可见分光光度法测定人尿中痕量1-萘酚

目的：建立一种可见分光光度法单独测尿中痕量1-萘酚的新方法。方法：在碱性介质中,1-蔡酚（1-NAP）与氯霉素作用生成蓝色物质导致体系的吸光度增加,2-NAP不干扰测定。结果：该蓝色产物的最大吸收波长Amax=472．0nm,其吸光度与1-NAP摩尔浓度在7．64×10^-7mol／L-6．31×10^-4mol／L范围内线性关系良好,线性回归方程为△A=0．3146C＋0．0239,相关系数r=0．9973,检出限为2．29×10^-7mol／L,相对标准偏差RSD为3．25％-6．42％,加标回收率为95．3％-105．7％。结论：本方法灵敏、简单、快速、易于推广,用于人尿中1-NAP含量的单独测定结果满意。     

Extraction and determination of opium alkaloids in urine samples using dispersive liquid-liquid microextraction followed by high-performance liquid chromatography

A simple, rapid and sensitive method based on dispersive liquid-liquid microextraction (DLLME) combined with high-performance liquid chromatography-ultraviolet detection (HPLC-UV) was used to determine opium alkaloids in urine samples. Some effective parameters on extraction were studied and optimized. Under the optimum conditions, enrichment factors and recoveries for different opiates are in the range of 63.0-104.5 and 31.5-52.2%, respectively. The calibration graphs are linear in the range of 0.50-500 μg L(-1) and limit of detections (LODs) are in the range of 0.2-10 μg L(-1). The relative standard deviations (RSDs) for 200 μg L(-1) of morphine, codeine and thebaine, 5.0 μg L(-1) of papaverine and 10.0 μg L(-1) of noscapine in diluted urine sample are in the range of 2.8-6.1% (n=7). The relative recoveries of urine samples spiked with alkaloids are 84.3-106.0%. The obtained results show that DLLME combined with HPLC-UV is a fast and simple method for the determination of opium alkaloids in urine samples.     

Simultaneous determination of urinary tryptophan, tryptophan-related metabolites and creatinine by high performance liquid chromatography with ultraviolet and fluorimetric detection

A high performance liquid chromatography method with ultraviolet and fluorimetric detection has been developed for the simultaneous determination of urinary creatinine (Cr), tryptophan (Trp) and three Trp-related metabolites including kynurenine (Kyn), kynurenic acid (Kyna) and 5-hydroxyindole-3-acetic acid (5-HIAA). Samples were pretreated by centrifugation after a freeze-thaw cycle to remove protein and other precipitates. Separation was achieved by an Agilent HC-C18 (2) analytical column and a gradient elution program with a constant flow rate 1mL/min at an ambient temperature. Total run time was 30 min. Cr, Kyn and Kyna were measured by a variable wavelength detector at wavelengths 258 nm, 365 nm and 344 nm respectively. Trp and 5-HIAA were measured by a fluorescence detector with an excitation wavelength of 295 nm and an emission wavelength of 340 nm. This allowed the determination of Kyn/Cr, Kyna/Cr, Trp/Cr and 5-HIAA/Cr concentration ratios in a single run on the same urine sample. Good linear responses were found with correlation coefficient (r)>0.999 for all analytes within the concentration range of physiological level. The limit of detection of the developed method was: Cr, 0.0002 g/L; Kyn, 0.1 μmol/L; Kyna, 0.04 μmol/L; Trp, 0.02 μmol/L and 5-HIAA, 0.01 μmol/L. Recoveries from spiked human urine were: Cr, 93.0-106.4%; Kyn, 97.9-106.9%; Kyna, 98.5-105.6%; Trp, 96.7-105.2% and 5-HIAA, 96.1-99.7%. CVs of repeatability and intermediate precision of all analytes were less than 5%. This method has been applied to the analysis of urine samples from normal subjects.     

Simultaneous determination of a new anticancer agent (NB-506) and its active metabolite in human plasma and urine by high-performance liquid chromatography with ultraviolet detection

A high-performance liquid chromatographic method with ultraviolet detection has been developed to quantify NB-506 and its active metabolite in human plasma and urine. This method is based on solid-phase extraction, thereby allowing the simultaneous measurement of the drug and metabolite with the limit of quantification of 0.01 μg/ml in plasma and 0.1 μg/ml in urine. Standard curves for the compounds were linear in the concentration ranges investigated. The range for the drug in plasma was 0.01–2.5 μg/ml, and for the metabolite 0.01–1 μg/ml. In urine, the range for both compounds was 0.1–10 μg/ml. The method was validated and applied to the assay of plasma and urinary samples from phase I studies.     

Dalbavancin: Quantification in human plasma and urine by a new improved high performance liquid chromatography-tandem mass spectrometry method

Dalbavancin is a novel second-generation lipoglycopeptide antibiotic with activity against broad range of Gram-positive pathogens. In order to determine the pharmacokinetics (PK) of dalbavancin in pediatric patients, a new High Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS) bioanalytical method has been developed for quantification of dalbavancin in plasma and in urine. The plasma method was validated for dalbavancin in the linear range from 0.5 μg/mL to 500 μg/mL using 50 μL of K(2) EDTA plasma. For dalbavancin spiked in urine, non-specific binding (NSB) of the drug to polypropylene (PP) urine collection containers was observed. The loss amounted to about 10% per transfer. After successfully establishing the collection/sampling procedure for urine by addition of Triton X-100 to the collection vessels (with a purpose of preventing NSB), the method was validated for dalbavancin in the range from 0.05 μg/mL to 50 μg/mL, using 100 μL of urine. These methods were used to quantify dalbavancin in plasma and urine of hospitalized children in a pediatric dalbavancin PK study. Eighteen percent of the total number of plasma study samples was reassayed for incurred samples reproducibility (ISR) and all the reassayed dalbavancin concentrations were within the ± 20% limits. For urine, all the collected samples were reassayed for ISR and the original dalbavancin concentration was confirmed within the ± 20% limits for 17 (94%) samples; the one remaining urine sample had its reassayed concentration confirmed within ± 25% of the original result.     

Determination of muscarine in human urine by electrospray liquid chromatographic-mass spectrometric

A liquid chromatography-mass spectrometry based method for determination of muscarine in human urine was developed and validated. The method involved a solid phase extraction of muscarine from urine using Strata X-CW column. Separation of muscarine was achieved within 16.0 min on a reversed phase Gemini C18 analytical column (150 mm × 2.0mm i.d., 5 μm) with a mobile phase consisted of aqueous 8 mmol/L heptafluorobutyric acid and acetonitrile in a gradient mode. Mass spectrometric detection was performed at m/z 174 and m/z 216 for muscarine and acetylmuscarine (internal standard), respectively. The linearity was satisfactory with a coefficient of determination (R(2)) 0.9993 at concentration range from 0.3 ng/mL to 2.0 μg/mL, LOD and LOQ for muscarine was 0.09 ng/mL and 0.3 ng/mL, respectively. The found out recoveries of muscarine were 96% or 95% for concentration 0.3 ng/mL and 0.2 μg/mL or 2.0 μg/mL, respectively. The precision in the intra-assay-study varied from 0.48% to 1.39% and in the inter-assay-study from 2.39% to 5.49%. The accuracy ranged from -3.3% to -6%. The validation results demonstrated that the method fulfilled satisfactory requirements for precision and accuracy across the calibration curve. The applicability of the method has been demonstrated by analyzing clinical urine samples. The method offers the fast objective identification of intoxication by muscarine and can become a routine screening alternative to more difficult microscopic examination of spores in the gastric content in clinical practice.     

Simple and rapid quantification of gadolinium in urine and blood plasma samples by means of total reflection X-ray fluorescence (TXRF)

A simple and rapid method to determine gadolinium (Gd) concentrations in urine and blood plasma samples by means of total reflection X-ray fluorescence (TXRF) was developed. With a limit of detection (LOD) of 100 μg L(-1) in urine and 80 μg L(-1) in blood plasma and a limit of quantification (LOQ) of 330 μg L(-1) in urine and 270 μg L(-1) in blood plasma, it allows analyzing urine samples taken from magnetic resonance imaging (MRI) patients during a period of up to 20 hours after the administration of Gd-based MRI contrast agents by means of TXRF. By parallel determination of the urinary creatinine concentration, it was possible to monitor the excretion kinetics of Gd from the patient's body. The Gd concentration in blood plasma samples, taken immediately after an MRI examination, could be determined after rapid and easy sample preparation by centrifugation. All measurements were validated with inductively coupled plasma mass spectrometry (ICP-MS). TXRF is considered to be an attractive alternative for fast and simple Gd analysis in human body fluids during daily routine in clinical laboratories.     

Capillary electrophoretic determination of DNA damage markers: content of 8-hydroxy-2'-deoxyguanosine and 8-nitroguanine in urine

A sensitive and low-cost analytical method has been developed to determine 8-hydroxy-2'-deoxyguanosine (8-OHdG) and 8-nitroguanine (8-NO(2)Gua) based on capillary electrophoresis with amperometric detection (CE-AD) after solid phase extraction (SPE). Under optimized condition, these two markers were well separated from other components coexisting in urine, exhibiting a linear calibration over the concentration range of 0.1-50.0 μg/mL with the detection limits ranging from 0.02 to 0.06 μg/mL. The relative standard deviations (RSDs) were in the range of 0.1-2.1% for peak area, 0.1-1.5% for migration time, respectively. The average recovery and RSD were within the range of 100.0-108.0% and 0.1-1.7%, respectively. It was found that the urinary contents of 8-OHdG and 8-NO(2)Gua in cancer patients were significantly higher than those in healthy ones.     

Use of Hantzsch reaction for quantitation of milnacipran as a magic treatment for fibromyalgia syndrome in human plasma and urine

A new fluorimetric procedure is described for analysis of milnacipran in its bulk, tablet dosage forms, as well as in biological human samples such as plasma and urine. The suggested method relies on the construction of a derivative with strong fluorescence called dihydropyridine derivative. This derivative resulted from the interaction of the primary amino group in the studied drug and acetylacetone/formaldehyde in McIlvaine buffer (pH 5). The fluorescent dihydropyridine derivative was measured at 470 nm. Influences of experimental variables namely pH, reagent concentration and temperature were examined and optimized. The calibration curve showed linearity over the range of 0.15–1.25 μg ml^?1 of milnacipran with an R² value of 0.9998. The detection limit was 0.02 μg ml^?1 and the determination limit was 0.07 μg ml^?1. The developed procedure was successfully used in the assay of the studied drug in Avermilan® tablets with excellent selectivity. In addition, the reaction was applied to estimate the drug in spiked human plasma and urine with mean percentage recoveries of 100.04 ± 1.61 and 99.78 ± 0.81% for urine and plasma, respectively.     

Iodine Nutrition During Pregnancy in Toronto,Canada

ObjectiveTo evaluate the status of iodine nutrition among pregnant women presenting for routine antenatal care in Toronto, Canada, as determined by the median urine iodine concentration (UIC) of this population.MethodsA cross-sectional, observational study was conducted involving 142 pregnant women recruited from four low-risk antenatal outpatient clinics in Toronto, Canada. Subjects completed a questionnaire and provided a spot urine sample for the measurement of iodine concentration.ResultsMean maternal age was 33.8 ± 4.3 years. Mean gestational age was 29.3 ± 7.8 weeks. The median UIC was 221 μg/L (interquartile range, 142 to 397 μg/L). Six women (4.2%) had urine iodine levels <50 μg/L, and 36 women (25.4%) had levels between 50 and 150 μg/L.ConclusionThis cohort of primarily Caucasian, well-educated, and relatively affluent pregnant women in Toronto, Canada, are iodine sufficient, perhaps due to universal salt iodization and/or other dietary and lifestyle factors. (Endocr Pract. 2013;19:206-211)     

Quantification of acetaminophen (paracetamol) in human plasma and urine by stable isotope-dilution GC-MS and GC-MS/MS as pentafluorobenzyl ether derivative

We report on the quantitative determination of acetaminophen (paracetamol; NAPAP-d(0)) in human plasma and urine by GC-MS and GC-MS/MS in the electron-capture negative-ion chemical ionization (ECNICI) mode after derivatization with pentafluorobenzyl (PFB) bromide (PFB-Br). Commercially available tetradeuterated acetaminophen (NAPAP-d(4)) was used as the internal standard. NAPAP-d(0) and NAPAP-d(4) were extracted from 100-μL aliquots of plasma and urine with 300 μL ethyl acetate (EA) by vortexing (60s). After centrifugation the EA phase was collected, the solvent was removed under a stream of nitrogen gas, and the residue was reconstituted in acetonitrile (MeCN, 100 μL). PFB-Br (10 μL, 30 vol% in MeCN) and N,N-diisopropylethylamine (10 μL) were added and the mixture was incubated for 60 min at 30 °C. Then, solvents and reagents were removed under nitrogen and the residue was taken up with 1000 μL of toluene, from which 1-μL aliquots were injected in the splitless mode. GC-MS quantification was performed by selected-ion monitoring ions due to [M-PFB](-) and [M-PFB-H](-), m/z 150 and m/z 149 for NAPAP-d(0) and m/z 154 and m/z 153 for NAPAP-d(4), respectively. GC-MS/MS quantification was performed by selected-reaction monitoring the transition m/z 150 → m/z 107 and m/z 149 → m/z 134 for NAPAP-d(0) and m/z 154 → m/z 111 and m/z 153 → m/z 138 for NAPAP-d(4). The method was validated for human plasma (range, 0-130 μM NAPAP-d(0)) and urine (range, 0-1300 μM NAPAP-d(0)). Accuracy (recovery, %) ranged between 89 and 119%, and imprecision (RSD, %) was below 19% in these matrices and ranges. A close correlation (r>0.999) was found between the concentrations measured by GC-MS and GC-MS/MS. By this method, acetaminophen can be reliably quantified in small plasma and urine sample volumes (e.g., 10 μL). The analytical performance of the method makes it especially useful in pediatrics.     

玉竹多糖对酒精诱导的HepG2细胞损伤的保护作用及其机制*

目的: 探究玉竹多糖对酒精诱导HepG2细胞损伤的保护作用及潜在的分子机制。方法: 通过噻唑蓝(MTT)法筛选酒精处理HepG2细胞的合适浓度和玉竹多糖干预浓度后,将HepG2细胞按照不同干预浓度(200 μg/L、400 μg/L和600 μg/L)的玉竹多糖分组,并设未添加玉竹多糖的空白组,预处理1 h后,再用4%酒精处理24 h,每组设置3个复孔,检测细胞内丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)活性,检测细胞内活性氧(ROS)、丙二醛(MDA)、谷胱甘肽(GSH)、白介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)水平,检测细胞Kelch 样环氧氯丙烷相关蛋白-1(Keap1)、磷酸化核因子E2相关因子 2(p-Nrf2)、磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶-1(NQO1)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和半胱氨酸天冬氨酸蛋白酶3(cleaved-caspase-3)蛋白表达。结果: 与4%酒精处理后的HepG2细胞对比,各浓度玉竹多糖的干预能有效下调酒精诱导HepG2细胞内ALT和AST活性(P＜0.05);200 μg/L浓度玉竹多糖组的IL-1β和TNF-α水平明显下降(P＜ 0.05),而GSH水平明显上升(P＜0.01);400 μg/L和600 μg/L浓度玉竹多糖组的ROS、MDA、IL-1β和TNF-α水平明显下降(P＜0.05或P＜ 0.01),而GSH水平明显上升(P＜0.01)。玉竹多糖在有效上调酒精诱导HepG2细胞内p-Nrf2和NQO1蛋白表达的同时也下调Bax/ Bcl-2指数(P＜0.05),抑制Keap1和cleaved-caspase-3蛋白表达(P＜0.05)。结论: 玉竹多糖能通过调控Nrf2/ Keap1通路改善酒精诱导HepG2细胞氧化应激损伤,从而降低HepG2细胞炎症指数和细胞凋亡水平,其中400 μg/L和600 μg/L玉竹多糖的干预效果较好。     

Probably never human. Review of chimpanzee material culture: Implications for human evolution,by W.C. McGrew. Cambridge,U.K., Cambridge University Press, 1992, 277 pp, $79.95, cloth

Pregnanediol-3α-glucuronide (PdG) was measured in the urine of six Goeldi's monkeys during pregnancy and the postpartum period. A stress-free, non-invasive urine sampling technique permitted frequent collection of urine from members of the breeding group. A comparison of the periovulatory profiles of PdG and estrone conjugates revealed close agreement. The day of ovulation was defined as that immediately preceding a 2-4 day period with two consecutive urine samples for which the PdG content was in excess of 0.20 μg/mg Cr and 0.40 μg/mg Cr, respectively. In urine samples collected from parturition to the next ovulation, 70.9% of the PdG-values were below 0.20 μg/mg Cr, whereas 99.2% of the urinary PdG concentrations measured during pregnancy were greater than this “threshold concentration”. A conception cycle was therefore defined as one in which the concentration of urinary PdG remained above 0.20 μg/mg Cr in all urine samples collected between day 1 and day 20 after ovulation. Gestation length was 151.5 ± 1.6 days (mean ± SEM, n = 6; range 147-157 days). The postpartum ovulation occurred 22.6 ± 4.7 days (mean ± SEM, n = 9; range 11-53 days) following birth. With the exception of two non-conception postpartum cycles observed in one female, with inter-ovulatory intervals of 26 and 27 days, postpartum ovulation resulted in conception, giving a 77.8% conception rate for nine observed cycles. The simple and rapid radioimmunoassay used in this study requires 5 h from urine collection to the final result, hence permitting daily monitoring of a large sample of females. It thus has important potential for conservation breeding programs and for other scientific investigations carried out with this endangered primate species. © 1994 Wiley-Liss, Inc.     

Determination of the antihypertensive agent 1-(2-aminoethyl)-3-(2,6-Dichlorophenyl)thiourea in plasma and urine by high-performance liquid chromatography

A rapid, sensitive and specific normal-phase (adsorption) high-performance liquid chromatographic (HPLC) assay was developed for the determination of 1-(2-aminoethyl)-3-(2,6-dichlorophenyl)thiourea [I] in plasma and urine. The assay involves the extraction of the compound into methylene chloride from plasma or urine buffered to pH 10, and the HPLC analysis of the residue dissolved in methylene chloride—methanol—heptane (85:10:5). A 10-μm silica gel column was used with methylene chloride—methanol—heptane—ammonium hydroxide (85:10:5:0.1) as the eluting solvent. The effluent was monitored at 254 nm and quantitation was based on the peak height vs. concentration technique. The assay has a recovery of 64.5 ± 4.5% (S.D.) from plasma and 96.0 ± 6.3% (S.D.) from urine in the concentration range of 0.1–2 μg per ml and 2–40 μg per 0.1 ml of plasma and urine, respectively, with a limit of detection of 0.05–0.1 μg [I] per ml of plasma using a 1-ml specimen and 0.1 μg per ml urine using a 0.1-ml specimen, respectively. The assay was applied to the determination of plasma levels and urinary excretion of the compound [I] in dog following the oral administration of 28.8 mg of [I] · maleate per kg body weight.The HPLC assay was also used to determine the stability of [I] and for the measurement of a potential degradation product, clonidine [II] [2-(2,6-dichlorophenylamino)-2-imidazoline] in pooled human plasma stored at ?17°C, and pooled human urine stored at ?17°C and ?90°C, respectively.     

Determination of isometamidium residues in animal-derived foods by liquid chromatography-tandem mass spectrometry

In this paper, a new determination method for isometamidium residues in animal-derived foods was developed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Isometamidium residues in bovine tissues and milk were extracted with the mixed solution of acetonitrile and 0.25 mol/L of ammonium formate-methanol (v/v, 1:1), concentrated and degreased, and determined by LC-MS/MS with quantification by external standard method. The results showed that the peak area of chromatogram was linearly related to the concentration of isometamidium in the range of 1-100 μg/L, and the limits of detection (LOD) and quantification (LOQ) were 0.05 μg/kg and 5 μg/kg, respectively. The average recoveries of spiked samples were in the range of 73.8-93.9% with relative standard deviations ranged from 2.3% to 7.5%. This method is simple, accurate and suitable for the identification and quantification for isometamidium in animal-derived foods.     

Effect of nickel on red blood cell parameters and on serum vitamin B12, folate and homocysteine concentrations during pregnancy with and without anemia

BackgroundResearch to date suggests that nickel affects not only the metabolism of vitamin B12 but also folates and thus may affect hematopoiesis processes.ObjectiveThe aim of the study was to examine the relationship of nickel (Ni) status to red blood cell (RBC) parameters and serum vitamin B12, folate and homocysteine concentrations in the course of normal pregnancy and in pregnant women with anemia.MethodsThe study included fifty-three pregnant women recruited to the study from the Lower Silesia region of Poland, 17 % of whom developed anemia. Nickel concentration was determined in urine, whole blood and food samples by atomic absorption spectrometry. At the same time as the food and urine samples were taken, blood was also collected for the determination of RBC parameters and serum vitamin B12, homocysteine and folate concentrations.ResultsThe median reported Ni intake, and the urinary and whole blood nickel contents for the studied pregnant women for the first trimester were respectively – 162.46 μg/day, 3.98 μg/L and 3.32 μg/L; for the second trimester – 110.48 μg/day, 6.86 μg/L and 1.04 μg/L; and for the third trimester – 132.20 μg/day, 3.41 μg/L and 0.70 μg/L. With regard to Ni concentration in whole blood (p = 0.0204) and in urine (p = 0.0003), the differences in the values for individual trimesters were statistically significant. The whole blood Ni level was significantly higher (9.28 vs 3.62 μg/L, p = 0.0114), while the concentration of homosysteine was significantly lower (4.09 vs 5.04 μmol/L, p = 0.0165) in pregnant women with anemia compared to those without anemia. The whole blood Ni concentration was negatively correlated with almost all RBC parameters in non-anemic pregnant women.ConclusionsNi status changes with the development of normal pregnancy, and in the case of anemia, an increase in Ni concentration in whole blood is observed. The demonstrated correlations between the Ni status in pregnant women and RBC parameters as well as serum vitamin B12 and folate concentrations suggest that nickel is associated with the methionine–folate cycle, iron homeostasis and bacterial synthesis of vitamin B12 in humans.     

甘草提取物通过调控miR-212-5p/BCL2L2 基因抑制甲状腺肿瘤细胞的增殖、迁移和侵袭

目的探讨甘草提取物GL-1对甲状腺肿瘤细胞增殖、迁移和侵袭的影响及其分子机制。方法以10、20、30 μg/mL GL-1处理甲状腺肿瘤细胞CAL-62,或在CAL-62细胞中转染miR-212-5p mimics、anti-miR-212-5p、si-BCL2L2、pcDNA-BCL2L2。其中转染pcDNA-BCL2L2细胞并以30 μg/mL GL-1处理。噻唑蓝比色法 (MTT)检测CAL-62细胞增殖,Transwell小室法检测CAL-62细胞迁移和侵袭,实时定量PCR (qPCR)检测CAL-62细胞中miR-212-5p表达,Western blot检测相关蛋白Bcl-2样蛋白2 (BCL2L2)、细胞周期蛋白D1 (Cyclin D1)和基质金属蛋白酶-2 (MMP-2)表达。生物学信息预测miR-212-5p的下游靶基因,双荧光素酶基因报告实验进一步验证。数据采用单因素方差分析、Tukey’s事后检验和t检验。结果与对照组相比,10、20、30 μg/mL浓度GL-1降低CAL-62细胞24、48、72 h的细胞活性 (P < 0.05),并呈剂量、时间依赖性。与对照组相比,10、20、30 μg/mL浓度GL-1干预后,CAL-62细胞侵袭数[(143.56±14.22)个、(100.32±10.23)个、(68.23±6.49)个比(189.65±15.23)个]、迁移数[(198.56±14.35)个、(141.35±12.58)个、(89.56±8.95)个比 (295.36±17.56)个]和BCL2L2蛋白表达量 (0.76±0.08、0.51±0.06、0.24±0.02比1.00±0.12)均降低 (P 均< 0.05),而miR-212-5p水平 (1.61±0.11、1.99±0.13、2.28±0.15比1.00±0.07)升高(P < 0.05),并呈剂量依赖性。过表达miR-212-5p和沉默BCL2L2表达在24、48、72 h时CAL-62细胞活性、细胞迁移数、侵袭数和Cyclin D1、MMP-2蛋白表达量降低 (P < 0.05)。生物学信息预测和双荧光素酶基因报告实验证实BCL2L2是miR-212-5p的靶基因。过表达miR-212-5p抑制BCL2L2蛋白水平,沉默miR-212-5p促进BCL2L2蛋白表达 (P < 0.05)。过表达BCL2L2可逆转GL-1对CAL-62细胞增殖、迁移、侵袭及Cyclin D1、MMP-2蛋白表达的抑制作用。结论 GL-1通过miR-212-5p/BCL2L2抑制甲状腺肿瘤细胞的增殖、迁移和侵袭。     

高效氯氰菊酯对草鱼肝胰脏和肾脏溶菌酶活性的影响

研究了暴露在不同高效氯氰菊酯浓度下的草鱼Ctenopharyngodon idella肝胰脏和肾脏溶菌酶(LSZ)的活性变化.实验中高效氯氰菊酯浓度设5组,分别为0 μg/L、0.5μg/L、1.0 μg/L、3.0μg/L和5.0 μg/L,每组分别于1d、5d和12 d取样,测定肝胰脏和肾脏溶菌酶活性.结果显示,肝胰脏LSZ活性在暴露1d、5d、12 d时,各处理组均显著下降(P ＜0.05,P ＜0.01).肾脏LSZ活性在暴露1d、5d时,0.5μg/L、1.0 μg/L和3.0 μg/L组显著上升(P＜0.05,P＜0.01),5.0 μg/L组显著下降(P＜0.01);暴露12d时,0.5μg/L、1.0 μg/L组显著上升(P＜0.05),3.0μg/L和5.0 μg/L组显著下降(P＜0.05,P＜0.01).表明高效氯氰菊酯对草鱼肝胰脏和肾脏具有明显的毒害作用.     

A rapid capillary electrophoresis with electrochemiluminescence method for the assay of human urinary proline and hydroxyproline.

A novel simultaneous determination method for free and total proline (Pro) and hydroxyproline (Hyp) in human urine was developed, based on capillary electrophoresis (CE) with electrochemiluminescence (ECL) detection, using tris-(2,2'-bipyridyl) ruthenium(II). Experimental conditions, such as the Ru(bpy)(3)2+ concentration, detection potentials, buffer concentration and pH in CE or in the ECL cell, injection voltage and time were investigated in detail. Under optimized conditions, the linear range, detection limit and sample recoveries for the method were 0.01-2 mmol/L (correlation coefficient, 0.9970), 4 micromol/L and 96.4-101.2% in human urine, respectively. The results show that the method has potential applications in monitoring the level of Pro and Hyp in body fluids from patients with bone disease, tumours or chronic uraemia.     

Monitoring of mycotoxin biomarkers in the Czech Republic

AFM₁ was determined in 72 (72%) samples of human urine, range 19-6064 pg/g creatinine, mean 367 pg/g creatinine, median 158 pg/g creatinine and 90% percentile 755 pg/g creatinine in 1997. AFM₁ was determined in 46 (43.8%) samples of human urine, range 21-19219 pg/g creatinine, mean 414 pg/g creatinine, median 96 pg/g creatinine and 90% percentile 415 pg/g creatinine in 1998. OTA was determined in 2077 (94.2%) samples of human serum, range 0.1–13.7 μg/L, mean 0.28 μg/L, median 0.2 μg/L and 90% percentile 0.5 μg/L in 1994–2002. OTA was determined in 12 (40%) samples of human kidneys, range 0.1–0.2 μg/kg, mean 0.07 μg/kg, and median 0.05 μg/kg in 2001. Presented at the 26^th Mykotoxin-Workshop in Herrsching, Germany, May 17–19, 2004.