Publications by authors named "Hongying Zhu"

42 Publications

Metabolomic profiling of single enlarged lysosomes.

Nat Methods 2021 Jul 14;18(7):788-798. Epub 2021 Jun 14.

Institute on Aging and Brain Disorders, the First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, Hefei National Laboratory for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei, China.

Lysosomes are critical for cellular metabolism and are heterogeneously involved in various cellular processes. The ability to measure lysosomal metabolic heterogeneity is essential for understanding their physiological roles. We therefore built a single-lysosome mass spectrometry (SLMS) platform integrating lysosomal patch-clamp recording and induced nano-electrospray ionization (nanoESI)/mass spectrometry (MS) that enables concurrent metabolic and electrophysiological profiling of individual enlarged lysosomes. The accuracy and reliability of this technique were validated by supporting previous findings, such as the transportability of lysosomal cationic amino acids transporters such as PQLC2 and the lysosomal trapping of lysosomotropic, hydrophobic weak base drugs such as lidocaine. We derived metabolites from single lysosomes in various cell types and classified lysosomes into five major subpopulations based on their chemical and biological divergence. Senescence and carcinoma altered metabolic profiles of lysosomes in a type-specific manner. Thus, SLMS can open more avenues for investigating heterogeneous lysosomal metabolic changes during physiological and pathological processes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41592-021-01182-8DOI Listing
July 2021

Impact of inappropriate empirical antibiotic treatment on clinical outcomes of urinary tract infections caused by Escherichia coli: a retrospective cohort study.

J Glob Antimicrob Resist 2021 Jun 9;26:148-153. Epub 2021 Jun 9.

Jiangxi Provincial Key Laboratory of Medicine, Clinical Laboratory of the Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, People's Republic of China. Electronic address:

Objectives: We aimed to determine the clinical impact of inappropriate empirical antibiotic treatment (IEAT) compared with appropriate empirical antibiotic treatment (AEAT) in hospitalised patients with urinary tract infections (UTIs) caused by Escherichia coli (E. coli).

Methods: This retrospective cohort study included adult patients with a primary diagnosis of UTI who were treated with empirical antibiotics at a tertiary hospital in southern China over a 2-year period. Clinical data of patients who received IEAT were compared with those of patients receiving AEAT. We used multivariable logistic regression to identify the predictors for receiving IEAT and the risk factors affecting clinical outcomes.

Results: A total of 213 patients were enrolled (median age, 61 years), of whom 103 (48.4%) received IEAT. IEAT was associated with empirical use of fluoroquinolones, male sex and age-adjusted Charlson comorbidity index (aCCI) score >6. Hospital length of stay (LOS) was longer for patients who received IEAT than for those who received AEAT (13.6 ± 8.6 days vs. 10.8 ± 7.9 days; P = 0.008). IEAT was an independent risk factor for longer LOS along with aCCI score ≥2, lung disease and cardiac disease.

Conclusion: Empirical use of fluoroquinolones for UTIs should be avoided, especially in male patients with aCCI score >6. Improved empirical antimicrobial therapy may have a beneficial impact in reducing bacterial resistance and healthcare costs by decreasing the LOS. Therefore, interventions to promote in-depth antibiotic stewardship programmes in China are needed.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jgar.2021.05.016DOI Listing
June 2021

Phytotoxicity and nutrition behavior of process water obtained from the hydrothermal carbonization of poultry litter and its effect on lettuce germination and growth.

Environ Sci Pollut Res Int 2021 Jun 9. Epub 2021 Jun 9.

School of Petroleum Engineering and Environmental Engineering, Yan'an University, Yan'an City, 716000, Shaanxi, China.

Nine hydrothermal carbonization process waters (PWs) of poultry litter were prepared at 180, 220, and 260 °C for 1, 4, and 8 h, respectively. They were characterized with pH, EC (electric conductivity), DOC (dissolved organic carbon), TN (total nitrogen), NH-N, NO-N, etc. After diluted according to TN, the PWs were supplied as liquid nitrogen fertilizers and their phytotoxic and nutrition effects on lettuce germination and growth were studied. The results showed that the PWs from short time (1 h) were with low DOC/TN and DOC/NH-N and high NH-N/TN. Compared with the PWs from long time 4 and 8 h, they provided more NH-N and less DOC and resulted in lettuce with relatively high germination index (GI), dry biomass, and low antioxidant enzyme activities. Especially, the PW from 220 °C and 1 h significantly enhanced the dry weight by 196.3% relative to negative control of nitrogen deficiency. However, all the PWs led lettuce to an unhealthy condition, which decreased GI and the chlorophyll content and increased antioxidant enzyme activities. Furthermore, it was confirmed by linear regression that the ratios of DOC/TN, NH-N/TN, and DOC/NH-N were the determining indexes for evaluating the phytotoxicity and nutrition behavior of the PWs as liquid nitrogen fertilizers.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s11356-021-14697-6DOI Listing
June 2021

Brain ethanol metabolism by astrocytic ALDH2 drives the behavioural effects of ethanol intoxication.

Nat Metab 2021 03 22;3(3):337-351. Epub 2021 Mar 22.

Laboratory for Integrative Neuroscience, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD, USA.

Alcohol is among the most widely used psychoactive substances worldwide. Ethanol metabolites such as acetate, thought to be primarily the result of ethanol breakdown by hepatic aldehyde dehydrogenase 2 (ALDH2), contribute to alcohol's behavioural effects and alcoholism. Here, we show that ALDH2 is expressed in astrocytes in the mouse cerebellum and that ethanol metabolism by astrocytic ALDH2 mediates behavioural effects associated with ethanol intoxication. We show that ALDH2 is expressed in astrocytes in specific brain regions and that astrocytic, but not hepatocytic, ALDH2 is required to produce ethanol-derived acetate in the mouse cerebellum. Cerebellar astrocytic ALDH2 mediates low-dose ethanol-induced elevation of GABA levels, enhancement of tonic inhibition and impairment of balance and coordination skills. Thus, astrocytic ALDH2 controls the production, cellular and behavioural effects of alcohol metabolites in a brain-region-specific manner. Our data indicate that astrocytic ALDH2 is an important, but previously under-recognized, target in the brain to alter alcohol pharmacokinetics and potentially treat alcohol use disorder.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s42255-021-00357-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8294184PMC
March 2021

Correlation between monoamine neurotransmitter and cytokine levels and the occurrence of post-traumatic stress disorder among operating room nurses.

Ann Palliat Med 2020 Nov;9(6):3947-3956

Department of Operating Theater, The Fourth People's Hospital of Haikou, Haikou, China.

Background: The aim of the present study was to investigate the correlation between monoamine neurotransmitter and cytokine levels and the occurrence of post-traumatic stress disorder (PTSD) among operating room nurses.

Methods: A total of 131 nursing staff were selected and assigned into the PTSD, non-PTSD, and control group. Enzyme-linked immunosorbent assay was applied to determine the monoamine neurotransmitters in plasma and serum cytokines. Receiver-operating characteristic curve analysis was conducted to assess the sensitivity and specificity of neurotransmitters and cytokines in the clinical detection of PTSD among operating room nurses. Addenbrooke's Cognitive Examination-Revised and the Connor-Davidson Resilience Scale were used to evaluate the correlation between neurotransmitter and cytokine levels and the clinical characteristics of operating room nurses with PTSD.

Results: Our study found that the monoamine neurotransmitters and cytokines among nurses in the PTSD group were significantly higher than those in the non-PTSD and control group. Neurotransmitter and cytokine levels as clinical predictors of PTSD among operating room nurses have good sensitivity and specificity, and were negatively correlated with cognitive function and resilience.

Conclusions: The findings of the present study confirm that monoamine neurotransmitter and cytokine levels are correlated with the occurrence of PTSD among operating room nurses.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.21037/apm-20-1829DOI Listing
November 2020

A Retrospective Analysis of Risk Factors and Patient Outcomes of Bloodstream Infection with Extended-Spectrum β-Lactamase-Producing in a Chinese Tertiary Hospital.

Infect Drug Resist 2020 24;13:4289-4296. Epub 2020 Nov 24.

Jiangxi Provincial Key Laboratory of Medicine, Clinical Laboratory of the Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, People's Republic of China.

Objective: The present study assessed risk factors and patient outcomes of bloodstream infection (BSI) caused by extended-spectrum β-lactamase (ESBL)-producing ().

Methods: A retrospective study was performed to analyze risk factors and patient outcomes of BSI caused by extended-spectrum β-lactamase-producing (ESBL-EC) in one Chinese tertiary hospital over a 7.5-year period. The clinical characteristics of patients infected with ESBL-producing and non-ESBL-producing were compared. Predictors of 30-day mortality in patients with BSI were also identified in our study.

Results: The results of drug sensitivity showed that quinolones, aminoglycosides, -lactam/-lactamase inhibitor combinations (BLICs) and trimethoprim/sulfamethoxazole exhibited significant differences between the ESBL and non-ESBL groups. Of the 963 patients with BSI, 57.6% developed ESBL-EC. Multivariate analysis showed that biliary tract infection (BTI) [P<0.001,OR (95% CI):1.798 (1.334-2.425)], urinary tract obstructive disease [P=0.001,OR (95% CI):2.106 (1.366-3.248)], surgery within 3 months [P=0.002,OR (95% CI):1.591 (1.178-2.147)], hospitalization within 3 months [P<0.001,OR (95% CI):2.075 (1.579-2.725)], ICU admission [P=0.011,OR (95% CI):1.684 (1.124-2.522)] and history of cephalosporin use [P=0.006,OR (95% CI):3.097 (1.392-6.891)] were statistically significant. In mortality analysis, aCCI>2 [P=0.016,OR (95% CI): 2.453 (1.179-5.103)], gastrointestinal catheterization [P=0.004, OR (95% CI): 2.525 (1.333-4.782)] were significantly associated with 30-day mortality. According to Kaplan-Meier survival analysis, we found that in SOFA<2 group and SOFA≥2 group, the mortality rate of patients treated with BLICs were lower than that of carbapenems(P<0.05).

Conclusion: This study showed that BTI, urinary tract obstructive disease, surgery within 3 months, hospitalization within 3 months, ICU admission and cephalosporin exposure were independent risk factors for the emergence of ESBL-EC BSI. Analysis of risk factors for 30-day mortality revealed that the factors independently associated with a higher risk of mortality were aCCI>2, gastrointestinal catheterization. Compared to carbapenems, the BLICs had preferable effect to treat patients with ESBL-EC BSI. Notably, patients with severe illness were inlcined to use carbapenems, which affected the analysis results. Therefore, we suggest that BLICs could be recommended to treat mild patients with ESBL-EC bacteremia.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.2147/IDR.S269989DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7699446PMC
November 2020

Ultra-sensitive and rapid detection of nucleic acids and microorganisms in body fluids using single-molecule tethering.

Nat Commun 2020 09 22;11(1):4774. Epub 2020 Sep 22.

Scanogen Inc., Baltimore, MD, 21244, USA.

Detection of microbial nucleic acids in body fluids has become the preferred method for rapid diagnosis of many infectious diseases. However, culture-based diagnostics that are time-consuming remain the gold standard approach in certain cases, such as sepsis. New culture-free methods are urgently needed. Here, we describe Single MOLecule Tethering or SMOLT, an amplification-free and purification-free molecular assay that can detect microorganisms in body fluids with high sensitivity without the need of culturing. The signal of SMOLT is generated by the displacement of micron-size beads tethered by DNA probes that are between 1 and 7 microns long. The molecular extension of thousands of DNA probes is determined with sub-micron precision using a robust and rapid optical approach. We demonstrate that SMOLT can detect nucleic acids directly in blood, urine and sputum at sub-femtomolar concentrations, and microorganisms in blood at 1 CFU mL (colony forming unit per milliliter) threefold faster, with higher multiplexing capacity and with a more straight-forward protocol than amplified methodologies. SMOLT's clinical utility is further demonstrated by developing a multiplex assay for simultaneous detection of sepsis-causing Candida species directly in whole blood.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41467-020-18574-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7508858PMC
September 2020

Ketamine delays progression of oxidative and damaged cataract through regulating HMGB-1/NF-κB in lens epithelial cells.

Immunopharmacol Immunotoxicol 2018 Aug 15;40(4):303-308. Epub 2018 Aug 15.

a Department of Ophthalmology , The Second Affiliated Hospital of Zhejiang Chinese Medical University , Hangzhou , Zhejiang , China.

Objective: Lens epithelial cell (LEC) membrane damage is one of the pathogenesis of cataract. High mobility group box-1 (HMGB-1) and nuclear factor-κB (NF-κB) play vital roles in a variety of diseases, such as inflammation. Ketamine has numerous pharmacological effects that can inhibit inflammation. However, its role in cataract rats LECs has not yet been elucidated.

Materials And Methods: LECs were isolated from SD rats and cultured in vitro. The cells were randomly divided into three groups, including the control group, cataract model group induced by HO, and ketamine group treated by 10 mM ketamine under HO environment. LECs proliferation was assessed by MTT assay. LECs apoptosis was evaluated by Caspase-3 activity detection. NF-κB mRNA and protein expressions were tested by real-time PCR and Western blot. HMGB-1 expressions in cells and supernatant were detected by real-time PCR and ELISA. TNF-α and IL-1β secretions were detected by ELISA.

Results: In HO model group, the LECs proliferation was significantly inhibited, the caspase-3 activity significantly increased, HMGB-1 mRNA and secretion significantly enhanced, NF-κB mRNA and protein levels significantly elevated, compared to the Control group (p < .05). While the TNF-α and IL-1β secretions significantly up-regulated in HO model group compared to the Control group (p < .05). Ketamine significantly promoted the LECs proliferation, significantly reduced the caspase-3 activity, and significantly declined the HMGB expression compared to HO model group (p < .05). The NF-κB mRNA and protein levels were significantly decreased, TNF-α and IL-1β secretions were significantly decreased in the Ketamine group compared with the model group (p < .05).

Conclusions: Ketamine delays the progression of oxidative and damaged cataract by regulating HMGB-1/NF-κB expression, inhibiting TNF-α, IL-1β, and apoptosis, and promoting cell proliferation.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/08923973.2018.1478851DOI Listing
August 2018

Nontargeted Metabolomics for Phenolic and Polyhydroxy Compounds Profile of Pepper ( L.) Products Based on LC-MS/MS Analysis.

Molecules 2018 Aug 9;23(8). Epub 2018 Aug 9.

Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, China.

In the present study, nontargeted metabolomics was used to screen the phenolic and polyhydroxy compounds in pepper products. A total of 186 phenolic and polyhydroxy compounds, including anthocyanins, proanthocyanidins, catechin derivatives, flavanones, flavones, flavonols, isoflavones and 3---coumaroyl quinic acid -hexoside, quinic acid (polyhydroxy compounds), etc. For the selected 50 types of phenolic compound, except malvidin 3,5-diglucoside (malvin), l-epicatechin and 4'-hydroxy-5,7-dimethoxyflavanone, other compound contents were present in high contents in freeze-dried pepper berries, and pinocembrin was relatively abundant in two kinds of pepper products. The score plots of principal component analysis indicated that the pepper samples can be classified into four groups on the basis of the type pepper processing. This study provided a comprehensive profile of the phenolic and polyhydroxy compounds of different pepper products and partly clarified the factors responsible for different metabolite profiles in ongoing studies and the changes of phenolic compounds for the browning mechanism of black pepper.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/molecules23081985DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6222777PMC
August 2018

Moderate UV Exposure Enhances Learning and Memory by Promoting a Novel Glutamate Biosynthetic Pathway in the Brain.

Cell 2018 06 17;173(7):1716-1727.e17. Epub 2018 May 17.

Hefei National Laboratory for Physical Sciences at the Microscale, Neurodegenerative Disorder Research Center, School of Life Sciences, University of Science and Technology of China, 230026 Hefei, China; Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, 200031 Shanghai, China. Electronic address:

Sunlight exposure is known to affect mood, learning, and cognition. However, the molecular and cellular mechanisms remain elusive. Here, we show that moderate UV exposure elevated blood urocanic acid (UCA), which then crossed the blood-brain barrier. Single-cell mass spectrometry and isotopic labeling revealed a novel intra-neuronal metabolic pathway converting UCA to glutamate (GLU) after UV exposure. This UV-triggered GLU synthesis promoted its packaging into synaptic vesicles and its release at glutamatergic terminals in the motor cortex and hippocampus. Related behaviors, like rotarod learning and object recognition memory, were enhanced after UV exposure. All UV-induced metabolic, electrophysiological, and behavioral effects could be reproduced by the intravenous injection of UCA and diminished by the application of inhibitor or short hairpin RNA (shRNA) against urocanase, an enzyme critical for the conversion of UCA to GLU. These findings reveal a new GLU biosynthetic pathway, which could contribute to some of the sunlight-induced neurobehavioral changes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.cell.2018.04.014DOI Listing
June 2018

Contribution of Polyphenol Oxidation, Chlorophyll and Vitamin C Degradation to the Blackening of Piper nigrum L.

Molecules 2018 Feb 9;23(2). Epub 2018 Feb 9.

Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences (CATAS), Wanning 571533, China.

Black pepper ( L.) is the most widely used spice in the world. Blackening is considered to be beneficial and important in the processing of black pepper because it contributes to its color and flavor. The purpose of this paper is to investigate polyphenol oxidation as well as the chlorophyll and vitamin C (VC) degradation in the blackening of L. Black pepper was produced by four methods, and changes in polyphenols, chlorophyll and VC were studied by high performance liquid chromatography (HPLC) and ultraviolet-visible and visible (UV-Vis) spectrophotometry. The results show that polyphenol oxidase activity significantly decreased during the preparation of black pepper, and the concentrations of phenolic compounds, VC, and chlorophyll a and b also significantly decreased. Polyphenol oxidation and chlorophyll and VC degradation contribute to the blackening. A crude extract of phenolic compounds from black pepper was prepared by the system solvent method. The greater the polarity of the extraction solvent, the higher the extraction rates of the phenolic compounds and the total phenol content. Pepper phenolic compounds were analyzed by HPLC analysis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/molecules23020370DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6017850PMC
February 2018

Long non‑coding nuclear paraspeckle assembly transcript 1 acts as prognosis biomarker and increases cell growth and invasion in cervical cancer by sequestering microRNA‑101.

Mol Med Rep 2018 Feb 29;17(2):2771-2777. Epub 2017 Nov 29.

Department of Obstetrics and Gynecology, Xianyang Caihong Hospital, Xianyang, Shaanxi 712021, P.R. China.

Emerging studies have focused on the essential role of long non‑coding RNAs (lncRNAs) in cervical carcinogenesis. A recent study has indicated that nuclear paraspeckle assembly transcript 1 (NEAT1) is highly expressed in the human cervical tissue. However, whether NEAT1 is involved in cervical cancer remains to be elucidated. Reverse transcription‑quantitative polymerase chain reaction results demonstrated that the expression of NEAT1 was higher in cervical cancer cells/tissues compared with that in normal human keratinocytes/tissues. Patients with higher NEAT1 level had poorer clinical characteristics and a shorter survival time compared with those that exhibited lower NEAT1 expression levels. In vitro, flow cytometery analysis revealed that transfection with NEAT1 small interfering RNA retarded cervical cancer cell (Caski and HeLa) growth by decreasing the percentage of S phase in the cell cycle and inducing cell apoptosis. In addition, the colony formation assay, wound healing assay and matrigel invasion assay results indicated that downregulation of NEAT1 inhibited colony formation, cell migration and invasion. Further investigation using the luciferase reporter assay revealed that the expression of mircoRNA‑101 (miR‑101) target gene Fos was positively associated with NEAT1 expression due to NEAT1‑competitive molecular sequestering of miR‑101 via base pairing. Furthermore, reduction of miR‑101 expression by inhibitor transfection reversed the effect of NEAT1 siRNA on cervical cancer cells. To conclude, the present data indicated that NEAT1 promoted cervical cancer progression by targeting miR‑101.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3892/mmr.2017.8186DOI Listing
February 2018

Single-neuron identification of chemical constituents, physiological changes, and metabolism using mass spectrometry.

Proc Natl Acad Sci U S A 2017 03 21;114(10):2586-2591. Epub 2017 Feb 21.

CAS Key Laboratory of Urban Pollutant Conversion, School of Chemistry and Materials Science, University of Science and Technology of China, Hefei, Anhui 230026, China;

The use of single-cell assays has emerged as a cutting-edge technique during the past decade. Although single-cell mass spectrometry (MS) has recently achieved remarkable results, deep biological insights have not yet been obtained, probably because of various technical issues, including the unavoidable use of matrices, the inability to maintain cell viability, low throughput because of sample pretreatment, and the lack of recordings of cell physiological activities from the same cell. In this study, we describe a patch clamp/MS-based platform that enables the sensitive, rapid, and in situ chemical profiling of single living neurons. This approach integrates modified patch clamp technique and modified MS measurements to directly collect and detect nanoliter-scale samples from the cytoplasm of single neurons in mice brain slices. Abundant possible cytoplasmic constituents were detected in a single neuron at a relatively fast rate, and over 50 metabolites were identified in this study. The advantages of direct, rapid, and in situ sampling and analysis enabled us to measure the biological activities of the cytoplasmic constituents in a single neuron, including comparing neuron types by cytoplasmic chemical constituents; observing changes in constituent concentrations as the physiological conditions, such as age, vary; and identifying the metabolic pathways of small molecules.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1073/pnas.1615557114DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5347563PMC
March 2017

Calling Biomarkers in Milk Using a Protein Microarray on Your Smartphone.

PLoS One 2015 26;10(8):e0134360. Epub 2015 Aug 26.

RIKILT Wageningen UR, Wageningen, The Netherlands; Laboratory of Organic Chemistry, Wageningen University, Wageningen, The Netherlands.

Here we present the concept of a protein microarray-based fluorescence immunoassay for multiple biomarker detection in milk extracts by an ordinary smartphone. A multiplex immunoassay was designed on a microarray chip, having built-in positive and negative quality controls. After the immunoassay procedure, the 48 microspots were labelled with Quantum Dots (QD) depending on the protein biomarker levels in the sample. QD-fluorescence was subsequently detected by the smartphone camera under UV light excitation from LEDs embedded in a simple 3D-printed opto-mechanical smartphone attachment. The somewhat aberrant images obtained under such conditions, were corrected by newly developed Android-based software on the same smartphone, and protein biomarker profiles were calculated. The indirect detection of recombinant bovine somatotropin (rbST) in milk extracts based on altered biomarker profile of anti-rbST antibodies was selected as a real-life challenge. RbST-treated and untreated cows clearly showed reproducible treatment-dependent biomarker profiles in milk, in excellent agreement with results from a flow cytometer reference method. In a pilot experiment, anti-rbST antibody detection was multiplexed with the detection of another rbST-dependent biomarker, insulin-like growth factor 1 (IGF-1). Milk extract IGF-1 levels were found to be increased after rbST treatment and correlated with the results obtained from the reference method. These data clearly demonstrate the potential of the portable protein microarray concept towards simultaneous detection of multiple biomarkers. We envisage broad application of this 'protein microarray on a smartphone'-concept for on-site testing, e.g., in food safety, environment and health monitoring.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0134360PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4550345PMC
May 2016

Humidity independent mass spectrometry for gas phase chemical analysis via ambient proton transfer reaction.

Anal Chim Acta 2015 Mar 18;867:67-73. Epub 2015 Feb 18.

Department of Chemistry, University of Science and Technology of China, Hefei, Anhui 230026, PR China. Electronic address:

In this work, a humidity independent mass spectrometric method was developed for rapid analysis of gas phase chemicals. This method is based upon ambient proton transfer reaction between gas phase chemicals and charged water droplets, in a reaction chamber with nearly saturate humidity under atmospheric pressure. The humidity independent nature enables direct and rapid analysis of raw gas phase samples, avoiding time- and sample-consuming sample pretreatments in conventional mass spectrometry methods to control sample humidity. Acetone, benzene, toluene, ethylbenzene and meta-xylene were used to evaluate the analytical performance of present method. The limits of detection for benzene, toluene, ethylbenzene and meta-xylene are in the range of ∼0.1 to ∼0.3 ppbV; that of benzene is well below the present European Union permissible exposure limit for benzene vapor (5 μg m(-3), ∼1.44 ppbV), with linear ranges of approximately two orders of magnitude. The majority of the homemade device contains a stainless steel tube as reaction chamber and an ultrasonic humidifier as the source of charged water droplets, which makes this cheap device easy to assemble and facile to operate. In addition, potential application of this method was illustrated by the real time identification of raw gas phase chemicals released from plants at different physiological stages.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.aca.2015.02.043DOI Listing
March 2015

Opto-fluidics based microscopy and flow cytometry on a cell phone for blood analysis.

Methods Mol Biol 2015 ;1256:171-90

Electrical Engineering Department, University of California, Los Angeles, CA, 90095, USA,

Blood analysis is one of the most important clinical tests for medical diagnosis. Flow cytometry and optical microscopy are widely used techniques to perform blood analysis and therefore cost-effective translation of these technologies to resource limited settings is critical for various global health as well as telemedicine applications. In this chapter, we review our recent progress on the integration of imaging flow cytometry and fluorescent microscopy on a cell phone using compact, light-weight and cost-effective opto-fluidic attachments integrated onto the camera module of a smartphone. In our cell-phone based opto-fluidic imaging cytometry design, fluorescently labeled cells are delivered into the imaging area using a disposable micro-fluidic chip that is positioned above the existing camera unit of the cell phone. Battery powered light-emitting diodes (LEDs) are butt-coupled to the sides of this micro-fluidic chip without any lenses, which effectively acts as a multimode slab waveguide, where the excitation light is guided to excite the fluorescent targets within the micro-fluidic chip. Since the excitation light propagates perpendicular to the detection path, an inexpensive plastic absorption filter is able to reject most of the scattered light and create a decent dark-field background for fluorescent imaging. With this excitation geometry, the cell-phone camera can record fluorescent movies of the particles/cells as they are flowing through the microchannel. The digital frames of these fluorescent movies are then rapidly processed to quantify the count and the density of the labeled particles/cells within the solution under test. With a similar opto-fluidic design, we have recently demonstrated imaging and automated counting of stationary blood cells (e.g., labeled white blood cells or unlabeled red blood cells) loaded within a disposable cell counting chamber. We tested the performance of this cell-phone based imaging cytometry and blood analysis platform by measuring the density of red and white blood cells as well as hemoglobin concentration in human blood samples, which showed a good match to our measurement results obtained using a commercially available hematology analyzer. Such a cell-phone enabled opto-fluidics microscopy, flow cytometry, and blood analysis platform could be especially useful for various telemedicine applications in remote and resource-limited settings.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-4939-2172-0_12DOI Listing
September 2015

Cellphone-based detection platform for rbST biomarker analysis in milk extracts using a microsphere fluorescence immunoassay.

Anal Bioanal Chem 2014 Nov 28;406(27):6857-66. Epub 2014 Jun 28.

RIKILT Wageningen UR, P.O. Box 230, 6700 AE, Wageningen, The Netherlands.

Current contaminant and residue monitoring throughout the food chain is based on sampling, transport, administration, and analysis in specialized control laboratories. This is a highly inefficient and costly process since typically more than 99% of the samples are found to be compliant. On-site simplified prescreening may provide a scenario in which only samples that are suspect are transported and further processed. Such a prescreening can be performed using a small attachment on a cellphone. To this end, a cellphone-based imaging platform for a microsphere fluorescence immunoassay that detects the presence of anti-recombinant bovine somatotropin (rbST) antibodies in milk extracts was developed. RbST administration to cows increases their milk production, but is illegal in the EU and a public health concern in the USA. The cellphone monitors the presence of anti-rbST antibodies (rbST biomarker), which are endogenously produced upon administration of rbST and excreted in milk. The rbST biomarker present in milk extracts was captured by rbST covalently coupled to paramagnetic microspheres and labeled by quantum dot (QD)-coupled detection antibodies. The emitted fluorescence light from these captured QDs was then imaged using the cellphone camera. Additionally, a dark-field image was taken in which all microspheres present were visible. The fluorescence and dark-field microimages were analyzed using a custom-developed Android application running on the same cellphone. With this setup, the microsphere fluorescence immunoassay and cellphone-based detection were successfully applied to milk sample extracts from rbST-treated and untreated cows. An 80% true-positive rate and 95% true-negative rate were achieved using this setup. Next, the cellphone-based detection platform was benchmarked against a newly developed planar imaging array alternative and found to be equally performing versus the much more sophisticated alternative. Using cellphone-based on-site analysis in future residue monitoring can limit the number of samples for laboratory analysis already at an early stage. Therewith, the entire monitoring process can become much more efficient and economical.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00216-014-7984-4DOI Listing
November 2014

Screening of complicated matrixes with paper assisted ultrasonic spray ionization mass spectrometry.

J Am Soc Mass Spectrom 2014 Jun 25;25(6):935-42. Epub 2014 Mar 25.

Department of Chemistry, University of Science and Technology of China, Hefei, Anhui, 230026, People's Republic of China.

To analyze compounds in complicated matrixes using mass spectrometry, we describe a novel ambient ionization approach, termed paper assisted ultrasonic spray ionization (PAUSI). The ionization process is based on the ultrasonic vibration of the piezoelectric ceramic disk, on which the samples are placed. Porous materials are utilized to generate fine initial droplet, which could alleviate matrix effect during ionization process for complicated matrix. PAUSI was evaluated as an attractive tool to screen analytes from complicated matrixes, such as (1) bovine serum with NaCl 150 g/L, (2) viscous samples, and (3) biological fluid, without any sample preparation. Moreover, it provides great advantage in simplifying the mass spectrometry analysis process, and the ionization device is inexpensive and easy to operate.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s13361-014-0862-7DOI Listing
June 2014

A high-affinity CDR-grafted antibody against influenza A H5N1 viruses recognizes a conserved epitope of H5 hemagglutinin.

PLoS One 2014 18;9(2):e88777. Epub 2014 Feb 18.

Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China ; Shanghai Institute of Immunology, Shanghai Jiaotong University School of Medicine, Shanghai, China.

Highly pathogenic avian influenza (HPAI) H5N1 virus infection is still a potential threat to public health worldwide. While vaccines and antiviral drugs are currently under development, neutralizing antibodies could offer an alternative strategy to prevent and treat H5N1 virus infection. In the present study, we had developed a humanized antibody against H5N1 viruses from mouse-derived hybridoma in order to minimize its immunogenicity for potential clinical application. The humanized antibody hH5M9 was generated by transferring the mouse complementarity determining region (CDR) residues together with four key framework region (FR) residues onto the FR of the human antibody. This humanized antibody exhibited high affinity and specificity comparable to the parental mouse or chimeric counterpart with broad and strong neutralization activity against all H5N1 clades and subclades except for Egypt clades investigated. Furthermore, through epitope mapping we identified a linear epitope on the top region of hemagglutinin (HA) that was H5N1 specific and conserved. Our results for the first time reported a humanized antibody against H5N1 viruses by CDR grafting method. With the expected lower immunogenicity, this humanized antibody was expected to be more efficacious than murine or human-mouse chimeric antibodies for future application in humans.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0088777PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3928294PMC
December 2014

Separation and characterization of sucrose esters from Oriental tobacco leaves using accelerated solvent extraction followed by SPE coupled to HPLC with ion-trap MS detection.

J Sep Sci 2013 Aug 3;36(15):2486-95. Epub 2013 Jul 3.

Department of Chemistry, University of Science and Technology of China, Anhui, PR China.

Sucrose esters (SEs) were successfully extracted from Oriental tobacco leaves using a new methodology based on accelerated solvent extraction followed by hydrophilic-lipophilic balanced cartridge cleanup step. The SEs were detected by HPLC with ion-trap MS detection using an electrospray interface operated in the positive ion mode. This methodology combines the high efficiency of extraction provided by a pressurized fluid and the highly sensitive characterization offered by ion-trap MS. Under the optimized conditions, 14 SEs were first identified among a total of 23 SEs found in Oriental tobacco leaves. Under the same conditions, only four new SEs were extracted by using traditional ultrasound-assisted extraction and liquid-solid extraction methods. The present method might be potentially useful in high-efficiency extraction and sensitive characterization of SEs from complex matrices such as tobacco leaves.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/jssc.201300294DOI Listing
August 2013

Wide-field fluorescent microscopy and fluorescent imaging flow cytometry on a cell-phone.

J Vis Exp 2013 Apr 11(74). Epub 2013 Apr 11.

Electrical Engineering Department, University of California, Los Angeles, USA.

Fluorescent microscopy and flow cytometry are widely used tools in biomedical research and clinical diagnosis. However these devices are in general relatively bulky and costly, making them less effective in the resource limited settings. To potentially address these limitations, we have recently demonstrated the integration of wide-field fluorescent microscopy and imaging flow cytometry tools on cell-phones using compact, light-weight, and cost-effective opto-fluidic attachments. In our flow cytometry design, fluorescently labeled cells are flushed through a microfluidic channel that is positioned above the existing cell-phone camera unit. Battery powered light-emitting diodes (LEDs) are butt-coupled to the side of this microfluidic chip, which effectively acts as a multi-mode slab waveguide, where the excitation light is guided to uniformly excite the fluorescent targets. The cell-phone camera records a time lapse movie of the fluorescent cells flowing through the microfluidic channel, where the digital frames of this movie are processed to count the number of the labeled cells within the target solution of interest. Using a similar opto-fluidic design, we can also image these fluorescently labeled cells in static mode by e.g. sandwiching the fluorescent particles between two glass slides and capturing their fluorescent images using the cell-phone camera, which can achieve a spatial resolution of e.g. - 10 μm over a very large field-of-view of - 81 mm(2). This cell-phone based fluorescent imaging flow cytometry and microscopy platform might be useful especially in resource limited settings, for e.g. counting of CD4+ T cells toward monitoring of HIV+ patients or for detection of water-borne parasites in drinking water.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3791/50451DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3654336PMC
April 2013

Graphene oxide-dispersed pristine CNTs support for MnO2 nanorods as high performance supercapacitor electrodes.

ChemSusChem 2013 Mar 18;6(3):474-80. Epub 2013 Feb 18.

Department of Chemistry, University of Science and Technology of China, Hefei, Anhui, 230026, PR China.

A MnO2 -CNT-graphene oxide (MCGO) nanocomposite is fabricated using graphene oxide (GO) as a surfactant to directly disperse pristine carbon nanotubes (CNTs) for the subsequent deposition of MnO2 nanorods. The resulting MCGO nanocomposite is used as a supercapacitor electrode that shows ideal capacitive behavior (i.e., rectangular-shaped cyclic voltammograms), large specific capacitance (4.7 times higher than that of free MnO2 ) even at high mass loading (3.0 mg cm(-2) ), high energy density (30.4-14.2 Wh kg(-1) ), large power density (2.6-50.5 kW kg(-1) ), and still retains approximately 94 % of the initial specific capacitance after 1000 cycles. The advanced capacity, rate capability, and cycling stability may be attributed to the unique architecture, excellent ion wettability of GO with enriched oxygen-containing functional groups, high conductivity of CNTs, and their synergistic effects when combined with the other components. The results suggest that the MnO2 -CNT-GO hybrid nanocomposite architecture is very promising for next generation high-performance energy storage devices.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/cssc.201200709DOI Listing
March 2013

Cost-effective and rapid blood analysis on a cell-phone.

Lab Chip 2013 Apr;13(7):1282-8

Electrical Engineering Department, University of California, Los Angeles, CA 90095, USA.

We demonstrate a compact and cost-effective imaging cytometry platform installed on a cell-phone for the measurement of the density of red and white blood cells as well as hemoglobin concentration in human blood samples. Fluorescent and bright-field images of blood samples are captured using separate optical attachments to the cell-phone and are rapidly processed through a custom-developed smart application running on the phone for counting of blood cells and determining hemoglobin density. We evaluated the performance of this cell-phone based blood analysis platform using anonymous human blood samples and achieved comparable results to a standard bench-top hematology analyser. Test results can either be stored on the cell-phone memory or be transmitted to a central server, providing remote diagnosis opportunities even in field settings.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1039/c3lc41408fDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3594636PMC
April 2013

Optical imaging techniques for point-of-care diagnostics.

Lab Chip 2013 Jan 9;13(1):51-67. Epub 2012 Oct 9.

Electrical Engineering Department, University of California, Los Angeles, CA 90095, USA.

Improving access to effective and affordable healthcare has long been a global endeavor. In this quest, the development of cost-effective and easy-to-use medical testing equipment that enables rapid and accurate diagnosis is essential to reduce the time and costs associated with healthcare services. To this end, point-of-care (POC) diagnostics plays a crucial role in healthcare delivery in both developed and developing countries by bringing medical testing to patients, or to sites near patients. As the diagnosis of a wide range of diseases, including various types of cancers and many endemics, relies on optical techniques, numerous compact and cost-effective optical imaging platforms have been developed in recent years for use at the POC. Here, we review the state-of-the-art optical imaging techniques that can have a significant impact on global health by facilitating effective and affordable POC diagnostics.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1039/c2lc40864cDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3510351PMC
January 2013

Effect of FTY720 on some physiological indexes of Non-Obese Diabetic (NOD) Mice.

Int J Mol Sci 2012 18;13(5):6129-37. Epub 2012 May 18.

College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310035, China; E-Mail:

The studies were performed to investigate the physiological characteristics of non-obese diabetic (NOD) mice treated with FTY720. At the age of 12 weeks, each mouse was fed with FTY720 or physiological saline once a day for 10 weeks running, and their blood glucose, weight, anti-GAD antibody and organ indexes were determined. No mouse in group FTY720 (NOD mice treated with FTY720) showed diabetic symptoms. The average content of serum anti-GAD antibody in group FTY720 decreased 48.75% (P < 0.01). It was concluded that the spleen, kidney and liver of NOD mice treated with FTY720 shriveled significantly in the progression of diabetes (P < 0.01 or P < 0.05). The body weight of group FTY720 mice was slightly lower than that of the model control (MC) group and these two groups both had less body weight than the normal control (NC) group (P < 0.01). The result of tests of anti-GAD antibody suggested that FTY720 treatment could suppress the anti-GAD response.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/ijms13056129DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3382803PMC
October 2015

Quantum dot enabled detection of Escherichia coli using a cell-phone.

Analyst 2012 Jun 7;137(11):2541-4. Epub 2012 Mar 7.

Electrical Engineering Department, University of California, Los Angeles, 90095, USA.

We report a cell-phone based Escherichia coli (E. coli) detection platform for screening of liquid samples. In this compact and cost-effective design attached to a cell-phone, we utilize anti-E. coli O157:H7 antibody functionalized glass capillaries as solid substrates to perform a quantum dot based sandwich assay for specific detection of E. coli O157:H7 in liquid samples. Using battery-powered inexpensive light-emitting-diodes (LEDs) we excite/pump these labelled E. coli particles captured on the capillary surface, where the emission from the quantum dots is then imaged using the cell-phone camera unit through an additional lens that is inserted between the capillary and the cell-phone. By quantifying the fluorescent light emission from each capillary tube, the concentration of E. coli in the sample is determined. We experimentally confirmed the detection limit of this cell-phone based fluorescent imaging and sensing platform as ∼5 to 10 cfu mL(-1) in buffer solution. We also tested the specificity of this E. coli detection platform by spiking samples with different species (e.g., Salmonella) to confirm that non-specific binding/detection is negligible. We further demonstrated the proof-of-concept of our approach in a complex food matrix, e.g., fat-free milk, where a similar detection limit of ∼5 to 10 cfu mL(-1) was achieved despite challenges associated with the density of proteins that exist in milk. Our results reveal the promising potential of this cell-phone enabled field-portable and cost-effective E. coli detection platform for e.g., screening of water and food samples even in resource limited environments. The presented platform can also be applicable to other pathogens of interest through the use of different antibodies.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1039/c2an35071hDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3683133PMC
June 2012

Optofluidic on-chip tomography.

Annu Int Conf IEEE Eng Med Biol Soc 2011 ;2011:8463-6

Electrical Engineering Department, University of California, Los Angeles, CA 90095, USA.

The first demonstration of optofluidic tomography is presented. Using partially coherent illumination, holograms of objects are recorded at multiple viewing angles, as they flow through a microfluidic channel placed directly on the top of an opto-electronic sensor array. These lensfree holograms are then digitally processed to compute pixel super-resolved tomograms of micro-objects to achieve sectional opto-fluidic imaging on a chip.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1109/IEMBS.2011.6092088DOI Listing
May 2012

Wide-field fluorescent microscopy on a cell-phone.

Annu Int Conf IEEE Eng Med Biol Soc 2011 ;2011:6801-4

Electrical Engineering Department, University of California, Los Angeles, CA 90095, USA.

We demonstrate wide-field fluorescent imaging on a cell-phone, using compact and cost-effective optical components that are mechanically attached to the existing camera unit of the cell-phone. Battery powered light-emitting diodes (LEDs) are used to side-pump the sample of interest using butt-coupling. The pump light is guided within the sample cuvette to excite the specimen uniformly. The fluorescent emission from the sample is then imaged with an additional lens that is put in front of the existing lens of the cell-phone camera. Because the excitation occurs through guided waves that propagate perpendicular to the detection path, an inexpensive plastic color filter is sufficient to create the dark-field background needed for fluorescent imaging. The imaging performance of this light-weight platform (~28 grams) is characterized with red and green fluorescent microbeads, achieving an imaging field-of-view of ~81 mm(2) and a spatial resolution of ~10 μm, which is enhanced through digital processing of the captured cell-phone images using compressive sampling based sparse signal recovery. We demonstrate the performance of this cell-phone fluorescent microscope by imaging labeled white-blood cells separated from whole blood samples as well as water-borne pathogenic protozoan parasites such as Giardia Lamblia cysts.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1109/IEMBS.2011.6091677DOI Listing
August 2012

Optofluidic fluorescent imaging cytometry on a cell phone.

Anal Chem 2011 Sep 2;83(17):6641-7. Epub 2011 Aug 2.

Electrical Engineering Department, University of California, Los Angeles, Los Angeles, California 90095, USA.

Fluorescent microscopy and flow cytometry are widely used tools in biomedical sciences. Cost-effective translation of these technologies to remote and resource-limited environments could create new opportunities especially for telemedicine applications. Toward this direction, here we demonstrate the integration of imaging cytometry and fluorescent microscopy on a cell phone using a compact, lightweight, and cost-effective optofluidic attachment. In this cell-phone-based optofluidic imaging cytometry platform, fluorescently labeled particles or cells of interest are continuously delivered to our imaging volume through a disposable microfluidic channel that is positioned above the existing camera unit of the cell phone. The same microfluidic device also acts as a multilayered optofluidic waveguide and efficiently guides our excitation light, which is butt-coupled from the side facets of our microfluidic channel using inexpensive light-emitting diodes. Since the excitation of the sample volume occurs through guided waves that propagate perpendicular to the detection path, our cell-phone camera can record fluorescent movies of the specimens as they are flowing through the microchannel. The digital frames of these fluorescent movies are then rapidly processed to quantify the count and the density of the labeled particles/cells within the target solution of interest. We tested the performance of our cell-phone-based imaging cytometer by measuring the density of white blood cells in human blood samples, which provided a decent match to a commercially available hematology analyzer. We further characterized the imaging quality of the same platform to demonstrate a spatial resolution of ~2 μm. This cell-phone-enabled optofluidic imaging flow cytometer could especially be useful for rapid and sensitive imaging of bodily fluids for conducting various cell counts (e.g., toward monitoring of HIV+ patients) or rare cell analysis as well as for screening of water quality in remote and resource-poor settings.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1021/ac201587aDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3263930PMC
September 2011

Optofluidic Tomography on a Chip.

Appl Phys Lett 2011 Apr 20;98(16):161109. Epub 2011 Apr 20.

Using lensfree holography we demonstrate optofluidic tomography on a chip. A partially coherent light source is utilized to illuminate the objects flowing within a microfluidic channel placed directly on a digital sensor array. The light source is rotated to record lensfree holograms of the objects at different viewing directions. By capturing multiple frames at each illumination angle, pixel super-resolution techniques are utilized to reconstruct high-resolution transmission images at each angle. Tomograms of flowing objects are then computed through filtered back-projection of these reconstructed lensfree images, thereby enabling optical sectioning on-a-chip. The proof-of-concept is demonstrated by lensfree tomographic imaging of C. elegans.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1063/1.3548564DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3094459PMC
April 2011
-->