GenBank. Bioinformatics analysis showed that the ROP11 protein (Mr 57,020) consisted of the signal peptide (amino acids 1-26), 12 conservative domains, a serine/threonine protein kinase catalytic domain (amino acids 170-511), and two potential N-glycosylation sites.


Assuntos
Biologia Computacional , Proteínas de Protozoários/genética , Toxoplasma/genética , Clonagem Molecular , Expressão Gênica , Vetores Genéticos
20.
Sci Total Environ ; 857(Pt 2): 159465, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36257419

RESUMO

Swampy/septic odor caused by thioethers has become the main taste and odor (T&O) problem in drinking water of China. Improving its removal performance by commonly traditional water treatment process is significant. In our study, we have found that pre-oxidation could modify the background dissolved organic matter (DOM) properties and thus improve the coagulation performance of thioethers, increasing the coagulation removal rates by 1.5-3 times. Particularly, after pre-ozonation only protein-like substances remained, and thioethers removal was 1.5 times higher than that after pre-chlorination (only coagulation not including oxidation). Compared with humic acid (HA), the thioethers compounds removal efficiencies under bovine serum albumin (BSA) as background DOM was increased by 0.3-3 times. Through Freundlich model analysis, the binding strength of BSA (KF = 20.712, at 298 K) to dimethyl disulfide (DMDS) was enhanced by 60 % compared to HA (KF = 12.778, at 298 K). According to thermodynamic parameters, the binding effect between HA/BSA and thioethers compounds was mainly van der Waals forces and hydrogen bond. BSA with more amino structure and oxygen groups was more easily to adsorb DMDS through hydrogen bond and thus achieved better coagulation performance. Therefore, pre-ozonation combined with coagulation was suggested to be more suitable for thioethers compounds control.


Assuntos
Ozônio , Poluentes Químicos da Água , Purificação da Água , Sulfetos , Substâncias Húmicas/análise , Oxirredução , Ozônio/análise , Poluentes Químicos da Água/análise
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