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1.
Genome Res ; 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-32024662

RESUMO

Massively parallel sequencing of the polyadenylated RNAs has played a key role in delineating transcriptome complexity, including alternative use of an exon, promoter, 5' or 3' splice site or polyadenylation site, and RNA modification. However, reads derived from the current RNA-seq technologies are usually short and deprived of information on modification, compromising their potential in defining transcriptome complexity. Here, we applied a direct RNA sequencing method with ultralong reads using Oxford Nanopore Technologies to study the transcriptome complexity in Caenorhabditis elegans We generated approximately six million reads using native poly(A)-tailed mRNAs from three developmental stages, with average read lengths ranging from 900 to 1100 nt. Around half of the reads represent full-length transcripts. To utilize the full-length transcripts in defining transcriptome complexity, we devised a method to classify the long reads as the same as existing transcripts or as a novel transcript using sequence mapping tracks rather than existing intron/exon structures, which allowed us to identify roughly 57,000 novel isoforms and recover at least 26,000 out of the 33,500 existing isoforms. The sets of genes with differential expression versus differential isoform usage over development are largely different, implying a fine-tuned regulation at isoform level. We also observed an unexpected increase in putative RNA modification in all bases in the coding region relative to the UTR, suggesting their possible roles in translation. The RNA reads and the method for read classification are expected to deliver new insights into RNA processing and modification and their underlying biology in the future.

2.
AAPS PharmSciTech ; 21(3): 85, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31997020

RESUMO

Nanoparticles (NPs) containing the hydrophilic drug salidroside (Sal) and the hydrophobic drug tamoxifen (Tam) were prepared using a triblock copolymer poly(lactic-co-glycolic acid) (PLGA)-poly(ethylene glycol) (PEG)-PLGA to achieve synergism in the treatment of breast cancer. The double emulsion (w/o/w) method was used to prepare Sal-Tam NPs with an average particle size of 275.3 ± 44.0 nm, a polydispersity index of 0.302 ± 0.102, and a zeta potential of - 6.98 ± 2.99. The entrapment efficiency of the hydrophilic and hydrophobic components was 32.63% ± 0.73% and 49.18% ± 3.04%, respectively. On differential scanning calorimetry, the NPs showed the amorphous nature of both Sal and Tam. The sustained release of Sal and Tam from the NPs was significantly prolonged under physiological conditions (pH 7.4). The CCK-8 assay using the 4T1 cell line revealed a 1.7-fold decrease in the IC50 value for Sal-Tam NPs when compared with free Tam. The in vivo anti-tumor effect was assessed in BALB/c mice, and the results demonstrated that these NPs decreased the tumor volume compared with saline and showed high anti-tumor activity. A pharmacokinetic study showed significant enhancement of the bioavailability of Tam in Sal-Tam NPs compared with free Tam in suspension. The intracellular and mitochondrial anti-oxidative effect of Sal was thought to be attributed to the promising anti-tumor effect of drug co-delivery. This study confirmed that the use of Sal-Tam NPs may be a promising approach in breast cancer therapy.

3.
Biomed Chromatogr ; 34(2): e4757, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31755125

RESUMO

Er-Zhi-Wan (EZW) is a traditional Chinese medicine with many clinical applications and used as a health product in East Asia. Five active ingredients (salidroside, specnuezhenide, nuezhenoside, luteolin, and oleanolic acid) were screened out from EZW to develop an in vitro rapid evaluation method for the classification of in vivo drug absorption behavior by biopharmaceutics classification system (BCS). Ultra-performance liquid chromatography was used for quantitative analysis. Solubility and permeability were assayed by equilibrium solubility and multiple models: everted rat intestinal sac model, cultured Caco-2 cells, octanol-water partition coefficient (LogP) method. The BCS properties of drugs were predicted using software applications, and the correlations of measured and predicted values of factors affecting oral drug absorption were calculated. The results were verified by measuring the absolute bioavailability of the active ingredients. Salidroside, specnuezhenide, and nuezhenoside were classified as BCS class III drugs, and luteolin was classified as a BCS class III/I drug because of the difference in LogP and intestinal permeability. Oleanolic acid was classified as a BCS class II/IV drug in acidic media and BCS class I/III drug in other media. Overall, EZW may be classified as a BCS class III drug, and permeability was identified as the primary factor limiting absorption. The results provide a novel method for the evaluation of the in vivo absorption of oral traditional Chinese medicines.

4.
Int J Biol Macromol ; 143: 873-880, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31712148

RESUMO

Poor solubility and stability are limitation factors of flavonoids application. Polysaccharides from herbs have been proven to be functional foods and potential promising natural supplements with large molecular weight and complex structures. Astragalus polysaccharides (APS) are the main components of Astragalus membranaceus (Fisch.) and novel synergistic pharmacological effects between Astragalus polysaccharides and flavonoids have been reported. However, the general effects of polysaccharides when co-administered with flavonoids are currently unknown. Herein, the influences of APS on aqueous solubilities and stabilities of fifteen flavonoids were systematically investigated, and the mechanism of effects of polysaccharides on flavonoids was further considered. The results showed that APS could significantly enhance the solubilities and stabilities of the flavonoids, with solubilization effect improved by 68.88-fold for quercetin and negatively correlated with the aqueous solubility of the flavonoid itself. A phase solubility study and Differential scanning calorimetry characterization indicated that APS could form complexes with flavonoids at 1:1 ratio with K values ranging from 1491 to 55,395 L·mol-1, a tendency to improved solubilization at higher association constant values was also observed. Those findings could provide the basis for a new approach to solving the problems of poor solubility and stability of flavonoids through the application of natural macromolecules.

5.
J Ethnopharmacol ; 244: 112148, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31400507

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: As one of the new drugs of traditional Chinese medicine, Sanye Tablet is employed as a hypolipidemic in the traditional medicine, but the biopharmaceutical properties of the drug is still unclear. AIM OF THE STUDY: Through the study of biopharmaceutical properties, the classical biopharmaceutics classification system (BCS) can be used to classify and predict the in vivo absorption properties. On this basis, the biopharmaceutical properties closely related to traditional Chinese medicine preparations are added and a modified BCS model is established to predict and judge the absorption degree of traditional Chinese medicine compound. MATERIALS AND METHODS: Representative components of Sanye Tablet were selected and subjected to different in vitro tests. The experimental results were compared with the results of the BCS to evaluate the accuracy and applicability to Sanye Tablet. We take parameters of dissolution and stability based on product characteristics into account. A "modified-BCS" was developed and the results of the improved method and the classic method were compared. Also the ability of each classification system to predict and determine the extent of absorption of the Chinese herbal compound was investigated based on the absolute bioavailability of representative components. RESULTS: For classic BCS, the five representative components (except for nuciferine) are all class III, nuciferine is class I/II obtained by Caco-2 cell assay and class III/IV obtained by everted gut sac assay. For modified BCS, paeoniflorin is class III, rutin, hyperoside and salvianolic acid B are class III/IV, and nuciferine is class I/II based on Caco-2 cell assay, class III/IV based on everted gut sac assay. Nuciferine is the best of the five components, with absolute bioavailability reaching 61.91% based on in vivo bioavailability test. CONCLUSIONS: The five representative components (except for nuciferine) are all class III/IV, which correlates well with the absolute bioavailability results and demonstrates that they are poorly absorbed substances. The correlation between the classification results obtained using the "modified-BCS" and absorption in the body is better than the correlation obtained using the classic method, suggesting that the improved BCS is more suitable for the characterization of Sanye Tablet. These results indicate that the oral formulation of Sanye Tablet is a BCS III/IV drug.

6.
Molecules ; 24(13)2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31269661

RESUMO

Scutellaria baicalensis Georgi (SBG) is not just as a traditional herbal medicine but also a popular functional food in China and other Asian countries. A sensitive simple strategy was developed for the first time to analyze SBG from eight different geographical sources using high-performance liquid chromatography (HPLC) coupled with multivariate chemometric methods. Two unsupervised pattern recognition models, hierarchical cluster analysis (HCA) and principal components analysis (PCA), and a supervised pattern recognition model, partial least squares discriminant analysis (PLS-DA), were used to analyze the chemical compositions and physical traits of SBG. The important chemical markers baicalin, baicalein, and wogonoside were analyzed quantitatively and with PLS-DA. These methods distinguished rotten xylem (kuqin) and strip types (tiaoqin) of SBG and found that the thickness of the slice had a significant impact on the classification of SBG. Two classes of strip types were identified: one as the uncut pharmaceutical, which was sectioned with a thickness >3 mm; the other as a thin-sectioned strip type, with a thickness of <2 mm. This fingerprinting technique coupled to a chemometric analysis was used for the simultaneous quantitation of three components (chemical markers) of SBG, and greatly simplified the complicated identification of the multiple components of this plant relative to traditional methods. The strategy can clearly distinguish between kuqin and tiaoqin of SBG, and suggests that the thickness of the slice can be used as the basis for evaluation of SBG. These data provide a theoretical basis and scientific evidence for the development and utilization of SBG.


Assuntos
Análise de Componente Principal , Scutellaria baicalensis/química , Xilema/química , Análise por Conglomerados , Análise Discriminante , Flavanonas/análise , Flavonoides/análise , Glucosídeos/análise , Análise dos Mínimos Quadrados
7.
Genetics ; 212(3): 801-813, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31064822

RESUMO

Hybrid male progeny from interspecies crosses are more prone to sterility or inviability than hybrid female progeny, and the male sterility and inviability often demonstrate parent-of-origin asymmetry. However, the underlying genetic mechanism of asymmetric sterility or inviability remains elusive. We previously established a genome-wide hybrid incompatibility (HI) landscape between Caenorhabditis briggsae and C. nigoni by phenotyping a large collection of C. nigoni strains each carrying a C. briggsae introgression. In this study, we systematically dissect the genetic mechanism of asymmetric sterility and inviability in both hybrid male and female progeny between the two species. Specifically, we performed reciprocal crosses between C . briggsae and different C. nigoni strains that each carry a GFP-labeled C. briggsae genomic fragment referred to as introgression, and scored the HI phenotypes in the F1 progeny. The aggregated introgressions cover 94.6% of the C. briggsae genome, including 100% of the X chromosome. Surprisingly, we observed that two C. briggsae X fragments that produce C. nigoni male sterility as an introgression rescued hybrid F1 sterility in males fathered by C. briggsae Subsequent backcrossing analyses indicated that a specific interaction between the X-linked interaction and one autosome introgression is required to rescue the hybrid male sterility. In addition, we identified another two C. briggsae genomic intervals on chromosomes II and IV that can rescue the inviability, but not the sterility, of hybrid F1 males fathered by C. nigoni, suggesting the involvement of differential epistatic interactions in the asymmetric hybrid male fertility and inviability. Importantly, backcrossing of the rescued sterile males with C. nigoni led to the isolation of a 1.1-Mb genomic interval that specifically interacts with an X-linked introgression, which is essential for hybrid male fertility. We further identified three C. briggsae genomic intervals on chromosome I, II, and III that produced inviability in all F1 progeny, dependent on or independent of the parent-of-origin. Taken together, we identified multiple independent interacting loci that are responsible for asymmetric hybrid male and female sterility, and inviability, which lays a foundation for their molecular characterization.


Assuntos
Caenorhabditis/genética , Infertilidade Masculina/genética , Cromossomo X/genética , Animais , Caenorhabditis/fisiologia , Epistasia Genética , Feminino , Hibridização Genética , Masculino
8.
J Food Drug Anal ; 27(1): 365-372, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30648592

RESUMO

A strategy combining chemical UPLC-MS profiling and multivariate classification techniques has been used for the comparison of raw and processed Aconiti Lateralis Radix. UPLC-MS was used to identify 18 characteristic compounds, which were selected for discrimination of the raw and two processed products (Heishunpian and Baifupian). Chemometric analyses, including the combination of a heat map and hierarchical cluster analysis (HCA) and principal component analysis (PCA), were used to visualize the discrimination of raw and two processed products. HCA and PCA provided a clear discrimination of raw Aconiti Lateralis Radix, Heishunpian and Baifupian. Finally, the counter-propagation artificial neural network (CP-ANN) was applied to confirm the results of HCA, PCA and to explore the effect of 18 compounds on samples differentiation and the rationality of processing. The results showed that this strategy could be successfully used for comparison of raw and two processed products of Aconiti Lateralis Radix, which could be used as a general procedure to compare herbal medicines and related processed products to elaborate the rationality of processing from the perspective of chemical composition.

9.
Nat Prod Res ; 33(14): 2113-2115, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29888952

RESUMO

In this study, the appearance and texture of E. rutaecarpa were linked with the chemical constituents to explore methods of classification of E. rutaecarpa. The Chemometrics such as Hierarchical cluster analysis (HCA), principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) models were used for analysis. According to the models, samples of E. rutaecarpa were divided into three categories based on their source: Evodia, Stone Tiger and Sparse Evodia. The Evodia category could be subdivided into two categories, one representing large fruits with a greater degree of cracking and the other representing large fruits with little or no cracking. The method provided by this study combines chemometrics with HPLC fingerprints, which can provide a basis and reference for the identification of E. rutaecarpa and enables establishment of a grade standard.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Evodia/química , Frutas/química , Cromatografia Líquida de Alta Pressão/estatística & dados numéricos , Análise por Conglomerados , Plantas Medicinais/química , Análise de Componente Principal
10.
Molecules ; 24(1)2018 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-30591701

RESUMO

Puerariae Lobatae Radix (PLR), a well-known herbal medicine, is the root of Pueraria lobata (Willd.) Ohwi and has been employed for the treatment and prevention of cardiovascular and cerebrovascular diseases. The purpose of this study was to compare the associated-extraction efficiency of six cyclodextrins (CDs) on five flavonoids in PLR, namely puerarin, daidzein, daidzin, genistein and genistin, which are the major secondary metabolites, and exhibit low water solubility. The six CDs applied were ß-cyclodextrin (ß-CD), γ-cyclodextrin (γ-CD), hydroxypropyl-ß-cyclodextrin (HP-ß-CD), hydroxypropyl-γ-cyclodextrin (HP-γ-CD), carboxymethyl-ß-cyclodextrin (CM-ß-CD), and sulfobutyl ether ß-cyclodextrin (SBE-ß-CD). They can be grouped into one of the following three categories: traditional cyclodextrins (ß-CD and γ-CD), water-soluble cyclodextrin derivatives (HP-ß-CD and HP-γ-CD) and ionic cyclodextrin derivatives (SBE-ß-CD and CM-ß-CD). High-performance liquid chromatography (HPLC) was used to analyze the five flavonoids in the original aqueous extracts (OAE) in the presence or absence of various CDs. The associated-extraction efficiency of the various CDs followed the ranking: SBE-ß-CD > HP-ß-CD > CM-ß-CD > HP-γ-CD > γ-CD > ß-CD. It was clear that SBE-ß-CD presented the highest associated-extraction capability, and it was used to extract the four flavonoids from three PLR products, including raw product, stir- fried product, and product simmered with wheat bran. The results showed that SBE-ß-CD could improve the extraction capability of flavonoids, both from the raw product and in processed products of PLR. In conclusion, CDs, especially SBE-ß-CD, have a promising application for the associated-extraction of flavonoids from PLR.


Assuntos
Ciclodextrinas/isolamento & purificação , Flavonoides/análise , Pueraria/química , Cromatografia Líquida de Alta Pressão , Ciclodextrinas/química , Análise de Componente Principal , beta-Ciclodextrinas/química
11.
Molecules ; 23(9)2018 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-30201911

RESUMO

Phellodendri Chinensis Cortex (PCC) and Phellodendri Amurensis Cortex (PAC) are increasingly being used as traditional herbal medicines, but they are often mistaken for each other. In this study, the fingerprints of PCC from six different geographical sources were obtained by high-performance liquid chromatography, and multivariate chemometric methods were used for comprehensive analysis. Two unsupervised pattern recognition models (principal component analysis and hierarchical cluster analysis) and a supervised pattern recognition model (partial least squares discriminant analysis) were established on the basis of the chemical composition and physical traits of PCC and PAC. PCC and PAC were found to be distinguishable by these methods. The PCC category was divisible into two categories, one with more crude cork and a maximum thickness of ~1.5 mm, and the other with less net crude cork and a maximum thickness of 0.5 mm. According to the model established by partial least squares discriminant analysis (PLS-DA), the important chemical marker berberine hydrochloride was obtained and analyzed quantitatively. From these results combined with chemometric and content analyses, the preliminary classification standards for phellodendron were established as three grades: superior, first-order and mixed. Compared with the traditional identification methods of thin layer chromatography identification and microscopic identification, our method for quality evaluation is relatively simple. It provides a basis and reference for identification of PCC and enables establishment of grade standards. It also could be applied in quality control for compound preparations containing PCC.


Assuntos
Medicamentos de Ervas Chinesas/química , Phellodendron/química , Análise de Componente Principal , Berberina/análise , Cromatografia Líquida de Alta Pressão , Análise por Conglomerados , Análise Discriminante , Geografia , Análise dos Mínimos Quadrados
12.
J Pharm Biomed Anal ; 160: 323-329, 2018 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-30114610

RESUMO

The public concern and increasing awareness of herbal medicines calls for methods available to assure quality and authenticity. In the present study, a novel strategy combing 1H nuclear magnetic resonance (1H NMR) spectral analyses coupled with chemometrics was developed and validated to allow comprehensive analysis and rapid authentication of herbal medicines. Polygoni Multiflori Radix, a widely used herbal medicine and a kind of popular functional food, was taken as an example. Characteristic profiling of 1H NMR fingerprints was achieved by genetic algorithm and a counter-propagation artificial neural network. Multivariate classification methods were employed to evaluate and validate the performance of the developed characteristic profiling of 1H NMR fingerprints. The results well showed that the proposed method improved the predictive ability of the PLS-DA, and presented equivalent performance to the original spectrum in other classification methods. In conclusion, the developed method is an accurate, sensitive and rapid authentication of Polygoni Multiflori Radix, which could be a useful tool for rapid authentication and quality control of herbal medicines.


Assuntos
Espectroscopia de Ressonância Magnética/métodos , Compostos Fitoquímicos/análise , Polygonum/química , Análise dos Mínimos Quadrados , Plantas Medicinais/química , Análise de Componente Principal
13.
Int J Biol Macromol ; 119: 677-682, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30067956

RESUMO

2,3,5,4'-Tetrahydroxy-stilbene-2-O-ß-d-glucoside (THSG) is a bioactive polyhydroxystilbene compound obtained from Polygonum multiflorum Thunb. Although it has broad therapeutic potential, the application of THSG is limited due to its poor stability in vitro, poor absorption in the intestine, and short-term storage in tissues. Polysaccharides from herbal medicines have been proven to be functional foods and potential natural plant derived drugs. However, the effects of polysaccharides when co-administered with other active ingredients are currently unknown. The present study investigated the influences of Ophiopogon japonicus polysaccharides (OJPs) on THSG. Results showed that OJPs notably enhanced aqueous solubility, and stability of THSG, but slightly decreased the permeability of THSG. In addition, Tmax, Cmax, and AUC(0-tn) of THSG were 3.5 fold, 1.45 fold and 2.32 fold higher for THSG-OJP. Thus, OJPs could potentially be used to improve the biopharmaceutical properties and prolong the pharmacological effects of THSG. This finding could provide a reference point for further applications of polysaccharides from herbal medicines.


Assuntos
Glucosídeos/farmacologia , Glucosídeos/farmacocinética , Ophiopogon/química , Polissacarídeos/farmacologia , Estilbenos/farmacologia , Estilbenos/farmacocinética , Interações de Medicamentos , Glucosídeos/química , Permeabilidade , Solubilidade , Estilbenos/química
14.
DNA Res ; 25(6): 577-586, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30085012

RESUMO

Mitochondrial genome (mtDNA) carries not only well-conserved protein coding, tRNA and rRNA genes, but also highly variable non-coding regions (NCRs). However, the NCRs show poor conservation across species, making their function and evolution elusive. Identification and functional characterization of NCRs across species would be critical for addressing these questions. To this end, we devised a computational pipeline and performed de novo assembly and annotation of mtDNA from 19 Caenorhabditis species using next-generation sequencing (NGS) data. The mtDNAs for 14 out of the 19 species are reported for the first time. Comparison of the 19 genomes reveals species-specific sampling of partial displacement-loop (D-loop) sequence as a novel NCR inserted into a unique tRNA cluster, suggesting an important role of the D-loop and the tRNA cluster in shaping NCR evolution. Intriguingly, RNA-Seq analysis suggests that a novel NCR resulting from a recent duplication of NADH dehydrogenase subunit 5 (ND5) could be utilized as a 3' UTR for up-regulation of its upstream gene. The expression analysis shows a species- and sex-specific expression of mitochondrial genes encoded by mtDNA and nucleus, respectively. Our analyses provide important insights into the function and evolution of mitochondrial NCRs and pave the way for further studying the function and evolution of mitochondrial genome.


Assuntos
Caenorhabditis/genética , Duplicação Gênica , Genoma Mitocondrial , NADH Desidrogenase/genética , Animais , DNA Mitocondrial/química , Evolução Molecular , Feminino , Regulação da Expressão Gênica , Genômica , Masculino , Subunidades Proteicas/genética , RNA de Transferência/genética , Software
15.
Artigo em Inglês | MEDLINE | ID: mdl-30050583

RESUMO

Renal injury is among the adverse drug reactions (ADRs) caused by herbal medicine products (HMPs). Traditional Chinese medicines (TCMs) have been practiced for over 2000 years in China and East Asia, and herbs are currently used worldwide for the treatment and prevention of chronic and acute disease. Operetta high content analysis (HCA, PerkinElmer, Waltham, MA, USA), which is an in vitro, sensitive, reproducible, multiparametric screening method, was used to evaluate the cytotoxicity of HMPs in cultures of HEK293 human embryo kidney cells. Cytotoxic results were validated by an animal-based subacute toxicity assay. The renal safety of 18 active pharmaceutical agents from 13 TCM herbs with known nephrotoxic potential was evaluated in HEK293 human embryonic kidney cells. A panel of five parameters, cell viability, nuclear area, nuclear roundness, mitochondrial mass, and mitochondrial membrane potential, was utilized to evaluate drug-induced renal mitochondrial and nuclear injury. HCA can be a useful tool for preclinical screening and postclinical evaluation of HMPs. The nephrotoxicity of diosbulbin B and other HMPs was evident at a concentration as low as 0.01 µM.

16.
Zhongguo Zhong Yao Za Zhi ; 43(3): 511-519, 2018 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-29600615

RESUMO

Nuciferin is the main active ingredients in Nelumbinis Folium, which was proved to have good hypolipidemic, antioxidative and anti-inflammatory bioactivities. Currently, in vivo pharmacokinetic studies of nuciferin showed different results based on different animal models. In vitro evaluation experiments were low-cost, stable and controllable. Biopharmaceutical classification system(BCS) was an effective and reliable in vitro simulation method to evaluate the bioavailability of oral drugs. It was a scientific framework for classifying drugs or active pharmaceutical ingredients(API) according to their solubility and impermeability in vitro. In the study, BCS was applied in an active ingredient in traditional Chinese medicine(TCM), which was consisted of numerous chemical components. To study the equilibrium solubility of nuciferine, ideal solution model, Ape blat model and polynomial model were adopted. The permeability was measured based on partition coefficient(logP) and distribution coefficient(logD). Besides, in vitro apparent permeabilities of Caco-2 cells and murine intestine tissues were evaluated. Nuciferine was classified as BCSⅠ, since it had a good solubility and permeability in all methods under acidic conditions. However, in neutral and alkaline environments, nuciferine was classified as BCSⅣ by using everted intestinal sac. It indicated that the species of experimental animals has a significant influence on the absorption of nuciferine. This experiment can provide data support to the prediction in a complex environment(medicinal materials and absorbed parts). The application of BCS on TCM ingredients provided a new in vitro method to evaluate and screen out the druggability of TCM ingredients.


Assuntos
Aporfinas/química , Produtos Biológicos/classificação , Nelumbo/química , Animais , Disponibilidade Biológica , Biofarmácia , Células CACO-2 , Humanos , Absorção Intestinal , Camundongos , Permeabilidade , Folhas de Planta/química , Solubilidade
17.
Genetics ; 209(1): 37-49, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29567658

RESUMO

Intercellular signaling interactions play a key role in breaking fate symmetry during animal development. Identification of signaling interactions at cellular resolution is technically challenging, especially in a developing embryo. Here, we develop a platform that allows automated inference and validation of signaling interactions for every cell cycle of Caenorhabditis elegans embryogenesis. This is achieved by the generation of a systems-level cell contact map, which consists of 1114 highly confident intercellular contacts, by modeling analysis and is validated through cell membrane labeling coupled with cell lineage analysis. We apply the map to identify cell pairs between which a Notch signaling interaction takes place. By generating expression patterns for two ligands and two receptors of the Notch signaling pathway with cellular resolution using the automated expression profiling technique, we are able to refine existing and identify novel Notch interactions during C. elegans embryogenesis. Targeted cell ablation followed by cell lineage analysis demonstrates the roles of signaling interactions during cell division in breaking fate symmetry. Finally, we describe the development of a website that allows online access to the cell-cell contact map for mapping of other signaling interactions by the community. The platform can be adapted to establish cellular interactions from any other signaling pathway.


Assuntos
Ciclo Celular , Desenvolvimento Embrionário , Transdução de Sinais , Animais , Animais Geneticamente Modificados , Biomarcadores , Caenorhabditis elegans/embriologia , Caenorhabditis elegans/genética , Caenorhabditis elegans/metabolismo , Comunicação Celular , Linhagem da Célula , Proteínas de Drosophila/metabolismo , Dosagem de Genes , Ligação Proteica , Receptores Notch/metabolismo , Reprodutibilidade dos Testes , Transgenes
18.
J Food Drug Anal ; 26(1): 90-99, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29389593

RESUMO

Polygoni Multiflori Radix (PMR) is increasingly being used not just as a traditional herbal medicine but also as a popular functional food. In this study, multivariate chemometric methods and mass spectrometry were combined to analyze the ultra-high-performance liquid chromatograph (UPLC) fingerprints of PMR from six different geographical origins. A chemometric strategy based on multivariate curve resolution-alternating least squares (MCR-ALS) and three classification methods is proposed to analyze the UPLC fingerprints obtained. Common chromatographic problems, including the background contribution, baseline contribution, and peak overlap, were handled by the established MCR-ALS model. A total of 22 components were resolved. Moreover, relative species concentrations were obtained from the MCR-ALS model, which was used for multivariate classification analysis. Principal component analysis (PCA) and Ward's method have been applied to classify 72 PMR samples from six different geographical regions. The PCA score plot showed that the PMR samples fell into four clusters, which related to the geographical location and climate of the source areas. The results were then corroborated by Ward's method. In addition, according to the variance-weighted distance between cluster centers obtained from Ward's method, five components were identified as the most significant variables (chemical markers) for cluster discrimination. A counter-propagation artificial neural network has been applied to confirm and predict the effects of chemical markers on different samples. Finally, the five chemical markers were identified by UPLC-quadrupole time-of-flight mass spectrometer. Components 3, 12, 16, 18, and 19 were identified as 2,3,5,4'-tetrahydroxy-stilbene-2-O-ß-d-glucoside, emodin-8-O-ß-d-glucopyranoside, emodin-8-O-(6'-O-acetyl)-ß-d-glucopyranoside, emodin, and physcion, respectively. In conclusion, the proposed method can be applied for the comprehensive analysis of natural samples.


Assuntos
Cromatografia Líquida de Alta Pressão , Gastrópodes/química , Gastrópodes/classificação , Metabolômica , Animais , Espectrometria de Massas , Metabolômica/métodos , Estrutura Molecular
19.
Nucleic Acids Res ; 46(3): 1295-1307, 2018 02 16.
Artigo em Inglês | MEDLINE | ID: mdl-29325078

RESUMO

DNA recombination is required for effective segregation and diversification of genomes and for the successful completion of meiosis. Recent studies in various species hybrids have demonstrated a genetic link between DNA recombination and speciation. Consistent with this, we observed a striking suppression of recombination in the hybrids between two nematodes, the hermaphroditic Caenorhabditis briggsae and the gonochoristic C. nigoni. To unravel the molecular basis underlying the recombination suppression in their hybrids, we generated a C. nigoni genome with chromosome-level contiguity and produced an improved C. briggsae genome with resolved gaps up to 2.8 Mb. The genome alignment reveals not only high sequence divergences but also pervasive intra- and inter-chromosomal sequence re-arrangements between the two species, which are plausible culprits for the observed suppression. Comparison of recombination boundary sequences suggests that recombination in the hybrid requires extensive sequence homology, which is rarely seen between the two genomes. The new genomes and genomic libraries form invaluable resources for studying genome evolution, hybrid incompatibilities and sex evolution for this pair of model species.


Assuntos
Caenorhabditis/genética , Quimera/genética , Genoma , Organismos Hermafroditas/genética , Recombinação Genética , Animais , Sequência de Bases , Evolução Biológica , Caenorhabditis/classificação , Mapeamento Cromossômico , Cruzamentos Genéticos , Feminino , Biblioteca Genômica , Masculino , Meiose , Alinhamento de Sequência
20.
J Sci Food Agric ; 98(9): 3588-3594, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29315633

RESUMO

BACKGROUND: The highly selective capture of individual components of complex systems such as Chinese medicine extracts is a great challenge. With the rapid development of nanoscience, magnetic materials have attracted increased attention as suitable candidates for use in drug transport vehicles. RESULTS: In this work, amino-modified Fe3 O4 nanoparticles (MNP-NH2 ) have been synthesized and successfully used to selectively isolate luteolin, a type of flavonoid, from peanut shell, coupled with high-performance liquid chromatography. MNP-NH2 were characterized by transmission electron microscopy, scanning electron microscopy, Fourier transform infrared spectroscopy and vibrating sample magnetometer analysis, which showed that the amino functional groups were successfully bound to the surface of the magnetic particles. Adsorption of six different flavonoids by MNP-NH2 showed that the mechanism of adsorption was related to the number of ortho-phenolic hydroxyl groups. The optimized adsorption conditions were extraction for 40 min at 30 °C by single-factor experiments and the final elution conditions were determined to be 5 mL of 20% glacial acetic acid (methanol:water = 60:40) and ultrasonication for 40 min by L9 (34 ) orthogonal test. CONCLUSION: Based on these findings, we first created a method for separating luteolin from peanut shell extract by MNP-NH2 . This newly developed magnetic method of extracting chemical compounds from herbal foodstuffs and medicines provides new ideas for the utilization of a cash crop. © 2018 Society of Chemical Industry.


Assuntos
Arachis/química , Compostos Férricos/química , Flavonoides/isolamento & purificação , Nanopartículas/química , Nozes/química , Adsorção , Aminas/química , Cromatografia Líquida de Alta Pressão , Luteolina/isolamento & purificação , Fenômenos Magnéticos , Microscopia Eletrônica de Transmissão , Espectrofotometria Infravermelho
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