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MAIN CONCLUSION: The secondary metabolic conversion of monolignans to sesquilignans/dilignans was closely related to seed germination and seedling establishment in Arctium lappa. Arctium lappa plants are used as a kind of traditional Chinese medicines for nearly 1500 years, and so far, only a few studies have put focus on the key secondary metabolic changes during seed germination and seedling establishment. In the current study, a combined approach was used to investigate the correlation among secondary metabolites, plant hormone signaling, and transcriptional profiles at the early critical stages of A. lappa seed germination and seedling establishment. Of 50 metabolites in methonolic extracts of A. lappa samples, 35 metabolites were identified with LC-MS/MS and 15 metabolites were identified with GC-MS. Their qualitative properties were examined according to the predicted chemical structures. The quantitative analysis was performed for deciphering their metabolic profiles, discovering that the secondary metabolic conversion from monolignans to sesquilignans/dilignans was closely correlated to the initiation of A. lappa seed germination and seedling establishment. Furthermore, the critical transcriptional changes in primary metabolisms, translational regulation at different cellular compartments, and multiple plant hormone signaling pathways were revealed. In addition, the combined approach provides unprecedented insights into key regulatory mechanisms in both gene transcription and secondary metabolites besides many known primary metabolites during seed germination of an important traditional Chinese medicinal plant species. The results not only provide new insights to understand the regulation of key medicinal components of 'ARCTII FRUCTUS', arctiin and arctigenin at the stages of seed germination and seedling establishment, but also potentially spur the development of seed-based cultivation in A. lappa plants.
Assuntos
Arctium , Germinação , Lignanas , Sementes , Arctium/genética , Arctium/metabolismo , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Lignanas/metabolismo , Plântula/genética , Plântula/crescimento & desenvolvimento , Plântula/metabolismo , Regulação da Expressão Gênica de Plantas , Espectrometria de Massas em Tandem , Lignina/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Metabolismo SecundárioRESUMO
BACKGROUND: Recent research suggests that periodontitis can increase the risk of chronic obstructive pulmonary disease (COPD). In this study, we performed two-sample Mendelian randomization (MR) and investigated the causal effect of periodontitis (PD) on the genetic prediction of COPD. The study aimed to estimate how exposures affected outcomes. METHODS: Published data from the Gene-Lifestyle Interaction in the Dental Endpoints (GLIDE) Consortium's genome-wide association studies (GWAS) for periodontitis (17,353 cases and 28,210 controls) and COPD (16,488 cases and 169,688 controls) from European ancestry were utilized. This study employed a two-sample MR analysis approach and applied several complementary methods, including weighted median, inverse variance weighted (IVW), and MR-Egger regression. Multivariable Mendelian randomization (MVMR) analysis was further conducted to mitigate the influence of smoking on COPD. RESULTS: We chose five single-nucleotide polymorphisms (SNPs) as instrumental variables for periodontitis. A strong genetically predicted causal link between periodontitis and COPD, that is, periodontitis as an independent risk factor for COPD was detected. PD (OR = 1.102951, 95% CI: 1.005-1.211, p = 0.039) MR-Egger regression and weighted median analysis results were coincident with those of the IVW method. According to the sensitivity analysis, horizontal pleiotropy's effect on causal estimations seemed unlikely. However, reverse MR analysis revealed no significant genetic causal association between COPD and periodontitis. IVW (OR = 1.048 > 1, 95%CI: 0.973-1.128, p = 0.2082) MR Egger (OR = 0.826, 95%CI:0.658-1.037, p = 0.1104) and weighted median (OR = 1.043, 95%CI: 0.941-1.156, p = 0.4239). The results of multivariable Mendelian randomization (MVMR) analysis, after adjusting for the confounding effect of smoking, suggest a potential causal relationship between periodontitis and COPD (P = 0.035). CONCLUSION: In this study, periodontitis was found to be independent of COPD and a significant risk factor, providing new insights into periodontitis-mediated mechanisms underlying COPD development.
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Estudo de Associação Genômica Ampla , Análise da Randomização Mendeliana , Polimorfismo de Nucleotídeo Único , Doença Pulmonar Obstrutiva Crônica , Fumar , Humanos , Doença Pulmonar Obstrutiva Crônica/genética , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Fatores de Risco , Fumar/epidemiologia , Fumar/efeitos adversos , Periodontite/genética , Periodontite/epidemiologia , Índice de Gravidade de Doença , Predisposição Genética para Doença , Doenças Periodontais/genética , Doenças Periodontais/epidemiologiaRESUMO
The use of small interfering RNA (siRNA) in melanoma treatment remains limited owing to its biological properties. Herein, we developed a carrier system containing hyaluronic acid and protamine for siRNA delivery. Considering zeta potential and particle size as standards, the ratio of each component in liposome nanoparticles prepared was screened using the control variable method, and siRNA cationic liposome nanoparticles were prepared based on the optimal results obtained. The encapsulation rate of the cationic liposome nanoparticles was measured, and particle morphology was observed. B16F10 cells were treated with the nanoparticles; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, cell scratch experiments, and cell uptake experiments were performed to determine the effectiveness of the loaded siRNA. A mouse model was then established, and tumour tissues were subjected to haematoxylin-eosin staining. The inhibition of the survivin gene and protein expression were assessed using reverse transcription-polymerase chain reaction and western blotting, respectively. The results showed that the optimal mass ratio of hyaluronic acid (HA)-siRNA-to-protamine was 1.0; in the HA-siRNA-protamine complex containing 25 µg siRNA, the addition of 50 µL liposomes yielded optimal particles. And encapsulation rate was 85.07%. The nanoparticles demonstrated a significant inhibitory effect against melanoma cells; siRNA liposomes may inhibit tumour growth by down-regulating survivin. Survivin-siRNA cationic liposome nanoparticles could effectively inhibit the proliferation and migration of melanoma B16F10 cells in vitro and the proliferation of subcutaneous melanoma B16F10 cells, probably by inhibiting survivin mRNA and protein expression. Graphical abstract.
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Ácido Hialurônico/química , Melanoma/patologia , Nanopartículas/química , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/farmacologia , Animais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Química Farmacêutica , Relação Dose-Resposta a Droga , Portadores de Fármacos/química , Estabilidade de Medicamentos , Feminino , Lipossomos/química , Camundongos , Camundongos Endogâmicos C57BL , Tamanho da Partícula , Distribuição Aleatória , Propriedades de Superfície , Survivina/efeitos dos fármacosRESUMO
Agricultural plastic films have been proven highly advantageous, but they also cause pollution of plastic debris and associated chemicals. Phthalates (phthalic acid esters, PAEs), an important additive of agricultural films, can be released and contaminate the environment. Here, we analyzed the agricultural plastic usage and assessed plastic debris in China and developed a method to estimate PAE emissions from agricultural films. Additionally, the environmental fate of PAEs was evaluated using a fugacity-based multimedia model. The agricultural plastic film usage in China in 2017 was 2,528,600 tons. After agricultural film recycling and water erosion, the plastic debris amount was estimated as 465,016 tons. The water erosion process carried 4329 tons of plastic debris into the aquatic environment. During its lifetime, the agricultural film released a total of 91.5 tons of two typical types of PAEs. PAEs from the mulching film would mostly be removed through degradation, while those from the greenhouse film accumulate in vegetables. Populated regions exhibited more serious PAE pollution in vegetables but with no immediate health risks. The model was well evaluated using comparable measured concentrations and uncertainty analysis based on the Monte Carlo method. The findings from this study demonstrate the serious agricultural plastic pollution problem and associated PAE contamination in China.
Assuntos
Ácidos Ftálicos , Poluentes do Solo , China , Ésteres , Plásticos , Poluentes do Solo/análiseRESUMO
Stimulated emission depletion (STED) nanoscopy provides subdiffraction resolution while preserving the benefits of fluorescence confocal microscopy in live-cell imaging. However, there are several challenges for multicolor STED nanoscopy, including sophisticated microscopy architectures, fast photobleaching, and cross talk of fluorescent probes. Here, we introduce two types of nanoscale fluorescent semiconducting polymer dots (Pdots) with different emission wavelengths: CNPPV (poly[2-methoxy-5-(2-ethylhexyloxy)-1,4-(1-cyanovinylene-1,4-phenylene)]) Pdots and PDFDP (poly[{9,9-dihexyl-2,7-bis(1-cyanovinylene)fluorene}-alt-co-{2,5-bis (N,N'-diphenylamino)-1,4-phenylene}]) Pdots, for dual-color STED bioimaging and cellular tracking. Besides bright fluorescence, strong photostability, and easy bioconjugation, these Pdots have large Stokes shifts, which make it possible to share both excitation and depletion beams, thus requiring only a single pair of laser beams for the dual-color STED imaging. Long-term tracking of cellular organelles by the Pdots has been achieved in living cells, and the dynamic interaction of endosomes derived from clathrin-mediated and caveolae-mediated endocytic pathways has been monitored for the first time to propose their interaction models. These results demonstrate the promise of Pdots as excellent probes for live-cell multicolor STED nanoscopy.
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Rastreamento de Células/métodos , Corantes Fluorescentes/uso terapêutico , Polímeros/química , Pontos Quânticos/química , Humanos , LasersRESUMO
Study on the modified mechanism of wood cell walls, it is very important for improving treatment reagents, optimizing treatment technology, and enhancing wood density, mechanical properties, dimensional stability, and so on. Samples of plantation Chinese fir were treated gradually with synthesized water-soluble low-molecular-weight phenol formaldehyde (PF) resins under vacuum and pressure. The correlated physical and chemical properties of the treated and untreated reference samples were determined by X-ray diffractometer (XRD), Fourier transform infrared spectrometer (FTIR), and nuclear magnetic resonance spectrometer(NMR) (Using method of Cross Polarization/Magic Angle Spinning for continuous testing) with high precision and resolution. The results showed that, after treated with water-soluble low-molecular-weight PF resin, the average values of crystallinity from the treated samples were decreased obviously, and the average reduction rate was 12.67%, 11.91% and 6.26%, respectively. Comparing water-soluble, low-molecular-weight PF resin modified Chinese fir with untreated reference samples, no new chemical shifts and characteristic peaks of functional groups from esters, ethers, etc. were present by using FTIR and ¹³C NMR spectrum. It was considered that there was no distinct chemical reaction between the water-soluble low-molecular-weight PF resin and Chinese Fir cell walls. But water-soluble low-molecular-weight PF resin could enter into the structure relatively loose, large size spaces, relatively area large amorphous regions in cell walls of Chinese fir tracheids, and form physical filling, which resulting in the decreasing of relative crystallinity. This study has important reference value for the development of new wood modification reagents and the optimization of wood modification process. The findings also provide important theoretical foundation for further proving the modification mechanisms of wood cell walls and enriching the modified theories of wood cell walls.
Assuntos
Parede Celular , Cunninghamia/citologia , Formaldeído/química , Fenóis/química , Polímeros/química , Madeira , Espectroscopia de Ressonância Magnética , Peso MolecularRESUMO
Microplastics (MPs) are emerging contaminants found globally. However, their effects on soil-plant systems in salt-affected habitats remain unknown. Here, we examined the effects of polyethylene (PE) and polylactic acid (PLA) on soil properties, maize performance, and bacterial communities in soils with different salinity levels. Overall, MPs decreased soil electrical conductivity and increased NH4+-N and NO3--N contents. Adding NaCl alone had promoting and inhibitive effects on plant growth in a concentration-dependent manner. Overall, the addition of 0.2% PLA increased shoot biomass, while 2% PLA decreased it. Salinity increased Na content and decreased K/Na ratio in plant tissues (particularly roots), which were further modified by MPs. NaCl and MPs singly and jointly regulated the expression of functional genes related to salt tolerance in leaves, including ZMSOS1, ZMHKT1, and ZMHAK1. Exposure to NaCl alone had a slight effect on soil bacterial α-diversity, but in most cases, MPs increased ACE, Chao1, and Shannon indexes. Both MPs and NaCl altered bacterial community composition, although the specific effects varied depending on the type and concentration of MPs and the salinity level. Overall, PLA had more pronounced effects on soil-plant systems compared to PE. These findings bridge knowledge gaps in the risks of MPs in salt-affected habitats.
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Bactérias , Microplásticos , Microbiologia do Solo , Poluentes do Solo , Solo , Zea mays , Poluentes do Solo/toxicidade , Solo/química , Microplásticos/toxicidade , Zea mays/efeitos dos fármacos , Zea mays/crescimento & desenvolvimento , Bactérias/genética , Bactérias/efeitos dos fármacos , Bactérias/metabolismo , Bactérias/classificação , Cloreto de Sódio/toxicidade , Poliésteres , Salinidade , Polietileno , Microbiota/efeitos dos fármacosRESUMO
Cryopreservation of umbilical cord blood-derived mesenchymal stem cells (UCB-derived MSCs) is crucial step for its clinical applications in cell transplantation therapy. In the cryopreservation of MSCs, dimethyl sulfoxide has been widely used as a cryoprotectant (CPA). However, it has been proved that DMSO has toxic side effects to human body. In this study, DMSO-free CPA solutions which contained ethylene glycol (EG), 1, 2-propylene glycol (PG) and sucrose as basic CPAs, supplemented with polyvinyl alcohol (PVA) as an additive, were developed for the cryopreservation of UCB-derived MSCs. The cryopreservation of UCB-derived MSCs was achieved by vitrification via plunging into liquid nitrogen and by programmed freezing via an optical-DSC system respectively. The viability of thawed UCB-derived MSCs was tested by trypan blue exclusion assay. Results showed that the viability of thawed UCB-derived MSCs was enhanced from 71.2% to 95.4% in the presence of PVA for vitrification, but only < 10% to 45% of viability was found for programmed freezing. These results indicate that PVA exerts a beneficial effect on the cryopreservation of UCB-derived MSCs and suggest the vitrification in combination with the dimethyl sulfoxide free CPA solutions supplemented with PVA would be an efficient protocol for the cryopreservation of UCB-derived MSCs.
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Criopreservação , Sangue Fetal/citologia , Células-Tronco Mesenquimais/citologia , Sobrevivência Celular , Crioprotetores/farmacologia , Dimetil Sulfóxido/toxicidade , Etilenoglicol/farmacologia , Feminino , Sangue Fetal/efeitos dos fármacos , Feto , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Álcool de Polivinil/farmacologia , Propilenoglicol/farmacologia , Sacarose/farmacologia , Azul Tripano , Vitrificação/efeitos dos fármacosRESUMO
Periodontitis is a type of systemic immune inflammation that is caused by the complex infection of a variety of microorganisms in the subgingival plaque and the imbalance of the microbial ecological environment in the mouth. Periodontitis and chronic kidney disease (CKD) share many risk factors, such as obesity, smoking, and age. A growing body of data supports a strong correlation between periodontitis and kidney disease. Evidence supports the role of periodontal inflammation and elevated serum inflammatory mediators in renal atherosclerosis, renal deterioration, and end-stage renal disease (ESRD) development. Periodontitis is a risk factor for kidney disease. However, to our knowledge, there are few studies detailing the possible link between periodontitis and CKD. This review summarizes the possible mechanisms underlying periodontitis and CKD. More importantly, it highlights novel and potential pathogenic factors for CKD, including bacteria, pro-inflammatory mediators and oxidative stress. However, most research on the relationship between periodontitis and systemic disease has not determined causality, and these diseases are largely linked by bidirectional associations. Future research will focus on exploring these links to contribute to new treatments for CKD.
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Knowledge of the glass transition behavior of vitrification solutions is important for research and planning of the cryopreservation of biological materials by vitrification. This brief communication shows the analysis for the glass transition and glass stability of the multi-component vitrification solutions containing propanediol (PE), dimethyl sulfoxide (Me2SO) and polyvinyl alcohol (PVA) by using differential scanning calorimetry (DSC) during the cooling and subsequent warming between 25 and -150 degrees C. The glass formation of the solutions was enhanced by introduction of PVA. Partial glass formed during cooling and the fractions of free water in the partial glass matrix increased with the increasing of PVA concentration, which caused slight decline of glass transition temperature, T(g). Exothermic peaks of devitrification were delayed and broadened, which may result from the inhibition of ice nucleation or recrystallization of PVA.
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Crioprotetores/química , Transição de Fase , Varredura Diferencial de Calorimetria , Criopreservação/métodos , Dimetil Sulfóxido/química , Álcool de Polivinil/química , Propilenoglicóis/química , Temperatura , TermodinâmicaRESUMO
Coxsackievirus 16 (CA16) causes hand, foot, and mouth disease (HFMD) in young children and infants, and it can lead to fatal neurological complications. This study investigated antiviral effects of Siji Antiviral Mixture (SAM) on CA16 in neonatal mice and the protective effects of SAM on CA16 induced brain injuries. Neonatal BALB/c mice and SH-SY5Y cells were used and injected with CA16 stains to study the efficacy. ELISA and Western blotting were used to measure the cytokines levels and proteins expression. Genes transduction was also used to verify interaction mechanism. As the results shown, SAM could reduce the clinical scores at the beginning and delay disease development in vivo. Treatment with SAM decreased the levels of LDH, CK-MB, caspase 3 and Bax, ER stress, and inflammatory reaction induced by CA16 infection. Further siRNA transfection results showed that CA16 induced ER stress and inflammatory reaction through PERK/STAT3/NF-κB signaling and the protective effects of SAM might be through inhibiting PERK/STAT3/NF-κB signaling. HPLC analysis showed fingerprint profiles of SAM had 42 chromatographic peaks. Collectively, our study highlighted distinct roles of SAM in inhibiting CA16 infection and brain injury. The molecular mechanism of SAM might be through inhibiting PERK/STAT3/NF-κB signaling.
Assuntos
Antivirais/farmacologia , Lesões Encefálicas/metabolismo , NF-kappa B/fisiologia , Animais , Criança , Enterovirus , Humanos , Lactente , Camundongos , Camundongos Endogâmicos BALB C , Fator de Transcrição STAT3/metabolismo , eIF-2 Quinase/metabolismoRESUMO
OBJECTIVE: To study of exogenous substances on the germination rate of Platycodon grandiflorum and offer the basis for the standardized culture of P. grandiflorum. METHOD: At 25 degrees C, under darkness and lightness, do pretreatmeats on seeds by using GA3, H2O2, KMnO4, PEG at different concentrations. RESULT: The results indicated that the pretreatments with GA3 at a concentrating of 50-250 mg x L(-1) and 1%-2% of H2O2 could increase germination of P. grandiflorum seed at a certain degree. Contrariwise 0.1%-0.5% of KMnO4 and 15%-35% of PEG inhibited the germination. CONCLUSION: Pretreatment with 50-250 mg x L(-1)GA3 and H2O2 at a low cencontration could increase the seed germination of P. grandiflorum.
Assuntos
Germinação/fisiologia , Giberelinas/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Plantas Medicinais/crescimento & desenvolvimento , Platycodon/crescimento & desenvolvimento , Relação Dose-Resposta a Droga , Giberelinas/administração & dosagem , Peróxido de Hidrogênio/administração & dosagem , Peróxido de Hidrogênio/farmacologia , Luz , Plantas Medicinais/fisiologia , Platycodon/fisiologia , Polietilenoglicóis/farmacologia , Permanganato de Potássio/farmacologia , Sementes/crescimento & desenvolvimento , Sementes/fisiologiaRESUMO
The dehalogenation measurements could be carried out with the decomposition of plastic wastes simultaneously or successively. This paper reviewed the progresses in dehalogenation followed by thermochemical conversion of plastic-containing wastes for clean energy production. The pre-treatment method of MCT or HTT can eliminate the halogen in plastic wastes. The additives such as alkali-based metal oxides (e.g., CaO, NaOH), iron powders and minerals (e.g., quartz) can work as reaction mediums and accelerators with the objective of enhancing the mechanochemical reaction. The dehalogenation of waste plastics could be achieved by co-grinding with sustainable additives such as bio-wastes (e.g., rice husk), recyclable minerals (e.g., red mud) via MCT for solid fuels production. Interestingly, the solid fuel properties (e.g., particle size) could be significantly improved by HTT in addition with lignocellulosic biomass. Furthermore, the halogenated compounds in downstream thermal process could be eliminated by using catalysts and adsorbents. Most dehalogenation of plastic wastes primarily focuses on the transformation of organic halogen into inorganic halogen in terms of halogen hydrides or salts. The integrated process of MCT or HTT with the catalytic thermal decomposition is a promising way for clean energy production. The low-cost additives (e.g., red mud) used in the pre-treatment by MCT or HTT lead to a considerable synergistic effects including catalytic effect contributing to the follow-up thermal decomposition.
Assuntos
Fontes Geradoras de Energia , Halogenação , Incineração , Plásticos/química , Eliminação de Resíduos , Resíduos Sólidos/análise , Incineração/instrumentação , Eliminação de Resíduos/instrumentaçãoRESUMO
BACKGROUND AND OBJECTIVES: Glycopyrronium is a once-daily long-acting muscarinic antagonist for the maintenance treatment of patients with chronic obstructive pulmonary disease. This study assessed the pharmacokinetics of inhaled glycopyrronium 50 µg once-daily for 14 days in healthy Chinese subjects. METHODS: In this open-label study, 12 Chinese healthy subjects (six males and six females; mean age 23.1 years [range 18-26 years]) were enrolled and completed the study. Glycopyrronium in plasma was determined using validated liquid chromatography-mass spectrometry method with a lower limit of quantification of 1.5 pg/mL. Plasma pharmacokinetic parameters were determined on Day 1 after first dose and on Day 14 (steady state) after last dose using non-compartmental analysis. Trough pharmacokinetic samples (Days 5, 7, 10 and 12) were collected. Safety was also assessed. RESULTS: Glycopyrronium was rapidly absorbed into the systemic circulation after inhalation and its plasma concentrations decreased rapidly thereafter. Median time to reach maximum concentration (T max) was reached within 5 min after inhalation on both Days 1 and 14. Accumulation in the systemic exposure to glycopyrronium was observed from the time of first dose administration on Day 1 up to Day 14 and the observed accumulation ratio (R acc) values of area under the plasma drug concentration-time curve [AUC] from time 0 to 24 h post-dose (AUC0-24h) and maximum plasma drug concentration (C max) (Day 14/Day 1) were 2.77 and 1.59, respectively. The elimination half-life (T 1/2) was not reported. Mean effective half-life (T 1/2,acc) was 37.7 h. Pharmacokinetic steady state was reached after 5 days of daily dosing. One subject experienced dry mouth; otherwise glycopyrronium was well tolerated. CONCLUSIONS: Comparison of systemic exposure to glycopyrronium in Chinese versus the non-Chinese population did not indicate clinically relevant ethnic differences. Multiple inhaled doses of glycopyrronium were safe and well tolerated.
Assuntos
Glicopirrolato/farmacocinética , Antagonistas Muscarínicos/farmacocinética , Absorção Fisiológica , Administração por Inalação , Adolescente , Adulto , Área Sob a Curva , Povo Asiático , Disponibilidade Biológica , China , Feminino , Glicopirrolato/administração & dosagem , Glicopirrolato/efeitos adversos , Glicopirrolato/sangue , Meia-Vida , Humanos , Masculino , Antagonistas Muscarínicos/administração & dosagem , Antagonistas Muscarínicos/efeitos adversos , Antagonistas Muscarínicos/sangue , Adulto JovemRESUMO
BACKGROUND: Lecanicillium lecanii has been developed as a biopesticide and used in biological control of several agricultural insects. To improve fungal virulence, an optimised polyethylene glycol (PEG)-mediated protoplast transformation system was established for L. lecanii. Pr1A-like cuticle-degrading protease gene (Cdep1) from Beauveria bassiana was transferred into L. lecanii, and its resulting activity against Aphis gossypii was assessed. RESULTS: The optimised protoplast generation yielded 2.5 × 10(8) protoplasts g(-1) wet mycelium of fungi, and gave nearly 98% viability and 80% regeneration on plates. Protease activities were increased about fivefold in transformants expressing CDEP1. The median lethal concentration (LC50 ) for transformants expressing CDEP1 was twofold lower than that for the wild type (WT). The median survival time (LT50 ) for transformants expressing CDEP1 was also 14.2% shorter than that for WT, though no significant difference. There were no significant differences in conidial germination as colony growth and conidial yield on plates between transformants expressing CDEP1 and WT. The transformants expressing CDEP1 grew significantly quicker than WT in insects. The transformants expressing CDEP1 were lower in conidial yields on insect cadavers, but insignificantly different from WT. CONCLUSION: The PEG-mediated protoplast transformation system was effective for L. lecanii, and the expression of CDEP1 significantly enhanced fungal virulence against cotton aphids. © 2016 Society of Chemical Industry.
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Afídeos/microbiologia , Hypocreales/genética , Controle Biológico de Vetores/métodos , Polietilenoglicóis , Protoplastos , Animais , Beauveria/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Hypocreales/enzimologia , Hypocreales/patogenicidade , Peptídeo Hidrolases/genética , Peptídeo Hidrolases/metabolismo , Esporos Fúngicos/crescimento & desenvolvimento , VirulênciaRESUMO
Enterovirus 71 (EV71) infections can cause hand, foot and mouth disease (HFMD), meningoencephalitis, neonatal sepsis, and even fatal encephalitis in children. Unfortunately, there is currently no effective treatment for EV71 infection due to the lack of understanding of viral replication and infection; and viral infections have emerged as an imperative global hazard. Thus, it is extremely important to understand the mechanism of EV71 replication in order to prevent and control the diseases associated with EV71 infections. Early growth response-1 (EGR1) is a multifunctional transcription factor that regulates diverse biological functions, including inflammation, apoptosis, differentiation, tumorigenesis, and even viral infection. Here, we provide new insight into the role of EV71 infection in regulating EGR1 production; and reveal a novel mechanism by which EGR1 facilitates EV71 replication. We demonstrate that EV71 activates EGR1 expression during infection by stimulating the protein kinase A/protein kinase CÉ/phosphoinositide 3-kinase/Akt (PKA/PKCÉ/PI3K/Akt) cascade. We further reveal that EV71-activated EGR1, in turn, regulates the internal ribosomal entry site (IRES) of EV71 to enhance viral replication. In addition, EGR1 facilitates EV71 replication by binding directly to stem-loops I and IV of EV71 5'-untranslated region (5'UTR) with its first two zinc fingers. Moreover, EGR1 protein co-localizes with EV71 RNA in the cytoplasm of infected cells to facilitate viral replication. Our results reveal an important new role of EGR1 in viral infection, provide new insight into the novel mechanism underlying the regulation of EV71 replication, and suggest a potential application of EGR1 in the control of EV71 infection.
Assuntos
Proteína 1 de Resposta de Crescimento Precoce/fisiologia , Enterovirus Humano A/fisiologia , Genoma Viral , RNA Viral/metabolismo , Replicação Viral , Células Cultivadas , Proteína 1 de Resposta de Crescimento Precoce/metabolismo , Infecções por Enterovirus/genética , Infecções por Enterovirus/virologia , Regulação da Expressão Gênica , Células HEK293 , Humanos , Ligação Proteica , Replicação Viral/genéticaRESUMO
PURPOSE: To investigate the methods of isolation, culture and identification of BMMSCs derived from rabbit mandible. METHODS: BMMSCs were collected from rabbit mandible and isolated by density gradient centrifugation. Cells were adherently cultured in vitro, and P2 or P3 BMMSCs populations were collected and examined. Cell growth was observed by inverted microscopy; the propagation of BMMSCs were tested by MTT and a growth curve was drawn after statistical analysis; colony-forming unit-fibroblast(CFU-F) was detected by examination of colony formation; the potential of multi-directional differentiation into osteoblasts, adipocytes and skeletal muscle cells was estimated by pertinent methods; the surface marks of BMMSCs were detected by flow cytometry, the data was analysed using SPSS16.0 software package. RESULTS: The majority of adherent cells were long fusiform, and few were small triangle; the growth curve of BMMSCs showed that every passage experienced incubation period, log phase and platform period; the rate of colony formation was 37%. Growth of BMMSCs represented the appearance of CFU-F; BMMSCs after inducted differentiation showed osteogenic and adipogenic potential. The staining of mineralized nodules was positive by alizarin red S and the positive staining of oil red O appeared in lipid drops around cell nucleus. The staining of skeletal muscle cells was positive by desmin immunofluorescence; the cell surface marks assessed with flow cytometry indicated that these BMMSCs expressed CD90 and CD146 in high percentage (about 98.7% and 98.1%, respectively). CONCLUSIONS: The highly uniformed BMMSCs derived from rabbit mandible can be collected. These BMMSCs have the ability of self-replication and propagation, as well as potential of multi-directional differentiation in vitro.
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Células da Medula Óssea , Células-Tronco Mesenquimais , Animais , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Fibroblastos , Citometria de Fluxo , Mandíbula , CoelhosRESUMO
Given its essential role in the process of hepatitis C virus (HCV) replication, the viral NS3/4A serine protease is arguably the most thoroughly characterized HCV enzyme and the most intensively pursued anti-HCV target for drug development thus far. Recent data have demonstrated promise for the NS3 protease inhibitor boceprevir, which, when added to the standard of care peginterferon and ribavirin, improves sustained virological response while shortening duration of therapy in genotype-1-infected individuals. This review discusses the mechanism of action of boceprevir, its effects on HCV, and its viral resistance.
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Antivirais/uso terapêutico , Hepatite C/tratamento farmacológico , Prolina/análogos & derivados , Farmacorresistência Viral , Hepacivirus/fisiologia , Hepatite C/virologia , Humanos , Interferon alfa-2 , Interferon-alfa/uso terapêutico , Polietilenoglicóis/uso terapêutico , Prolina/uso terapêutico , Proteínas Recombinantes/uso terapêutico , Ribavirina/uso terapêutico , Proteínas não Estruturais Virais/antagonistas & inibidoresRESUMO
Enterovirus 71 (EV 71) is one of the commonest causative agents of hand, foot, and mouth disease (HFMD), which infects mainly young children. It has been associated with severe neurological complications worldwide, and has caused significant deaths in many provinces of China from March to May 2008. In this study, RNA interference (RNAi) was used as an antiviral agent to inhibit EV 71 replication in rhabdomyosarcoma (RD) cells. Three small interfering RNAs (siRNAs) targeting extremely conserved regions among multiple EV 71 strains in China could effectively block the replication of EV 71 strain Shzh-98. Combination transfection of these three siRNAs could produce a strong inhibitory effect not only in strain Shzh-98, but also in one epidemic strain Fuyang-0805 isolated from a child in the city of Fuyang with a clinical diagnosis of HFMD in 2008. These strategies and results suggest that RNAi has potential therapeutic use for the suppression of EV 71 infection in a broad spectrum of viral strains.