Publications by authors named "Ziyi He"

40 Publications

Circulating Natural Autoantibodies to HER2-Derived Peptides Performed Antitumor Effects on Oral Squamous Cell Carcinoma.

Front Pharmacol 2021 5;12:693989. Epub 2021 Nov 5.

Beijing Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing, China.

Natural autoantibodies play a crucial role in destruction of malignant tumors due to immune surveillance function. Epidermal growth factor receptor 2 (HER2) has been found to be highly expressed in a variety of epithelial tumors including oral squamous cell carcinoma (OSCC). The present study was thus undertaken to investigate the effect of anti-HER2 natural autoantibodies on OSCC. Compared with cancer-adjacent tissues, cancer tissues from OSCC patients exhibited higher HER2 expression especially in those with middle & advanced stage OSCC. Plasma anti-HER2 IgG levels examined with an enzyme-linked immunosorbent assay (ELISA) developed in-house showed differences between control subjects, individuals with oral benign tumor and patients with OSCC. In addition, anti-HER2 IgG-abundant plasma was screened from healthy donors to treat OSCC cells and to prepare for anti-HER2 intravenous immunoglobulin (IVIg). Both anti-HER2 IgG-abundant plasma and anti-HER2 IVIg could significantly inhibit proliferation and invasion of OSCC cells by inducing the apoptosis, and also regulate apoptosis-associated factors and epithelial-mesenchymal transition (EMT), respectively. Besides, the complement-dependent cytotoxicity (CDC) pathway was likely to contribute to the anti-HER2 IgG mediated inhibition of OSCC cells. After the HER2 gene was knocked down with HER2-specific siRNAs, the inhibitory effects on OSCC cell proliferation and apoptotic induction faded away. In conclusion, human plasma IgG, or IVIg against HER2 may be a promising agent for anti-OSCC therapy.
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http://dx.doi.org/10.3389/fphar.2021.693989DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8602057PMC
November 2021

Proteomic Distributions in CD34+ Microvascular Niche Patterns of Glioblastoma.

J Histochem Cytochem 2021 Nov 9:221554211058098. Epub 2021 Nov 9.

Department of Neurosurgery, Sanbo Brain Hospital, Capital Medical University, Beijing, China.

The poor clinical prognosis and microvascular patterns of glioblastoma (GBM) are of serious concern to many clinicians and researchers. However, very few studies have examined the correlation between microvascular niche patterns (MVNPs) and proteomic distribution. In this study, CD34 immunofluorescence staining and matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-IMS) technology were used to investigate the protein distributions in MVNPs. CD34+ microvascular phenotype could be divided into four types: microvascular sprouting (MS), vascular cluster (VC), vascular garland (VG), and glomeruloid vascular proliferation (GVP). Based on such characteristics, MVNPs were divided into two types by cluster analysis, namely, type I, comprising primarily MS and VC, and type II, comprising many VGs and GVPs. Survival analysis indicated the type of MVNPs to be an independent prognostic factor for progression-free and overall survival in GBM. MALDI-IMS results showed the peaks at m/z 1037 and 8960 to exhibit stronger ion signals in type II, while those at m/z 3240 and 3265 exhibited stronger ion signals in type I. The findings may assist future research on therapy and help predict prognosis in GBM. However, due to the limited number of studies, more well-designed studies are warranted to further verify our results.
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http://dx.doi.org/10.1369/00221554211058098DOI Listing
November 2021

Three-Month Pulmonary Function and Radiological Outcomes in COVID-19 Survivors: A Longitudinal Patient Cohort Study.

Open Forum Infect Dis 2021 Sep 14;8(9):ofaa540. Epub 2020 Nov 14.

National Clinical Research Center for Infectious Diseases, The Third People's Hospital of Shenzhen, The Second Affiliated Hospital of Southern University of Science and Technology, Shenzhen, Guangdong, China.

Background: This study aimed to investigate pulmonary function and radiological outcomes in a group of coronavirus disease 2019 (COVID-19) survivors.

Methods: One hundred seventy-two COVID-19 survivors in a follow-up clinic in a referral hospital underwent high-resolution computed tomography (CT) of the thorax and pulmonary function at 3 months after hospital discharge.

Results: The median duration from hospital discharge to radiological and pulmonary function test (interquartile range) was 90 (88-95) days. Abnormal pulmonary function was found in 11 (6.40%) patients, and abnormal small airway function (FEF) in 12 (6.98%). Six (3.49%) patients had obstructive ventilation impairment, and 6 (3.49%) had restrictive ventilatory impairment. No significant differences in lung function parameters were observed between the nonsevere and severe groups. Of 142 COVID-19 patients who underwent CT scan, 122 (85.91%) showed residual CT abnormalities and 52 (36.62%) showed chronic and fibrotic changes. The ground-glass opacities absorption in the lungs of severe cases was less satisfactory than that of nonsevere patients. The severe patients had higher CT scores than the nonsevere cases (2.00 vs 0.00;  < .001).

Conclusions: Of the COVID-19 survivors in our study, 6.40% still presented pulmonary function abnormality 3 months after discharge, which did not vary by disease severity during hospitalization; 85.91% of patients had abnormalities on chest CT, with fibrous stripes and ground-glass opacities being the most common patterns.
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http://dx.doi.org/10.1093/ofid/ofaa540DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7717449PMC
September 2021

Elevations of monocyte and neutrophils, and higher levels of granulocyte colony-stimulating factor in peripheral blood in lung cancer patients.

Thorac Cancer 2021 Oct 8;12(20):2680-2690. Epub 2021 Sep 8.

Department of Immunology, School of Basic Medical Sciences, NHC Key Laboratory of Medical Immunology, Peking University, Beijing, China.

Introduction: Immune cells and molecules are considered as clinical biomarkers and potential targets for immunotherapy. Analyses of the composition of peripheral blood cells hold promise for providing a basis for diagnosing and prognosis lung cancer. In this study, we assessed correlations between immune cell subset profiles in peripheral blood and disease prognosis in patients with lung cancer.

Methods: One hundred and thirteen patients with lung cancer and 99 age-matched healthy people were enrolled in this study. The percentage and cell count of monocytes, neutrophils, T cells, B cells, natural killer (NK), and NKT cells in peripheral blood were analyzed by flow cytometry or peripheral blood analyzer. Serum cytokines and colony-stimulating factors were detected by enzyme-linked immunosorbent assay (ELISA).

Results: A reduction in antitumor NK cells (p < 0.0001) and an increase in the protumor MDSCs (p < 0.0001) were observed in the lung cancer patients compared with the controls. Monocyte counts were significantly higher in lung cancer patients with histories of smoking (p < 0.05) or drinking (p < 0.01) than in patients with no relevant history or healthy controls. The number of neutrophils and the neutrophil-to-lymphocyte ratio (NLR) were particularly higher in patients with liver metastasis (p < 0.01) compared with no metastasis patients or healthy controls. Levels of the monocyte-derived cytokine interleukin-6 (p < 0.05), granulocyte colony-stimulating factor (G-CSF) (p < 0.0001), and granulocyte-macrophage colony-stimulating factor (GM-CSF) (p < 0.0001) were higher in patients than in controls. G-CSF levels decreased during the remission phase (p < 0.05), and positively correlated with carbohydrate antigen 19-9 (p < 0.05) and gene mutation (p < 0.05).

Conclusion: Monocyte and neutrophil counts were higher in peripheral blood in lung cancer patients than in controls, especially when patients had histories of smoking, drinking, and liver metastasis. Serum levels of G-CSF and GM-CSF were higher in lung cancer patients, and G-CSF levels positively correlated with disease severity.
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http://dx.doi.org/10.1111/1759-7714.14103DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8520797PMC
October 2021

Case Report: Drug-Induced Immune Haemolytic Anaemia Caused by Cefoperazone-Tazobactam/ Sulbactam Combination Therapy.

Front Med (Lausanne) 2021 12;8:697192. Epub 2021 Aug 12.

Department of Blood Transfusion, Dongguan Maternal and Child Health Hospital, Dongguan, China.

There has previously been a report of a patient developing haemolytic anaemia following exposure to cefoperazone. Another case has been reported involving the detection of cefoperazone-dependent antibodies in the absence of immune haemolytic anaemia. To date, no serological evidence has been reported to suggest that cefoperazone can lead to drug-induced immune haemolytic anaemia (DIIHA). This report aims to fill these gaps in knowledge by describing a case of DIIHA caused by cefoperazone-dependent antibodies. A 59-year-old man developed fatal haemolytic anaemia while receiving cefoperazone-tazobactam or cefoperazone-sulbactam for the treatment of a lung infection that occurred after craniocerebral surgery. This eventually led to renal function impairment. Prior to the discontinuation of cefoperazone treatment, the patient showed strong positive (4+) results for both anti-IgG and anti-C3d direct antiglobulin test (DAT), while cefoperazone-dependent IgM and IgG antibodies were detected. The patient's plasma and O-type RBCs were incubated with tazobactam or sulbactam solution at 37°C for 3 h, the results of DAT for anti-IgG and anti-C3d were both positive. Forty-three days after the discontinuation of cefoperazone, the results of DAT for anti-IgG and anti-C3d were negative. Meanwhile incubation of the patient's fresh serum and his own RBCs with cefoperazone at 37°C, gave rise to mild haemolysis, and the results of DAT for both anti-IgG and anti-C3d were positive. It is suggested that cefoperazone-dependent antibodies can activate complement, and the non-immunologic protein adsorption effect of tazobactam or sulbactam can enhance IgG and complement binding to RBCs. This may promote the formation of immunocomplexes and complement activation, thereby aggravating haemolysis.
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http://dx.doi.org/10.3389/fmed.2021.697192DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8415779PMC
August 2021

3D microfluidic tumor models for biomimetic engineering of glioma niche and detection of cell morphology, migration and phenotype change.

Talanta 2021 Nov 13;234:122702. Epub 2021 Jul 13.

Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, MOE Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing, 100084, China; School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 100029, China. Electronic address:

In this work, an integrated 3-dimensional microfluidic device was developed for simulation of the immune microenvironment of glioma niche through the co-culture of three kinds of related cells. Glioma cells, endothelial cells and macrophages were co-cultured together in the microfluidic device, spatially separated by the design of a coffer structure and the use of hydrogel. This platform enabled separate monitoring of the morphology change and migration of cells, as well as molecular interactions between different kinds of cells. Tumor cells were found to exhibit EMT like shape change to become thinner, and sensitive perception and taxis toward macrophages. The influence of tumor cells and the microenvironment, macrophages would be re-educated and the phenotype could be changed from M1 (tumor-suppressive) to M2 (tumor-supportive), which could be validated through cytokines analysis. This 3D microfluidic tumor model provides a powerful tool for studying the biological properties of glioma niche.
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http://dx.doi.org/10.1016/j.talanta.2021.122702DOI Listing
November 2021

A portable droplet generation system for ultra-wide dynamic range digital PCR based on a vibrating sharp-tip capillary.

Biosens Bioelectron 2021 Nov 24;191:113458. Epub 2021 Jun 24.

C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, WV, 26506, USA. Electronic address:

Monodisperse droplet has been widely used as a versatile tool in different disciplines including biosensing. Existing methods still struggle to balance the droplet generation performance with system simplicity. Here we introduce a novel droplet generation scheme based on the acoustic streaming generated from a vibrating sharp-tip capillary. The unique fluid pattern enables efficient droplet generation without any external pressure sources. This method achieved real-time modulation of droplet size over an ultra-wide range (6.77-661 μm), high throughput (up to 5000 droplets/s), and good monodispersity (<4%) with a power consumption below 60 mW. This method has enabled a multi-volume digital PCR with a dynamic range of ~6 orders of magnitude and multiplexing capability. It has also enabled a simple protocol to produce cell-laden alginate microcapsules in variable sizes with excellent biocompatibility. Overall, the present method combines high performance with small footprint and portability, which will be especially valuable for droplet applications requiring variable droplet size and performed in resource-limited settings.
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http://dx.doi.org/10.1016/j.bios.2021.113458DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8376801PMC
November 2021

One-step enzyme kinetics measurement in 3D printed microfluidics devices based on a high-performance single vibrating sharp-tip mixer.

Anal Chim Acta 2021 Aug 24;1172:338677. Epub 2021 May 24.

C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, WV, USA. Electronic address:

Measuring enzyme kinetics is of great importance to understand many biological processes and improve biosensing and industrial applications. Conventional methods of measuring enzyme kinetics require to prepare a series of solutions with different substrate concentrations and measure the signal response over time with these solutions, leading to tedious sample preparation steps, high reagents/sample consumption, and difficulties in studying fast enzyme kinetics. Here we report a one-step assay to measure enzyme kinetics using a 3D-printed microfluidic device, which eliminates the steps of preparing and handling multiple solutions thereby simplifying the whole workflow significantly. The assay is enabled by a highly efficient vibrating sharp-tip mixing method that can mix multiple streams of fluids with minimal mixing length (∼300 μm) and time (as low as 3 ms), and a wide range of working flow rates from 1.5 μL/min to 750 μL/min. Owing to the high performance of the mixer, a series of experiments with different substrate concentrations are performed by simply adjusting the flow rates of reagents loaded from three inlets in one experiment run. The Michaelis-Menten kinetics of the horseradish peroxidase (HRP)-catalyzed reaction between HO and amplex red is measured in this system. The calculated Michaelis constant is consistent with the values from literature and conventional analysis methods. Due to the simplicity in fabrication and operation, rapid analysis, low power consumption (1.4-45.0 mW), and high temporal resolution, this method will significantly facilitate enzyme kinetics measurement, and offers great potential for optimizing enzyme based biosensing experiments and probing many biochemical processes.
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http://dx.doi.org/10.1016/j.aca.2021.338677DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8206521PMC
August 2021

Inhibitory effects of circulating natural autoantibodies to CD47-derived peptides on OSCC cells.

Oral Dis 2021 May 24. Epub 2021 May 24.

Beijing Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing, China.

Objectives: Natural autoantibodies serve as an important anti-tumorigenic component in the body. This study was thus designed to investigate whether circulating natural IgG autoantibodies against a cluster of differentiation 47 (CD47) could exert inhibitory effects on oral squamous cell carcinoma (OSCC).

Subjects And Methods: The expression levels of 13 tumor-targeted genes in three OSCC cell lines were analyzed by qPCR, and CD47 expression in OSCC tissues was also verified with IHC staining. An in-house ELISA was performed to analyze circulating anti-CD47 IgG levels in control subjects, oral benign tumor, and OSCC patients, and to detect anti-CD47 IgG-abundant plasma. Three OSCC cell lines were treated with anti-CD47 IgG-abundant and -deficient plasma, respectively, followed by the analysis of cell proliferation, apoptosis, and invasion/metastasis.

Results: The CD47 gene showed the highest expression among 13 genes detected in three OSCC cell lines; its expression was significantly higher in OSCC tissues than adjacent tissues. Plasma anti-CD47 IgG levels showed the differences between control subjects, oral benign tumor, and OSCC patients. Anti-CD47 IgG-abundant plasma could evidently reduce cell viability via suppressing p-AKT expression and inducing cell apoptosis and inhibit the invasion of all three OSCC cell lines.

Conclusions: Natural autoantibodies against CD47 may be a potential agent for OSCC immunotherapy.
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http://dx.doi.org/10.1111/odi.13922DOI Listing
May 2021

A case of hyperhaemolysis syndrome in a pregnant Chinese woman with β-thalassemia during perinatal transfusion.

Transfus Med 2021 Feb 16;31(1):24-29. Epub 2020 Dec 16.

Blood Transfusion Research Center, Dongguan City Blood Station, Dongguan, China.

Objectives: To report a case of hyperhaemolysis syndrome (HHS) that occurred during perinatal blood transfusion in a pregnant Chinese woman with β-thalassemia to deepen the understanding of HHS and the risk of transfusion therapy for patients with thalassemia.

Background: Most HHS cases occur in people with sickle cell disease. So far, no cases of HHS have been reported in the Chinese population. Here, we report a pregnant Chinese women with β-thalassemia experiencing HHS.

Methods: The patient received ABO- and RhD-matched red blood cell transfusion from six blood donors in four perinatal transfusions. Haemoglobinuria and lower haemoglobin levels compared to those before transfusion were observed after each transfusion, and the lactate dehydrogenase was consistently elevated. The blood samples were collected at different time points during the hospitalisation for direct antiglobulin test (DAT), antibody screening test and acid elution test. The antigens of six blood donors were identified, and the cross-matching tests were repeated using the blood sample of the patient with specific irregular antibodies after the last transfusion.

Results: The DAT of the patient was negative for anti-IgG and positive (1+) for anti-C3d, and no red blood cell antibodies were detected in the eluent before, between and after transfusions. Before and between transfusions, blood samples were negative for red blood cell irregular antibodies, whereas IgM anti-P and IgG anti-Jk were detected in blood samples the next day after the last transfusion. In the six donors, two were negative for P and Jk , one was positive for P and negative for Jk , and three were negative for P and positive for Jk . The tentative cross-matching tests using the indirect antiglobulin method in saline showed that only agglutination occurred in the blood samples of the patient collected after last transfusion and the three Jk -positive blood donors.

Discussion: The clinical manifestations and laboratory test results suggested that HHS occurred in this patient with β-thalassemia after each transfusion. Clinicians should be aware that HHS can occur with compatible blood transfusion.
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http://dx.doi.org/10.1111/tme.12748DOI Listing
February 2021

Composable Microfluidic Plates (cPlate): A Simple and Scalable Fluid Manipulation System for Multiplexed Enzyme-Linked Immunosorbent Assay (ELISA).

Anal Chem 2021 01 16;93(3):1489-1497. Epub 2020 Dec 16.

C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, West Virginia 26506, United States.

Enzyme-linked immunosorbent assay (ELISA) is the gold standard method for protein biomarkers. However, scaling up ELISA for multiplexed biomarker analysis is not a trivial task due to the lengthy procedures for fluid manipulation and high reagent/sample consumption. Herein, we present a highly scalable multiplexed ELISA that achieves a similar level of performance to commercial single-target ELISA kits as well as shorter assay time, less consumption, and simpler procedures. This ELISA is enabled by a novel microscale fluid manipulation method, composable microfluidic plates (cPlate), which are comprised of miniaturized 96-well plates and their corresponding channel plates. By assembling and disassembling the plates, all of the fluid manipulations for 96 independent ELISA reactions can be achieved simultaneously without any external fluid manipulation equipment. Simultaneous quantification of four protein biomarkers in serum samples is demonstrated with the cPlate system, achieving high sensitivity and specificity (∼ pg/mL), short assay time (∼1 h), low consumption (∼5 μL/well), high scalability, and ease of use. This platform is further applied to probe the levels of three protein biomarkers related to vascular dysfunction under pulmonary nanoparticle exposure in rat's plasma. Because of the low cost, portability, and instrument-free nature of the cPlate system, it will have great potential for multiplexed point-of-care testing in resource-limited regions.
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http://dx.doi.org/10.1021/acs.analchem.0c03651DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8276162PMC
January 2021

Longitudinal Changes on Clinical Features in 28 Children With COVID-19 in Shenzhen, China.

Front Med (Lausanne) 2020 4;7:579406. Epub 2020 Nov 4.

Institute of Hepatology, Shenzhen Third People's Hospital, Shenzhen, China.

To investigate the clinical characteristics of children with coronavirus disease 2019 (COVID-19) and identify the occurrence of viral shedding of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) during follow-up. We retrospectively retrieved data from pediatric patients with COVID-19 from the Shenzhen Third People's Hospital in China. The dynamics of SARS-CoV-2 and antibodies against SARS-CoV-2 were analyzed during hospitalization and after discharge. From January 23 to March 15, 2020, a total of 28 pediatric patients were diagnosed with COVID-19 and were followed for at least 1 month. The median age was 7 years (IQR 3.5-10) and none of the children progressed to severe COVID-19 during hospitalization. Ten patients tested positive for SARS-CoV-2 1 month after discharge while four patients tested positive during the 2nd month after discharge. Only three of 12 children showed detectable immunoglobulin-M (IgM) on day 5, 18, and 21 after illness onset, respectively. COVID-19 disease was relatively mild among children while a number did test positive after discharge from the hospital. Public health initiatives should thus adapt control measures targeted toward children.
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http://dx.doi.org/10.3389/fmed.2020.579406DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672180PMC
November 2020

Probiotics treatment improves cognitive impairment in patients and animals: A systematic review and meta-analysis.

Neurosci Biobehav Rev 2021 01 4;120:159-172. Epub 2020 Nov 4.

Department of Behavioral Neurosciences, Science Research Center of Medical School, Shaoxing University, Shaoxing, Zhejiang, China; Laboratory of Forensic Toxicology, Judicial Identification Center of Shaoxing University, Shaoxing, Zhejiang, China.

The gut-brain axis has received considerable attention in recent years, and the "psychobiotics" concept indicates that probiotics have a potential positive effect on cognitive function. Therefore, the aim of this study was to quantitatively evaluate the influence of probiotics on cognition. We conducted a random-eff ;ects meta-analysis of 7 controlled clinical trials and 11 animals studies to evaluate the eff ;ects of probiotics on cognitive function. Probiotics supplementation enhanced cognitive function in both human (0.24 [0.05-0.42]; I = 0 %) and animal studies (0.90 [0.47-1.34]; I = 74 %). Subgroup analyses indicated that the effects of probiotics on cognitively impaired individuals (0.25 [0.05-0.45]; I = 0 %) were greater than those on healthy ones (0.15 [-0.30 to 0.60]; I = 0 %). Furthermore, compared with a multiple-probiotic supplement, a single strain of probiotics was more effective in humans. The meta-analysis provided some suggestions for probiotics intervention and tended to support a customized approach for different individuals to ameliorate cognitive disorders. Future additional clinical trials are necessary to evaluate therapeutic effect and influencing factors.
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http://dx.doi.org/10.1016/j.neubiorev.2020.10.027DOI Listing
January 2021

Mineralization of the herbicide swep by a two-strain consortium and characterization of a new amidase for hydrolyzing swep.

Microb Cell Fact 2020 Jan 7;19(1). Epub 2020 Jan 7.

Department of Microbiology, Key Lab of Microbiology for Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, China.

Background: Swep is an excellent carbamate herbicide that kills weeds by interfering with metabolic processes and inhibiting cell division at the growth point. Due to the large amount of use, swep residues in soil and water not only cause environmental pollution but also accumulate through the food chain, ultimately pose a threat to human health. This herbicide is degraded in soil mainly by microbial activity, but no studies on the biotransformation of swep have been reported.

Results: In this study, a consortium consisting of two bacterial strains, Comamonas sp. SWP-3 and Alicycliphilus sp. PH-34, was enriched from a contaminated soil sample and shown to be capable of mineralizing swep. Swep was first transformed by Comamonas sp. SWP-3 to the intermediate 3,4-dichloroaniline (3,4-DCA), after which 3,4-DCA was mineralized by Alicycliphilus sp. PH-34. An amidase gene, designated as ppa, responsible for the transformation of swep into 3,4-DCA was cloned from strain SWP-3. The expressed Ppa protein efficiently hydrolyzed swep and a number of other structural analogues, such as propanil, chlorpropham and propham. Ppa shared less than 50% identity with previously reported arylamidases and displayed maximal activity at 30 °C and pH 8.6. Gly449 and Val266 were confirmed by sequential error prone PCR to be the key catalytic sites for Ppa in the conversion of swep.

Conclusions: These results provide additional microbial resources for the potential remediation of swep-contaminated sites and add new insights into the catalytic mechanism of amidase in the hydrolysis of swep.
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http://dx.doi.org/10.1186/s12934-020-1276-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6945715PMC
January 2020

A study of natural IgG antibodies against ATP-binding cassette subfamily C member 3 in oral squamous cell carcinoma.

J Cancer Res Ther 2019 ;15(4):921-926

Beijing Institution of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing, China.

Aims: ATP-binding cassette subfamily C member 3 (ABCC3) is involved in multidrug resistance and is overexpressed in some solid tumors. Recent work revealed an increase in circulating anti-ABCC3 antibodies in lung and esophageal cancers. This in vitro study was undertaken to investigate the effects of the natural IgG antibody against the ABCC3-derived peptide antigen on proliferation of oral squamous cell carcinoma (OSCC) cells and augment the development of efficient and effective treatments in patients with OSCC.

Subjects And Methods: An in-house enzyme-linked immunosorbent assay was applied to detect anti-ABCC3 IgG antibody in human plasma. Two OSCC cell lines, CAL27 and SCC15, were cultured with 20% plasma either positive or negative for anti-ABCC3 IgG. Cell proliferation was quantified by the CCK-8 method, and cell apoptosis and cell cycle distribution were analyzed by flow cytometry. The expression of the ABCC3 gene in the cell lines was analyzed by reverse transcriptase quantitative real-time polymerase chain reaction.

Results: The results showed that plasma anti-ABCC3 IgG significantly inhibited the proliferation of CAL27 cells but not SCC15 cells, although ABCC3 was expressed in both cell lines. The proportion of apoptotic cells was significantly higher in CAL27 cells treated with anti-ABCC3 IgG-positive plasma than in those treated with IgG-negative plasma. Cell cycle progression was arrested in CAL27 cells treated with anti-ABCC3 IgG-positive plasma.

Conclusions: Our data suggest that human plasma anti-ABCC3 IgG may be a promising agent in anti-OSCC therapy, although further studies are needed to arrive at a definitive conclusion.
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http://dx.doi.org/10.4103/jcrt.JCRT_150_18DOI Listing
February 2020

Acoustofluidic enzyme-linked immunosorbent assay (ELISA) platform enabled by coupled acoustic streaming.

Anal Chim Acta 2019 Nov 1;1079:129-138. Epub 2019 Jun 1.

C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, WV, USA. Electronic address:

Bead-based ELISA in microfluidics is a promising platform for reducing the reagent consumption and improving assay throughput due to its fast binding kinetics and low reagent consumption. Current microfluidic bead-based immunoassay mainly relies on magnetic and centrifugal forces to manipulate microparticles to complete an assay protocol. Here we report an acoustic streaming-based microfluidic method that can perform all the fluid and particle operations of bead-based ELISA. A series of sharp-edge structures are used to generate a long-range coupled acoustic streaming that enables simultaneous particle trapping and active molecular exchange in a dead-end microchannel. The device achieved >99% of molecular exchange in 4 min, while maintaining a particle trapping efficiency of 85%. This acoustofluidic-based method demonstrates all three key operations in a bead-based immunoassay: (1) Beads "immobilization"; (2) Active mixing of fluid for bead/target binding; (3) Active molecular exchange for reagent loading and washing. Finally, on-chip quantitative detection of biomarker IL-6 with a limit of detection ∼50 pg/mL is achieved using this platform including an enzymatic signal amplification step. The small footprint, simple setup, and continuous flow operation of the acoustic streaming-based method makes it an attractive platform for continuous flow bead-based immunoassay.
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http://dx.doi.org/10.1016/j.aca.2019.05.073DOI Listing
November 2019

In Situ Monitoring of Fluid Shear Stress Enhanced Adherence of Bacteria to Cancer Cells on Microfluidic Chip.

Anal Chem 2019 05 15;91(9):5973-5979. Epub 2019 Apr 15.

Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, MOE Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology , Tsinghua University , Beijing 100084 , China.

Mechanosensing mechanisms for surface recognition by bacteria play an important role in inflammation and phagocytosis. Here, we describe a set of DNA probes for revealing microbe adherence to cancer cells under fluid shear stress. DNA probes modified with a biotin group, an azido group, and hexadecanoic acid were indiscriminately anchored to the cell surface, acting as indicators for the membrane proteins, cell-surface carbohydrate, and phospholipids. When cancer cells were exposed to bacteria in fluid, enhanced accumulation of membrane proteins was indicated by the strong fluorescence aggregation, meanwhile the weakened accumulation of cell-surface carbohydrate and phospholipids indication was indicated by attenuated fluorescence. Further research demonstrates that this mechanosensing strategy was applicable to different bacterial-cancer cell interactions. This study not only uncovered new cellular mechanotransduction mechanisms, but also provided a versatile method that enabled in situ and dynamic indication of cancer cell responses to mechanical stimuli.
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http://dx.doi.org/10.1021/acs.analchem.9b00394DOI Listing
May 2019

Capillary Vibrating Sharp-Edge Spray Ionization (cVSSI) for Voltage-Free Liquid Chromatography-Mass Spectrometry.

J Am Soc Mass Spectrom 2019 May 21;30(5):824-831. Epub 2019 Feb 21.

C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, WV, USA.

Here, we report a continuous flow-based ionization method, capillary vibrating sharp-edge spray ionization (cVSSI), that nebulizes liquid sample directly at the outlet of a capillary without using high-speed nebulization gas or a high electrical field. cVSSI is built upon the recently reported VSSI principle which nebulizes bulk liquid using vibrating sharp-edges. By attaching a short piece of fused silica capillary on top of the vibrating glass slide in VSSI, liquid is nebulized at the outlet of the capillary as the result of the vibration. Utilizing standard 360-μm OD/100-μm ID capillary, cVSSI works with a wide range of flow rates from 1 μL/min to 1 mL/min. The power consumption is as low as 130 mW. ESI-like MS spectra are obtained for small molecules, peptides, and proteins. Five orders of magnitude linear response for acetaminophen solution is achieved with a limit of detection (LOD) of 3 nM. cVSSI is also demonstrated to be compatible with LC-MS analysis. Two LC-MS applications are demonstrated with cVSSI: (1) separation and detection of a mixture of small molecules and (2) bottom-up proteomics using a protein digest. A mixture of nine common metabolites was appropriately separated and detected using LC-cVSSI-MS. In the bottom-up experiment, 78 peptides were detected using LC-cVSSI-MS/MS with a protein coverage of 100% for cytochrome c, which is comparable with the coverage obtained using LC-ESI-MS. cVSSI offers a means of interfacing LC or other continuous flow-based applications to mass spectrometers with the salient features of voltage-free, flexibility, small footprint, and low power consumption.
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http://dx.doi.org/10.1007/s13361-019-02147-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6560627PMC
May 2019

Live imaging of cell membrane-localized MT1-MMP activity on a microfluidic chip.

Chem Commun (Camb) 2018 Oct;54(81):11435-11438

Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, MOE Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing, China.

We designed an enzyme-activatable probe for real time in situ tracking of MT1-MMP activity. The MT1-MMP responses of endothelial cells were investigated under the regulation of shear stress and biochemical factor on a microfluidic chip. This strategy can be beneficial to evaluate membrane protein responses to external stimuli and to engineer cells for basic and clinical research.
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http://dx.doi.org/10.1039/c8cc07117aDOI Listing
October 2018

Evaluating nanomedicine with microfluidics.

Nanotechnology 2018 Dec 14;29(49):492001. Epub 2018 Sep 14.

C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, WV, United States of America.

Nanomedicines are engineered nanoscale structures that have an extensive range of application in the diagnosis and therapy of many diseases. Despite the rapid progress in and tremendous potential of nanomedicines, their clinical translational process is still slow, owing to the difficulty in understanding, evaluating, and predicting their behavior in complex living organisms. Microfluidic techniques offer a promising way to resolve these challenges. Carefully designed microfluidic chips enable in vivo microenvironment simulation and high-throughput analysis, thus providing robust platforms for nanomedicine evaluation. Here, we summarize the recent developments and achievements in microfluidic methods for nanomedicine evaluation, categorized into four sections based on their target systems: single cell, multicellular system, organ, and organism levels. Finally, we provide our perspectives on the challenges and future directions of microfluidics-based nanomedicine evaluation.
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http://dx.doi.org/10.1088/1361-6528/aae18aDOI Listing
December 2018

Phagocytic intracellular digestion in amphioxus ().

Proc Biol Sci 2018 06;285(1880)

State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, People's Republic of China.

The digestive methods employed by amphioxus ()-both intracellular phagocytic digestion and extracellular digestion-have been discussed since 1937. Recent studies also show that epithelial cells lining the digestive tract can express many immune genes. Here, in , using a special tissue fixation method, we show that some epithelial cells, especially those lining the large diverticulum protruding from the gut tube, phagocytize food particles directly, and can rely on this kind of phagocytic intracellular digestion to obtain energy throughout all stages of its life. Gene expression profiles suggest that diverticulum epithelial cells have functional features of both digestive cells and phagocytes. In starved , these cells accumulate endogenous digestive and hydrolytic enzymes, whereas, when sated, they express many kinds of immune genes in response to stimulation by phagocytized food particles. We also found that the distal hindgut epithelium can phagocytize food particles, but not as many. These results illustrate phagocytic intercellular digestion in , explain why digestive tract epithelial cells express typical immune genes and suggest that the main physiological function of the diverticulum is different from that of the vertebrate liver.
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http://dx.doi.org/10.1098/rspb.2018.0438DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6015868PMC
June 2018

Shear Stress-Enhanced Internalization of Cell Membrane Proteins Indicated by a Hairpin-Type DNA Probe.

Anal Chem 2018 05 9;90(9):5540-5545. Epub 2018 Apr 9.

Beijing Key Laboratory of Microanalytical Methods and Instrumentation, MOE Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Department of Chemistry , Tsinghua University , Beijing 100084 , China.

Shear stress is an important mechanical stimulus that plays a critical role in modulating cell functions. In this study, we investigated the regulating effects of shear stress on the internalization of cell membrane proteins in a microfluidic chip. A hairpin-type DNA probe was developed and indiscriminately anchored to the cell surface, acting as an indicator for the membrane proteins. When cells were exposed to shear stress generated from fluid cell medium containing external proteins, strong fluorescence was emanated from intracellular regions. With intensive investigation, results revealed that shear stress could enhance the specific cell endocytosis pathway and promote membrane protein internalization. This process was indicated by the enhanced intracellular fluorescence, generated from the internalized and mitochondria accumulated DNA probes. This study not only uncovered new cellular mechanotransduction mechanisms but also provided a versatile method that enabled in situ and dynamic indication of cell responses to mechanical stimuli.
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http://dx.doi.org/10.1021/acs.analchem.8b00755DOI Listing
May 2018

A dual-functional microfluidic chip for on-line detection of interleukin-8 based on rolling circle amplification.

Biosens Bioelectron 2018 Apr 12;102:652-660. Epub 2017 Dec 12.

Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, MOE Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing 100084, People's Republic of China. Electronic address:

Interleukin 8 (IL-8), also known as C-X-C motif ligand 8(CXCL8), is a proinflammatory chemokine functioned in neutrophil chemotaxis and activation. And it plays an important role in the process of glioma stem-like cell vascularization in the latest research. Herein, a dual-function microfluidic biosensor based on rolling circle amplification (RCA) was fabricated for cell culture and online IL-8 detection. A microfluidic chip was designed with two high passages connected by the vertical channels. One of the channels with immobilized capture antibody was prepared for IL-8 detection and another channel for cell culture. Immunoassays were achieved by a sandwich structure consisting of antibodies, IL-8, and aptamers. Signal amplification was mainly due to RCA and biotin-streptavidin linkage. The linear range for IL-8 was 7.5 -120pgmL in this assay. Moreover, the developed method was successfully applied to detect the IL-8 in tumor-derived endothelial cells (TDEC) and Human Umbilical Vein Endothelial cells (HUVEC) under chemical hypoxia condition. Semi-quantitative detection of IL-8 consumption in HUVEC cells in low oxygen condition was also achieved. These results were in statistical agreement with those obtained by commercial assay of enzyme-linked immunoassay kit (ELISA). The microfluidic chip based biosensor reported hereby has a large prospect in the basic research and clinical diagnosis of cancer stem cell.
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http://dx.doi.org/10.1016/j.bios.2017.12.017DOI Listing
April 2018

Evaluation of drug combination for glioblastoma based on an intestine-liver metabolic model on microchip.

Analyst 2017 Sep;142(19):3629-3638

State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.

An intestine-liver-glioblastoma biomimetic system was developed to evaluate the drug combination therapy for glioblastoma. A hollow fiber (HF) was embedded into the upper layer of the microfluidic chip for culturing Caco-2 cells to mimic drug delivery as an artificial intestine. HepG2 cells cultured in the bottom chamber of the chip acted as an artificial liver for metabolizing the drugs. The dual-drug combination to glioblastoma U251 cells was evaluated based on the intestine-liver metabolic model. The drugs, irinotecan (CPT-11), temozolomide (TMZ) and cyclophosphamide (CP), were used to dynamically stimulate the cells by continuous infusion into the intestine unit. After intestine absorption and liver metabolism, the prodrugs were transformed to active metabolites, which induced glioblastoma cells apoptosis. The anticancer activity of the CPT-11 and TMZ combination is significantly enhanced compared to that of the single drug treatments. Combination index (CI) values of the combination groups, CPT-11 and TMZ, CPT-11 and CP, and TMZ and CP, at half maximal inhibitory concentration were 0.137, 0.288, and 0.482, respectively. The results indicated that the CPT-11 and TMZ combination was superior to the CPT-11 and CP group as well as the TMZ and CP group towards the U251 cells. The metabolism mechanism of CPT-11 and TMZ was further studied by coupling with mass spectrometric analysis. The biomimetic model enables the performance of long-term cell co-culture, drug delivery, metabolism and real-time analysis of drug effects, promising systematic in vitro mimicking of physiological and pharmacological processes.
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http://dx.doi.org/10.1039/c7an00453bDOI Listing
September 2017

Study of antioxidant effects on malignant glioma cells by constructing a tumor-microvascular structure on microchip.

Anal Chim Acta 2017 07 16;978:1-9. Epub 2017 May 16.

Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, The Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing 100084, China. Electronic address:

In this work, a three-dimensional tumor-microvascular structure was simulated on a microfluidic chip for study of antioxidants effects on malignant glioma cells in vitro. The 3D hydrogel containing lumen was constructed to co-culture endothelial cells and glioma cells to mimic tumor microvascular environment. Macroporous gelatin transglutaminase (TG) hydrogel was prepared with biological and mechanical properties suitable for cells culture and nutrient refresh. To reform a vessel structure, U87 cells were dispersed in the TG-gelatin hydrogel and HUVEC cells were seeded in the lumen of hydrogel. Three typical antioxidants (α-lipoic acid, catechins and ascorbic acid) have been selected to research the antioxidant effects of glioma cells in the simulative tumor microenvironment. The results showed that the HUVEC cells formed vessel presented the transportation and penetrable functions for antioxidants from lumen to glioma cells. The antioxidants displayed higher selectivity to U87 cells than HUVEC cells and α-lipoic acid has a strong antioxidant capacity.
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http://dx.doi.org/10.1016/j.aca.2017.05.009DOI Listing
July 2017

Quantitative determination of VEGF165 in cell culture medium by aptamer sandwich based chemiluminescence assay.

Talanta 2017 Aug 25;171:197-203. Epub 2017 Apr 25.

Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, The Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing 100084, China. Electronic address:

In this work, we have developed a sensitive and selective chemiluminescence (CL) assay for vascular endothelial growth factor (VEGF165) quantitative detection based on two specific VEGF165 binding aptamers (Apt). VEGF is a predominant biomarker in cancer angiogenesis, and sensitive detection method of VEGF are highly demanded for both academic study and clinical diagnosis of multiple cancers. In our experiment, VEGF165 was captured in a sandwich structure assembled by two binding aptamers, one capture aptamer was immobilized on streptavidin-coated magnetic beads (MBs) and another VEGF-binding aptamer was labeled by biotin for further phosphatase conjunction. After Apt-VEGF-Apt sandwich was formed on MBs surface, alkaline phosphatase (ALP) was modified to the second aptamer to catalyze CL reaction. By applying 4-methoxy-4-(3-phosphatephenyl)-spiro-(1,2-dioxetane-3,2-adamantane) (AMPPD) as CL substrate, strong signal intensity was achieved. VEGF165 content as low as 1ng/mL was detected in standard spiked samples by our assay, and linear range of working curve was confirmed from 1 to 20ng/mL. Then our method was successfully applied for cell culture medium analysis and on-chip hypoxic HepG2-HUVEC co-culture model study with excellent accuracy equal to ELISA Kit. Our developed assay demonstrated an outstanding performance in VEGF165 quantification and may be further extended to clinical testing of important biomarkers as well as probing microchip cell culture model.
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http://dx.doi.org/10.1016/j.talanta.2017.04.057DOI Listing
August 2017

Gold nanoparticles modified porous silicon chip for SALDI-MS determination of glutathione in cells.

Talanta 2017 Jun 21;168:222-229. Epub 2017 Feb 21.

Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China. Electronic address:

The gold nanoparticles (Au NPs) modified porous silicon chip based surface assisted laser desorption/ionization mass spectrometry (SALDI-MS) was developed to capture and analyze glutathione (GSH) in cells. With silver-assisted chemical etching, Ag nanoparticles (Ag NPs) were generated and deposited on the silicon surface and the nanopores were etched on silicon substrate. Then Au NPs were in-situ synthesized on the ridges of silicon nanopores. This Ag-Au NPs modified porous silicon surface could specially capture and enrich thiol compounds through Au-S binding, and it could also function as matrix to assist ionization for SALDI-MS. The silicon chip was array patterned for high throughput SALDI-MS detection. GSH and cysteine could be distinguished without the interference from matrix signals. This approach was successfully applied to preconcentration and detection of GSH in Caco-2 cells. The GSH alterations in cells under drug stimulation were investigated. This invented silicon chip showed great potential for more efficient analysis of small thiol biomarkers in complex biological samples.
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http://dx.doi.org/10.1016/j.talanta.2017.02.041DOI Listing
June 2017

A DNA-directed covalent conjugation fluorescence probe for in vitro detection of functional matrix metalloproteinases.

Analyst 2017 Feb 23;142(4):634-640. Epub 2017 Jan 23.

Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, P. R. China.

Matrix metalloproteinases (MMPs) have been considered to contribute to the progression of tumorigenesis and tumor invasion; MMP-9 in particular, has been regarded as a priority target in cancer treatment due to its massive up-regulation in malignant tissues and its ability to degrade type IV collagen. In this work, we employed a DNA-directed covalent conjugation method to design a fluorescence probe for in vitro detection of functional matrix metalloproteinases, by which a nitrilotriacetic acid (NTA)-modified DNA probe can combine with the Zn in the active site of MMPs, and then a molecule beacon (MB) modified FITC and BHQ1 can open to bond with their complementary base, NTA-modified DNA. We can evaluate the amount of MMPs in the medium according to the fluorescence intensity. The detection procedure can be finished in 30 min with good selectivity, cheap reagents and easy preparation. All the results and the amount of secreted MMPs under three different cell culture conditions are in accordance with previous reports. Satisfactory results are obtained. Furthermore, owing to the importance of MMP-9, we designed an approach to achieve the desired selectivity and specificity of our work, using dual amplification for improving fluorescence intensity based on RCA to detect the amount of MMP-9.
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http://dx.doi.org/10.1039/c6an02339hDOI Listing
February 2017

DNA-mediated cell surface engineering for multiplexed glycan profiling using MALDI-TOF mass spectrometry.

Chem Sci 2016 Aug 5;7(8):5448-5452. Epub 2016 May 5.

Department of Chemistry , Beijing Key Laboratory of Microanalytical Methods and Instrumentation , Tsinghua University , Beijing 100084 , China . Email:

Glycans are crucial for many key biological processes and their alterations are often a hallmark of disease. Thus, multiplexed and sensitive analysis of glycans is of intense interest. Here we report a novel approach using DNA-mediated cell surface engineering for glycan profiling by MALDI-TOF mass spectrometry (MS). Following lectin binding, DNA amplification and hybridization, glycans on the cell surface are specifically labeled by short DNA probes, which can be facilely released, ionized and detected in MALDI-TOF MS. This strategy converts the analysis of glycans to the detection of DNA probes, overcoming the complicated composition and low ionization efficiency of glycans, enabling detection and facilitating multiplex analysis. The amplification procedure also improves the sensitivity. This approach has been applied to evaluate glycomic alterations in cancer cells and provided the intrinsic distribution of glycans in tissues using MALDI imaging mass spectrometry.
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http://dx.doi.org/10.1039/c6sc00215cDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6021752PMC
August 2016
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