Publications by authors named "Mengxi Wu"

53 Publications

Ultrasensitive Multiparameter Phenotyping of Rare Cells Using an Integrated Digital-Molecular-Counting Microfluidic Well Plate.

Small 2021 Jun 25:e2101743. Epub 2021 Jun 25.

Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA.

Integrated microfluidic cellular phenotyping platforms provide a promising means of studying a variety of inflammatory diseases mediated by cell-secreted cytokines. However, immunosensors integrated in previous microfluidic platforms lack the sensitivity to detect small signals in the cellular secretion of proinflammatory cytokines with high precision. This limitation prohibits researchers from studying cells secreting cytokines at low abundance or existing at a small population. Herein, the authors present an integrated platform named the "digital Phenoplate (dPP)," which integrates digital immunosensors into a microfluidic chip with on-chip cell assay chambers, and demonstrates ultrasensitive cellular cytokine secretory profile measurement. The integrated sensors yield a limit of detection as small as 0.25 pg mL for mouse tumor necrosis factor alpha (TNF-α). Each on-chip cell assay chamber confines cells whose population ranges from ≈20 to 600 in arrayed single-cell trapping microwells. Together, these microfluidic features of the dPP simultaneously permit precise counting and image-based cytometry of individual cells while performing parallel measurements of TNF-α released from rare cells under multiple stimulant conditions for multiple samples. The dPP platform is broadly applicable to the characterization of cellular phenotypes demanding high precision and high throughput.
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http://dx.doi.org/10.1002/smll.202101743DOI Listing
June 2021

Effective Enrichment Strategy Using Boronic Acid-Functionalized Mesoporous Graphene-Silica Composites for Intact N- and O-Linked Glycopeptide Analysis in Human Serum.

Anal Chem 2021 05 20;93(17):6682-6691. Epub 2021 Apr 20.

The Fifth People's Hospital, Fudan University, and the Shanghai Key Laboratory of Medical Epigenetics, the International Co-laboratory of Medical Epigenetics and Metabolism, Ministry of Science and Technology, Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China.

The heterogeneity and low abundance of protein glycosylation present challenging barriers to the analysis of intact glycopeptides, which is key to comprehensively understanding the role of glycosylation in an organism. Efficient and specific enrichment of intact glycopeptides could help greatly with this problem. Here, we propose a new enrichment strategy using a boronic acid (BA)-functionalized mesoporous graphene-silica composite (denoted as [email protected]) for isolating intact glycopeptides from complex biological samples. The merits of this composite, including high surface area and synergistic effect from size exclusion functionality of mesoporous material, hydrophilic interaction of silica, and the reversible covalent binding with BA, enable the effective and specific enrichment of both intact N- and O-glycopeptides. The results from the enrichment performance of the strategy evaluated by standard glycoproteins and the application to global N- and O-glycosylation analyses in human serum indicate the robustness and potential of the strategy for intact glycopeptide analysis.
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http://dx.doi.org/10.1021/acs.analchem.0c05482DOI Listing
May 2021

Inhibition of CXCR4 ameliorates hypoxia-induced pulmonary arterial hypertension in rats.

Am J Transl Res 2021 15;13(3):1458-1470. Epub 2021 Mar 15.

Department of Anesthesiology, The Affiliated Wuxi People's Hospital of Nanjing Medical University Wuxi, Jiangsu Province, China.

Pulmonary vascular remodeling due to aberrant proliferation and migration of pulmonary artery smooth muscle cells (PASMCs) is the main characteristic of pulmonary arterial hypertension (PAH). CXCR4 is a specific stem cell surface receptor of cytokine CXCL12 which could regulate homing of hematopoietic progenitor cells and their mobilization. There is evidence that bone marrow-derived CXCR4 proangiogenic cell accumulation take an important part in the development of pulmonary arterial hypertension; however, the underlying mechanisms still remain unknown. Here, we explored the expression profile of CXCR4 both in hypoxia rats and PAH patients by measuring proliferation and migration of PASMCs. We performed western blot analysis to detect downstream molecules. We demonstrated that CXCR4 expression level was increased in both rats exposed to chronic hypoxia and PAH patients in reconstructed pulmonary arterioles. The inhibition of CXCR4 expression slowed down the process of hypoxic-PAH by reducing the mean right ventricular systolic pressure, right ventricular hypertrophy, and pulmonary vascular remodeling in vivo experimental mode. CXCR4 overexpression and inhibition regulated the cell growth of PASMCs in hypoxia condition, which are the critical cellular events in vascular disease. Furthermore, activation of β-catenin signaling and upregulation of CXCR4 could be blocked by AMD3100 both in vivo and vitro. Taken together, inhibition of CXCR4 expression could downregulate β-catenin, reduced pulmonary artery smooth muscle cell proliferation, and ameliorated pulmonary vascular remodeling in hypoxia rats. These findings suggest that CXCL12/CXCR4 is critical in driving PAH and uncover a correlation between β-catenin dependent signaling.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8014346PMC
March 2021

OGP: A Repository of Experimentally Characterized O-Glycoproteins to Facilitate Studies on O-Glycosylation.

Genomics Proteomics Bioinformatics 2021 Feb 10. Epub 2021 Feb 10.

Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China; The Fifth People's Hospital, Fudan University, and the Shanghai Key Laboratory of Medical Epigenetics, the International Co-laboratory of Medical Epigenetics and Metabolism, Ministry of Science and Technology, Fudan University, Shanghai 200032, China; NHC Key Laboratory of Glycoconjugates Research (Fudan University), Shanghai 200032, China. Electronic address:

Numerous studies on cancer, biopharmaceuticals, and clinical trials have necessitated comprehensive and precise analysis of protein O-glycosylation. However, the lack of updated and convenient databases deters the storage of and reference to emerging O-glycoprotein data. To resolve this issue, an O-glycoprotein repository named OGP was established in this work. It was constructed with a collection of O-glycoprotein data from different sources. OGP contains 9354 O-glycosylation sites and 11,633 site-specific O-glycans mapping to 2133 O-glycoproteins, and it is the largest O-glycoprotein repository thus far. Based on the recorded O-glycosylation sites, an O-glycosylation site prediction tool was developed. Moreover, an OGP-based website is already available (http://www.oglyp.org/). The website comprises four specially designed and user-friendly modules: statistical analysis, database search, site prediction, and data submission. The first version of OGP repository and the website allow users to obtain various O-glycoprotein-related information, such as protein accession numbers, O-glycosylation sites, glycopeptide sequences, site-specific glycan structures, experimental methods, and potential O-glycosylation sites. O-glycosylation data mining can be performed efficiently on this website, which will greatly facilitate related studies. In addition, the database is accessible from OGP website (http://www.oglyp.org/download.php).
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http://dx.doi.org/10.1016/j.gpb.2020.05.003DOI Listing
February 2021

Recent advances in software tools for more generic and precise intact glycopeptide analysis.

Mol Cell Proteomics 2021 Feb 5:100060. Epub 2021 Feb 5.

The Fifth People's Hospital of Fudan University and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China; NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai, 200032, China; Department of Chemistry, Fudan University, Shanghai, 200043, China; The Shanghai Key Laboratory of Medical Epigenetics and the International Co-laboratory of Medical Epigenetics and Metabolism, Ministry of Science and Technology, Fudan University, Shanghai, 200032, China. Electronic address:

Intact glycopeptide identification has long been known as a key and challenging barrier to the comprehensive and accurate understanding the role of glycosylation in an organism. Intact glycopeptide analysis is a blossoming field that has received increasing attention in recent years. Mass spectrometry (MS)-based strategies and relative software tools are major drivers that have greatly facilitated the analysis of intact glycopeptides, particularly intact N-glycopeptides. This manuscript provides a systematic review of the intact glycopeptide identification process using mass spectrometry data generated in shotgun proteomic experiments, which typically focus on N-glycopeptide analysis. Particular attention is paid to the software tools that have been recently developed in the last decade for the interpretation and quality control of glycopeptide spectra acquired using different MS strategies. The review also provides information about the characteristics and applications of these software tools, discusses their advantages and disadvantages, and concludes with a discussion of outstanding tools.
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http://dx.doi.org/10.1074/mcp.R120.002090DOI Listing
February 2021

Enhancing vigilance for cerebral air embolism after pneumonectomy: a case report.

BMC Pulm Med 2021 Jan 7;21(1):16. Epub 2021 Jan 7.

Department of Thoracic Surgery, Shenzhen Hospital, Southern Medical University, No.1333 Xinhu Road, Baoan District, Shenzhen, 518101, Guangdong, China.

Background: Vascular air embolism (VAE) is a rare but important complication that has not been paid enough attention to in the medical process such as surgery and anesthesia.

Case Presentation: We report for the first time that a 54-year-old male patient with central lung cancer developed severe complications of CAE after right pneumonectomy. After targeted first-aid measures such as assisted breathing, mannitol dehydration and antibiotic treatment, the patient gradually improved. The patient became conscious at discharge after 25 days of treatment but left limb was left with nerve injury symptoms.

Conclusion: We analyzed the possible causes of CAE in this case, and the findings from this report would be highly useful as a reference to clinicians.
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http://dx.doi.org/10.1186/s12890-020-01358-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7788539PMC
January 2021

An ultrafast and highly efficient enrichment method for both N-Glycopeptides and N-Glycans by bacterial cellulose.

Anal Chim Acta 2020 Dec 8;1140:60-68. Epub 2020 Oct 8.

Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China; The Fifth People's Hospital, Fudan University, and the Shanghai Key Laboratory of Medical Epigenetics, the International Co-laboratory of Medical Epigenetics and Metabolism, Ministry of Science and Technology, Fudan University, Shanghai, 200032, China; NHC Key Laboratory of Glycoconjugates Research (Fudan University), Shanghai, 200032, China. Electronic address:

A powerful and fast glycopeptide/glycan enrichment method is critical for the efficiency and throughput of mass spectrometry (MS)-based glycoproteomic and glycomic analyses, especially for large-scale sample analysis. Here, we report an ultrafast and effective method for both intact N-glycopeptide and N-glycan enrichment and apply it to human serum samples. In this method, a natural hydrophilic material, bacterial cellulose (BC), was adopted and fully optimized for enrichment. This method offers the following advantages: (i) The enrichment material has natural hydrophilicity and is low-cost, biocompatible, biodegradable and easily accessible; (ii) the whole enrichment procedure is remarkably simple and fast. It takes only 10 min for intact glycopeptides/glycans to be easily purified from mixtures; (iii) the specificity of this method is over 94% for both glycan and glycopeptide enrichment; and (iv) the outstanding specificity of this technique enables high isolation efficiency for the enrichment of both intact glycopeptides and glycans. A total of 36 N-glycans and 31 N-glycopeptides were identified from human immunoglobulin G (IgG). The glycan and glycopeptide absorption capacity of BC was as high as 333 μg/mg and 250 μg/mg (IgG/BC) respectively. The selectivity for glycan and glycopeptide enrichment reached 1:100 (IgG/bovine serum albumin (BSA), molar ratio) and 1:200 (maltoheptaose (DP7)/BSA, molar ratio), respectively. Furthermore, a total of 159 N-glycans and 523 N-glycopeptides were identified in human serum by using this method. Overall, the BC-based enrichment method we present here provides an ultrafast and highly efficient method for the enrichment of both N-glycopeptides and N-glycans in complex samples and shows great potential in large-scale glycoproteomic and glycomic analyses.
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http://dx.doi.org/10.1016/j.aca.2020.10.006DOI Listing
December 2020

Correction: High-throughput cell focusing and separation via acoustofluidic tweezers.

Lab Chip 2020 09 19;20(18):3470. Epub 2020 Aug 19.

Department of Mechanical Engineering and Material Science, Duke University, Durham, NC 27707, USA.

Correction for 'High-throughput cell focusing and separation via acoustofluidic tweezers' by Mengxi Wu et al., Lab Chip, 2018, 18, 3003-3010, DOI: .
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http://dx.doi.org/10.1039/d0lc90091eDOI Listing
September 2020

Correction: Separating extracellular vesicles and lipoproteins via acoustofluidics.

Lab Chip 2020 09 19;20(18):3472. Epub 2020 Aug 19.

Department of Mechanical Engineering and Material Science, Duke University, Durham, NC 27707, USA.

Correction for 'Separating extracellular vesicles and lipoproteins via acoustofluidics' by Mengxi Wu et al., Lab Chip, 2019, 19, 1174-1182, DOI: .
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http://dx.doi.org/10.1039/d0lc90093aDOI Listing
September 2020

Development of a Computational Tool for Automated Interpretation of Intact -Glycopeptide Tandem Mass Spectra from Single Proteins.

Anal Chem 2020 05 21;92(9):6777-6784. Epub 2020 Apr 21.

Precise and automated analysis of site-specific -glycosylation on single proteins is crucial for comprehensive characterization of some important glycoproteins, such as tumor biomarkers and recombinant drug proteins. Mass spectrometry has been proven to be a powerful technique for protein sequencing and -glycosylation analysis. However, challenges remain in developing computational tools for intact -glycopeptide analysis, which has greatly hindered the development of mass-spectrometry-based -glycosylation analysis. Herein, an integrated strategy together with a dedicated automated computational tool termed AOGP was developed for intact -glycopeptide analysis on single proteins. AOGP utilized sequencing for -glycans and a database search strategy for peptide backbones. The false discovery rate (FDR) of the identification results was controlled and validated by a mixed Gaussian distribution estimation method. AOGP exhibited superior performance in identifying intact -glycopeptides of the human erythropoietin with a total of 188 -glycopeptide spectra reported under 1% FDR. AOGP is developed in Python, is fully open-sourced, and is equipped with a user-friendly interface. Such an easy-operating and robust tool would greatly facilitate -glycosylation analysis on single proteins in tumor biomarker and recombinant drug protein development.
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http://dx.doi.org/10.1021/acs.analchem.0c01091DOI Listing
May 2020

Source-oriented characterization of single particles from in-port ship emissions in Guangzhou, China.

Sci Total Environ 2020 Jul 24;724:138179. Epub 2020 Mar 24.

Institute of Mass Spectrometer and Atmospheric Environment, Jinan University, Guangzhou 510632, China; Guangdong Provincial Engineering Research Center for On-Line Source Apportionment System of Air Pollution, Guangzhou 510632, China.

In this work, we analyzed freshly emitted particles from ship exhaust in the Guangzhou port region before and after the implementation of a clean fuel policy. We used a single particle aerosol mass spectrometer (SPAMS) to measure the changes in the chemical compositions of single particles and evaluate the role of V as a tracer for ship emissions. Particles from high sulfur fuel (S) oil (HS) combustion ships consisted of 54.8% elemental carbon-vanadium-sulfate (EC-V-S) and 25.0% vanadium-sulfate (V-S) particles, while particles from low S oil (LS) combustion ships were composed of 38.7% organic carbon-sulfate (OC-S) and 28.6% elemental and organic carbon (ECOC) particles. The sulfate-containing particles exhibited a moderate decrease from 95% in HS emissions to 78% in LS emissions, which still suggests the dominant role of sulfate in LS emissions after the implementation of a clean fuel policy. The V-containing particles showed a sharp decrease from 67% in HS emissions to 14% in LS emissions along with the decrease in the relative peak area (RPA) of V, suggesting a remarkable reduction in V in ship exhaust. The count of V-containing particles in urban Guangzhou in June 2017 was generally ten times lower than that in June 2016, which was in accordance with the sharp decrease in V-containing particles in LS emissions rather than in HS emissions. Despite the decrease in V in source-oriented ship emitted particles, the ubiquitous distribution of V in particles from lower S combustion ships suggests V is still effective as a tracer of ship emissions in port regions after the implementation of the clean fuel policy. Furthermore, the particles from LS emissions were investigated in comparison to those from gasoline vehicles (GV), diesel vehicles (DV) and coal combustion (CC) sources to better resolve ship-related particles in port regions.
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http://dx.doi.org/10.1016/j.scitotenv.2020.138179DOI Listing
July 2020

A disposable acoustofluidic chip for nano/microparticle separation using unidirectional acoustic transducers.

Lab Chip 2020 04 20;20(7):1298-1308. Epub 2020 Mar 20.

Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA.

Separation of nano/microparticles based on surface acoustic waves (SAWs) has shown great promise for biological, chemical, and medical applications ranging from sample purification to cancer diagnosis. However, the permanent bonding of a microchannel onto relatively expensive piezoelectric substrates and excitation transducers renders the SAW separation devices non-disposable. This limitation not only requires cumbersome cleaning and increased labor and material costs, but also leads to cross-contamination, preventing their implementation in many biological, chemical, and medical applications. Here, we demonstrate a high-performance, disposable acoustofluidic platform for nano/microparticle separation. Leveraging unidirectional interdigital transducers (IDTs), a hybrid channel design with hard/soft materials, and tilted-angle standing SAWs (taSSAWs), our disposable acoustofluidic devices achieve acoustic radiation forces comparable to those generated by existing permanently bonded, non-disposable devices. Our disposable devices can separate not only microparticles but also nanoparticles. Moreover, they can differentiate bacteria from human red blood cells (RBCs) with a purity of up to 96%. Altogether, we developed a unidirectional IDT-based, disposable acoustofluidic platform for micro/nanoparticle separation that can achieve high separation efficiency, versatility, and biocompatibility.
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http://dx.doi.org/10.1039/d0lc00106fDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7199844PMC
April 2020

Acoustic Cell Separation Based on Density and Mechanical Properties.

J Biomech Eng 2020 03;142(3)

Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708.

Density and mechanical properties (e.g., compressibility or bulk modulus) are important cellular biophysical markers. As such, developing a method to separate cells directly based on these properties can benefit various applications including biological research, diagnosis, prognosis, and therapeutics. As a potential solution, surface acoustic wave (SAW)-based cell separation has demonstrated advantages in terms of biocompatibility and compact device size. However, most SAW-reliant cell separations are achieved using an entangled effect of density, various mechanical properties, and size. In this work, we demonstrate SAW-based separation of cells/particles based on their density and compressibility, irrespective of their sizes, by manipulating the acoustic properties of the fluidic medium. Using our platform, SAW-based separation is achieved by varying the dimensions of the microfluidic channels, the wavelengths of acoustic signals, and the properties of the fluid media. Our method was applied to separate paraformaldehyde-treated and fresh Hela cells based on differences in mechanical properties; a recovery rate of 85% for fixed cells was achieved. It was also applied to separate red blood cells (RBCs) and white blood cells (WBCs) which have different densities. A recovery rate of 80.5% for WBCs was achieved.
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http://dx.doi.org/10.1115/1.4046180DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7104781PMC
March 2020

Acoustofluidic Salivary Exosome Isolation: A Liquid Biopsy Compatible Approach for Human Papillomavirus-Associated Oropharyngeal Cancer Detection.

J Mol Diagn 2020 01;22(1):50-59

Department of Mechanical Engineering and Materials Science, Duke University, Durham, North Carolina. Electronic address:

Previous efforts to evaluate the detection of human papilloma viral (HPV) DNA in whole saliva as a diagnostic measure for HPV-associated oropharyngeal cancer (HPV-OPC) have not shown sufficient clinical performance. We hypothesize that salivary exosomes are packaged with HPV-associated biomarkers, and efficient enrichment of salivary exosomes through isolation can enhance diagnostic and prognostic performance for HPV-OPC. In this study, an acoustofluidic (the fusion of acoustics and microfluidics) platform was developed to perform size-based isolation of salivary exosomes. These data showed that this platform is capable of consistently isolating exosomes from saliva samples, regardless of viscosity variation and collection method. Compared with the current gold standard, differential centrifugation, droplet digital RT-PCR analysis showed that the average yield of salivary exosomal small RNA from the acoustofluidic platform is 15 times higher. With this high-yield exosome isolation platform, we show that HPV16 DNA could be detected in isolated exosomes from the saliva of HPV-associated OPC patients at 80% concordance with tissues/biopsies positive for HPV16. Overall, these data demonstrated that the acoustofluidic platform can achieve high-purity and high-yield salivary exosome isolation for downstream salivary exosome-based liquid biopsy applications. Additionally, HPV16 DNA sequences in HPV-OPC patients are packaged in salivary exosomes and their isolation will enhance the detection of HPV16 DNA.
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http://dx.doi.org/10.1016/j.jmoldx.2019.08.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6943372PMC
January 2020

Acoustofluidic Synthesis of Particulate Nanomaterials.

Adv Sci (Weinh) 2019 Oct 27;6(19):1900913. Epub 2019 Aug 27.

Department of Mechanical Engineering and Materials Science Duke University Durham NC 27708 USA.

Synthesis of nanoparticles and particulate nanomaterials with tailored properties is a central step toward many applications ranging from energy conversion and imaging/display to biosensing and nanomedicine. While existing microfluidics-based synthesis methods offer precise control over the synthesis process, most of them rely on passive, partial mixing of reagents, which limits their applicability and potentially, adversely alter the properties of synthesized products. Here, an acoustofluidic (i.e., the fusion of acoustic and microfluidics) synthesis platform is reported to synthesize nanoparticles and nanomaterials in a controllable, reproducible manner through acoustic-streaming-based active mixing of reagents. The acoustofluidic strategy allows for the dynamic control of the reaction conditions simply by adjusting the strength of the acoustic streaming. With this platform, the synthesis of versatile nanoparticles/nanomaterials is demonstrated including the synthesis of polymeric nanoparticles, chitosan nanoparticles, organic-inorganic hybrid nanomaterials, metal-organic framework biocomposites, and lipid-DNA complexes. The acoustofluidic synthesis platform, when incorporated with varying flow rates, compositions, or concentrations of reagents, will lend itself unprecedented flexibility in establishing various reaction conditions and thus enable the synthesis of versatile nanoparticles and nanomaterials with prescribed properties.
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http://dx.doi.org/10.1002/advs.201900913DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6774021PMC
October 2019

Multiple genes recruited from hormone pathways partition maize diterpenoid defences.

Nat Plants 2019 10 16;5(10):1043-1056. Epub 2019 Sep 16.

Section of Cell and Developmental Biology, University of California San Diego, La Jolla, CA, USA.

Duplication and divergence of primary pathway genes underlie the evolution of plant specialized metabolism; however, mechanisms partitioning parallel hormone and defence pathways are often speculative. For example, the primary pathway intermediate ent-kaurene is essential for gibberellin biosynthesis and is also a proposed precursor for maize antibiotics. By integrating transcriptional coregulation patterns, genome-wide association studies, combinatorial enzyme assays, proteomics and targeted mutant analyses, we show that maize kauralexin biosynthesis proceeds via the positional isomer ent-isokaurene formed by a diterpene synthase pair recruited from gibberellin metabolism. The oxygenation and subsequent desaturation of ent-isokaurene by three promiscuous cytochrome P450s and a new steroid 5α reductase indirectly yields predominant ent-kaurene-associated antibiotics required for Fusarium stalk rot resistance. The divergence and differential expression of pathway branches derived from multiple duplicated hormone-metabolic genes minimizes dysregulation of primary metabolism via the circuitous biosynthesis of ent-kaurene-related antibiotics without the production of growth hormone precursors during defence.
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http://dx.doi.org/10.1038/s41477-019-0509-6DOI Listing
October 2019

Acoustofluidic separation of cells and particles.

Microsyst Nanoeng 2019 3;5:32. Epub 2019 Jun 3.

1Department of Mechanical Engineering and Material Science, Duke University, Durham, NC 27708 USA.

Acoustofluidics, the integration of acoustics and microfluidics, is a rapidly growing research field that is addressing challenges in biology, medicine, chemistry, engineering, and physics. In particular, acoustofluidic separation of biological targets from complex fluids has proven to be a powerful tool due to the label-free, biocompatible, and contact-free nature of the technology. By carefully designing and tuning the applied acoustic field, cells and other bioparticles can be isolated with high yield, purity, and biocompatibility. Recent advances in acoustofluidics, such as the development of automated, point-of-care devices for isolating sub-micron bioparticles, address many of the limitations of conventional separation tools. More importantly, advances in the research lab are quickly being adopted to solve clinical problems. In this review article, we discuss working principles of acoustofluidic separation, compare different approaches of acoustofluidic separation, and provide a synopsis of how it is being applied in both traditional applications, such as blood component separation, cell washing, and fluorescence activated cell sorting, as well as emerging applications, including circulating tumor cell and exosome isolation.
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http://dx.doi.org/10.1038/s41378-019-0064-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6545324PMC
June 2019

Wave number-spiral acoustic tweezers for dynamic and reconfigurable manipulation of particles and cells.

Sci Adv 2019 May 31;5(5):eaau6062. Epub 2019 May 31.

Department of Mechanical Engineering and Material Science, Duke University, Durham, NC 27708, USA.

Acoustic tweezers have recently raised great interest across many fields including biology, chemistry, engineering, and medicine, as they can perform contactless, label-free, biocompatible, and precise manipulation of particles and cells. Here, we present wave number-spiral acoustic tweezers, which are capable of dynamically reshaping surface acoustic wave (SAW) wavefields to various pressure distributions to facilitate dynamic and programmable particle/cell manipulation. SAWs propagating in multiple directions can be simultaneously and independently controlled by simply modulating the multitone excitation signals. This allows for dynamic reshaping of SAW wavefields to desired distributions, thus achieving programmable particle/cell manipulation. We experimentally demonstrated the multiple functions of wave number-spiral acoustic tweezers, among which are multiconfiguration patterning; parallel merging; pattern translation, transformation, and rotation; and dynamic translation of single microparticles along complex paths. This wave number-spiral design has the potential to revolutionize future acoustic tweezers development and advance many applications, including microscale assembly, bioprinting, and cell-cell interaction research.
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http://dx.doi.org/10.1126/sciadv.aau6062DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6544454PMC
May 2019

A multi-parallel N-glycopeptide enrichment strategy for high-throughput and in-depth mapping of the N-glycoproteome in metastatic human hepatocellular carcinoma cell lines.

Talanta 2019 Jul 8;199:254-261. Epub 2019 Feb 8.

The Fifth People's Hospital of Shanghai and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, People's Republic of China; Department of Chemistry, Fudan University, Shanghai 200433, People's Republic of China; NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai 200433, People's Republic of China. Electronic address:

N-glycosylation is deeply involved in many biological processes, and approximately 50% of mammalian proteins are predicted to be glycosylated. Many large-scale studies have been carried out to reveal the glycosylation status involved in different physiological pathologies across species. However, the lack of a highly specific and high-throughput N-glycosylated enrichment method not only results in extended time requirements but also limits the depth of mapping when handling a large number of samples. In this study, we firstly optimized traditional zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) enrichment and found that using of 70% acetonitrile (ACN), 0.1% trifluoroacetic acid (TFA) as the enrichment buffer, 2800 g as the washing speed and 600 μL as the washing volume achieved the best specificity, which is higher than 75%. On this basis, we developed a multi-parallel enrichment strategy assisted by a filter-coated 96-well plate, which achieved high specificity and high throughput simultaneously. This strategy allowed us to enrich large numbers of fractionated samples from hepatocellular carcinoma (HCC) cell lines in less than 2 h. Its good specificity helped us achieve in-depth mapping of the N-glycoproteome in metastatic HCC cell lines. A total of 5466 N-glycosites from 2383 glycoproteins were identified, among which 1900 N-glycosites were unannotated in UniProt. The in-depth glycoproteome mapping provides insight into the N-glycosylation status in HCC cell lines with differences in metastatic potential and contributes to biomarker discovery.
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http://dx.doi.org/10.1016/j.talanta.2019.02.010DOI Listing
July 2019

Separating extracellular vesicles and lipoproteins via acoustofluidics.

Lab Chip 2019 03;19(7):1174-1182

Department of Mechanical Engineering and Material Science, Duke University, Durham, NC 27707, USA.

Extracellular vesicles (EVs) and lipoproteins are abundant and co-exist in blood. Both have been proven to be valuable as diagnostic biomarkers and for therapeutics. However, EVs and lipoproteins are both on the submicron scale and overlap in size distributions. Conventional methods to separate EVs and lipoproteins are inefficient and time-consuming. Here we present an acoustofluidic-based separation technique that is based on the acoustic property differences of EVs and lipoproteins. By using the acoustofluidic technology, EVs and subgroups of lipoproteins are separated in a label-free, contact-free, and continuous manner. With its ability for simple, rapid, efficient, continuous-flow isolation, our acoustofluidic technology could be a valuable tool for health monitoring, disease diagnosis, and personalized medicine.
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http://dx.doi.org/10.1039/c8lc01134fDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6453118PMC
March 2019

Plastic-based acoustofluidic devices for high-throughput, biocompatible platelet separation.

Lab Chip 2019 01;19(3):394-402

Department of Mechanical Engineering and Material Science, Duke University, Durham, NC 27707, USA.

Platelet separation is a crucial step for both blood donation and treatment of essential thrombocytosis. Here we present an acoustofluidic device that is capable of performing high-throughput, biocompatible platelet separation using sound waves. The device is entirely made of plastic material, which renders the device disposable and more suitable for clinical use. We used this device to process undiluted human whole blood, and we demonstrate a sample throughput of 20 mL min-1, a platelet recovery rate of 87.3%, and a red/white blood cell removal rate of 88.9%. We preserved better platelet function and integrity for isolated platelets than those which are isolated using established methods. Our device features advantages such as rapid fabrication, high throughput, and biocompatibility, so it is a promising alternative to existing platelet separation approaches.
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http://dx.doi.org/10.1039/c8lc00527cDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6366625PMC
January 2019

Spatiotemporal filtering of high harmonics in solids.

Opt Lett 2018 Nov;43(21):5339-5342

We study the macroscopic spatial and temporal properties of harmonic radiation generated by a model solid in the interaction with an intense, focused laser beam. We show that different temporal contributions to the harmonic yield can be separated in the spatial domain because they lead to radiation with different divergences, similar to what is observed in gas-phase harmonic generation. We show that applying a spatial filter in the far field results in a temporal separation of the two contributions upon refocusing, which yields spatially collimated harmonics, a spectrum with well-resolved peaks, and a subcycle time profile of the harmonic radiation with only one burst per half-cycle.
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http://dx.doi.org/10.1364/OL.43.005339DOI Listing
November 2018

RETRACTED: The interplay between reactive oxygen and nitrogen species contributes in the regulatory mechanism of the nitro-oxidative stress induced by cadmium in Arabidopsis.

J Hazard Mater 2018 02 6;344:1007-1024. Epub 2017 Dec 6.

College of Landscape Architecture, Sichuan Agricultural University, Chengdu, Sichuan 611130, China. Electronic address:

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor, after consultation with the corresponding author Dr. Shiliang Liu due to image issues. The article reused several images from the author's paper published in Environmental Pollution 239 (2018) 53-68 (which has been retracted due to image issues): Figures 1c, 1d, 2a, 2b, 2c, 4a, 9a and 9b. The article also plagiarized part of a paper from other authors that had appeared in Plant Physiology, 150, 229-243 (2009). The images that were reused were Fig 5 a, 5c, 5e and 5 g. This was brought to the editors’ attention via a letter to the editor. One of the conditions of submission of a paper for publication is that authors declare explicitly that their work is original and has not appeared in a publication elsewhere. Re-use of any data should be appropriately cited. As such this article represents a severe abuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process.
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http://dx.doi.org/10.1016/j.jhazmat.2017.12.004DOI Listing
February 2018

Standing Surface Acoustic Wave (SSAW)-Based Fluorescence-Activated Cell Sorter.

Small 2018 10 31;14(40):e1801996. Epub 2018 Aug 31.

Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC, 27708, USA.

Microfluidic fluorescence-activated cell sorters (μFACS) have attracted considerable interest because of their ability to identify and separate cells in inexpensive and biosafe ways. Here a high-performance μFACS is presented by integrating a standing surface acoustic wave (SSAW)-based, 3D cell-focusing unit, an in-plane fluorescent detection unit, and an SSAW-based cell-deflection unit on a single chip. Without using sheath flow or precise flow rate control, the SSAW-based cell-focusing technique can focus cells into a single file at a designated position. The tight focusing of cells enables an in-plane-integrated optical detection system to accurately distinguish individual cells of interest. In the acoustic-based cell-deflection unit, a focused interdigital transducer design is utilized to deflect cells from the focused stream within a minimized area, resulting in a high-throughput sorting ability. Each unit is experimentally characterized, respectively, and the integrated SSAW-based FACS is used to sort mammalian cells (HeLa) at different throughputs. A sorting purity of greater than 90% is achieved at a throughput of 2500 events s . The SSAW-based FACS is efficient, fast, biosafe, biocompatible and has a small footprint, making it a competitive alternative to more expensive, bulkier traditional FACS.
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http://dx.doi.org/10.1002/smll.201801996DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6291339PMC
October 2018

High-throughput cell focusing and separation via acoustofluidic tweezers.

Lab Chip 2018 09;18(19):3003-3010

Department of Mechanical Engineering and Material Science, Duke University, Durham, NC 27707, USA.

Separation of particles and cells is an important function in many biological and biomedical protocols. Although a variety of microfluidic-based techniques have been developed so far, there is clearly still a demand for a precise, fast, and biocompatible method for separation of microparticles and cells. By combining acoustics and hydrodynamics, we have developed a method which we integrated into three-dimensional acoustofluidic tweezers (3D-AFT) to rapidly and efficiently separate microparticles and cells into multiple high-purity fractions. Compared with other acoustophoresis methods, this 3D-AFT method significantly increases the throughput by an order of magnitude, is label-free and gently handles the sorted cells. We demonstrate not only the separation of 10, 12, and 15 micron particles at a throughput up to 500 μl min-1 using this 3D-AFT method, but also the separation of erythrocytes, leukocytes, and cancer cells. This 3D-AFT method is able to meet various separation demands thus offering a viable alternative with potential for clinical applications.
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http://dx.doi.org/10.1039/c8lc00434jDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6203445PMC
September 2018

Circulating Tumor Cell Phenotyping via High-Throughput Acoustic Separation.

Small 2018 08 3;14(32):e1801131. Epub 2018 Jul 3.

Department of Mechanical Engineering and Material Science, Duke University, Durham, NC, 27708, USA.

The study of circulating tumor cells (CTCs) offers pathways to develop new diagnostic and prognostic biomarkers that benefit cancer treatments. In order to fully exploit and interpret the information provided by CTCs, the development of a platform is reported that integrates acoustics and microfluidics to isolate rare CTCs from peripheral blood in high throughput while preserving their structural, biological, and functional integrity. Cancer cells are first isolated from leukocytes with a throughput of 7.5 mL h , achieving a recovery rate of at least 86% while maintaining the cells' ability to proliferate. High-throughput acoustic separation enables statistical analysis of isolated CTCs from prostate cancer patients to be performed to determine their size distribution and phenotypic heterogeneity for a range of biomarkers, including the visualization of CTCs with a loss of expression for the prostate specific membrane antigen. The method also enables the isolation of even rarer, but clinically important, CTC clusters.
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http://dx.doi.org/10.1002/smll.201801131DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6105522PMC
August 2018
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