4类尿液生物标记物联合血清指标在急性肾损伤中的应用价值分析

目的分析4类尿液生物标记物(尿N-乙酰-β-D-氨基葡萄糖苷酶、肝脏型尿脂肪酸结合蛋白、尿富半胱氨酸蛋白61、尿胎球蛋白A)联合血清指标(血肌酐、血尿素氮)在急性肾损伤中的应用价值。方法选取我院2014年3月-2016年3月收治的98例急性肾损伤患者作为观察组,另选同期105例健康体检者为对照组,在征得两组受试者知情同意下对其4类尿液生物标记物和血清指标进行检测与比较。结果观察组4类尿液生物标记物、血清指标均高于对照组,差异有统计学意义(P0.05);观察组内4类尿液生物标记物、血清指标相比较,3期2期1期且差异有统计学意义(P0.05);4类尿液生物标记物联合血清指标诊断急性肾损伤ROC曲线下面积0.974、95%CI:0.945-1.000,较4类尿液生物标记物和血清指标单用更具敏感性。结论 4类尿液生物标记物联合血清指标可提高急性肾损伤的诊断效果,利于病情的早期监测,值得在临床工作中推广使用。     

实验性胃溃疡大鼠尿液的代谢组学研究

目的探讨大鼠实验性胃溃疡发生与自愈过程中尿液代谢表型的变化,揭示其生物学特征。方法 40只Wistar雄性大鼠完全随机分为5组：对照组与模型1天、3天、7天1、0天组,每组8只;采用乙酸性胃溃疡模型,运用气相色谱-质谱（GC-MS）技术获得各组大鼠尿液的代谢物谱,应用主成分分析（PCA）及偏最小二乘法判别分析（PLS-DA）进行模式识别,寻找表征不同代谢状态的生物标记物,根据这些物质水平的变化阐明相关机制。结果各组尿液代谢物谱明显不同;PCA得分图显示各组区分良好,不同时间模型组沿一定的轨迹向对照组靠拢;PLS-DA获得了15种生物标记物,包括一些有机酸、脂肪酸与氨基酸;不同组的标记物水平呈动态变化。结论大鼠胃溃疡发生与自愈过程中出现了能量与物质代谢异常,以及神经调节紊乱。     

代谢组学的研究进展及其在肾病综合征中的应用前景

系列“组学”技术的发展推动了基础研究水平的提高和深入。代谢组学作为一门新兴学科,着重研究的是生物整体、器官或组织的内源性代谢物质的代谢途径及其所受内在或外在因素影响下代谢整体的变化轨迹来反映某种病理生物过程。目前,代谢组学已广泛应用于生物学和医学相关的很多领域,在肝脏、心脏、肿瘤、肾脏等疾病中也有较深入的研究。     

基于质谱技术的外泌体蛋白质组学在肿瘤诊断中的研究进展

外泌体是一类参与多种生理病理过程的细胞间信息交流载体,其脂质双分子层结构能有效维护所携带的生物信息分子的稳定性。因此,外泌体相对于血清、血浆等更加适合于组学研究,如蛋白质组学、转录组学及代谢组学等。随着质谱技术的不断发展,蛋白质作为诸多生物功能的直接执行者受到越来越多的关注,尤其是外泌体内的蛋白质及其生物功能。外泌体用于蛋白质组学的研究有助于阐明肿瘤的发生和发展机制,进而寻找特异性的肿瘤标志物和可用于精准治疗的靶点。     

诊断蛋白质组学及其在肺癌生物标记物中的研究进展

寻找肿瘤特异性标记物．以期对恶性肿瘤进行早期诊断是蛋白质组学一项重要的任务。新的疾病状态生物标志物的发现。有助于加深对疾病发生发展过程的理解。找到更多的药物靶标。这就是“诊断蛋白质组学”的研究内容。蛋白质组研究技术可以简单分为两大类。即以双向凝胶电泳技术为代表的蛋白质组分离技术和以质谱技术为核心的蛋白质组鉴定技术。本文就目前常用的诊断蛋白质组学研究技术以及其在肺癌生物标记物研究中的应用作一综述。[第一段]     

蛋白质组学在某些癌症早期检测中的研究进展

寻找理想的表型生物标记物 ,阐明疾病发生发展过程中起关键作用的蛋白质分子 ,以用于癌症的早期检测诊断及预后评估一直是癌症研究领域的热点和难点。由于癌症的演变是多阶段、多因素、多基因参与的复杂病理过程 ,早期又缺乏用于筛查分离关键分子的有效技术手段 ,致使癌症检测标记物的发展一直比较迟缓。近几年蛋白质组学的起步和发展为癌症早期检测研究注入活力。蛋白质组 (Proteome)指细胞或组织基因组编码的全部蛋白质[1] 。蛋白质组学是通过对细胞或组织整体水平蛋白质属性的研究 (表达水平、转录后修饰、翻译后加工、相互作用等 ) ,在…     

代谢组学在强直性脊柱炎研究中的应用

代谢组学是继基因组学、转录组学和蛋白质组学之后兴起的系统生物学的一个新的分支，它是通过考察生物体系（细胞、组织或生物体）受刺激或扰动后（如将某个特定的基因变异或环境变化后）代谢产物图谱及其动态变化，研究生物体系的代谢网络的一种技术。与其他三种组学研究的DNA、RNA和蛋白质等生物大分子不同，代谢组学是对生物体系中的小分子化合物进行定性定量研究。近年来，代谢组学相关技术发展迅速，     

妊娠期糖尿患者的尿液代谢组学研究

目的应用代谢组学研究方法,对妊娠期糖尿病(GDM)患者的尿液代谢物进行差异性筛选,鉴定妊娠期糖尿病的生物标志物。方法采用超高效液相色谱-质谱联用技术对15例妊娠期糖尿患者与15例正常孕妇的尿液进行研究,分析GDM患者的尿液代谢的改变。结果多元统计分析的结果发现,妊娠期糖尿患者的尿液代谢与正常孕妇差异明显,尿液中发现并鉴定出17个差异代谢物及相应的代谢通路,它们分别涉及甘氨酸、丝氨酸和苏氨酸代谢、精氨酸和脯氨酸代谢,组氨酸代谢,色氨酸代谢,糖酵解和糖质新生等多个代谢通路。结论这些标志物的发现为深入理解GDM患者发病机制提供理论依据,液相色谱-质谱联用结合多变量分析的尿液代谢组学研究技术在产科疾病的早期诊断以及疾病机制研究中展现了巨大的潜在价值。     

蛋白质组学在消化系统疾病中的应用进展

蛋白质组学已经在消化系统疾病中有了较为广泛的开展和研究,主要集中于早期新标记物的寻找,并在疾病的病因、病理生理及预后、新药靶方面做了许多相关的研究。现对蛋白质组学在消化系统疾病中的应用进展综述如下。1蛋白质组学的研究内容和研究方法蛋白质组的概念首先是由Wasinger等提出,指基因组所表达的全部蛋白质。蛋白质组学是指应用各种技术手段来研究蛋白质组的一门新型学科,其目的是从整体的角度分析细胞内动态变化的蛋白质组成成分、表达水平与修饰状态,了解蛋白质之间的相互作用和联系,揭示蛋白质功能和细胞生命活动规律。蛋白质组…     

定量蛋白质组学技术筛选前列腺癌患者尿液中差异表达蛋白价值

目的探讨同位素标记相对和绝对定量蛋白质组学技术在含有前列腺液的尿液样本中筛选前列腺癌差异表达蛋白质的价值。方法良性前列腺增生患者10例(良性增生组)、高级别前列腺上皮内瘤变患者10例(上皮内瘤变组)、前列腺癌患者10例(前列腺癌组),取3组患者前列腺按摩后的前段尿液标本,并将3组患者尿液样本进行等体积混合,应用同位素标记相对和绝对定量技术联合液相串联质谱分析技术进行蛋白质鉴定和相对定量;检索Gene Ontology Database(GO数据库),对筛查得到的蛋白质所参与生物学过程及分子功能进行生物信息学分析。结果含有3组前列腺液的尿液混合样本中共鉴定到蛋白质2 370种;相对于良性增生组,前列腺癌组患者表达差异倍数在2倍以上的蛋白质共52个,其中表达上调蛋白20个,表达下调蛋白32个;相对于良性增生组,上皮内瘤变组患者表达差异蛋白质共60个,其中表达上调蛋白22个,表达下调蛋白38个;鉴定蛋白质主要参与细胞过程、单器官过程和生物功能调节等生物功能,及结合功能、催化活性、酶调节活性等分子功能。结论定量蛋白质组学技术有助于鉴定前列腺癌相关的尿液差异表达蛋白质。     

尿红细胞3种检测方法的结果差异及影响因素分析

目的探讨3种方法（镜检与尿干化学法及UF-50全自动尿沉渣分析仪）测定尿红细胞结果的吻合性及其影响因素。方法采用镜检法和尿干化学法及UF-50全自动尿沉渣分析仪对同一尿液标本进行尿液红细胞测定。结果3种方法检出红细胞阳性率分别是：UF-50全自动尿沉渣分析仪法为29．4％，尿干化学法为23．2％，镜检法为21．6％。UF-50全自动尿沉渣分析仪法明显高于尿干化学法和镜检法。3种方法阳性率不尽相同，差异有统计学意义。结论3种方法测定尿红细胞结果有一定的差异，联合检测和结合临床资料综合分析才能更好地确定尿液中红细胞的有无及其来源（肾性与非肾性）。     

三种方法联合检测尿液有形成分的比较和评价

探讨Sysmex UF-50尿流式细胞仪、干化学分析仪及尿沉渣镜检三种方法检测尿液中红细胞（RBC）、白细胞（WBC）和管型（CAST）的应用及影响因素。方法对640例病人的尿液分别用UF-50尿沉渣分析仪、干化学分析仪及尿沉渣镜检三种方法进行检测,并对结果进行比较。结果 UF-50检测RBC、WBC和CAST的阳性率分别为：29.38%、24.38%和8.75%。干化学法检测RBC和WBC的阳性率是33.90%和15.1%;尿沉渣镜检法检测RBC、WBC和CAST的阳性率分别为：19.22%、20.94%和2.03%。结论 UF-50尿流式细胞仪和干化学分析仪不能取代尿沉渣镜检,建议三种方法联合应用,减少检验误差,提高尿液分析质量。     

UF-1000i尿液沉渣分仪析检测尿红细胞的影响因素

目的：探讨UF-1000i尿液沉渣分仪析检测尿红细胞（Red Blood Cell, RBC）影响因素。方法对住院病人400例晨尿标本同时进行UF-1000i尿液沉渣分仪析检测和尿沉渣显微镜检查,分析两者结果的差异。结果400例受检标本中,UF-1000i检测RBC阳性144例,阳性率36.0%,显微镜检测的RBC阳性131例,阳性率32.8%,尿沉渣分析仪RBC检测结果与镜检结果比较存在假阳性。两种方法检测RBC的差异有统计学意义（χ2=6.85714,P＆lt;0.01）。结论当尿液中细菌集簇分布、酵母菌污染、存在草酸钙结晶等可引起UF-1000i尿液沉渣分析仪尿RBC检测结果假阳性,应通过显微镜镜检来纠正其对尿RBC检测的影响。     

Screening for microalbuminuria in patients with diabetes mellitus: frozen storage of urine samples decreases their albumin content

The influence of storage on urinary albumin concentration was prospectively studied with use of overnight urine specimens (Albustix negative) from 73 diabetic patients. From each urine sample four aliquots were taken. One was stored at 4 degrees C and assayed within two weeks, the other three were stored at -20 degrees C and assayed within two weeks and after two and six months. Albumin concentration was measured with laser immunonephelometry. The detection limit, 1 mg/L, suffices for the screening of diabetic patients for microalbuminuria. After storage for two and six months at -20 degrees C, significantly lower albumin concentrations were found. The difference was mainly caused by lower concentrations found in urine samples in which a precipitate had formed, which was the case in 22 and 25 samples, respectively. Thus, freezing of urine samples for determination of low concentrations of albumin may yield falsely low results. Urine samples are best stored at 4 degrees C and assayed within two weeks.     

Solving a myth: Does boric acid stabilize aldosterone in urine at typical clinical laboratory storage conditions?

Aldosterone is produced by the adrenal gland and plays an important role in blood pressure regulation and electrolyte hemostasis. Clinically, measurement of urine aldosterone provides evidence for the diagnosis of hyper- and hypo-aldosteronism. Urine specimen that is collected in consecutive 24 h is preferred, which mitigates the risk of misdiagnosis due to large diurnal variation in aldosterone secretion. Preservatives such as boric acid are routinely added to the collection containers prior to urine collection. However, little is known of the effectiveness of these preservatives on stabilizing aldosterone in urine. In the current study, we examined the stability of urine aldosterone under typical clinical laboratory storage conditions with and without the supplementation of boric acid. Our result demonstrated that the addition of boric acid is unnecessary.     

Critical study of common conditions of storage of glucocorticoids and catecholamines in 24-h urine collected during resting and exercising conditions

BACKGROUND: Except immediate freezing of the samples, no practical method has been validated for preservation of glucocorticoids and catecholamines in 24-h urine collection. Furthermore, the influence of urine storage at bladder temperature during periods of different lengths and the effect of prior exercise on preservation of these hormones in the bladder have not been investigated until now. METHODS: Ten healthy volunteers collected their urine both after a resting and after an exercise session. Urine was aliquoted into tubes which were stored during 24 h in the presence or in the absence of preservatives and at different temperatures. Two samples were stored either 3 or 9 h at 37 degrees C (bladder temperature) without additive. RESULTS AND CONCLUSIONS: When collecting 24-h urine samples for glucocorticoids determination, sample can be stored at room temperature during the 24-h collection period without compromising glucocorticoids preservation. When collecting 24-h urine samples for catecholamines determination, samples have to be chilled without preservative during the whole of the collection period. If the samples have to be stored at room temperature, HCl should be used. Moreover, we report for the first time that catecholamines can be degraded in the bladder and therefore that subjects should urinate every 3 h during either a resting or an exercising day.     

Mass spectrometry-based hepcidin measurements in serum and urine: analytical aspects and clinical implications

BACKGROUND: Discovery of the central role of hepcidin in body iron regulation has shed new light on the pathophysiology of iron disorders. Information is lacking on newer analytical approaches to measure hepcidin in serum and urine. Recent reports on the measurement of urine and serum hepcidin by surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) necessitate analytical and clinical evaluation of MS-based methodologies. METHODS: We used SELDI-TOF MS, immunocapture, and tandem MS to identify and characterize hepcidin in serum and urine. In addition to diagnostic application, we investigated analytical reproducibility and biological and preanalytical variation for both serum and urine on Normal Phase 20 and Immobilized Metal Affinity Capture 30 ProteinChip arrays. We obtained samples from healthy controls and patients with documented iron-deficiency anemia, inflammation-induced anemia, thalassemia major, and hereditary hemochromatosis. RESULTS: Proteomic techniques showed that hepcidin-20, -22, and -25 isoforms are present in urine. Hepcidin-25 in serum had the same amino acid sequence as hepcidin-25 in urine, whereas hepcidin-22 was not detected in serum. The interarray CV was 15% to 27%, and interspot CV was 11% to 13%. Preliminary studies showed that hepcidin-25 differentiated disorders of iron metabolism. Urine hepcidin is more affected by multiple freeze-thaw cycles and storage conditions, but less influenced by diurnal variation, than is serum hepcidin. CONCLUSION: SELDI-TOF MS can be used to measure hepcidin in both serum and urine, but serum requires a standardized sampling protocol.     

不同放置条件下放置时间对临床尿液样本检测结果的影响

目的:探讨临床尿液样本不同放置条件下的放置时间对检测结果的影响,以提高对尿液标本分析前的质量控制。方法:随机选取 33 名该院门诊就医病人,收集其随机尿液样本,分别检测尿生化、尿常规、尿流式相关检测指标。采用配对 t 检验和配对秩和检验进行统计分析。结果:在室温放置条件下,尿钠(Na)在 4h 显著升高(P <0.05);尿肌酐(CREA)、尿酸(UA)、白细胞平均前项散射光强度(WBC-MFsc)在 6h 出现明显变化(P <0.05),其中 CREA、UA显著升高,WBC-MFsc 显著下降;尿总蛋白(TP)在8h 显著升高(P <0.05),其余各检测指标在室温放置 8h 无显著性变化(P >0.05)。在 4℃冰箱保存条件下,尿钾(K)在 6h显著降低(P <0.05),尿 UA 在 8h显著降低(P <0.05);其余各检测指标在 8h 无显著性变化(P >0.05)。结论:不同放置条件下的放置时间对某些尿液检测结果可产生影响,在实际工作中应规范尿液标本检测流程。     

静脉滴注头孢唑啉钠对尿糖的影响

目的观察静脉输入头孢唑啉钠后对尿糖定性的影响。方法将40例血糖正常患者随机分成2组,监测输注头孢唑啉钠后2、4、6 h的尿糖变化。结果静滴头孢唑啉钠组对尿糖定性在2 h内有显著影响(P<0.01)。结论静脉使用头孢唑啉钠同时又必须监测尿糖患者选择监测尿糖时间宜在点滴头孢唑啉钠2 h以后。     

Influence of biological variations and sample handling on measured microalbuminuria in diabetic patients.

Five immunochemical assays for determining low concentrations of albumin were investigated. These were a radioimmunoassay (RIA); turbidimetric immunoassays (TIA) both according to end-point measuring principle on a Cobas Fara and Hitachi 717 analysers, and according to kinetic measuring principle on a Turbitimer instrument; and a nephelometric immunoassay (NIA). All achieved the analytical goal necessary for optimal patient care. The correlations between the albumin concentrations measured with the different techniques were very good. In vitro glycation of albumin did not influence albumin concentrations measured by the five assays. Urine albumin excretion measured over 3 consecutive days showed considerable day-to-day variation. This was highest for spot-urine specimens and significantly lower for 24 h and timed-overnight samples. Variation of storage temperature (room temperature, 4 degrees C, -20 degrees C), time (up till 3 months), and pH (within the range pH 5-8) of the urine samples did not change significantly the measured albumin concentrations. Different sample preparations (vortex-mixing, centrifugation, and thawing) had no influence on the measured albumin concentration. In conclusion, a maximum standardization of the collection of timed-overnight urine samples for screening and 24 h urine samples for confirmation of microalbuminuria during 3 consecutive days is more crucial than the choice of the immunological technique.