1.Preparation of an anti-cotinine monoclonal antibody and its application in immunological detection.
Yajing LEI ; Lifang ZHOU ; Anxing WANG ; Shuqing CHEN
Journal of Zhejiang University. Medical sciences 2020;49(6):765-771
OBJECTIVE:
To prepare monoclonal antibody against cotinine (COT) and to establish immunoassay for detecting COT in human urinary samples.
METHODS:
BALB/c mice were immunized with synthesized cotinine-bovine serum albumin (COT-BSA) to screen monoclonal antibody with technique of cell fusion. The monoclonal antibody was used for the indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and colloidal gold immunochromatographic strip assay for the detection of COT in human urine.
RESULTS:
The monoclonal antibody against COT was identified by ic-ELISA with a 50%inhibitive concentration (IC
CONCLUSIONS
The ic-ELISA and colloidal gold immunochromatographic strip assay using the prepared monoclonal antibody against COT have been proved to be reliable for the rapid detection of COT in human urines, which may be used for monitoring of environmental tobacco smoke.
Animals
;
Antibodies, Monoclonal
;
Cotinine/urine*
;
Enzyme-Linked Immunosorbent Assay
;
Gold Colloid
;
Humans
;
Mice
;
Mice, Inbred BALB C
;
Urinalysis/methods*
2.Clinical significance of microscopic hematuria and hydronephrosis in ureteral calculi patients visiting emergency department
Jaemin LEE ; Sang Chan JIN ; Woo Ik CHOI ; Wonho JUNG ; Ki Ho KIM ; Young Jin SEO ; Kyung Seop LEE
Journal of the Korean Society of Emergency Medicine 2019;30(1):77-82
OBJECTIVE: This study examined the clinical significance of microscopic hematuria and grade of hydronephrosis in ureteral calculi patients visiting the emergency department (ED). METHODS: The EDs of two medical centers were reviewed retrospectively from August 1, 2012 to July 31, 2017. The total ureteral calculi patients were 2,069 and 1,593 patients were analyzed after exclusion. The normal range of microscopic hematuria in both medical centers was 0–5/high power field (HPF) of red blood cell in urinalysis. Therefore, microscopic hematuria more than 5/HPF was defined as positive. The size of the stone, grade of hydronephrosis (normal, mild, moderate, and severe) and location of ureteral calculi in the non-contrast abdomen and pelvis computed tomography (CT) was measured by one person at each medical center using same method. The patients with or without microscopic hematuria and grade of hydronephrosis were then compared. RESULTS: The median size of the ureteral calculi patients without and with microscopic hematuria was 5.4 mm (range, 3.8–9.0 mm) and 4.0 mm (range, 3.0–5.8 mm) (P < 0.001). The patients grouped as normal to mild, and moderate to severe hydronephrosis, without microscopic hematuria was 191 (16.5%) and 91 (20.9%), respectively. On the other hand, microscopic hematuria was 966 (83.5%) and 345 (79.1%) (P=0.042). According to the location of ureteral calculi, with and without microscopic hematuria was 81.9% and 18.1% in the upper ureter, 82.9% and 17.1% in the mid ureter, and 82.6% and 17.4% in the lower ureter, respectively (P=0.935). CONCLUSION: The median size of the stone and grade of hydronephrosis were related to microscopic hematuria but the location of the ureteral calculi was not related. Therefore, in cases without microscopic hematuria in suspected ureteral calculi, clinicians should check the abdomen and pelvis CT for an accurate diagnosis and treatment of ureteral calculi.
Abdomen
;
Diagnosis
;
Emergencies
;
Emergency Service, Hospital
;
Erythrocytes
;
Hand
;
Hematuria
;
Humans
;
Hydronephrosis
;
Methods
;
Pelvis
;
Reference Values
;
Retrospective Studies
;
Ureter
;
Ureteral Calculi
;
Urinalysis
4.Normalisation of urinary biomarkers to creatinine for clinical practice and research--when and why.
Kai Wen Aaron TANG ; Qi Chun TOH ; Boon Wee TEO
Singapore medical journal 2015;56(1):7-10
Acute kidney injury (AKI) and chronic kidney disease (CKD) are major health problems. Urinary biomarkers have both diagnostic and prognostic utility in AKI and CKD. However, how biomarker excretion rates should be reported, especially whether they should be normalised to urinary creatinine concentration (uCr), is controversial. Some studies suggest that normalisation to uCr may be inappropriate at times, as urinary creatinine excretion rate may vary greatly, depending on the situation. Notably, recent studies suggest that while normalisation of values to UCr may be valid for the evaluation of CKD and prediction of AKI sequelae and occurrences, it could be inappropriate for the diagnosis of AKI, or in the presence of certain acute kidney disease states.
Acute Kidney Injury
;
urine
;
Biomarkers
;
urine
;
Creatinine
;
urine
;
Glomerular Filtration Rate
;
Humans
;
Kidney
;
physiopathology
;
Nephrology
;
methods
;
standards
;
Renal Insufficiency, Chronic
;
urine
;
Treatment Outcome
;
Urinalysis
;
standards
5.1H-NMR based metabonomic approach to evaluate detoxification effect of vinegar-processed Euphorbia kansui.
Yu-Mei LIU ; Rong-Rong HUI ; Cui-Cui HE ; Jin-Ao DUAN ; Jian-Xin LI
China Journal of Chinese Materia Medica 2014;39(2):322-326
Euphorbia kansui (EK) is a toxic herbal drug, and often used after vinegar-processing to reduce its toxicity. In present study, a 1H-NMR based metabonomic approach was used to evaluate the detoxification effect of vinegar-processed EK. The water extracts of EK and VEK were administered orally to male SD rats at doses of 9 g x kg(-1) x d(-1) for 1 week, respectively, and one more week observation was further conducted. The control group was orally given with saline. Histopathological studies of liver samples on the 8th and 15th day were conducted, and the metabolites of rat urine and liver were analysed by 1H-NMR. Histopathological studies of liver samples from EK and VEK treated rats showed no negative impacts. In metabonomic analyses of urines, changes of metabolites indicated liver damages, kidney lesions and imbalance of gut microbes in the second week. VEK-treated rats showed a quite lower toxicity compared with EK-treated ones. The present study revealed that the metabonomic approach might be helpful for the evaluation of toxicity of EK and detoxic effect of VEK.
Acetic Acid
;
chemistry
;
Animals
;
Chemistry, Pharmaceutical
;
Drugs, Chinese Herbal
;
chemistry
;
pharmacokinetics
;
toxicity
;
Euphorbia
;
chemistry
;
Inactivation, Metabolic
;
Liver
;
drug effects
;
metabolism
;
Magnetic Resonance Spectroscopy
;
Male
;
Metabolomics
;
methods
;
Rats
;
Rats, Sprague-Dawley
;
Urinalysis
6.Usefulness of Urine Cytology as a Routine Work-up in the Detection of Recurrence in Patients With Prior Non-Muscle-Invasive Bladder Cancer: Practicality and Cost-Effectiveness.
Bong Gi OK ; Yoon Seob JI ; Young Hwii KO ; Phil Hyun SONG
Korean Journal of Urology 2014;55(10):650-655
PURPOSE: To investigate the usefulness of urine cytology in the detection of tumor recurrence in terms of practicality and cost-effectiveness. MATERIALS AND METHODS: We retrospectively analyzed 393 patients who underwent transurethral resection of bladder tumor (TURBT) for non-muscle-invasive bladder cancer (NMIBC) from January 2010 to June 2013. All patients underwent cystoscopy, urine cytology, urinalysis, and computed tomography (CT) at 3 and 6 months after TURBT. In 62 cases, abnormal bladder lesions were identified on cystoscopy within 6 months. Suspicious lesions were confirmed pathologically by TURBT or biopsy. Patients were grouped by modalities: group I, urine cytology; group II, CT; group III, urinalysis; group IV, urine cytology plus CT; group V, urine cytology plus urinalysis; group VI, CT plus urinalysis; group VII, combination of all three modalities. Each group was compared by cost per cancer detected. RESULTS: Forty-nine patients were confirmed to have tumor recurrence and 13 patients were confirmed to have inflammation by pathology. The overall tumor recurrence rate was 12.5% (49/393) and recurrent cases were revealed as NMIBC. Sensitivity in group I (24.5%) was lower than in group II (55.1%, p=0.001) and group III (57.1%, p<0.001). However, in group VII (77.6%), the sensitivity was statistically similar to that of group VI (75.5%, p=0.872). Under the Korean insurance system, total cost per cancer detected for group VII was almost double that of group VI (p=0.041). CONCLUSIONS: Routine urine cytology may not be useful for follow-up of bladder cancer in terms of practicality and cost-effectiveness. Application of urine cytology needs to be adjusted according to each patient.
Aged
;
Aged, 80 and over
;
Cost-Benefit Analysis
;
Cystoscopy/economics
;
Cytodiagnosis/economics/methods
;
Female
;
Health Care Costs/*statistics & numerical data
;
Humans
;
Male
;
Middle Aged
;
Neoplasm Recurrence, Local/*diagnosis/economics/pathology
;
Neoplasm Staging
;
Republic of Korea
;
Retrospective Studies
;
Sensitivity and Specificity
;
Tomography, X-Ray Computed/economics
;
Urinalysis/economics/methods
;
Urinary Bladder Neoplasms/*diagnosis/economics/pathology/surgery
;
Urine/*cytology
7.Study on a new urine analysis core module based on semi-reflection mirror.
Longcong CHEN ; Gaiqin LIU ; Nan HU ; Ruiying ZHANG ; Qifeng JIANG ; Bin GAO ; Xingliang XIONG
Journal of Biomedical Engineering 2014;31(6):1288-1293
A new urine analysis core module based on high performance 32-bit microprocessor and high precision color sensor was presented. A novel optical structure and a specific circuit were applied to improve measurement precision and temperature was used to compensate for results in this core module. The information of urine test peice, such as all original data and color RGB value, reflectivity, semi-quantitative level, etc. can be output. The results showed that the measuring precision was about 95% or above with ideal stability and reliability using this presented core module, which can be conveniently applied in various urine analyzers, and can greatly decrease the cost of urine analyzers in development and production.
Color
;
Equipment Design
;
Microcomputers
;
Reproducibility of Results
;
Temperature
;
Urinalysis
;
instrumentation
;
methods
8.Paper-Based Analytical Device for Quantitative Urinalysis.
Seong Geun JEONG ; Jongmin KIM ; Jin Oh NAM ; Young Shin SONG ; Chang Soo LEE
International Neurourology Journal 2013;17(4):155-161
Paper-based analytical devices are fluidic chips fabricated with extremely inexpensive materials, namely paper, thereby allowing their use as a zero-cost analytical device in third-world countries that lack access to expensive diagnostic infrastructures. The aim of this review is to discuss: (1) microfluidic paper-based analytical devices (microPADs) for quantitative analysis, (2) fabrication of two- or three-dimensional microPADs, (3) analytical methods of microPADs, and (4) our opinions regarding the future applications of microPADs for quantitative urinalysis.
Developing Countries
;
Methods
;
Microfluidics
;
Urinalysis*
9.Determination of volatile nitrosamines in urine through gas chromatography-mass spectrometry.
Run-zheng GUO ; Yan-jian WAN ; Chun-jiang WU ; Yan ZHANG ; Qi-hui HUANG ; Hai-xia LI ; Xu WANG ; Zhong-hou CAO ; Yu-hua MAO ; Jie SHEN ; Wei XIA ; Yuan-yuan LI ; Shun-qing XU
Chinese Journal of Preventive Medicine 2013;47(3):270-273
OBJECTIVETo establish a detection method based on gas chromatography-mass spectrometry (GC-MS) for concentrations of volatile nitrosamine compounds in urine, and apply it to the test of real samples.
METHODSTarget compounds dichloromethane in urine samples was extracted with dichloromethane through liquid-liquid extraction, then the dichloromethane extract was filtrated, evaporated with nitrogen at 40°C to dryness, and the volume was set with 0.2 ml dichloromethane. Analysis of nine volatile nitroso-compounds were performed with GC-MS under selected ion monitoring mode, external reference method was used for quantification, and the detection limit, repeatability and sensitivity were evaluated. In addition, nine volatile nitroso-compounds of 92 urine samples in a town of Anhui province were measured.
RESULTSA good linear range of 2 - 200 ng/ml (with correlation coefficient 0.9985 - 0.9999) were obtained for the above mentioned nine kinds of analyte, and the lowest examination concentration was 0.05 - 0.50 ng/ml. The addition standard recoveries were 68%-102% with the RSD of 0.4% - 5.5% (n = 3). The detection limits were 0.001 - 0.013 ng/ml urine. The detection rate of N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N-nitrosodi-n-propylamine (NDPA), N-nitrosopyrrolidine (NPYR), N-nitrosomorpholine (NMOR), N-nitrosopiperidine (NPIP), N-nitrosodi-n-butylamine (NDBA) and N-nitrosodiphenylamine (NDPhA) were 71% (65), 74% (68), 65% (60), 80% (73), 92% (85), 78% (72), 76% (70), 87% (80), 98% (90), respectively, with the results (0.27 ± 0.12), (0.75 ± 0.29), (0.06 ± 0.02), (0.16 ± 0.07), (23.66 ± 5.18), (1.01 ± 0.35), (0.38 ± 0.11), (2.47 ± 0.52) and (15.13 ± 3.48) nmol/g creatinine.
CONCLUSIONSA gas chromatography-mass spectrometry detect method was developed for low level volatile nitrosamines in urine samples.
Gas Chromatography-Mass Spectrometry ; Humans ; Nitrosamines ; urine ; Urinalysis ; methods ; Volatile Organic Compounds ; urine
10.Simultaneous determination of melamine and cyanuric acid residues in water and urine by solid phase extraction-hydrophilic interaction liquid chromatography-electrospray tandem mass spectrometry method.
Hong-he LIU ; Li-sha MAO ; Li KANG ; Gui-hua LIU ; Chao-qiong PENG
Chinese Journal of Preventive Medicine 2013;47(7):641-647
OBJECTIVEWe aimed to establish a sensitive quantified method for the simultaneous determination of melamine and cyanuric acid residues in water and urine by hydrophilic interaction liquid chromatography coupled with electrospray tandem mass spectrometry (HILIC-ESI-MS/MS) with the pretreatment of hydrophilic functional silica gel and cation exchange resin mixed solid phase extraction column(MCT), and to investigate the melamine and cyanuric acid residues in 501 water and 216 urine from several province and city.
METHODSAbout 100 ml water (or 10 ml urine) was adjusted to pH 3.0 with concentrated hydrochloric acid, and then mixed with the internal standard solution((15)N3-melamine and (15)N3-(13)C3 -cyanuric acid) and 100 ml acetonitrile (10 ml for urine). The solution was cleaned with MCT solid-phase extraction column, and eluted once by 3 ml methanol and twice by 2.5 ml methanol (containing 5% ammonia water). The effluent was collected and dried by N2 flow at 40 °C, and then diluted to 2 mmol/L ammonium acetate containing 90% volume fraction acetonitrile. The completely dissolved solution was then filtered with 0.22 µm organic membrane; and the filtrate was detected by high performance liquid chromatography-tandem mass spectrometry and quantified with internal standards. The repeatability and sensitivity of the assay were evaluated. Then we detected the melamine and cyanuric acid residues in 501 water and 216 urine samples collected from several province and city.
RESULTSBy the quantification of internal standard (15)N3-melamine and (15)N3-(13)C3-cyanuric acid, the melamine and cyanuric acid were linear in the range of 2.0-1000.0 µg/L with correlation coefficient of 0.9998 and 0.9997. The detection limits of the method were separately 0.4 ng/L (melamine) and 0.3 ng/L (cyanuric acid) for water, and 4.0 ng/L (melamine) and 3.0 ng/L (cyanuric acid) for urine. The average recovery rate was around 95.3%-100.1% with the relative standard deviation (RSD) was <4.02%. Out of the 501 water samples, melamine was detected out in 19.9% (100/501) and cyanuric acid was detected out in 5.2% (26/501). The content was around 0.03-5.00 g/L. Melamine or cyanuric acid was detected out in 24.5% of the urine samples (53/216), with the content around 0.01-1.00 g/L.
CONCLUSIONThe established method of solid phase extraction-hydrophilic interaction liquid chromatography-electrospray tandem mass spectrometry can satisfy the requirement for detection of melamine and cyanuric acid residues in all sorts of water and urine. Meanwhile, the two substances widely existed in water and Chinese population.
Chromatography, Liquid ; methods ; Environmental Monitoring ; methods ; Humans ; Mass Spectrometry ; Solid Phase Extraction ; methods ; Triazines ; analysis ; urine ; Urinalysis ; methods

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