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1.
Anal Chem ; 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31565925

RESUMO

The emergence and ongoing spread of multidrug-resistant (MDR) bacteria is a major global public health threat. MDR has extensive-ly combated the potency of antibiotics. Development of new antibiotics requires several years with prohibitive cost that will not last. An alternative solution is to re-combine failed antibiotics, which has been proven to be not only cost effective, but also potent. However, selection of the optimal combinations of these chemicals through conventional trial-and-error methods is challenging and slow, since M candidates with N doses lead to NM possible combinations. Herein, we present a artificial intelligence (AI) guided chemical combination optimization technique, namely Streamlined Rapid Identification of Combinatorial Therapies (STRICT), which is phenotype based and can efficiently learn and identify the optimal drug-combinations with minimal experimental efforts. With the guidance of STRICT, we successfully identified potent combinations of five antibiotics from 26 antibiotics that are indi-vidually ineffective at inhibiting an artificially induced strain of MDR bacteria. Rather than examine millions of tests, STRICT ac-complished this task with only 120 carefully selected tests. Our results indicate that STRICT is a powerful platform to identify effi-cacious multi-antibiotic combinations for the treatment of MDR bacteria. The AI-guided platform introduced here is an effective tool for drug repurposing, beneficial towards large-scale drug screening for other disease models, and also has a broad application in chemical combination optimization to deliver a desired endpoint for a complex system.

2.
Antiviral Res ; 171: 104598, 2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31513822

RESUMO

As antiviral drug resistance develops and new viruses emerge there is a pressing need to develop strategies to rapidly develop antiviral therapeutics. Here we use phospho-specific flow cytometry to assess perturbations of many different cellular signaling pathways during treatment with drug combinations that are highly effective in blocking Herpes simplex virus type 1 (HSV-1) infection. We discovered two antiviral drug combinations act on distinct signaling pathways, either STAT1 or S6 phosphorylation, to block HSV-1 infection. We focused on upregulation of S6 phosphorylation by HSV-1 infection, and our subsequent finding that ribavirin antagonizes this upregulation of S6 phosphorylation. We go on to show that the S6 kinase inhibitor SL0101 blocks HSV-1 replication in vitro and in an in vivo animal model of HSV-1 infection. Overall, we have used an unbiased analysis of cellular signaling pathways during treatment by antiviral drug combinations to discover a novel antiviral drug target against HSV-1 infection. The outcomes of the approach we present highlight the importance of analyzing how antiviral drugs modulate cellular and pathogen-induced signaling as a method to discover new drug therapy targets.

3.
SLAS Technol ; : 2472630319867903, 2019 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-31381466

RESUMO

Typography-like templates for polydimethylsiloxane (PDMS) microfluidic chips using a fused deposition modeling (FDM) three-dimensional (3D) printer are presented. This rapid and fast proposed scheme did not require complicated photolithographic fabrication facilities and could deliver resolutions of ~100 µm. Polylactic acid (PLA) was adopted as the material to generate the 3D-printed units, which were then carefully assembled on a glass substrate using a heat-melt-curd strategy. This craft of bonding offers a cost-effective way to design and modify the templates of microfluidic channels, thus reducing the processing time of microfluidic chips. Finally, a flexible microfluidic chip to be employed for cell-based drug screening was developed based on the modularized 3D-printed templates. The lithography-free, typography-like, 3D-printed templates create a modularized fabrication process and promote the prevalence of integrated microfluidic systems with minimal requirements and improved efficiency.

4.
Nanomedicine ; 21: 102047, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31271877

RESUMO

Drug delivery nanocarriers based on magnetic nanoparticles have attracted increasing attention due to their potential applications in magnetic resonance imaging, photodynamic therapy and targeted drug delivery. Herein, we have fabricated the multifunctional co-loaded magnetic nanocapsules (MNCPs) using a microemulsion process for enhancing targeted magnetic resonance imaging and in vivo photodynamic therapy. MNCPs were synthesized by co-loading Co@Mn magnetic nanoparticles and chlorin e6 into the matrix of an amphiphilic polymer, and further surface covalently coupled with target molecules. This work demonstrates that MNCPs have uniform sizes (dc: ~150 nm), favorable biocompatibility, long-term stability, excellent T2 relaxation values, and high drug loading efficiency. These advantages offer MNCPs successfully applied in targeted magnetic resonance imaging, real-time fluorescent labeling, and photodynamic therapy. The research results will contribute to rationally design novel nano-platform and provide a promising approach for further clinical integration of diagnosis and treatment in the near future.

5.
Front Immunol ; 10: 1571, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31354723

RESUMO

Colon cancer (CC) is one of the leading causes of cancer related mortality. Research over past decades have profoundly enhanced our understanding of immunotherapy, a major clinical accomplishment, and its potential role toward treating CC. However, studies investigating the expression of these immune checkpoints, such as epithelial cell adhesion molecule (EpCAM), programmed death-1 (PD-1), and programmed death-ligand 1 (PD-L1), by peripheral blood mononuclear cells (PBMCs) is lacking. Here, high-dimensional mass cytometry (CyTOF) is used to investigate immune alterations and promising immunotherapeutic targets expression by PBMCs of CC patients. Results reveal that expression of EpCAM and PD-L1 on CD4+ T cells significantly increased in patients with CC, compared with age- and sex- matching healthy controls and patients with colonic polyps. These differences are also validated in an independent patient cohort using flow cytometry. Further analysis revealed that EpCAM+ CD4+ T cells are PD-L1+ CCR5+ CCR6+. Immunofluorescence staining results demonstrate that the increase of EpCAM+ CD4+ T cells is also observed in tumor tissues, rather than para-cancerous tissues. To ascertain the functional disorders of the identified cell subset, phosphorylated signaling protein levels are assessed using imaging mass cytometry. Increases in pp38 MAPK and pMAPKAPK2 are observable, indicating abnormal activation of pp38 MAPK-pMAPKAPK2 signaling pathway. Results in this study indicate that EpCAM+ CD4+ T cells may play a role in CC development. Detailed knowledge on the functionality of EpCAM+ CD4+ T cells is of high translational relevance.

6.
Nanomedicine ; 20: 102019, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31125676

RESUMO

How to eradicate Helicobacter pylori (H. pylori) in vivo with antibiotic resistance owns tremendous clinical requirement. Herein, gold nanostars were conjugated with acid-sensitive cis-aconitic anhydride modified anti-H. pylori polyclonal antibodies, resultant pH sensitive gold nanostars@H. pylori-antibodies nanoprobes (GNS@Ab) were employed for the theranostics of H. pylori in vivo. Photoacoustic imaging confirmed that prepared GNS@Ab could target actively H. pylori in the stomach. GNS@Ab nanoprobes could kill H. pylori in vivo in model animals under NIR laser irradiation, all GNS@Ab nanoprobes could be excreted out of gut within 7 days after oral administration. Gastric local lesion caused by H. pylori restored to normal status within one month. GNS@Ab nanoprobes within therapeutic doses did not damage intestinal bacteria imbalance. Forty clinical specimens of H. pylori with antibiotic resistance were verified validity of GNS@Ab nanoprobes. Prepared oral pH-sensitive GNS@Ab nanoprobes own clinical translational potential in the theranostics of H. pylori in near future.

7.
Anal Chem ; 91(12): 7524-7530, 2019 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-31117398

RESUMO

The facile and economical identification of pathogenic bacteria, especially their antibiotic-resistance, is crucial in the realm of human health and safety. The presence of Escherichia coli ( E. coli) is considered as an indicator of water contamination and is closely related to human health. Herein, inspired by the biocatalysis of bacterial surfaces, we developed a simple and cost-effective colorimetric- and electrochemical-based bioassay that is capable of analyzing both the presence of E. coli and its relative level of antibiotic resistance. In this approach, p-benzoquinone is used as a redox mediator to monitor the bacterial concentration and specifically distinguish E. coli from four other common clinical bacteria, namely, Staphylococcus aureus ( S. aureus), Enterococcus faecalis ( E. faecalis), Salmonella pullorum ( S. pullorum), and Streptococcus mutans ( S. mutans). A visible color change, captured with a smartphone using a "light box", without relying on any complex instruments, can reflect the concentration of bacteria. The accurate quantification of E. coli was investigated with an electrochemical system in the concentration ranges of 1.0 × 103 to 1.0 × 109 CFU/mL. We further demonstrated the capability of the presented biosensor in identifying drug-resistant bacteria with two artificially induced antibiotic-resistant bacteria. Therefore, the presented bioassay is not only capable of detecting E. coli with high sensitivity and specificity but also provides a rapid solution to evaluate E. coli antibiotic resistance.

8.
PLoS One ; 14(5): e0215607, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31075149

RESUMO

BACKGROUND: Shorter, more effective treatments for tuberculosis (TB) are urgently needed. While many TB drugs are available, identification of the best regimens is challenging because of the large number of possible drug-dose combinations. We have found consistently that responses of cells or whole animals to drug-dose stimulations fit a parabolic response surface (PRS), allowing us to identify and optimize the best drug combinations by testing only a small fraction of the total search space. Previously, we used PRS methodology to identify three regimens (PRS Regimens I-III) that in murine models are much more effective than the standard regimen used to treat TB. However, PRS Regimens I and II are unsuitable for treating drug-resistant TB and PRS Regimen III includes an experimental drug. Here, we use PRS methodology to identify from an expanded pool of drugs new highly effective near-universal drug regimens comprising only approved drugs. METHODS AND FINDINGS: We evaluated combinations of 15 different drugs in a human macrophage TB model and identified the most promising 4-drug combinations. We then tested 14 of these combinations in Mycobacterium tuberculosis-infected BALB/c mice and chose for PRS dose optimization and further study the two most potent regimens, designated PRS Regimens IV and V, consisting of clofazimine (CFZ), bedaquiline (BDQ), pyrazinamide (PZA), and either amoxicillin/clavulanate (AC) or delamanid (DLM), respectively. We then evaluated the efficacy in mice of optimized PRS Regimens IV and V, as well as a 3-drug regimen, PRS Regimen VI (CFZ, BDQ, and PZA), and compared their efficacy to PRS Regimen III (CFZ, BDQ, PZA, and SQ109), previously shown to reduce the time to achieve relapse-free cure in mice by 80% compared with the Standard Regimen (isoniazid, rifampicin, PZA, and ethambutol). Efficacy measurements included early bactericidal activity, time to lung sterilization, and time to relapse-free cure. PRS Regimens III-VI all rapidly sterilized the lungs and achieved relapse-free cure in 3 weeks (PRS Regimens III, V, and VI) or 5 weeks (PRS Regimen IV). In contrast, mice treated with the Standard Regimen still had high numbers of bacteria in their lungs after 6-weeks treatment and none achieved relapse-free cure in this time-period. CONCLUSIONS: We have identified three new regimens that rapidly sterilize the lungs of mice and dramatically shorten the time required to achieve relapse-free cure. All mouse drug doses in these regimens extrapolate to doses that are readily achievable in humans. Because PRS Regimens IV and V contain only one first line drug (PZA) and exclude fluoroquinolones and aminoglycosides, they should be effective against most TB cases that are multidrug resistant (MDR-TB) and many that are extensively drug-resistant (XDR-TB). Hence, these regimens have potential to shorten dramatically the time required for treatment of both drug-sensitive and drug-resistant TB. If clinical trials confirm that these regimens dramatically shorten the time required to achieve relapse-free cure in humans, then this radically shortened treatment has the potential to improve treatment compliance, decrease the emergence of drug resistance, and decrease the healthcare burden of treating both drug-sensitive and drug-resistant TB.

9.
Proc Inst Mech Eng H ; 233(7): 683-694, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31113284

RESUMO

Single-cell analysis serves as an important approach to study cell functions and interactions. Catering to the demand of Big Data Era, fast reactions for single cells and paralleled high-throughput analysis have become an urgent need. Microdroplet in microfluidics has advantages of modularity and integrity, as well as high throughput and sensitivity, which present great potential in the field of single-cell analysis. This review is carried out on three aspects to introduce microdroplet chips for single-cell analysis: droplet formation, droplet detection and practical functions. Structures of droplet formation are categorized into three types, including T-shaped channel, flow-involved channel and three-dimensional micro-vortice. The detection methods, including fluorescence, Raman spectroscopy, mass spectroscopy and electrochemical detection, are summarized from applications. Both pros and cons for existing techniques are reviewed and discussed. The functions of microdroplets-on-chip cover cell culture, nucleic acid test and cell identification. For each field, principles/mechanisms and/or schematic images are laconically introduced. Microdroplet in microfluidics has become a major research direction in single-cell analysis. With updated methods of droplet formation such as inertial ordering and micro-vortice, microdroplets-based biochips will expect high throughput detection and high-accuracy trace detection for clinical diagnosis in the near future.

10.
SLAS Technol ; 24(4): 408-419, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30856358

RESUMO

Mass cytometry (CyTOF) is a critical cell profiling tool in acquiring multiparameter proteome data at the single-cell level. A major challenge in CyTOF analysis is sample-to-sample variance arising from the pipetting process, staining variation, and instrument sensitivity. To reduce such variations, cell barcoding strategies that enable the combination of individual samples prior to antibody staining and data acquisition on CyTOF are often utilized. The most prevalent barcoding strategy is based on a binary scheme that cross-examines the existence or nonexistence of certain mass signals; however, it is limited by low barcoding efficiency and high cost, especially for large sample size. Herein, we present a novel barcoding method for CyTOF application based on mass ratiometry. Different mass tags with specific fixed ratios are used to label CD45 antibody to achieve sample barcoding. The presented method exponentially increases the number of possible barcoded samples with the same amount of mass tags compared with conventional methods. It also reduces the overall time for the labeling process to 40 min and avoids the need for expensive commercial barcoding buffer reagents. Moreover, unlike the conventional barcoding process, this strategy does not pre-permeabilize cells before the barcoding procedure, which offers additional benefits in preserving surface biomarker signals.

11.
J Pharm Biomed Anal ; 168: 44-54, 2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-30784889

RESUMO

Tuberculosis is one of the top concerns in the world and acutely threatens human health. A new potent candidate regimen containing pyrazinamide (PZA), ethambutol (EMB), protionamide (PTO) and clofazimine (CFZ) was proposed by Parabolic Response Surface/Feedback System Control (FSC/PRS) system and showed excellent outcomes in vitro and vivo studies. Here, a convenient liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneously determination of four compounds in beagle dog plasma. The plasma samples, 50 µL for each, were pretreated by methanol on 96-well format plates and a further dilution step was designed to reduce predictable matrix effect and lessen the burden of subsequent analysis. The chromatographic separation was achieved on an Agilent SB-Aq column (4.6 mm × 150 mm, 5 µm) at 30 °C by a gradient elution within 6 min. The mobile phase was a mixture of 0.2% formic acid-5 mM ammonium acetate aqueous solution (phase A) and 0.2% formic acid methanol (phase B) with a total flow rate of 1 mL/min. The 30% of post-column eluant was injected into mass spectrometer, equipped with electrospray ionization (ESI) source under positive mode and multiple-reaction monitoring (MRM). This quantification method was proved to be satisfied in selectivity, accuracy, precision, linearity (r2 > 0.998), recovery, matrix effect and stability. Under the specialized conditions, the calibration curves ranged from 20 to 5000 ng/mL for PZA, 1 to 500 ng/mL for EMB, 1 to 500 ng/mL for PTO, and 1 to 200 ng/mL for CFZ. The quantitative accuracy was further assessed under different degrees of hemolyses in detail. This method was proved to be robust and efficient, and successfully applied to the pharmacokinetic study of the new regimen in Beagle dogs.


Assuntos
Antituberculosos/análise , Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem/métodos , Animais , Antituberculosos/farmacocinética , Calibragem , Clofazimina/análise , Clofazimina/farmacocinética , Cães , Etambutol/análise , Etambutol/farmacocinética , Protionamida/análise , Protionamida/farmacocinética , Pirazinamida/análise , Pirazinamida/farmacocinética , Reprodutibilidade dos Testes
12.
Talanta ; 197: 304-309, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30771940

RESUMO

Rapid and sensitive detection of live bacteria is crucial in the realm of clinical diagnosis, food industry and environmental quality control. A portable, feasible and cost-effective platform which enables rapid and accurate live bacteria detection is still challenging. Herein, we present a Bacterial Inhibition of GOX-catalyzed Reaction (BIGR) method for rapid and broad-spectrum detection of live bacteria, which results in a visible color change without any complex instrumentations. We validated this strategy with five common clinical bacteria, namely Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans and Salmonella pullorum. This method precludes the interference of dead bacteria. Only several microliters of samples and reagents are required in this assay and the overall analysis time is less than 20 min. In a further demonstration, the presented method is successfully applied for detection of ascites samples from infected mice. Our results suggest that this method serves as a rapid and dose-dependent visual detection of pathogens in the clinical and daily life.


Assuntos
Colorimetria , Enterococcus faecalis/isolamento & purificação , Escherichia coli/isolamento & purificação , Glucose Oxidase/antagonistas & inibidores , Salmonella/isolamento & purificação , Staphylococcus aureus/isolamento & purificação , Streptococcus mutans/isolamento & purificação , Animais , Biocatálise , Enterococcus faecalis/metabolismo , Escherichia coli/metabolismo , Glucose Oxidase/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Imagem Óptica , Salmonella/metabolismo , Staphylococcus aureus/metabolismo , Streptococcus mutans/metabolismo
13.
Biosens Bioelectron ; 129: 175-181, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30710755

RESUMO

White blood cells (WBCs) isolated from peripheral blood have been verified as important biomarkers for the diagnosis, treatment and prognosis of cancer. However, it's still under challenge to acquire high-purity WBCs, even by taking advantage of current microfluidic technology. Considering the universality of clinical magnetic activated cell sorting (MACS) method, new developments on microfluidic chip in combination of magnetic cells separation technologies may provide a fascinating approach for high-purity WBCs sorting and widely clinical application. Here, we present a flyover style microfluidic chip which has been elaborately embedded with two-stage magnetic separation in continuous flow for WBCs sorting. Immunomagnetic micro/nano-particles (IMNPs) labeled WBC (WBC@IMNPs) were sequentially separated by a lateral magnetic force and a vertical magnetic force, and the final separation purity of WBCs reached up to 93 ±â€¯1.67% at a flow rate of 20 µL min-1. Furthermore, the WBCs viability was up to 97.5 ±â€¯1.8%. Consequently, this novel flyover style microfluidic-chip with magnetic separation technology has been successfully demonstrated as cut-in-edge method for high-purity WBCs sorting, and obviously it's easy to extend for other types of cells sorting under great potential application in biomedical fields.


Assuntos
Separação Imunomagnética/instrumentação , Dispositivos Lab-On-A-Chip , Leucócitos/citologia , Animais , Sobrevivência Celular , Desenho de Equipamento , Campos Magnéticos , Camundongos Endogâmicos BALB C , Níquel/química
14.
JCI Insight ; 4(6)2019 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-30747724

RESUMO

Psoriasis (PS) is a systemic, immune-mediated inflammatory disorder. However, the whole lymphocyte compartment and the potential pathologies of PS have not been fully characterized. In the present study, we examined whole lymphocyte subsets and signal transduction proteins using high-dimensional single-cell mass cytometry and a bioinformatics pipeline for an in-depth characterization of the immune cell subsets and protein profiles involved in pathways in the peripheral blood of patients with PS. We identified 15 major immune cell populations in T cell lineages and characterized various CD3+CD4+ Th and CD3+CD8+ T cytotoxic cell populations simultaneously across 24 leukocyte markers and 7 proteins related to the signal transduction pathways. High-dimensional analysis identified 3 new subsets that are abundant in PS peripheral blood, resembling CD3-CD4+ lymphoid tissue inducer cells, Tc17 cells, and CD8+CXCR3+ Tregs. We confirmed the CD3-CD4+ cells, and their features and functions, in an independent PS cohort. The use of single-cell mass cytometry allows systemic-level characterization of lymphocyte subpopulations and dysregulated signaling pathways in the blood of patients with PS, identifying abnormalities of different immune cell subsets. We validated that the CD3-CD4+ cells had elevated OX40 and decreased FRA2 expression, which were positively associated with the PS area and severity index.

15.
Food Chem ; 277: 624-631, 2019 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-30502195

RESUMO

A rapid and fast detection of trace amounts of melamine in milk is reported by using Gold Nano Spheres embedded monolith conjugates. Monolith was synthesized by the polymerization of Glycidyl Methacrylate (GMA) and Ethylene Dimethacrylate(EDMA) (cross linker and functional monomer), Cyclohexanol (Porogen formation) and 2, 2-Dimethoxy-2-phenyl-acetophenone (photo-initiator) on gold coated silicon wafer. In order to gauge the influence of monolith on SERS signal activity, three shapes of gold nanoparticles namely Gold Nano Spheres (GNSs), Gold Nanorods (GNRs) and Triangular Gold Nanoprisms (GNPrs) were immobilized on monolithic surface and analyzed by the signal molecule Rhodamine (R6G). The Raman Enhancement efficiency of the above three shapes incorporated with monolith was monitored by calculating their maximum enhancement factors. Among three morphologies, Gold Nano Spheres integrated in GMA-EDMA Monolith Sensor (GNS@GEMS) was found more effective for detection of R6G than two others and was therefore projected in analysis of melamine sensing in commercial milk. A linear relationship of regression model (R2 = 0.99) was observed between melamine SERS intensity (at 710 cm-1) and varied concentrations (6.5 < mg/L < 0.125). The lower limit of detection (LOD) and limit of quantification (LOQ) for melamine was determined 0.11 mg/L and 0.38 mg/L respectively. The time window for detection of melamine was 10 min and hence our method is compatible with the FDA's tolerance limit in USA and China (1 mg/L).


Assuntos
Ouro/química , Leite/química , Nanosferas/química , Polímeros/química , Análise Espectral Raman/métodos , Triazinas/análise , Animais , Limite de Detecção , Porosidade , Rodaminas/química
16.
Anal Chem ; 90(20): 11760-11763, 2018 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-30216046

RESUMO

Rapid and portable PCR detection is essential for screening sexually transmitted infections regularly. We developed an infrared mediated RNA isothermal RT-PCR (IR-MERIT PCR) platform and its compatible multichamber microfluidic chip for simultaneous amplification and testing (SAT) detection. This microfluidic chip integrates RNA extraction, micropump, and multitarget detection function onto the same chip. By utilizing IR-light-emitting diode (LED) as heat source, this platform can fulfill isothermal amplification within 70 min.

17.
Proc Natl Acad Sci U S A ; 115(41): 10275-10280, 2018 10 09.
Artigo em Inglês | MEDLINE | ID: mdl-30249664

RESUMO

Exosomes play an important role in numerous cellular processes. Fundamental study and practical use of exosomes are significantly constrained by the lack of analytical tools capable of physical and biochemical characterization. In this paper, we present an optical approach capable of imaging single exosomes in a label-free manner, using interferometric plasmonic microscopy. We demonstrate monitoring of the real-time adsorption of exosomes onto a chemically modified Au surface, calculating the image intensity, and determining the size distribution. The sizing capability enables us to quantitatively measure the membrane fusion activity between exosomes and liposomes. We also report the recording of the dynamic interaction between exosomes and antibodies at the single-exosome level, and the tracking of hit-stay-run behavior of exosomes on an antibody-coated surface. We anticipate that the proposed method will contribute to clinical exosome analysis and to the exploration of fundamental issues such as the exosome-antibody binding kinetics.


Assuntos
Exossomos , Processamento de Imagem Assistida por Computador/métodos , Interferometria/métodos , Adsorção , Anticorpos/química , Calibragem , Linhagem Celular , Desenho de Equipamento , Exossomos/química , Exossomos/metabolismo , Humanos , Interferometria/instrumentação , Lipossomos/análise , Lipossomos/química , Fusão de Membrana , Microscopia de Fluorescência/métodos , Nanopartículas , Ressonância de Plasmônio de Superfície/métodos , Propriedades de Superfície
18.
Front Physiol ; 9: 491, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29780330

RESUMO

Aim: Combined use of herbal medicines in patients underwent dual antiplatelet therapy (DAPT) might cause bleeding or thrombosis because herbal medicines with anti-platelet activities may exhibit interactions with DAPT. In this study, we tried to use a feedback system control (FSC) optimization technique to optimize dose strategy and clarify possible interactions in combined use of DAPT and herbal medicines. Methods: Herbal medicines with reported anti-platelet activities were selected by searching related references in Pubmed. Experimental anti-platelet activities of representative compounds originated from these herbal medicines were investigated using in vitro assay, namely ADP-induced aggregation of rat platelet-rich-plasma. FSC scheme hybridized artificial intelligence calculation and bench experiments to iteratively optimize 4-drug combination and 2-drug combination from these drug candidates. Results: Totally 68 herbal medicines were reported to have anti-platelet activities. In the present study, 7 representative compounds from these herbal medicines were selected to study combinatorial drug optimization together with DAPT, i.e., aspirin and ticagrelor. FSC technique first down-selected 9 drug candidates to the most significant 5 drugs. Then, FSC further secured 4 drugs in the optimal combination, including aspirin, ticagrelor, ferulic acid from DangGui, and forskolin from MaoHouQiaoRuiHua. Finally, FSC quantitatively estimated the possible interactions between aspirin:ticagrelor, aspirin:ferulic acid, ticagrelor:forskolin, and ferulic acid:forskolin. The estimation was further verified by experimentally determined Combination Index (CI) values. Conclusion: Results of the present study suggested that FSC optimization technique could be used in optimization of anti-platelet drug combinations and might be helpful in designing personal anti-platelet therapy strategy. Furthermore, FSC analysis could also identify interactions between different drugs which might provide useful information for research of signal cascades in platelet.

19.
Biomicrofluidics ; 12(2): 024109, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29576839

RESUMO

Portable Polymerase Chain Reaction (PCR) devices combined with microfluidic chips or lateral flow stripes have shown great potential in the field of point-of-need testing (PoNT) as they only require a small volume of patient sample and are capable of presenting results in a short time. However, the detection for multiple targets in this field leaves much to be desired. Herein, we introduce a novel PCR platform by integrating a bottom-well microfluidic chip with an infra-red (IR) excited temperature control method and fluorescence co-detection of three PCR products. Microfluidic chips are utilized to partition different samples into individual bottom-wells. The oil phase in the main channel contains multi-walled carbon nanotubes which were used as a heat transfer medium that absorbs energy from the IR-light-emitting diode (LED) and transfers heat to the water phase below. Cyclical rapid heating and cooling necessary for PCR are achieved by alternative power switching of the IR-LED and Universal Serial Bus (USB) mini-fan with a pulse width modulation scheme. This design of the IR-LED PCR platform is economic, compact, and fully portable, making it a promising application in the field of PoNT. The bottom-well microfluidic chip and IR-LED PCR platform were combined to fulfill a three-stage thermal cycling PCR for 40 cycles within 90 min for Human Papilloma Virus (HPV) detection. The PCR fluorescent signal was successfully captured at the end of each cycle. The technique introduced here has broad applications in nucleic acid amplification and PoNT devices.

20.
Anal Chem ; 90(7): 4397-4405, 2018 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-29537252

RESUMO

Circulating tumor cells (CTCs) are rare cells that detach from a primary or metastasis tumor and flow into the bloodstream. Intact and viable tumor cells are needed for genetic characterization of CTCs, new drug development, and other research. Although separation of CTCs using spiral channel with two outlets has been reported, few literature demonstrated simultaneous isolation of different types of CTCs from human blood using cascaded inertial focusing microfluidic channel. Herein, we introduce a cascaded microfluidic device consisting of two spiral channels and one zigzag channel designed with different fluid fields, including lift force, Dean drag force, and centrifugal force. Both red blood cells (RBCs)-lysed human blood spiked with CTCs and 1:50 diluted human whole blood spiked with CTCs were tested on the presented chip. This chip successfully separated RBCs, white blood cells (WBCs), and two different types of tumor cells (human lung cancer cells (A549) and human breast cancer cells (MCF-7)) simultaneously based on their physical properties. A total of 80.75% of A549 and 73.75% of MCF-7 were faithfully separated from human whole blood. Furthermore, CTCs gathered from outlets could propagate and remained intact. The cell viability of A549 and MCF-7 were 95% and 98%, respectively. The entire separating process for CTCs from blood cells could be finished within 20 min. The cascaded microfluidic device introduced in this study serves as a novel platform for simultaneous isolation of multiple types of CTCs from patient blood.

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