Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 81
Filtrar
Filtros adicionais











País/Região como assunto
Intervalo de ano
1.
Front Immunol ; 10: 1647, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31379845

RESUMO

Background: Antibody-dependent cellular cytotoxicity (ADCC), which mainly mediated by natural killer (NK) cells, may play a critical role in human immunodeficiency virus type-1 (HIV-1) disease progression. However, the potential mechanisms that affecting NK-mediated ADCC response are still not well-elucidated. Methods: Antigen-antibody complex model of Ab-opsonized P815 cells was adopted to induce a typical non-specific ADCC response. The capacities of HIV-1 specific NK-ADCC were measured by using the combination model of gp120 protein and plasma of HIV-1 elite controllers. The levels of plasma cytokine were measured by ELISA. Anti-IL-2 blocking antibody was used to analyze the impact of activated CD56+ T cells on NK-ADCC response. Results: IL-2, IL-15, IFN-α, and IFN-ß could effectively enhance the non-specific and HIV-1-specific NK-ADCC responses. Compared with healthy controls, HIV-1-infected patients showed decreased plasma IL-2 levels, while no differences of plasma IFN-α, IL-15, and IFN-ß were presented. IL-2 production was detected from CD56+ T cells activated through antibody-dependent manner. The capability of NK-ADCC could be weakened by blocking IL-2 secretion from activated CD56+ T cells. Although no difference of frequencies of CD56+ T cells was found between HIV-1-infected patients and healthy controls, deficient IL-2 secretion from activated CD56+ T were found in chronic HIV-1 infection. Conclusions: The impaired ability of activated CD56+ T cells to secreting IL-2 might contribute to the attenuated NK cell-mediated ADCC function in HIV-1 infection.

2.
Int J Mol Sci ; 20(8)2019 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-31010074

RESUMO

As the most widely-used single cell whole genome amplification (WGA) approach, multiple displacement amplification (MDA) has a superior performance, due to the high-fidelity and processivity of phi29 DNA polymerase. However, chimeric reads, generated in MDA, cause severe disruption in many single-cell studies. Herein, we constructed ChimeraMiner, an improved chimeric read detection pipeline for analyzing the sequencing data of MDA and classified the chimeric sequences. Two datasets (MDA1 and MDA2) were used for evaluating and comparing the efficiency of ChimeraMiner and previous pipeline. Under the same hardware condition, ChimeraMiner spent only 43.4% (43.8% for MDA1 and 43.0% for MDA2) processing time. Respectively, 24.4 million (6.31%) read pairs out of 773 million reads, and 17.5 million (6.62%) read pairs out of 528 million reads were accurately classified as chimeras by ChimeraMiner. In addition to finding 83.60% (17,639,371) chimeras, which were detected by previous pipelines, ChimeraMiner screened 6,736,168 novel chimeras, most of which were missed by the previous pipeline. Applying in single-cell datasets, all three types of chimera were discovered in each dataset, which introduced plenty of false positives in structural variation (SV) detection. The identification and filtration of chimeras by ChimeraMiner removed most of the false positive SVs (83.8%). ChimeraMiner revealed improved efficiency in discovering chimeric reads, and is promising to be widely used in single-cell sequencing.


Assuntos
Quimera/genética , Biologia Computacional/métodos , Software , Genoma Humano , Humanos , Análise de Célula Única , Sequenciamento Completo do Genoma
3.
Gene ; 699: 145-154, 2019 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-30876822

RESUMO

BACKGROUND: Detecting heteroplasmic variations in the mitochondrial genome can help identify potential pathogenic possibilities, which is significant for disease prevention. The development of next-generation sequencing changed the quantification of mitochondrial DNA (mtDNA) heteroplasmy from scanning limited recorded points to the entire mitochondrial genome. However, due to the presence of nuclear mtDNA homologous sequences (nuMTs), maximally retaining real variations while excluding falsest heteroplasmic variations from nuMTs and sequencing errors presents a dilemma. RESULTS: Herein, we used an improved method for detecting low-frequency mtDNA heteroplasmic variations from whole genome sequencing data, including point variations and short-fragment length alterations, and evaluated the effect of this method. A two-step alignment was designed and performed to accelerate data processing, to obtain and retain the true mtDNA reads and to eliminate most nuMTs reads. After analyzing whole genome sequencing data of K562 and GM12878 cells, ~90% of heteroplasmic point variations were identified in MitoMap. The results were consistent with the results of an amplification refractory mutation system qPCR. Many linkages of the detected heteroplasmy variations were also discovered. CONCLUSIONS: Our improved method is a simple, efficient and accurate way to mine mitochondrial low-frequency heteroplasmic variations from whole genome sequencing data. By evaluating the highest misalignment possibility caused by the remaining nuMTs-like reads and sequencing errors, our procedure can detect mtDNA heteroplasmic variations whose heteroplasmy frequencies are as low as 0.2%.


Assuntos
Genoma Mitocondrial/genética , Mitocôndrias/genética , Mutação/genética , Linhagem Celular Tumoral , DNA Mitocondrial/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Células K562 , Análise de Sequência de DNA/métodos , Sequenciamento Completo do Genoma/métodos
4.
J Biomed Nanotechnol ; 15(5): 1106-1111, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30890240

RESUMO

In recent years, liquid-cell transmission electron microscopy (LC-TEM) has been developed as in-situ, dynamic characterization of nanoscale materials in the liquid, although the state-of-art focus is heavy materials such as metals, and alloys. Herein we present a practical and stable liquid cell fabricated with standard micro-electro-mechanical (MEM) processes on silicon wafers. The liquid cell is universal for commonly used TEM holder, which may not only keep the liquid available for several weeks, and it has been also proven protective factor from electron beam damage when characterizing biomolecules such as proteins and DNAs, which are typical light-element molecules. DNA polymerase has been successfully characterized in the physiological state (unlabeled in PBS buffer), providing single molecular resolution, the dynamic structural evolution of the molecules and the complex interactions. Although more understandings of this technique has to be explored in the future, as we have pointed out in the manuscript, this work has illustrated that the LC-TEM can also become a potential and promising strategy, besides to the cryo-TEM technique, in the high-resolution characterization of biomolecules, which may benefit relative researches and industry, such as molecular and structural biology, ecology, pharmacology and environmental sciences.


Assuntos
Nanotecnologia , DNA , DNA Polimerase Dirigida por DNA , Microscopia Eletrônica de Transmissão , Silício
5.
Nanotechnology ; 30(26): 26LT01, 2019 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-30836332

RESUMO

Freestanding graphene films are desired to be widely applied in biosensor fabrication due to their distinctive physical properties and improved performance. Chemical vapor deposition has been developed to efficiently fabricate large-area graphene. However, some of the fabricated graphene films might break or be contaminated in the current transferring step using polymers. A stable and high-quality transfer method is needed. Herein, we report on an advanced transfer method of large-area graphene film which uses fullerene as a supporting substrate. Unlike polymers, which are commonly eliminated by being dissolved in an organic solution, fullerene can be easily removed by evaporation in a vacuum because it has a different heat stability to graphene. By using the improved transferring method, the percentage of integrated freestanding films after transferring was increased from 60.7% to 93.4%. The vacuum is beneficial in terms of keeping the brittle freestanding films intact. Graphene films transferred using fullerene showed an advanced flatness and a simplicial elementary composition in comparison to those transferred using polymers. Even through there is trace residue, this stable allotrope of graphene is considered to have almost no impact on biomolecule sensing. These advantages make the fullerene transferring method an attractive candidate for fabricating large-area freestanding graphene films, especially for using in the field of biochemistry analysis and biosensors.

6.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 37(1): 76-80, 2019 Feb 01.
Artigo em Chinês | MEDLINE | ID: mdl-30854824

RESUMO

OBJECTIVE: We aim to examine teenagers with varying vertical facial skeletal types with near-normal occlusion. We further aim to identify and study mandibular morphology and dentition characteristics to establish normal ranges and variations for future clinical reference. METHODS: According to the results of the case studies, 42 adolescents with near-normal occlusion were divided into three groups, namely, low- (7 cases), average- (23 cases) and high-angle (12 cases) groups. We used Invivo 5 software for Digital Imaging and Communications in Medicine (DICOM) data to calculate the cant of occlusal plane, axis corner of L6, ∠L1/MP, ∠L6/MP, Balkwill angle and Bonwill triangle of each group. RESULTS: Markedly, the finding shows that the cant of occlusal plane and axis corner of L6 in the low-angle group were smaller than those of the other two groups. In the average-angle group, ∠L1/MP was larger than that of the high-angle group. Lastly, in the high-angle group, ∠L6/MP was smaller than those of the two other groups. On the one hand, these differences were considered statistically significant (P<0.05). On the other hand, other measurements show that these differences were considered statistically non-significant (P>0.05). CONCLUSIONS: In the low-angle group, the parallelisation of the occlusal plane tends to be more obvious compared with the two other groups. In the coronal section of the low-angle group, the axis of the mandibular first molar is up-right, whereas it is distally tilted in the sagittal section of the high-angle group. Furthermore, a number of differences are noted in the adult groups. Factors, such as aging and development in the craniofacial region, lead to changes in functional occlusion.


Assuntos
Cefalometria , Oclusão Dentária , Dentição , Adolescente , Humanos , Mandíbula , Dente
7.
Nanoscale ; 10(37): 17933-17941, 2018 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-30226245

RESUMO

Whole genome amplification (WGA) has laid the foundation for investigating complex genomic alteration with single-cell or even single-molecule resolution. Coupled with sequencing-based copy number variation (CNV) analysis, it promotes understanding of the nature of commonly existing genetic heterogeneity by constructing the sequencing profiles for every single cell. However, prevailing methods only provide insights into limited aspects due to their intrinsic technical challenges. Their output data, as a result, fails to render comprehensive information (which is) concerned. Here, we describe the CNV detection analysis based on micro-channel multiple displacement amplification (µcMDA), a protocol able to provide optimized amplification uniformity while inheriting the advantages of MDA chemistry. We demonstrate the analysis of both the normal diploid YH-1 cell line and the aneuploid K562 cancer cell line. In the detection of simulated CNVs ranging from 300 kb to 2 Mb, µcMDA can respectively increase the detection rates of copy number loss and gain by 28.8% and 40.2% on average, using only 0.2× sequencing data. When detecting the inherent CNVs in tumor cells, the resolution of CNV recognition can be improved to 250 kb. Starting from either superabundant template copies or minute single-cell-level input, this easily accessible approach is capable of providing quantitatively reliable coverage as well as more robust GC-content regression for CNV detection.

8.
Oncol Lett ; 16(4): 4863-4870, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30250552

RESUMO

Gastric cancer is a highly heterogeneous disease and the second leading cause of cancer-associated mortality. However, the genomic basis of gastric cancer is not completely understood and the underlying genetic heterogeneity has not been well studied. In the present study, 1,021 genes were sequenced and the somatic mutations of 45 formalin-fixed, paraffin-embedded gastric adenocarcinoma samples were assessed using next-generation sequencing technologies. In the present study, a median sequencing coverage depth of 708-fold was achieved. Somatic genomic alterations were detected in 37/45 patients (82.4%) and the most frequent genetic alterations identified were tumor protein P53 (TP53) gene mutations. Mutations in MLL4, ERBB3, FBXW7, MLL3, MTOR, NOTCH1, PIK3CA, KRAS, ERBB4 and EGFR were also detected. Patients with TP53 mutations had a higher number of somatic mutations, and the total number of somatic mutations was weakly correlated with patient age. These results provided data on the intratumoral heterogeneity of gastric cancer and may be used in order to develop personalized cancer therapy.

9.
Bioanalysis ; 10(15): 1177-1180, 2018 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-30136900
10.
J Biomol Struct Dyn ; : 1-10, 2018 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-30058436

RESUMO

Bromodomain-containing protein 9 (BRD9) has been employed as a potential target for anticancer drugs in recent years. In this work, molecular docking, molecular dynamics (MD) simulations, binding free energy calculations, and per residue energy decomposition approaches were performed to elucidate the different binding modes between four pyridinone-like scaffold inhibitors and BRD9 bromodomain. Analysis results indicate that non-polar contribution mainly deriving from van der Waals energy is a critical impact on binding affinity of inhibitors against BRD9. Some key residues Phe44, Phe47, Val49, and Ile53 (at ZA loop) enhance the binding energy of inhibitors in BRD9 by means of providing hydrophobic interactions. Moreover, it is observed that BRD9 is anchored by the formation of a stable hydrogen bond between the carbonyl of the inhibitors and the residue Asn100 (at BC loop), and a strong π-π stacking interaction formed between the residue Tyr106 (at BC loop) and the inhibitors. The existence of dimethoxyphenyl structure and the aromatic ring merged to pyridinone scaffold are useful to enhance the BRD9 binding affinity. These findings should guide the rational design of more prospective inhibitors targeting BRD9. Communicated by Ramaswamy H. Sarma.

11.
J Biomol Struct Dyn ; : 1-33, 2018 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-30044186

RESUMO

Tuberculosis (TB) is an infectious disease that causes a number of deaths, and the development of new, safer and more adequate TB inhibitors/drugs has become a necessity as well as a great challenge. Mycobacterial DNA gyrase B subunit has been identified to be one of the potentially underexploited drug targets in the field of anti-tubercular drug discovery. To design the novel and potent Mycobacterium tuberculosis inhibitors, we performed molecular modeling studies that combined the 3D-QSAR, molecular docking, molecular dynamics simulations, and binding free energy calculations. 48 quinoline-aminopiperidine inhibitors which act on DNA gyrase B subunit were used for constructing 3D-QSAR models. The results showed that the best CoMFA model had the high performance with q2 = 0.643, r2 = 0.947, while the best CoMSIA model yielded q2 = 0.536, r2 = 0.948. The contour map was in good agreement with the docking and molecular dynamics simulations which strongly demonstrated that the molecular modeling was reliable. Based on this information, several potential compounds were designed and their inhibitory activities were also verified by the accomplished models and ADME/T predictions. We hope that our research could bring new ideas to facilitate the development of novel inhibitors with higher inhibitory activity for TB.

12.
Int J Biol Macromol ; 118(Pt A): 1149-1156, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-30001602

RESUMO

As an attractive therapeutic target for non-small-cell lung cancer (NSCLC), anaplastic lymphoma kinase (ALK) has got increased attention, and the selectivity of ALK inhibitors is an enormous challenge. Recently, 2,4-Diarylaminopyrimidines with high inhibitory activity over InsR/IGF1R were reported as ALK inhibitors, which harboring phosphine oxide moiety. In this work, it is the first time to reveal that the incorporation of dimethylphosphine oxide moiety and the smaller active pocket of ALK is key factor in the selectivity of inhibitor 11q toward ALK over IGF1R/InsR. The results of molecular simulation indicate that the subtle change in the binding pocket of ALK is mainly associated with the flexibility of P-loop and the own residues K1150 and D1270. The replacement of the dimethylphosphine oxide and methylpiperazine of inhibitor 11q would alter the major inhibitory effects of binding and activation. The results further combined 3D-QSAR can not only profile the binding mechanism between the 2,4-Diarylaminopyrimidines inhibitors and ALK, but also supply the useful information for the rational design of a more potential small molecule inhibitor bound to ALK receptor.


Assuntos
Desenho de Drogas , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Inibidores de Proteínas Quinases/química , Pirimidinas/química , Receptores Proteína Tirosina Quinases/antagonistas & inibidores , Quinase do Linfoma Anaplásico , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/enzimologia , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/enzimologia , Receptores Proteína Tirosina Quinases/química
14.
Int J Pharm ; 543(1-2): 169-178, 2018 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-29567198

RESUMO

Hemoglobin (Hb)-loaded mesoporous silica nanoparticles (MSNs) coated with a lipid bilayer (LB-MSNs) were investigated as an erythrocyte mimic. MSNs with a large average pore size (10 nm) act as a rigid core and provide a protective environment for Hb encapsulated inside the pores. The colloidal stability of Hb-loaded MSNs was enhanced upon the application of a lipid bilayer, through fusion of PEGylated liposomes onto the exterior surface of Hb-loaded MSNs. The morphology and mesostructure of the MSNs were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and surface area analysis. The Hb loading capacity (mg/g) in MSNs was studied by thermogravimetric analysis (TGA). UV-Vis absorption spectroscopy revealed that Hb inside MSNs had an identical, but slightly broadened peak in the Soret region compared to free Hb. Furthermore the encapsulated Hb exhibits similar peroxidase-like activity in catalyzing the oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) with hydrogen peroxide. The introduction of a supported lipid bilayer (LB) demonstrated the potential to prevent premature Hb release (the burst release decreased from 25.50 ±â€¯0.33% to 6.73 ±â€¯0.83%) and increased the colloidal stability of the Hb-loaded MSNs (hydrodynamic diameter remained ∼250 nm for at least one week). The in vivo systemic circulation and biodistribution of LB-MSNs were studied in optically transparent zebrafish embryos, revealing that LB-MSNs have the potential to act as an erythrocyte mimic in transfusion therapy.


Assuntos
Hemoglobinas/administração & dosagem , Bicamadas Lipídicas/administração & dosagem , Nanopartículas/administração & dosagem , Dióxido de Silício/administração & dosagem , Animais , Animais Geneticamente Modificados/metabolismo , Circulação Sanguínea , Bovinos , Liberação Controlada de Fármacos , Embrião não Mamífero/metabolismo , Eritrócitos , Hemoglobinas/química , Bicamadas Lipídicas/química , Nanopartículas/química , Porosidade , Dióxido de Silício/química , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Peixe-Zebra/metabolismo
15.
Molecules ; 23(2)2018 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-29401641

RESUMO

The co-existence of wild-type and mutated mitochondrial DNA (mtDNA) molecules termed heteroplasmy becomes a research hot point of mitochondria. In this review, we listed several methods of mtDNA heteroplasmy research, including the enrichment of mtDNA and the way of calling heteroplasmic variations. At the present, while calling the novel ultra-low level heteroplasmy, high-throughput sequencing method is dominant while the detection limit of recorded mutations is accurate to 0.01% using the other quantitative approaches. In the future, the studies of mtDNA heteroplasmy may pay more attention to the single-cell level and focus on the linkage of mutations.


Assuntos
Biologia Computacional/métodos , DNA Mitocondrial/genética , Genoma Mitocondrial , Mutação INDEL , Mitocôndrias/genética , Polimorfismo de Nucleotídeo Único , Centrifugação com Gradiente de Concentração , DNA Mitocondrial/isolamento & purificação , DNA Mitocondrial/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase
16.
Adv Mater ; 30(24): e1703658, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29389041

RESUMO

DNA encodes the genetic information; recently, it has also become a key player in material science. Given the specific Watson-Crick base-pairing interactions between only four types of nucleotides, well-designed DNA self-assembly can be programmable and predictable. Stem-loops, sticky ends, Holliday junctions, DNA tiles, and lattices are typical motifs for forming DNA-based structures. The oligonucleotides experience thermal annealing in a near-neutral buffer containing a divalent cation (usually Mg2+ ) to produce a variety of DNA nanostructures. These structures not only show beautiful landscape, but can also be endowed with multifaceted functionalities. This Review begins with the fundamental characterization and evolutionary trajectory of DNA-based artificial structures, but concentrates on their biomedical applications. The coverage spans from controlled drug delivery to high therapeutic profile and accurate diagnosis. A variety of DNA-based materials, including aptamers, hydrogels, origamis, and tetrahedrons, are widely utilized in different biomedical fields. In addition, to achieve better performance and functionality, material hybridization is widely witnessed, and DNA nanostructure modification is also discussed. Although there are impressive advances and high expectations, the development of DNA-based structures/technologies is still hindered by several commonly recognized challenges, such as nuclease instability, lack of pharmacokinetics data, and relatively high synthesis cost.

17.
Phys Chem Chem Phys ; 20(9): 6034-6039, 2018 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-29344597

RESUMO

Confining protein crystallization to a millimetre size was achieved within 0.5 h after stopping 1 h intense trapping laser irradiation, which shows excellent performance in spatial and temporal controllability compared to spontaneous nucleation. A continuous-wave near-infrared laser beam is tightly focused into a glass/solution interfacial layer of a supersaturated buffer solution of hen egg-white lysozyme (HEWL). The crystallization is not observed during laser trapping, but initiated by stopping the laser irradiation. The generated crystals are localized densely in a circular area with a diameter of a few millimetres around the focal spot and show specific directions of the optical axes of the HEWL crystals. To interpret this unique crystallization, we propose a mechanism that nucleation and the subsequent growth take place in a highly concentrated domain consisting of HEWL liquid-like clusters after turning off laser trapping.


Assuntos
Lasers , Muramidase/química , Animais , Galinhas , Cristalização/métodos
18.
Sci Total Environ ; 618: 1254-1267, 2018 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-29089134

RESUMO

Massive partial sequencing of 16S rRNA genes has become the predominant tool used for studying microbial ecology. However, determining which hypervariable regions and primer sets should be used for screening microbial communities requires extensive investigation if controversial results are to be avoided. Here, the performances of different variable regions of the 16S rRNA gene on bacterial diversity studies were evaluated in silico with respect to the SILVA non-redundant reference database (SILVA SSU Ref 123NR), and subsequently verified using samples from Lake Taihu in China, a eutrophic lake. We found that the bacterial community composition results were strongly impacted by the different V regions. The results show that V1-V2 and V1-V3 regions were the most reliable regions in the full-length 16S rRNA sequences, while most V3 to V6 regions (including V3, V4, V3-V4, V5, V4-V5, V6, V3-V6, V4-V6, and V5-V6) were more closely aligned with the SILVA SSU Ref 123NR database. Overall, V4 was the most prominent V region for achieving good domain specificity, higher coverage and a broader spectrum in the Bacteria domain, as confirmed by the validation experiments. S-D-Bact-0564-a-S-15/S-D-Bact-0785-b-A-18 is, therefore, a promising primer set for surveying bacterial diversity in eutrophic lakes.


Assuntos
Bactérias/genética , Monitoramento Ambiental , Eutrofização , Lagos/microbiologia , Bactérias/classificação , China , Genes Bacterianos , Variação Genética , RNA Ribossômico 16S , Análise de Sequência de DNA
19.
Pharmacol Res ; 129: 491-499, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29175550

RESUMO

The smoothened (SMO) receptor, an essential signal transducer in the Hedgehog pathway, was targeted with antagonists to suppress the tumor. It is interesting that SMO D473H mutation confers resistance on inhibitor LDE-225 rather than LEQ-506. In this paper, the binding modes of them against the wild type and mutant SMO receptors were identified to gain insights into the resistant and non-resistant factors, based on a comprehensive protocol involving molecular docking, molecular dynamic simulations, free energy calculation and decomposition. A comparison of resistant LDE-225 and non-resistant LEQ-506 indicates that the volume of the binding cavity decreases seriously in the mutant complex with resistant LDE-225. In addition, the D473H mutation disrupts the hydrogen bond network with residues R400 and Q477, which results in the TM6 conformation inward. Owing to the absence of the hydrogen bond, residues R400 and Q477 make weak contributions to LDE-225. However, the D473H mutation along with TM6 conformational change has no effect on non-resistant LEQ-506. Finally, the resistance ascribes to adverse interaction between the greater polarity of mutant residue H473 and the nonpolar phenmethyl of LDE-225. The elaborate insights into structural and energetic mechanism of drug resistance provide an effective strategy to design rationally non-resistant antagonists.

20.
Biomed Res Int ; 2017: 8184160, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28904972

RESUMO

The characteristics of tongue coating are very important symbols for disease diagnosis in traditional Chinese medicine (TCM) theory. As a habitat of oral microbiota, bacteria on the tongue dorsum have been proved to be the cause of many oral diseases. The high-throughput next-generation sequencing (NGS) platforms have been widely applied in the analysis of bacterial 16S rRNA gene. We developed a methodology based on genus-specific multiprimer amplification and ligation-based sequencing for microbiota analysis. In order to validate the efficiency of the approach, we thoroughly analyzed six tongue coating samples from lung cancer patients with different TCM types, and more than 600 genera of bacteria were detected by this platform. The results showed that ligation-based parallel sequencing combined with enzyme digestion and multiamplification could expand the effective length of sequencing reads and could be applied in the microbiota analysis.


Assuntos
Bactérias/genética , RNA Ribossômico 16S/genética , Dermatopatias/microbiologia , Língua/microbiologia , Adulto , Idoso , Bactérias/classificação , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Dermatopatias/genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA