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1.
Arch Microbiol ; 206(4): 183, 2024 Mar 19.
Artículo en Inglés | MEDLINE | ID: mdl-38502272

RESUMEN

This study aimed to reveal that the effect of biosurfactant on the dispersion and degradation of crude oil. Whole genome analysis showed that Pseudomonas aeruginosa GB-3 contained abundant genes involved in biosurfactant synthesis and metabolic processes and had the potential to degrade oil. The biosurfactant produced by strain GB-3 was screened by various methods. The results showed that the surface tension reduction activity was 28.6 mN·m-1 and emulsification stability was exhibited at different pH, salinity and temperature. The biosurfactant was identified as rhamnolipid by LC-MS and FTIR. The fermentation conditions of strain GB-3 were optimized by response surface methodology, finally the optimal system (carbon source: glucose, nitrogen source: ammonium sulfate, C/N ratio:16:1, pH: 7, temperature: 30-35 °C) was determined. Compared with the initial fermentation, the yield of biosurfactant increased by 4.4 times after optimization. In addition, rhamnolipid biosurfactant as a dispersant could make the dispersion of crude oil reach 38% within seven days, which enhanced the bioavailability of crude oil. As a biostimulant, it could also improve the activity of indigenous microorganism and increase the degradation rate of crude oil by 10-15%. This study suggested that rhamnolipid biosurfactant had application prospect in bioremediation of marine oil-spill.


Asunto(s)
Petróleo , Pseudomonas aeruginosa , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/metabolismo , Tensoactivos/química , Glucolípidos/química , Petróleo/metabolismo
2.
Environ Sci Pollut Res Int ; 31(7): 10802-10817, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38212565

RESUMEN

Bioremediation is a sustainable and pollution-free technology for crude oil-contaminated soil. However, most studies are limited to the remediation of shallow crude oil-contaminated soil, while ignoring the deeper soil. Here, a high-efficiency composite microbial agent MAB-1 was provided containing Bacillus (naphthalene and pyrene), Acinetobacter (cyclohexane), and Microbacterium (xylene) to be synergism degradation of crude oil components combined with other treatments. According to the crude oil degradation rate, the up-layer (63.64%), middle-layer (50.84%), and underlying-layer (54.21%) crude oil-contaminated soil are suitable for bioaugmentation (BA), biostimulation (BS), and biostimulation+bioventing (BS+BV), respectively. Combined with GC-MS and carbon number distribution analysis, under the optimal biotreatment, the degradation rates of 2-ring and 3-ring PAHs in layers soil were about 70% and 45%, respectively, and the medium and long-chain alkanes were reduced during the remediation. More importantly, the relative abundance of bacteria associated with crude oil degradation increased in each layer after the optimal treatment, such as Microbacterium (2.10-14%), Bacillus (2.56-12.1%), and Acinetobacter (0.95-12.15%) in the up-layer soil; Rhodococcus (1.5-6.9%) in the middle-layer soil; and Pseudomonas (3-5.4%) and Rhodococcus (1.3-13.2%) in the underlying-layer soil. Our evaluation results demonstrated that crude oil removal can be accelerated by adopting appropriate bioremediation approach for different depths of soil, providing a new perspective for the remediation of actual crude oil-contaminated sites.


Asunto(s)
Microbiota , Petróleo , Contaminantes del Suelo , Biodegradación Ambiental , Suelo , Contaminantes del Suelo/análisis , Petróleo/metabolismo , Microbiología del Suelo , Hidrocarburos/metabolismo
3.
Environ Res ; 223: 115465, 2023 04 15.
Artículo en Inglés | MEDLINE | ID: mdl-36773642

RESUMEN

Bioaugmentation (BA) of oil-contaminated soil by immobilized microorganisms is considered to be a promising technology. However, available high-efficiency microbial agents remain very limited. Therefore, we prepared a SA/GO/C5 immobilized gel pellets by embedding the highly efficient crude oil degrading bacteria Bacillus C5 in the SA/GO composite material. The optimum preparation conditions of SA/GO/C5 immobilized gel pellets were: SA 3.0%, GO 25.0 µg/mL, embedding amount of C5 6%, water bath temperature of 50°C, CaCl2 solution concentration 3% and cross-linking time 20 h. BA experiments were carried out on crude oil contaminated soil to explore the removal effect of SA/GO/C5 immobilized pellets. The results showed that the SA/GO/C5 pellets exhibited excellent mechanical strength and specific surface area, which facilitated the attachment and growth of the Bacillus C5. Compared with free bacteria C5, the addition of SA/GO/C5 significantly promoted the removal of crude oil in soil, reaching 64.92% after 30 d, which was 2.1 times the removal rate of C5. The addition of SA/GO/C5 promoted the abundance of soil exogenous Bacillus C5 and indigenous crude oil degrading bacteria Alcanivorax and Marinobacter. In addition, the enrichment of hydrocarbon degradation-related functional abundance was predicted by PICRUSt2 in the SA/GO/C5 treatment group. This study demonstrated that SA/GO/C5 is an effective method for remediating crude oil-contaminated soil, providing a basis and option for immobilized microorganisms bioaugmentation to remediate organic contaminated soil.


Asunto(s)
Bacillus , Microbiota , Petróleo , Contaminantes del Suelo , Bacillus/metabolismo , Biodegradación Ambiental , Petróleo/metabolismo , Hidrocarburos , Contaminantes del Suelo/análisis , Bacterias/metabolismo , Suelo/química , Microbiología del Suelo
4.
J Hazard Mater ; 447: 130808, 2023 04 05.
Artículo en Inglés | MEDLINE | ID: mdl-36669400

RESUMEN

Biodegradation is one of the safest and most economical methods for the elimination of toxic chlorophenols and crude oil from the environment. In this study, aerobic degradation of the aforementioned compounds by composite microbial agent B-Cl, which consisted of Bacillus B1 and B2 in a 3:2 ratio, was analyzed. The biodegradation mechanism of B-Cl was assessed based on whole genome sequencing, Fourier transform infrared spectroscopy and gas chromatographic analyses. B-Cl was most effective at reducing Cl- concentrations (65.17%) and crude oil biodegradation (59.18%) at 7 d, which was when the content of alkanes ≤ C30 showed the greatest decrease. Furthermore, adding B-Cl solution to soil significantly decreased the 2,4-DCP and oil content to below the detection limit and by 80.68%, respectively, and reconstructed of the soil microbial into a system containing more CPs-degrading (exaA, frmA, L-2-HAD, dehH, ALDH, catABE), aromatic compounds-degrading (pcaGH, catAE, benA-xylX, paaHF) and alkane- and fatty acid-degrading (alkB, atoB, fadANJ) microorganisms. Moreover, the presence of 2,4-DCP was the main hinder of the observed effects. This study demonstrates the importance of adding B-Cl solution to determine the interplay of CPs with microbes and accelerating oil degradation, which can be used for in-situ bioremediation of CPs and oil-contaminated soil.


Asunto(s)
Microbiota , Petróleo , Contaminantes del Suelo , Biodegradación Ambiental , Bacterias/genética , Bacterias/metabolismo , Petróleo/metabolismo , Microbiología del Suelo , Alcanos/metabolismo , Suelo , Contaminantes del Suelo/metabolismo
5.
Biomed Pharmacother ; 158: 114066, 2023 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-36528915

RESUMEN

Lingguizhugan Decoction (LGZGD) is a classical traditional Chinese medicine prescription. Our previous studies found that disorders of lipid metabolism were reversed by LGZGD in heart failure (HF) mice. This study aimed to reveal the regulation of lipid metabolism of LGZGD. A mice model of HF was established by intraperitoneal injection of doxorubicin. The components of LGZGD were identified with the UHPLC-QTOF-MS method. The regulation of lipid metabolism by LGZGD was detected by serum lipidomics and heart tissue proteomics. Molecular docking was further performed to screen active components. A total of 78 compounds in LGZGD were identified. Results of lipidomics showed that 37 lipids illustrated a significant recovery trend to normal after the treatment of LGZGD. Results of proteomics demonstrated that 55 proteins were altered by the administration of LGZGD in HF mice. After enrichment analysis, the Prakg2/Ucp2/Plin1 axis on the Apelin pathway plays a vital role in HF treatment by LGZGD. Nine active components exhibited the outstanding ability of binding to the apelin receptor with MM-GBSA value lower than -60 Kcal/mol. In conclusion, all results combined together revealed that multi-component in the LGZGD had beneficial effects on the HF through ameliorating lipid disorders, which provides a novel insight into the cardioprotective effects of LGZGD and its clinical application.


Asunto(s)
Medicamentos Herbarios Chinos , Insuficiencia Cardíaca , Ratones , Animales , Lipidómica/métodos , Metabolismo de los Lípidos , Proteómica , Simulación del Acoplamiento Molecular , Insuficiencia Cardíaca/tratamiento farmacológico , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico
6.
Environ Sci Pollut Res Int ; 30(13): 35670-35682, 2023 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-36538219

RESUMEN

The groundwater polluted by an agricultural hormone site was taken as the research object, and a total of 7 groundwater samples were collected at different locations in the plant. The main pollutants in the research area were determined to be extractable petroleum hydrocarbons (C10-C40); 1,2-dichloroethane; 1,1,2-trichloroethane; carbon tetrachloride; vinyl chloride, and chloroform; the maximum content of these pollutants can reach 271 mg/L, 1.68 × 107 µg/L, 1.56 × 104 µg/L, 9.53 × 104 µg/L, 6.58 × 104 µg/L, and 4.81 × 104 µg/L, respectively. Aiming at the problems of groundwater pollution in this area, two sets of oxidation experiments have been carried out. The addition of NaHSO3 modified Fenton oxidation system was used in this contaminated water, which enhanced (2.2 ~ 46.7%) chemical oxygen demand (COD) removal rate. The highest removal rate of extractable petroleum hydrocarbons (C10-C40) can reach 99%. And the degradation rate of chlorinated hydrocarbon pollutants can reach 99% to 100%, which almost achieved the purpose of complete removal. In the NaHSO3 modified Fenton oxidation system, the addition of NaHSO3 accelerates the cycle of Fe3+/Fe2+ and ensures the continuous existence of Fe2+ in the reaction system, thereby producing more ·OH and further oxidizing and degrading organic pollutants. Our work has provided important insights for this economically important treatment of this type water body and laid the foundation for the engineering of this method.


Asunto(s)
Contaminantes Ambientales , Agua Subterránea , Petróleo , Contaminantes Químicos del Agua , Contaminantes Químicos del Agua/análisis , Peróxido de Hidrógeno/química , Oxidación-Reducción , Agua Subterránea/química , Contaminación del Agua , Hidrocarburos/química , Agua
7.
Chemosphere ; 313: 137367, 2023 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-36427578

RESUMEN

The chitosan-biochar composite is a clean and environmentally friendly immobilized microorganisms carrier. In this study, the chitosan-biochar composite as a carrier to immobilize a compound microbial agent contained Pseudomonas aeruginosa and Bacillus licheniformis, and investigated its role in the remediation of oil-contaminated soil. When using 1% (v/v) acetic acid, 3% (m/v) chitosan solution, 0.1% biochar, 4% (v/v) NaOH solution, freeze-drying 6 h, the optimal chitosan-biochar composite material could be obtained. The specific surfacearea of the material increased to 1.725 m2/g and the average pore size also increased from 130.2260 nm to 165.2980 nm after the addition of biochar through the analysis of specific surface area and pore size, which enlarged the contact area of microorganisms and crude oil with the material. SEM showed that the bacterial successfully adhered to the surface and internal of the material. Using FTIR, the results showed that the synthesis of composite carrier material was the covalent combination of -NH2 on chitosan and -COOH on biochar, forming a new chemical bond -NH-CO-. After 60 days of remediation of oil-contaminated soil, the removal rate of crude oil by chitosan-biochar composite immobilized microorganism method was 45.82%, which was 21.26% higher than that of natural remediation. Simultaneously, several oil-degrading bacteria increased at genus level, including Nocardioides (26.79%-33.09%), Bacillus (3.01%-4.10%), Dietzia (1.84%-5.56%), Pseudomonas (0-0.78%), among which Pseudomonas belongs to exogenous bacteria. The results indicated that the chitosan-biochar composite material has high application value in removing crude oil, and further provides a new strategy for bioremediation of oil-contaminated soil.


Asunto(s)
Quitosano , Petróleo , Contaminantes del Suelo , Biodegradación Ambiental , Suelo/química , Carbón Orgánico/química , Bacterias , Contaminantes del Suelo/análisis , Microbiología del Suelo
8.
Biodegradation ; 33(5): 461-476, 2022 10.
Artículo en Inglés | MEDLINE | ID: mdl-35729449

RESUMEN

In order to enhance the degradation effect of microorganisms on crude oil in the existence of chlorophenol compounds, oil-degrading bacteria C4 (Alcaligenes faecails), C5 (Bacillus sp.) and 2,4-dichlorophenol (2,4-DCP) degrading bacteria L3 (Bacillus marisflavi), L4 (Bacillus aquimaris) were isolated to construct a highly efficient consortium named (C4C5 + L3L4). When the compound bacteria agent combination by VC4: VC5: VL3: VL4 = 1:2:2:1, the crude oil degradation efficiency of 7 days was stable at 50.63% ~ 55.43% under different conditions. Degradation mechanism was analyzed by FTIR, GC-MS and IC technology and the following conclusions showed that in the system of adding consortium (C4C5 + L3L4), the heavy components were converted into saturated and unsaturated components. The bacterial consortium could first degrade medium and long chain alkanes into short chain hydrocarbons and then further degrade. And the dechlorination efficiency of 2,4-DCP in the degradation system reached 73.83%. The results suggested that the potential applicability and effectiveness of the selected bacteria consortium for the remediation of oil-contaminated water or soil with the existence of chlorophenol compound.


Asunto(s)
Clorofenoles , Petróleo , Contaminantes del Suelo , Bacterias/metabolismo , Biodegradación Ambiental , Clorofenoles/metabolismo , Hidrocarburos/metabolismo , Petróleo/análisis , Microbiología del Suelo , Contaminantes del Suelo/metabolismo
9.
J Pharm Biomed Anal ; 216: 114811, 2022 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-35576890

RESUMEN

Tripterygium wilfordii genus preparations (TWGP) are widely used in traditional Chinese medicines for the treatment of autoimmune diseases and immune diseases with definite therapeutic effects but high toxicity. The aim of this study is to identify and compare chemical compounds in three types of commercial TWGP using UHPLC-QTOF-MS/MS and molecular networking (MN) technology. First, the mass fragmentation pathways of 10 compounds were investigated, which included two sesquiterpene alkaloids, four diterpenoids, and four triterpenoids. The chemical compounds were then identified using UHPLC-QTOF-MS/MS and a conventional database. Following that, molecular network technology was used to further identify the GNPS platform. Finally, metabonomic data analysis was used to compare 92 commercial TWGP samples from 13 manufacturers. A total of 103 compounds were identified, with the molecular network detecting 40 of them. Moreover, the quality of commercial Tripterygium glycoside tablets varies greatly and 26 compounds, including triptolide, wilforine, wilforgine, and demethylzeylasteral, were discovered to be the main differential compounds in tripterygium glycosides tablets. This was the first time MN technology was used for compound analysis in TWGP, laying the foundation for classifying effective and toxic substances and TWGP quality control.


Asunto(s)
Medicamentos Herbarios Chinos , Tripterygium , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/química , Glicósidos , Comprimidos/química , Espectrometría de Masas en Tándem , Tripterygium/química
10.
J Ethnopharmacol ; 295: 115366, 2022 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-35551974

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: As a classical formula of traditional Chinese medicine (TCM), Lingguizhugan Decoction (LGZGD) has been used for treating heart failure (HF) because it has an efficiency of yang-warming and fluid-dispersing. However, the pharmacodynamic material basis of LGZGD responsible for the therapeutic benefits is not well understood. AIM OF THE STUDY: The aim of this study was to elucidate the pharmacodynamic material basis of LGZGD by an integrated approach. MATERIALS AND METHODS: Following oral administration of LGZGD in mice, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) was used to identify prototype substances. A heart failure (HF) model was established, followed by an untargeted metabolomics study to determine potential targets of LGZGD. The network pharmacology method was performed to screen substances that interacted with potential targets of LGZGD treating HF. Molecular docking technology was applied to further screen substances based on binding energy. Cell viability assays were conducted to verify pharmacodynamic effects of selected substances. RESULTS: In all, forty-two prototype substances were identified in the blood, urine, and fecal samples of mice. A total of fifty-five differential metabolites were identified using heart tissue untargeted metabolomics. Twenty-five substances of LGZGD were screened relating to thirty-three targets treating HF. Twenty-two substances were filtered according to their binding energy using molecular docking technology. Cell experiments revealed cinnamaldehyde, glycyrrhetinic acid, kaempferol, daidzein, caffeic acid, and catechin could significantly improve the survival rate of H9c2 cells, which might be the pharmacodynamic material basis of LGZGD. CONCLUSIONS: A scientific approach that integrated in vivo substances identification, metabolomics, network pharmacology, molecular docking, and cell pharmacodynamic assay has been developed to study the pharmacodynamic material basis of LGZGD in the treatment of HF.


Asunto(s)
Medicamentos Herbarios Chinos , Insuficiencia Cardíaca , Animales , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Insuficiencia Cardíaca/tratamiento farmacológico , Ratones , Simulación del Acoplamiento Molecular , Espectrometría de Masas en Tándem/métodos
11.
Int J Mol Sci ; 22(8)2021 Apr 09.
Artículo en Inglés | MEDLINE | ID: mdl-33918544

RESUMEN

Soybean (Glycine max) oil is one of the most widely used vegetable oils across the world. Breeding of soybean to reduce the saturated fatty acid (FA) content, which is linked to cardiovascular disease, would be of great significance for nutritional improvement. Acyl-acyl carrier protein thioesterases (FATs) can release free FAs and acyl-ACP, which ultimately affects the FA profile. In this study, we identified a pair of soybean FATB coding genes, GmFATB1a and GmFATB1b. Mutants that knock out either or both of the GmFATB1 genes were obtained via CRISPR/Cas9. Single mutants, fatb1a and fatb1b, showed a decrease in leaf palmitic and stearic acid contents, ranging from 11% to 21%. The double mutant, fatb1a:1b, had a 42% and 35% decrease in palmitic and stearic acid content, displayed growth defects, and were male sterility. Analysis of the seed oil profile revealed that fatb1a and fatb1b had significant lower palmitic and stearic acid contents, 39-53% and 17-37%, respectively, while that of the unsaturated FAs were the same. The relative content of the beneficial FA, linoleic acid, was increased by 1.3-3.6%. The oil profile changes in these mutants were confirmed for four generations. Overall, our data illustrate that GmFATB1 knockout mutants have great potential in improving the soybean oil quality for human health.


Asunto(s)
Sistemas CRISPR-Cas , Ácidos Grasos/metabolismo , Técnicas de Inactivación de Genes , Marcación de Gen , Glycine max/genética , Glycine max/metabolismo , Tioléster Hidrolasas/deficiencia , Expresión Génica , Estudios de Asociación Genética , Ingeniería Genética , Humanos , Mutación , Fenotipo , Proteínas de Plantas/genética , Aceite de Soja/genética , Aceite de Soja/metabolismo
12.
Discov Med ; 29(158): 145-157, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33007190

RESUMEN

Coronavirus disease 2019 (COVID-19), a newly identified acute respiratory disease caused by a strain of novel coronavirus (SARS-CoV-2), has become a worldwide pandemic. From December 2019 to present, millions of cases have been reported, bringing unprecedented pressure on both health and epidemic prevention services in every country. As frontline healthcare workers, ophthalmologists face an increased threat of viral infection, not only because of close contact with patients during examinations or operations, but also due to evidence showing that ocular fluids such as tears or conjunctival secretions may carry the virus. The risk that healthcare workers face is emphasized by the loss of our colleagues who have sacrificed themselves in combating the virus. As a result, it is necessary to have a comprehensive understanding of the threats that we face. In the first part of this review, we start by explaining the structure of SARS-CoV-2 and examining its transmission and means of infection. Next, we summarize the latest scientific advancements of epidemiology, clinical presentations, and current treatments of COVID-19. In the second half of the review, we emphasize the ocular transmission, symptomatic manifestations, and the essential knowledge in an ophthalmology clinic setting. As the pandemic of COVID-19 continues to pose a threat to global health, we hope that this review makes a contribution to combating COVID-19.


Asunto(s)
Betacoronavirus/patogenicidad , Infecciones por Coronavirus/complicaciones , Oftalmopatías/virología , Neumonía Viral/complicaciones , Antivirales/uso terapéutico , Betacoronavirus/efectos de los fármacos , Betacoronavirus/inmunología , COVID-19 , Prueba de COVID-19 , Técnicas de Laboratorio Clínico/métodos , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/terapia , Infecciones por Coronavirus/transmisión , Reposicionamiento de Medicamentos , Oftalmopatías/diagnóstico , Oftalmopatías/inmunología , Oftalmopatías/terapia , Humanos , Inmunización Pasiva/métodos , Factores Inmunológicos/uso terapéutico , Medicina Tradicional China/métodos , Pandemias , Neumonía Viral/epidemiología , Neumonía Viral/terapia , Neumonía Viral/transmisión , SARS-CoV-2 , Sueroterapia para COVID-19
13.
J Trace Elem Med Biol ; 58: 126438, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-31760326

RESUMEN

BACKGROUND: Observational studies have suggested that selenium levels might associate with the risk of cardio-metabolic diseases, but how circulating selenium is related to dyslipidemia remains inconclusive. OBJECTIVES: To investigate the association of circulating selenium levels with lipid profiles and dyslipidemia among US adults. METHODS: Using the data collected from the National Health and Nutrition Examination Survey (NHANES 1999-2006), we performed multivariate logistic regression to examine the association of circulating selenium levels (in quartiles) with total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), non-HDL-C, and atherogenic index (AI). RESULTS: We included 2903 adults (49.3 % male) (average age: 61.9) for analysis. Circulating selenium had non-linear association with TC, LDL-C, HDL-C, and AI (all p < 0.05). When comparing with the lowest quartile, subjects with the highest quartile of circulating selenium (>147.00 µg/L) had the higher odds of elevated TG (OR: 1.75, 95% CI = 1.14, 2.68), TC (OR: 2.47, 95% CI = 1.62, 3.76), LDL-C (OR: 2.52, 95% CI = 1.60, 3.96), non-HDL-C (OR: 2.17, 95% CI = 1.41, 3.33), AI (OR: 1.20, 95% CI = 0.73, 1.97) and low-HDL-C (OR: 2.10, 95% CI = 1.19, 3.72). Similar patterns were observed in subgroup analysis. CONCLUSIONS: Higher circulating selenium levels had non-linear association with lipid profiles and the increased odds of dyslipidemia.


Asunto(s)
Dislipidemias/sangre , Encuestas Nutricionales , Selenio/sangre , Femenino , Humanos , Lípidos/sangre , Masculino , Persona de Mediana Edad , Análisis Multivariante , Oportunidad Relativa
14.
Biomed Chromatogr ; 31(4)2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-27644052

RESUMEN

A novel, reliable and reproducible UPLC-MS/MS method was developed and validated to determine simultaneously the 10 bioactive constituents (baicalin, baicalein, wogonoside, wogonin, luteoloside, dictamnine, fraxinellone, obacunone, geniposidic acid and glycyrrhizic acid) in Jixingshizhen (JXSZ) preparation. Briefly, chromatographic separation was achieved on a Waters BEH C18 column with gradient elution employing a mobile phase composed of acetonitrile and 0.1% formic acid at a flow rate of 0.3 mL/min. All analytes containing internal standard (verapamil) were detected without interference in the multiple reaction monitoring mode with positive electrospray ionization. Further, a comprehensive validation of the method was rigorously executed according to the guidelines of the International Conference on Harmonization and Chinese Pharmacopoeia (2015 Edition). The results indicated that the validated method afforded desired linearity, precision, accuracy, sensitivity and stability. At length, the newly established method was successfully applied to quantify the 10 effective ingredients in JXSZ granules from different production batches, indicating the proposed method in this paper was particularly preferable for the routine analysis of JXSZ preparation as well as the quality control, particularly in situations where high sample throughput and fast analytical speed are required.


Asunto(s)
Cromatografía Liquida/métodos , Medicamentos Herbarios Chinos/análisis , Espectrometría de Masas en Tándem/métodos , Fraccionamiento Químico , Cromatografía Liquida/normas , Medicamentos Herbarios Chinos/química , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masas en Tándem/normas
15.
Proc Natl Acad Sci U S A ; 113(11): 2868-73, 2016 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-26929348

RESUMEN

A central challenge to the development of protein-based therapeutics is the inefficiency of delivery of protein cargo across the mammalian cell membrane, including escape from endosomes. Here we report that combining bioreducible lipid nanoparticles with negatively supercharged Cre recombinase or anionic Cas9:single-guide (sg)RNA complexes drives the electrostatic assembly of nanoparticles that mediate potent protein delivery and genome editing. These bioreducible lipids efficiently deliver protein cargo into cells, facilitate the escape of protein from endosomes in response to the reductive intracellular environment, and direct protein to its intracellular target sites. The delivery of supercharged Cre protein and Cas9:sgRNA complexed with bioreducible lipids into cultured human cells enables gene recombination and genome editing with efficiencies greater than 70%. In addition, we demonstrate that these lipids are effective for functional protein delivery into mouse brain for gene recombination in vivo. Therefore, the integration of this bioreducible lipid platform with protein engineering has the potential to advance the therapeutic relevance of protein-based genome editing.


Asunto(s)
Técnicas de Inactivación de Genes , Genes Sintéticos , Ingeniería Genética/métodos , Lípidos/química , Nanopartículas , Animales , Proteínas Bacterianas/administración & dosificación , Proteínas Bacterianas/genética , Proteína 9 Asociada a CRISPR , Sistemas CRISPR-Cas , Ceramidas/química , Colesterol/química , Portadores de Fármacos , Endocitosis , Endonucleasas/administración & dosificación , Endonucleasas/genética , Endosomas/metabolismo , Genes Reporteros , Proteínas Fluorescentes Verdes/biosíntesis , Proteínas Fluorescentes Verdes/genética , Células HeLa , Humanos , Hipotálamo/metabolismo , Integrasas/administración & dosificación , Integrasas/genética , Lípidos/administración & dosificación , Lípidos/síntesis química , Proteínas Luminiscentes/biosíntesis , Proteínas Luminiscentes/genética , Ratones , Estructura Molecular , Nanopartículas/administración & dosificación , Nanopartículas/química , Nanopartículas/metabolismo , Nanopartículas/toxicidad , Fosfatidiletanolaminas/química , ARN/genética , Proteínas Recombinantes/biosíntesis , Recombinación Genética , Electricidad Estática , Relación Estructura-Actividad , Tálamo/metabolismo
16.
Zhen Ci Yan Jiu ; 39(6): 466-71, 2014 Dec.
Artículo en Chino | MEDLINE | ID: mdl-25632571

RESUMEN

OBJECTIVE: To observe the effect of acupuncture therapy on 14-3-3, Bcl-2 and Bax expression levels in the cerebral cortex in neonatal rats with hypoxic-ischemic brain damage(HIBD). METHODS: Timed pregnant Sprague-Dawley rat dams were delivered either vaginally (normal group), or by C-section (sham-operation group) or by C-section with 5 min of global anoxia (anoxia group), with 8 rats in each group. The rat pups of the anoxia group were randomly divided into model group and acupuncture group (n =8). Acupuncture stimulation of "Naosanzhen" "Niesanzhen" and "Zhisanzhen" acupoints was given begin- ning from the 14th day after birth, once daily for 7 consecutive days. All rat pups were killed by decapitation on day 21 after birth, and then 14-3-3, Bcl-2 and Bax immunoactivity (expression) in the cerebral cortex were detected by immunohistochemistry. RESULTS: In comparison with the normal group, the expression level of cerebral cortical 14-3-3 was significantly decreased, and that of Bax remarkably increased in the model group (P<0. 01, P<0. 05). Compared to the model group, cortical 14-3-3 and Bcl-2 expression levels were markedly up-regulated in the acupuncture group (P<0.01, P<0.05). Compared to the normal group, cortical 14-3-3 expression level was obviously lower, but Bax expression level significantly higher in the sham-operation group (P<0. 05, P<0. 01). No significant differences were found between the model and normal groups in the expression levels of Bcl-2, and between the acupuncture and model groups in the expression levels of Bax (P>0. 05). CONCLUSION: Acupuncture intervention can increase the expression of 14-3-3 and Bcl-2 in the cerebral cortex in HIBD rats.


Asunto(s)
Proteínas 14-3-3/análisis , Terapia por Acupuntura , Corteza Cerebral/química , Hipoxia-Isquemia Encefálica/terapia , Animales , Femenino , Hipoxia-Isquemia Encefálica/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Ratas , Ratas Sprague-Dawley , Proteína X Asociada a bcl-2/análisis
17.
Zhongguo Zhen Jiu ; 32(9): 807-9, 2012 Sep.
Artículo en Chino | MEDLINE | ID: mdl-23227688

RESUMEN

The sources of clinical efficacy of slow needle insertion is discussed. From the basic factors of slow needle insertion, the requirements of this manipulation, main points of 4 steps (to straight the needle, to press softly, to twirl the needle and to hold without brute force), keys of operation and its significance are analyzed. Slow needle insertion is not only beneficial to the management and regulation of Shen (spirit) and induction of qi in the process of the spirit unity of doctor and patient, but also accumulates the needling sensation among three levels of the superficial, middled and deep layers of an acupoint, which could promote the curative effect.


Asunto(s)
Terapia por Acupuntura/métodos , Terapia por Acupuntura/historia , Terapia por Acupuntura/instrumentación , China , Historia del Siglo XX , Historia del Siglo XXI , Historia Antigua , Humanos , Medicina en la Literatura , Agujas
18.
J Sep Sci ; 34(9): 1069-75, 2011 May.
Artículo en Inglés | MEDLINE | ID: mdl-21416601

RESUMEN

Headspace single drop microextraction (HS-SDME) coupled with microwave extraction (ME) was developed and applied to the extraction of the essential oil from dried Syzygium aromaticum (L.) Merr. et Perry and Cuminum cyminum L. The operational parameters, such as microdrop volume, microwave absorption medium (MAM), extraction time, and microwave power were optimized. Ten microliters of decane was used as the microextraction solvent. Ionic liquid and carbonyl iron powder were used as MAM. The extraction time was less than 7 min at the microwave power of 440 W. The proposed method was compared with hydrodistillation (HD). There were no obvious differences in the constituents of essential oils obtained by the two methods.


Asunto(s)
Fraccionamiento Químico/métodos , Cuminum/química , Myrtaceae/química , Aceites Volátiles/aislamiento & purificación , Aceites de Plantas/aislamiento & purificación , Fraccionamiento Químico/instrumentación , Microondas , Aceites Volátiles/análisis
19.
J Chromatogr B Analyt Technol Biomed Life Sci ; 878(28): 2707-11, 2010 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-20810325

RESUMEN

The arctiin and arctigenin in the fruit of Arctium lappa L. were extracted by matrix solid-phase dispersion (MSPD) and determined by high-performance liquid chromatography (HPLC) with fluorescence detection. The experimental conditions for the MSPD were optimized. Silica gel was selected as dispersion adsorbent and methanol as elution solvent. The calibration curve showed good relationship (r>0.9998) in the concentration range of 0.010-5.0µgmL(-1) for arctiin and 0.025-7.5µgmL(-1) for arctigenin. The recoveries were between 74.4% and 100%. The proposed method consumed less sample, time and solvent compared with conventional methods, including ultrasonic and Soxhlet extraction.


Asunto(s)
Arctium/química , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/química , Frutas/química , Furanos/análisis , Glucósidos/análisis , Lignanos/análisis , Extracción en Fase Sólida/métodos , Adsorción , Furanos/química , Glucósidos/química , Lignanos/química , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Espectrometría de Fluorescencia
20.
J Sep Sci ; 32(20): 3544-9, 2009 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-19764052

RESUMEN

Ionic liquid was used as microwave absorption medium and applied to the extraction of essential oils from dried fruits of the Illicium verum Hook. f. and Cuminum cyminum L. by microwave-assisted extraction. The extraction time is less than 15 min at the microwave power of 440 W. The constituents of essential oils obtained by the proposed method were compared with those obtained by hydrodistillation. There is no obvious difference in the constituents of essential oils obtained by the two methods.


Asunto(s)
Cuminum/química , Illicium/química , Líquidos Iónicos/química , Microondas , Aceites Volátiles/aislamiento & purificación , Aceites de Plantas/aislamiento & purificación , Absorción , Cromatografía de Gases y Espectrometría de Masas , Extractos Vegetales/química
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