Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 186
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Molecules ; 25(5)2020 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-32155919

RESUMO

Quantitative analysis of formaldehyde (HCHO, FA), especially at low levels, in various environmental media is of great importance for assessing related environmental and human health risks. A highly efficient and convenient FA detection method based on surface-enhanced Raman spectroscopy (SERS) technology has been developed. This SERS-based method employs a reusable and soft silver-coated TiO2 nanotube array (TNA) material, such as an SERS substrate, which can be used as both a sensing platform and a degradation platform. The Ag-coated TNA exhibits superior detection sensitivity with high reproducibility and stability compared with other SERS substrates. The detection of FA is achieved using the well-known redox reaction of FA with 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole (AHMT) at room temperature. The limit of detection (LOD) for FA is 1.21 × 10-7 M. In addition, the stable catalytic performance of the array allows the degradation and cleaning of the AHMT-FA products adsorbed on the array surface under ultraviolet irradiation, making this material recyclable. This SERS platform displays a real-time monitoring platform that combines the detection and degradation of FA.

2.
BMC Genomics ; 21(1): 216, 2020 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-32151239

RESUMO

BACKGROUND: All retroviruses, including human immunodeficiency virus (HIV), must integrate a DNA copy of their genomes into the genome of the infected host cell to replicate. Although integrated retroviral DNA, known as a provirus, can be found at many sites in the host genome, integration is not random. The adaption of linker-mediated PCR (LM-PCR) protocols for high-throughput integration site mapping, using randomly-sheared genomic DNA and Illumina paired-end sequencing, has dramatically increased the number of mapped integration sites. Analysis of samples from human donors has shown that there is clonal expansion of HIV infected cells and that clonal expansion makes an important contribution to HIV persistence. However, analysis of HIV integration sites in samples taken from patients requires extensive PCR amplification and high-throughput sequencing, which makes the methodology prone to certain specific artifacts. RESULTS: To address the problems with artifacts, we use a comprehensive approach involving experimental procedures linked to a bioinformatics analysis pipeline. Using this combined approach, we are able to reduce the number of PCR/sequencing artifacts that arise and identify the ones that remain. Our streamlined workflow combines random cleavage of the DNA in the samples, end repair, and linker ligation in a single step. We provide guidance on primer and linker design that reduces some of the common artifacts. We also discuss how to identify and remove some of the common artifacts, including the products of PCR mispriming and PCR recombination, that have appeared in some published studies. Our improved bioinformatics pipeline rapidly parses the sequencing data and identifies bona fide integration sites in clonally expanded cells, producing an Excel-formatted report that can be used for additional data processing. CONCLUSIONS: We provide a detailed protocol that reduces the prevalence of artifacts that arise in the analysis of retroviral integration site data generated from in vivo samples and a bioinformatics pipeline that is able to remove the artifacts that remain.

3.
Sci Rep ; 10(1): 3499, 2020 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-32103103

RESUMO

Reproductive isolation between different host populations is often based on intraspecific sex pheromone differences. The mechanisms underlying these differences have not been thoroughly elucidated to date. Previous studies suggested that Chilo suppressalis has differentiated into rice and water-oat host populations, and these two populations manifest clear differences in sex pheromone titer and mating rhythm. Hence, this moth is an ideal model to investigate the endogenous mechanisms of intraspecific reproductive isolation. Here, we identified a series of putative genes associated with sex pheromone biosynthesis based on the C. suppressalis pheromone gland transcriptome data. Transcripts of most genes were at higher level in the rice population. Then we obtained 11 pivotal differentially expressed genes (DEGs). The expression levels of these DEGs exhibited a distinct increase in the rice population. Moreover, we also observed the expression rhythm of these DEGs is discrepant between two host populations. Our study offers a new understanding to elucidate the mechanisms of intraspecific reproductive isolation.

4.
J Virol Methods ; 279: 113841, 2020 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-32105753

RESUMO

Japanese Encephalitis (JE) is an acute infectious disease that threatens both human and pig populations throughout Asia. JE is caused by the Japanese Encephalitis Virus (JEV), of which genotype III (GIII) had been the most prevalent strain throughout Asia, but recent studies have shown that genotype I (GI) has replaced GIII as the predominant version. Pigs and mosquitoes play a primary role in JEV transmission. However, a method for the rapid differentiation between JEV G I and G III remains unavailable. This study aimed to establish a rapid JEV genotyping method using novel duplex TaqMan RT-qPCR assay.specific primer and probes located in the PrM/M gene that were able to specifically differentiate GI and GIII JEV, was selected as the duplex TaqMan RT-qPCR target.The specificity, sensitivity and reproducibility test of this assay were validated. The sensitivity of the assay was 10 genomic RNA copies for both GI and GIII JEV in field mosquito and pig samples,and more sensitive than the current methods. In addition, the novel assay can be completed in less than 1 h. Therefore, This duplex TaqMan RT-qPCR assay is a promising tool for rapid differential detection and epidemiology of GI and GIII JEV strains in China. The results showed that co-circulation of GI and GIII infections with GI infection being more prevalent in pigs or mosquitoes in eastern China.

5.
Chemosphere ; 244: 125582, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32050352

RESUMO

As the most typical geological environment, limestone landforms are widespreading in the world and affect the waters that flow around them, which may also change the fate of organic contaminants in these waters. In this study, aquatic environment surrounding limestone was simulated with calcium carbonate, and the photolysis of tetracycline was evaluated under UV irradiation (30 µW/cm2). More tetracycline (up to 98%) was removed in 4 h in the presence of calcium carbonate while only 50% of tetracycline was eliminated in control experiment. The removal of tetracycline was greatly enhanced due to the major roles of alkaline pH and minor roles of Ca2+ and HCO3-/CO32-. In alkaline pH, tetracycline existed as TCs- with higher electronic density in the ring structures, which was more easily attacked by OH. Besides, it could also change the bond orbital energy to facilitate tetracycline absorbing more photon. Moreover, alkaline pH was beneficial to generate more OH and thus promote the indirect photolysis. In addition, alkaline pH also changed the degradation path of tetracycline and rapidly convert tetracycline to the byproducts with m/z 457 via hydroxylation and hydrogen abstraction. This work provides not only better understanding about the fate of tetracycline in aquatic environments but also new insights into the treatment of antibiotic-contaminated water.


Assuntos
Poluentes Químicos da Água/química , Antibacterianos , Carbonato de Cálcio , Cinética , Fotólise , Tetraciclina/análise , Tetraciclina/química , Raios Ultravioleta , Poluentes Químicos da Água/análise
6.
Bioorg Chem ; 95: 103541, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31927334

RESUMO

Twenty three undescribed withanolides, daturmetelides A-W (1-23), were isolated from 70% EtOH extract of the leaves of Datura metel L. The structural characterizations and relative configurations of 1-23 were elucidated by extensive spectroscopic analysis as well as by comparison with literature values. The absolute configurations of 1 and 3 were determined by X-ray crystallography. Bioassay results showed that 1 and 7 exhibited moderate inhibitory effects against NO production in lipopolysaccharide-stimulated RAW 264.7 cells (IC50 values of 13.74 µM and 13.92 µM, respectively). In addition, 1 and 7 showed significant anti-inflammatory activities against the production of TNF-α, IL-1ß, IL-6 and COX-2. Western blot analysis was further performed to reveal the mechanism of anti-inflammatory action via inhibition of the NF-κB activation.

7.
Sci Rep ; 10(1): 44, 2020 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-31913290

RESUMO

Low miR-96-5p expression is characteristic of many cancers but its role in breast cancer (BCa) remains poorly defined. Here, the role of miR-96-5p in BC development was assessed. We demonstrate that exogenously expressing miR-96-5p inhibits the proliferative, migratory and invasive capacity of BCa cells. Mechanistically, miR-96-5p in BCa cells was found to target and downregulate catenin delta 1 (CTNND1) leading to decreased ß-catenin expression, a loss of WNT11 signaling, reduced cyclin D1 levels and lower MMP7 expression. Exogenously expressing CTNND1 alleviated these effects. In summary, we are the first to reveal that miR-96-5p inhibits the proliferative, invasive and migratory phenotypes of BCa cells the targeting of CTNND1 and subsequent Wnt/ß-catenin signaling. These data highlight miR-96-5p as a novel target for BC treatment.

8.
Braz J Med Biol Res ; 53(2): e9106, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31994603

RESUMO

Reperfusion strategies in acute myocardial infarction (AMI) can cause a series of additional clinical damage, defined as myocardial ischemia/reperfusion (I/R) injury, and thus there is a need for effective therapeutic methods to attenuate I/R injury. miR-26a-5p has been proven to be an essential regulator for biological processes in different cell types. Nevertheless, the role of miR-26a-5p in myocardial I/R injury has not yet been reported. We established an I/R injury model in vitro and in vivo. In vitro, we used cardiomyocytes to simulate I/R injury using hypoxia/reoxygenation (H/R) assay. In vivo, we used C57BL/6 mice to construct I/R injury model. The infarct area was examined by TTC staining. The level of miR-26a-5p and PTEN was determined by bioinformatics methods, qRT-PCR, and western blot. In addition, the viability and apoptosis of cardiomyocytes were separately detected by MTT and flow cytometry. The targeting relationship between miR-26a-5p and PTEN was analyzed by the TargetScan website and luciferase reporter assay. I/R and H/R treatment induced myocardial tissue injury and cardiomyocyte apoptosis, respectively. The results showed that miR-26a-5p was down-regulated in myocardial I/R injury. PTEN was found to be a direct target of miR-26a-5p. Furthermore, miR-26a-5p effectively improved viability and inhibited apoptosis in cardiomyocytes upon I/R injury by inhibiting PTEN expression to activate the PI3K/AKT signaling pathway. miR-26a-5p could protect cardiomyocytes against I/R injury by regulating the PTEN/PI3K/AKT pathway, which offers a potential approach for myocardial I/R injury treatment.

9.
Steroids ; 156: 108583, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31982422

RESUMO

Five undescribed ergostane-type C28 sterols, daturmetesides A-E (1-5), were isolated from the leaves of Datura metel L. The chemical structures of these new compounds were characterized through extensive spectroscopic analysis and comparison with literatures. Among them, the absolute structures of daturmetesides A and C were unambiguously determined by X-ray crystallography. The anti-inflammatory effect of daturmetesides A-E was all tested by measuring nitric oxide production in lipopolysaccharide-activated RAW264.7 cells. Daturmetesides A, C and D moderatelylowered the NO production with IC50 values ranging from 17.05 ± 0.35 to 24.88 ± 0.93 µM.

10.
J Hazard Mater ; 384: 121187, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31648895

RESUMO

Persistent organic micropollutants have seriously damaged aquatic ecological equilibrium and affected human health. Conventional adsorbents are limited due to slow adsorption rate. Therefore, it's significant to integrate adsorbent into porous membrane to develop a highly efficient continuous filtration method for water purification. Herein, ß-cyclodextrin polymer (ß-CDP) composite porous membranes were prepared via convenient interfacial cross-linking. The membranes combined the adsorption ability of ß-CDP and the convective mass transport process of filtration membrane to quickly remove contaminants from water by flow-through adsorption. In optimized preparation conditions, the composite membrane exhibited a 100% of removal efficiency towards bisphenol A and a high treating capacity up to 440 mg m-2. The treating capacity kept nearly unchanged in acidic and neutral pH condition, while increased greatly with the addition of salts due to the salting-out effect. Also, the membrane could completely remove pollutants with ultrahigh flux up to 2500 L·m-2 h-1. In addition, the used membranes were fully regenerated by mild ethanol cleaning.

11.
Cell Signal ; 67: 109491, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31809873

RESUMO

Excessive circulating free fatty acids (FFA) cause insulin resistance in peripheral tissues by inhibiting the proximal insulin signaling pathway. White adipose tissue (WAT) is a primary source of FFA generation and release through triglyceride (TG) hydrolysis. Thus, reducing excessive lipolysis in adipocytes ameliorates whole-body insulin resistance in type 2 diabetes. Here, we found that a noninvasive photobiomodulation therapy (PBMT), decreased FFA generation and release in WATs from high-fat diet (HFD)-fed mice and diabetic db/db mice. Meanwhile, plasma FFA and TG levels were reduced in two mouse models after PBMT. PBMT promoted mitochondrial reactive oxygen species (ROS) generation, which inhibited phosphatase and tensin homologue (PTEN) and promoted protein kinase B (AKT) activation. Photoactivation of AKT inhibited the transcriptional activity of Forkhead box transcription factor O1 (FoxO1), reducing expression of lipolytic enzymes and FFA generation and release. Eliminating ROS elimination or inhibiting AKT blocked the effects of the laser therapy in vivo and in vitro. Taken together, PBMT suppresses FFA generation and release in insulin-resistant adipocytes, contributing to improvement of insulin resistance in mouse models of type 2 diabetes.

12.
Antiviral Res ; 174: 104673, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31812636

RESUMO

Japanese Encephalitis virus (JEV) is a zoonotic flavivirus that is the most significant etiological agent of childhood viral neurological infections. However, no specific antiviral drug is currently available to treat JEV infections. The JEV envelope (E) protein is a class II viral fusion protein that mediates host cell entry, making interference with the interaction between the E protein of JEV and its cognate receptors an attractive strategy for anti-JEV drug development. In this study, we identified a peptide derived from a phage display peptide library against the E protein of JEV, designated P1, that potentially inhibits in vitro and in vivo JEV infections. P1 inhibits JEV infection in BHK-21 cells with 50% inhibitory capacity at a concentration of 35.9 µM. The time-of-addition assay indicates that JEV replication is significantly inhibited during pre-infection and co-infection of P1 with JEV while post-infection treatments with P1 have very little impact on JEV proliferation, showing that P1 inhibits JEV infection at early stages and indicating the potential prophylactic effect of P1. We adapted an in vitro BiFC assay system and demonstrated that P1 interacts with JEV E proteins and blocks their entry into cells. We also evaluated the therapeutic efficacy of P1 in a lethal JEV mouse model exhibiting systemic and brain infections. Interestingly, P1 treatment protected C57BL/6 mice against mortality, markedly reduced the viral loads in blood and brain, and diminished the histopathological lesions in the brain cells. In addition to controlling systemic infection, P1 has a very low level of cytotoxicity and acts in a sequence-specific manner, as scrambled peptide sP1 does not show any antiviral activity. In conclusion, our in vitro and in vivo experimental findings show that P1 possesses antiviral activity against JEV infections, is safe to use, and has potential for further development as an antiviral treatment against JEV infections.

13.
Life Sci ; 242: 117212, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31884092

RESUMO

AIMS: Oxidative stress is an important risk factor in development and progression of type 2 diabetes. Resveratrol (RSV), as a natural antioxidant, reduces intracellular reactive oxygen species (ROS) and oxidative stress. MATERIALS AND METHODS: The study investigated the effects of RSV treatment on high-fat diet (HFD)-fed mice and muscle, adipose, and hepatic cells of insulin resistance. HFD-fed mice were treated with RSV for 10 weeks. Blood glucose, plasma triglyceride (TG), body weight and glucose-lipid metabolism of skeletal muscle, fat and liver were examined. We further assessed the metabolic regulation of RSV in C2C12 myotubes, 3T3-L1 adipocytes and HepG2 cells of insulin resistance. KEY FINDINGS: We found that RSV reduced blood glucose, plasma TG and body weight, ameliorated insulin resistance in HFD-fed mice. RSV reduced lipid accumulation and increased glycogen storage in muscle and hepatic cells, promoted lipolysis in adipocytes. We further found RSV reduced ROS levels in muscle, adipose, and hepatic cells of insulin resistance, contributing to improvement of metabolic abnormalities in HFD-fed mice. SIGNIFICANCE: The study reveals that RSV ameliorates metabolic disorders and insulin resistance in HFD-fed mice, which provides further demonstration in RSV-treated type 2 diabetes.


Assuntos
Antioxidantes/farmacologia , Dieta Hiperlipídica/efeitos adversos , Resistência à Insulina , Doenças Metabólicas/tratamento farmacológico , Resveratrol/farmacologia , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Animais , Glicemia/análise , Células Hep G2/efeitos dos fármacos , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Doenças Metabólicas/etiologia , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Triglicerídeos/sangue
14.
J Med Virol ; 92(2): 149-160, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31517388

RESUMO

Classical swine fever virus (CSFV) is a single-stranded RNA flavivirus that can cause serious diseases in porcine species, including symptoms of infarction, systemic hemorrhage, high fever, or depression. Viperin is an important interferon-inducible antiviral gene that has been shown to inhibit CSFV, but the exact mechanisms by which it is able to do so remain poorly characterized. In the present study, we determined that CSFV infection led to viperin upregulation in PK-15 cells (porcine kidney cell). When viperin was overexpressed in these cells, this markedly attenuated CSFV replication, with clear reductions in viral copy number after 12 to 48 hours postinfection. Immunofluorescence microscopy revealed that the viral NS5A protein colocalized with viperin in infected cells, and this was confirmed via confocal laser scanning microscopy using labeled versions of these proteins, and by co-immunoprecipitation which confirmed that NS5A directly interacts with viperin. When NS5A was overexpressed, this inhibited the replication of CSFV, and we determined that the radical SAM domain and N-terminal domain of viperin was critical for its ability to bind to NS5A, with the latter being most important for this interaction. Together, our in vitro results highlight a potential mechanism whereby viperin is able to inhibit CSFV replication. These results have the potential to assist future efforts to prevent or treat systemic CSFV-induced disease, and may also offer more general insights into the antiviral role of viperin in innate immunity.

15.
Biotechnol Lett ; 42(3): 375-387, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31872317

RESUMO

OBJECTIVES: Hydrogen sulfide (H2S) is involved in regulating cell apoptosis and proliferation. However, The effects and mechanism of H2S on the apoptosis of mammary epithelial cells that suffer from an inflammatory response remain unknown. RESULTS: An inflammatory cell model was used to explore whether exogenous H2S regulates lipopolysaccharides (LPS)-induced cell proliferation and apoptosis. We found that H2S affected cell viability, the inflammatory response and apoptosis in LPS-treated cells in a concentration-dependent manner. Moreover, exogenous H2S rescued LPS-induced cystathionine γ-lyase (CSE) inhibition and cystathionine ß-synthase (CBS) synthesis. Interestingly, in cells undergoing inflammation-induced apoptosis, H2S activated the PI3K/Akt and NFκB signal pathways both tested concentrations. Akt appeared to be a key crosstalk molecule that played a "bridge" role. CONCLUSIONS: H2S regulates LPS-induced inflammation and apoptosis by activating the PI3K/Akt/NFκB signaling pathway. Hence, NaHS may be clinically useful for preventing or treating mastitis.

16.
Int Immunopharmacol ; : 106027, 2019 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-31791672

RESUMO

BACKGROUND: Regulatory T (Treg) cells are known as immune regulators to decrease infarct volume and improve outcomes after ischemic stroke. Thus, the strategies for increasing Treg cells in ischemic brain may have the beneficial effects on stroke. In this study, we aim to examine the effect of Interleukin-33 (IL-33) on Treg cell expansion in mouse model of ischemic stroke. METHODS: Mice were subjected to 30 min of middle cerebral artery occlusion (MCAO) followed by 24 h, 48 h of 72 h of reperfusion. Recombinant mouse IL-33 (2 µg) was pre-treated intracerebroventricularly at 30 min prior to MCAO. The percentage of Treg cells in ischemic brain, related cytokines and transcription factors, the levels of ST2 receptor, amphiregulin (AREG), and epidermal growth factor receptor (EGFR) were measured. RESULTS: IL-33 treatment can increase the number of Foxp3+ Treg cells in the ischemic brain and the levels of IL-10 and TGF- ß1 in serum and brain tissues at MCAO 48 h and 72 h, but not at MCAO 24 h. In the Treg cells separated from ischemic brain tissue following MCAO treated by IL-33, the expression level of ST2 receptor was up-regulated. In addition, IL-33 may increase the mRNA level of transcription factor Foxp3. Correspondingly, IL-33 treatment also elevated the levels of AREG and EGFR at MCAO 48 h and 72 h. CONCLUSION: We speculated that intracerebroventricular IL-33 can activate the downstream Foxp3 via ST2 receptor to increase Treg proportions in the ischemic brain. The elevated Treg cells produce AREG to activate EGFR located in neurons, which contribute to better outcomes.

17.
ACS Omega ; 4(23): 20101-20108, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31815210

RESUMO

As a new analytical technology, surface-enhanced Raman scattering (SERS) has received increasing attention, and researchers have discovered the importance of SERS-active materials. Considerable effort has been made by researchers to develop multiperformance and multipurpose SERS-active substrates ranging from coinage metals to transition metals and semiconductor materials. SERS-active substrates are critical for obtaining accurate and reproducible spectral information. Among all the substrate materials, semiconductors are considered one of the most promising materials, as they exhibit high chemical stability, good biocompatibility, high carrier mobility, and good controllability during fabrication. Here, we provide an overview of SERS enhancement mechanisms based on semiconductor materials, such as inorganic semiconductors, metal/semiconductor composites, and organic semiconductors.

18.
Zhongguo Zhong Yao Za Zhi ; 44(18): 3985-3993, 2019 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-31872735

RESUMO

In order to determine the quality evaluation method for standard decoction of Coptidis Rhizoma,15 batches of standard decoction of Coptidis Rhizoma were prepared by using standardized process. Parameters such as traits,p H value,indicative component content,fingerprint similarity,composition transfer rate and dry extract rate were selected as the indexes for quality evaluation. Similarity evaluation and cluster analysis were performed for HPLC fingerprint of standard decoction,and mathematical model was used to study the correlation between dry extract rate,berberine content,berberine transfer rate in standard decoction and berberine content in decoction pieces. The results showed that the similarity of fingerprints was greater than 0. 99 for these 15 batches of standard decoctions of Coptidis Rhizoma. In cluster analysis,the standard decoctions of Coptidis Rhizoma from 4 producing areas were classified into 3 categories,consistent with the content determination results,indicating that there were quality differences among different producing areas.R2 in three linear regression mathematical models established was all greater than 0. 9,with significant difference. The validation of three batches of data showed that the models had good accuracy. Therefore,this model can be used to predict the quality of standard decoction prepared from different Coptidis Rhizoma pieces. In the standard decoction process established in this study,the integrity of the traditional process was greatly preserved,and the established quality evaluation method could be used to comprehensively examine the quality of the standard decoction,which can provide a demonstration for the related research of water extraction preparation containing Coptidis Rhizoma pieces.


Assuntos
Berberina/análise , Coptis/química , Medicamentos de Ervas Chinesas/análise , Controle de Qualidade , Cromatografia Líquida de Alta Pressão , Modelos Lineares , Rizoma/química
19.
Zhongguo Zhong Yao Za Zhi ; 44(18): 4026-4033, 2019 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-31872741

RESUMO

An analysis method was established by UPLC fingerprint and then applied to simultaneous determination of multiple compounds in Gardeniae Fructus from different areas in China. Samples were separated on a Waters Acquity UPLC BEH C18( 2. 1 mm×50 mm,1. 7 µm) column with 0. 1% formic acid-water and acetonitrile solution as gradient mobile phase at a flow rate of 0. 4 m L·min-1 at various wavelengths. The similarity of samples was over 0. 95 with ″Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine( 2012 edition) ″. The UPLC common fingerprints for 32 batches were established with 19 common peaks identified. The samples were divided into 3 groups analyzed by HCA and PCA. Five components were identified as the main compositions which caused the differences of chemical constituents in the samples from different areas with partial least squares discriminant analysis( PLS-DA). The content of the total components in each area was Zhejiang > Fujian > Jiangxi > Sichuan. This method was accurate and viable,could be used to evaluate the quality of Gardeniae Fructus.


Assuntos
Medicamentos de Ervas Chinesas/normas , Frutas/química , Gardenia/química , Compostos Fitoquímicos/análise , China , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/análise
20.
Proc Natl Acad Sci U S A ; 116(51): 25891-25899, 2019 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-31776247

RESUMO

Understanding HIV-1 persistence despite antiretroviral therapy (ART) is of paramount importance. Both single-genome sequencing (SGS) and integration site analysis (ISA) provide useful information regarding the structure of persistent HIV DNA populations; however, until recently, there was no way to link integration sites to their cognate proviral sequences. Here, we used multiple-displacement amplification (MDA) of cellular DNA diluted to a proviral endpoint to obtain full-length proviral sequences and their corresponding sites of integration. We applied this method to lymph node and peripheral blood mononuclear cells from 5 ART-treated donors to determine whether groups of identical subgenomic sequences in the 2 compartments are the result of clonal expansion of infected cells or a viral genetic bottleneck. We found that identical proviral sequences can result from both cellular expansion and viral genetic bottlenecks occurring prior to ART initiation and following ART failure. We identified an expanded T cell clone carrying an intact provirus that matched a variant previously detected by viral outgrowth assays and expanded clones with wild-type and drug-resistant defective proviruses. We also found 2 clones from 1 donor that carried identical proviruses except for nonoverlapping deletions, from which we could infer the sequence of the intact parental virus. Thus, MDA-SGS can be used for "viral reconstruction" to better understand intrapatient HIV-1 evolution and to determine the clonality and structure of proviruses within expanded clones, including those with drug-resistant mutations. Importantly, we demonstrate that identical sequences observed by standard SGS are not always sufficient to establish proviral clonality.

SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA