目的评估尿液中性粒细胞明胶酶相关脂质运载蛋白（urinary neutrophil gelatinase—associated lipocalin，uNGAL）对危重病患者急性肾损伤（AKI）的早期诊断作用。方法选择上海市宝山区中西医结合医院ICU162例危重病患者，收集患者一...目的评估尿液中性粒细胞明胶酶相关脂质运载蛋白（urinary neutrophil gelatinase—associated lipocalin，uNGAL）对危重病患者急性肾损伤（AKI）的早期诊断作用。方法选择上海市宝山区中西医结合医院ICU162例危重病患者，收集患者一般情况、急性生理与慢性健康状况评分Ⅱ（acute physiology and chronic health evaluation，APACHEⅡ）及序贯器官衰竭估计评分（sequential organ failure assessment，SOFA），以及检测0、24、48h血肌酐（SCr）和uNGAL。比较分析AKI与非AKI患者指标水平，应用ROC曲线评价uNGAL对危重病患者AKI诊断的敏感度和特异度。结果AKI患者与非AKI患者uNGAL、SCr水平比较差异有统计学意义（P〈0．05）。参考2012改善全球肾脏病预后组织指南，将AKI患者分为三期，随着分期的升高，患者uNGAL与SOFA评分均呈上升趋势。uNGAL的ROC曲线下面积为0．838，敏感度与特异度均优于SCr。结论uNGAL有助于早期诊断AKI，可能对AKI严重程度提供帮助。展开更多
Objective: To evaluate the value of ligase chainreaction(LCR) in the diagnosis of diplococcusgonorrhoeae in urine. Methods: LCR detection of the urine for Neisseriagonorrhoeae and bacteria culture of discharge was per...Objective: To evaluate the value of ligase chainreaction(LCR) in the diagnosis of diplococcusgonorrhoeae in urine. Methods: LCR detection of the urine for Neisseriagonorrhoeae and bacteria culture of discharge was per-formed simultaneously to 276 patients with urethritisor cervicitis seeking treatment in sex transmitted dis-eases (STDs) outpatient clinic. For specimens withdiscordant results, polymerase chain reaction wasconducted. The purpose was to detect the respectivesensitivity and specificity of bacteria culture and LCR. Results: 24 of 276(8.7%) patients had positive LCRresults and 21 of 276(7.6%) were positive for culture.5 specimens had discordant results from LCR andbacteria culture. The sensitivity and specificity of LCRin the diagnosis of gonorrhoeae were 92.3% and100% respectively. Conclusion: This study showed that LCR had ahigher sensitivity and specificity for the diagnosis ofgonorrhoeae from urine.展开更多
A high affinity polyclonal antibody-based enzyme linked immunosorbent assay （ELISA） was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution facto...A high affinity polyclonal antibody-based enzyme linked immunosorbent assay （ELISA） was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1：5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 μg/ml, and the detection limit was 0.02 μg/ml for the assay. The overall recoveries and the coefficients of variation （CVs） were in the range of 82%-127% and 3.5%-8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol （ranging from 0.2 to 10 μg/ml） were detected by the ELISA and liquid chromatography （LC） method, and good correlations were obtained between the two methods （R^2=0.9643）. We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an altemative for the conventional LC method for zeranol in bovine urine.展开更多
To evaluate blood concentration of melatonin and urinary excretion of its metabolite, 6-sulfatoxymelatonin （6-OHMS）, in functional dyspepsia （FD）. METHODS： Ninety individuals were enrolled in the study： 30 in ea...To evaluate blood concentration of melatonin and urinary excretion of its metabolite, 6-sulfatoxymelatonin （6-OHMS）, in functional dyspepsia （FD）. METHODS： Ninety individuals were enrolled in the study： 30 in each study group： patients with postprandial distress syndrome （PDS）, epigastric pain syndrome （EPS）, and controls. Blood samples were drawn at 02：00 and 09：00 h and 24-h urine collection was performed. Serum melatonin and urinary 6-OHMS concentrations were measured by enzyme-linked immunosorbent assay. RESULTS： Serum melatonin concentration at night and in the morning was significantly （P 〈 0.001） higher inPDS patients [at 02：00 h-93.3 pg/mL, quartile range （QR）： 79.8-116.2; at 09.00 h-14.3 pg/mL, QR： 7.06-19.0] than in EPS （57.2 pg/mL, QR： 42.6-73.1, 8.1 pg/mL, QR： 4.2-9.3） and control patients （57.7 pg/mL, QR： 51.2-62.5; 8.1 pg/mL, QR： 5.4-10.3）. A similar relationship was observed for urinary 6-OHMS excretion. Patients with severe PDS symptoms had a higher melatonin concentration than these with moderate syndromes, whereas patients with severe EPS had a lower urinary 6-OHMS excretion than patients with moderate symptoms.CONCLUSION： Evaluation of melatonin serum concentrations and 24-h urinary 6-OHMS excretion are useful methods for differential diagnosis of various clinical forms of FD.展开更多
Objective To investigate the effect of glycosylation at Asn302 of pro-urokinase （pro-UK） on the stability in culture supematant. Methods Nonglycosylated pro-UK was constructed by site-directed mutagenesis of Asn302...Objective To investigate the effect of glycosylation at Asn302 of pro-urokinase （pro-UK） on the stability in culture supematant. Methods Nonglycosylated pro-UK was constructed by site-directed mutagenesis of Asn302 to Ala302. The pro-UK mutant and native pro-UK were transfected into dhfr-CHO cells, and serum-free culture supematant was harvested and incubated at 4℃ and 37℃, respectively. The pro-UK activity in culture supematant was measured by the optical density （OD） increase with time （ 12 hours） at 405 nm. Without thermolysin activation, the percentage of single chain pro-UK was measured. Rcsults After 48 hours of incubation at 4℃, the activities of pro-UK mutant and native pro-UK decreased 3.7% and 2.9% respectively, and at 37℃ decreased 37.9% and 23.5%, respectively. The total activity of native pro-UK was significantly higher than that of nonglycosylated mutant at 37℃. The single-chain percentage of native pro-UK was higher than that of nonglycosylated mutant at both 4℃ and 37℃. Conclusion Higher temperature increases the proteolysis of pro-UK. The glycosylation site on Asn302 is beneficial to pro-UK stability in culture supematant.展开更多
文摘目的评估尿液中性粒细胞明胶酶相关脂质运载蛋白（urinary neutrophil gelatinase—associated lipocalin，uNGAL）对危重病患者急性肾损伤（AKI）的早期诊断作用。方法选择上海市宝山区中西医结合医院ICU162例危重病患者，收集患者一般情况、急性生理与慢性健康状况评分Ⅱ（acute physiology and chronic health evaluation，APACHEⅡ）及序贯器官衰竭估计评分（sequential organ failure assessment，SOFA），以及检测0、24、48h血肌酐（SCr）和uNGAL。比较分析AKI与非AKI患者指标水平，应用ROC曲线评价uNGAL对危重病患者AKI诊断的敏感度和特异度。结果AKI患者与非AKI患者uNGAL、SCr水平比较差异有统计学意义（P〈0．05）。参考2012改善全球肾脏病预后组织指南，将AKI患者分为三期，随着分期的升高，患者uNGAL与SOFA评分均呈上升趋势。uNGAL的ROC曲线下面积为0．838，敏感度与特异度均优于SCr。结论uNGAL有助于早期诊断AKI，可能对AKI严重程度提供帮助。
文摘Objective: To evaluate the value of ligase chainreaction(LCR) in the diagnosis of diplococcusgonorrhoeae in urine. Methods: LCR detection of the urine for Neisseriagonorrhoeae and bacteria culture of discharge was per-formed simultaneously to 276 patients with urethritisor cervicitis seeking treatment in sex transmitted dis-eases (STDs) outpatient clinic. For specimens withdiscordant results, polymerase chain reaction wasconducted. The purpose was to detect the respectivesensitivity and specificity of bacteria culture and LCR. Results: 24 of 276(8.7%) patients had positive LCRresults and 21 of 276(7.6%) were positive for culture.5 specimens had discordant results from LCR andbacteria culture. The sensitivity and specificity of LCRin the diagnosis of gonorrhoeae were 92.3% and100% respectively. Conclusion: This study showed that LCR had ahigher sensitivity and specificity for the diagnosis ofgonorrhoeae from urine.
基金Project supported by the National Natural Science Foundation of China (No. 30471155) the Agriculture Key Technologies R & D Program of Shanghai (No. (2003) 9-4), China
文摘A high affinity polyclonal antibody-based enzyme linked immunosorbent assay （ELISA） was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1：5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 μg/ml, and the detection limit was 0.02 μg/ml for the assay. The overall recoveries and the coefficients of variation （CVs） were in the range of 82%-127% and 3.5%-8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol （ranging from 0.2 to 10 μg/ml） were detected by the ELISA and liquid chromatography （LC） method, and good correlations were obtained between the two methods （R^2=0.9643）. We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an altemative for the conventional LC method for zeranol in bovine urine.
基金Supported by The Ministry of Science and Higher Education of Poland,project NN 402481937
文摘To evaluate blood concentration of melatonin and urinary excretion of its metabolite, 6-sulfatoxymelatonin （6-OHMS）, in functional dyspepsia （FD）. METHODS： Ninety individuals were enrolled in the study： 30 in each study group： patients with postprandial distress syndrome （PDS）, epigastric pain syndrome （EPS）, and controls. Blood samples were drawn at 02：00 and 09：00 h and 24-h urine collection was performed. Serum melatonin and urinary 6-OHMS concentrations were measured by enzyme-linked immunosorbent assay. RESULTS： Serum melatonin concentration at night and in the morning was significantly （P 〈 0.001） higher inPDS patients [at 02：00 h-93.3 pg/mL, quartile range （QR）： 79.8-116.2; at 09.00 h-14.3 pg/mL, QR： 7.06-19.0] than in EPS （57.2 pg/mL, QR： 42.6-73.1, 8.1 pg/mL, QR： 4.2-9.3） and control patients （57.7 pg/mL, QR： 51.2-62.5; 8.1 pg/mL, QR： 5.4-10.3）. A similar relationship was observed for urinary 6-OHMS excretion. Patients with severe PDS symptoms had a higher melatonin concentration than these with moderate syndromes, whereas patients with severe EPS had a lower urinary 6-OHMS excretion than patients with moderate symptoms.CONCLUSION： Evaluation of melatonin serum concentrations and 24-h urinary 6-OHMS excretion are useful methods for differential diagnosis of various clinical forms of FD.
文摘Objective To investigate the effect of glycosylation at Asn302 of pro-urokinase （pro-UK） on the stability in culture supematant. Methods Nonglycosylated pro-UK was constructed by site-directed mutagenesis of Asn302 to Ala302. The pro-UK mutant and native pro-UK were transfected into dhfr-CHO cells, and serum-free culture supematant was harvested and incubated at 4℃ and 37℃, respectively. The pro-UK activity in culture supematant was measured by the optical density （OD） increase with time （ 12 hours） at 405 nm. Without thermolysin activation, the percentage of single chain pro-UK was measured. Rcsults After 48 hours of incubation at 4℃, the activities of pro-UK mutant and native pro-UK decreased 3.7% and 2.9% respectively, and at 37℃ decreased 37.9% and 23.5%, respectively. The total activity of native pro-UK was significantly higher than that of nonglycosylated mutant at 37℃. The single-chain percentage of native pro-UK was higher than that of nonglycosylated mutant at both 4℃ and 37℃. Conclusion Higher temperature increases the proteolysis of pro-UK. The glycosylation site on Asn302 is beneficial to pro-UK stability in culture supematant.
