微流控芯片电泳在快速分离尿蛋白中的临床应用价值

目的:探讨并建立微流控芯片的电泳分离方法,用于快速分离尿蛋白。方法:利用照相平饭湿蚀刻法在5cm×5cm×0.1cm石英玻片上,刻制管道宽度为70μm、深度约40μm的十字形电泳泳道,以500V为进样电压、2000~2500V为分离电压、pH10.5的75mmol/L硼酸缓冲液为电泳液分离尿蛋白。结果:在2~3min完成电泳,纯牛白蛋白、人运铁蛋白、人IgG均可得到单一分离峰,其混合样品亦可得到较好分离。在38例尿蛋白阳性患者中,用该法检测发现溢出性蛋白尿3例、选择性蛋白尿9例、非选择性蛋白尿26例,8例健康对照未检出蛋白峰。结论:本法具有快速、简便、检测成本低的特点,依据电荷和分子量的不同,可将蛋白尿初步分为溢出性蛋白尿、选择性蛋白尿、非选择性蛋白尿,有利于临床的诊断和预后判断,有较好的应用前景。     

琼脂糖凝胶区带电泳应用于蛋白尿的分析

目的：运用琼脂糖凝胶区带电泳进行蛋白尿的分析。方法：采用血清琼脂糖凝胶区带电泳技术分析未经处理和处理后的蛋白尿样本。结果：通过对95例临床尿蛋白阳性标本进行琼脂糖凝胶区带电泳，可粗略区分出溢出性蛋白尿2例、选择性蛋白尿30例、非选择性蛋白尿63例，5例健康组未检出蛋白峰。结论：该法简便快速，在蛋白浓度较高时有很好的重复性，可用于尿蛋白的分析，经透析浓缩处理后的样本电泳效果更好。     

糖尿病肾病患者尿蛋白电泳测定及其临床意义

目的 对糖尿病肾病患者进行琼脂糖凝胶尿蛋白电泳测定,分析患者尿蛋白成分特点,探讨糖尿病肾病患者进行尿蛋白电泳的临床意义.方法 采用全自动琼脂糖凝胶电泳分析仪对20例正常对照组及60例糖尿病患者进行尿蛋白电泳分析.结果 60例糖尿病患者,24例尿蛋白定性阴性中有17例没有检出尿蛋白,1例肾小管性蛋白尿,6例为生理性蛋白尿.36例尿蛋白定性阳性患者生理性蛋白尿7例,4例肾小管性蛋白尿,15例肾小球性蛋白尿,10例混合性蛋白尿.结论 对糖尿病患者进行尿蛋白电泳测定,对糖尿病患者早期肾损伤诊断、判断肾损伤程度和部位有重要意义.     

SDA-AGE尿蛋白电泳在高血压早期肾损伤中的临床应用

目的:探讨SDS-AGE尿蛋白电泳在诊断高血压早期肾损伤中的临床应用。方法:应用十二烷基磺酸钠-琼脂糖凝胶(SDS-AGE)对55例高血压患者新鲜晨尿进行分析。结果:根据尿蛋白电泳图谱,可区分为生理性、肾小管性、肾小球性、混合性蛋白尿,55例病例中未检出7例,生理性蛋白尿8例,肾小管性蛋白尿19例,肾小球性蛋白尿16例,混合性蛋白尿5例。结论:SDS-AGE尿蛋白电泳,无需浓缩尿液,操作简单省时,经扫描可获得半定量结果,能正确反映尿液各蛋白组分的含量及变化,对高血压早期肾损伤诊断有较高的临床应用价值。     

非浓缩尿蛋白SDS-琼脂糖凝胶电泳技术及临床应用

目的对尿液中各种蛋白质进行分离,用于区分尿蛋白的类型。方法应用十二烷基磺酸钠-琼脂糖凝胶电泳(SDS-AGE)法进行非浓缩尿蛋白电泳,对40例已被临床确诊的肾脏疾病患者的尿液样本进行电泳分析,使尿蛋白按分子量大小进行分离。结果根据尿蛋白分子量大小可以将其区分为肾小球性、肾小管性、混合性、溢出性及生理性蛋白尿。其中肾小球性蛋白尿13例、肾小管性蛋白尿9例、混合性蛋白尿11例、溢出性蛋白尿2例、生理性蛋白尿5例。结论本法具有分离效果好,检测灵敏度和诊断符合率高,操作简便,速度快,胶片易于保存等优点。可以将尿蛋白按分子量大小进行区分,经EDC扫描后可获半定量结果,对肾脏疾病的受损部位和受损程度的判断具有较高的价值。     

尿蛋白电泳在肾脏疾病评估中的作用

【目的】了解尿蛋白电泳对肾脏疾病的评估作用。【方法】应用十二烷基磺酸钠一琼脂糖凝胶电泳技术分析肾脏疾病患者非浓缩尿液样本。【结果】100例肾脏疾病非浓缩尿液标本中，通过电泳扫描结果如下：37例（37％）为。肾小球性蛋白尿，10例为肾小管性蛋白尿（10％），53例为混合性蛋白尿（53％）。尿蛋白电泳结果与临床病理诊断结果相符。【结论】非浓缩尿蛋白电泳能早期敏感检测尿蛋白的组分变化，协助临床诊断肾脏病变的严重程度。     

肾脏疾病患儿尿选择性蛋白电泳测定的临床应用评价

报告了149例肾脏疾病患儿尿蛋白定性与尿蛋白电泳结果。18例急性肾小球肾炎患儿均为选择性蛋白尿;肾病组88例选择性蛋白尿占59.1％,非选择性蛋白尿占40.9％,定性“++”非选择性蛋白尿约占24.1％(7/29);肾病综合征患儿以非选择性蛋白尿为主,占56.7％,紫癜性肾炎患儿13例中84.6％为高度选择性蛋白尿。所有病例尿蛋白定性“±～+”者均为高度选择性蛋白尿,定性“++～+++”者与非选择性蛋白尿例数呈正相关,r=0.926,y=25x-19.1。     

SDS-AGE尿蛋白电泳在肾脏疾病诊断中的应用

目的：采用十二烷基磺酸钠—琼脂糖凝胶（SDS-AGE）电泳的方法检测肾脏疾病患者尿液中的蛋白成分以及分子量,以此来判断患者肾脏损伤的部位以及程度。方法：采用法国SEB IA半自动电泳分析仪对肾脏疾病患者尿液做SDS-AGE电泳,分析蛋白尿的成分以及分子量的大小。结果：129例尿蛋白电泳显示,16例为生理性蛋白尿,46例为肾小球性蛋白尿,28例为肾小管性蛋白尿,22例为混合性蛋白尿,17例未检出蛋白尿。结论：SDS-AGE尿蛋白电泳技术对患者无创伤,电泳结果有助于判断尿蛋白的来源,分析肾脏损伤的部位以及程度,对临床诊断肾脏疾病具有重要的意义。     

108例糖尿病肾病患者的尿蛋白电泳结果分析

目的 对尿液中的各种蛋白质进行分离,用于区分尿蛋白类型,以判断糖尿病肾损害的部位和病变程度.方法 应用十二烷基磺酸钠-琼脂糖凝胶进行非浓缩尿电泳,对108例糖尿病肾病患者尿液标本进行分析.结果 108例尿蛋白电泳显示:4例为肾小管性蛋白尿,10例为生理性蛋白尿,22例为肾小球性蛋白尿,72例为混合性蛋白尿.结论 对已经出现尿蛋白的糖尿病肾病患者做尿蛋白电泳,可以区分尿蛋白各种成分,了解肾脏损伤程度和病变程度.此方法 标本留取容易,无创伤性,检测敏感性好,胶片可保存,值得临床推广.     

SDS-AGE尿蛋白电泳用于鉴别肾性和非肾性蛋白尿临床分析

目的 探讨各种肾性蛋白尿与非肾性蛋白尿SDS-AGE尿蛋白电泳谱带的模式与区别.方法 收集各种肾性及非肾性蛋白尿标本作SDS-AGE蛋白电泳,扫描观察谱带特点,分析各蛋白尿成分的区别.结果 肾前溢出性蛋白尿特定组分(25KDA)含量占优势,25KDA/70KDA高达361±102,而33KDA/70KDA比值则明显较小,肾小管损伤性蛋白尿则相反;肾后性蛋白尿160KDA/70KDA和≥165KDA/70KDA比值高达95±37和19.7±8.4,与肾小球损伤性蛋白尿及混合损伤性蛋白尿有显著差异.结论 尿蛋白标本作SDS-AGE尿蛋白电泳利用琼脂糖凝胶的选择性成分及多孔,结合蛋白质相对分子量可区分尿中不同蛋白组分,对肾脏损害性质、程度、部位判断有临床指导意义.     

Comparison between a second generation automated multicapillary electrophoresis system with an automated agarose gel electrophoresis system for the detection of M-components

During the last decade, capillary electrophoresis (CE) has emerged as an interesting alternative to traditional analysis of serum, plasma and urine proteins by agarose gel electrophoresis. Initially there was a considerable difference in resolution between the two methods but the quality of CE has improved significantly. We thus wanted to evaluate a second generation of automated multicapillary instruments (Capillarys, Sebia, Paris, France) and the high resolution (HR) buffer for serum or plasma protein analysis with an automated agarose gel electrophoresis system for the detection of M-components. The comparison between the two systems was performed with patients samples with and without M-components. The comparison included 76 serum samples with M-components > 1 g/L. There was a total agreement between the two methods for detection of these M-components. When studying samples containing oligoclonal bands/small M-components, there were differences between the two systems. The capillary electrophoresis system detected a slightly higher number of samples with oligoclonal bands but the two systems found oligoclonal bands in different samples. When looking at resolution, the agarose gel electrophoresis system yielded a slightly better resolution in the alpha and beta regions, but it required an experienced interpreter to be able to benefit from the increased resolution. The capillary electrophoresis has shorter turn-around times and bar-code reader that allows positive sample identification. The Capillarys in combination with HR buffer gives better resolution of the alpha and beta regions than the same instrument with the beta1-beta2+ buffer or the Paragon CZE2000 (Beckman) which was the first generation of capillary electrophoresis systems.     

Detection of apoptosis with gel eletrophoresis combined with computerized gel analysis system]

OBJECTIVE: To explore a new method for detecting cellular apoptosis. METHODS: Using routine electrophoresis assisted by computerized gel documentation analysis system(GDAS), apoptosis of the macrophages treated with dexamethasone was qualitatively and then quantitatively detected. RESULTS: Apoptosis occurred in the macrophages treated with dexamethasone, and DNA fragmentation of the apoptotic macrophages was visualized on 1% agarose gel electrophoresis. The apoptosis rates as determined by computerized GDAS were consistent with the results obtained from flow cytometry that showed typical apoptosis peak. CONCLUSION: Agarose gel electrophoresis assisted by computerized GDAS was relatively simple and less costly in qualitative and quantitative detection of apoptosis with precision.     

Anomalous Urinary Proteins in Patients with Serum M-Components

Abnormal urinary proteins were investigated in 365 patients with serum M-components; 250 urines were available for study. All specimens were concentrated and processed by electrophoresis and immunoelectrophoresis. An overall frequency of Bence Jones proteinuria was found in 63%, whereas other M-components were detected in 18%. In patients with multiple myeloma, Bence Jones proteinuria occurred in 71%, while M-components in the form of whole molecules of immunoglobulins or as Fc and F'c fragments were observed in 20%. Bence Jones globulin was found in 60% of patients with IgG myeloma, 69% of those with IgA myeloma and 100% of patients with IgD myeloma and light chain disease. In regard to other diseases, Bence Jones proteinuria was found in 44% of patients with macroglobulinemia, in 60% of patients with nonplasmacytic neoplastic diseases and in 28% of patients with non-malignant conditions. Other M-components were detected in the urine of 13%, 14% and 6% respectively.

The detection of abnormal urinary proteins in patients with M-components in the serum is an important diagnostic tool and may have prognostic significance.

     

白蛋白百分含量不同时两种方法测定尿总蛋白结果的差异

目的：比较尿白蛋白百分含量不同时用邻苯三酚红钼法（PRM）和苄乙氯铵法（BEC）测定尿总蛋白结果的差异。方法：利用琼脂糖凝胶普通电泳分析尿蛋白组分,然后比较PRM法和BEC法测定94例实验标本的尿总蛋白的结果;同时总结2 154例阳性尿液的白蛋白百分含量。结果：白蛋白百分含量为96%～100%、81%～95%、66%～80%、51%～65%、31%～50%、10%～30%和〈10%的7组中,两法测定结果的相对偏差分别为10.0%、4.5%、6.6%、14.9%、21.7%、53.0%和65.2%,BEC法测定结果偏低。统计显示白蛋白含量大于66%时两法测定结果无明显差异,白蛋白小于65%时差异有显著性。在所有2 154例阳性尿液中,白蛋白含量为96%～100%的病例占45.0%,白蛋白含量为66%～95%的病例占45.2%,白蛋白含量〈65%的病例占9.8%。结论：临床上约10%蛋白阳性尿液的白蛋白含量小于65%,这些尿液采用PRM法和BEC法测定总蛋白时,结果差异有显著性。     

免疫固定技术检测尿本-周蛋白及其临床分析

目的 用免疫固定技术检测尿液本-周蛋白,并对阳性结果患者的临床诊断进行分析,探讨其在临床上的应用.方法 用非浓缩晨尿标本以免疫固定电泳法进行尿液本-周蛋白测定,以临床初次诊断及最终确诊对阳性结果进行分类.结果 在所有晨尿标本的检测中,有110例尿液本-周蛋白呈阳性结果,其中被确诊为M蛋白病有101例;肾淀粉样变性有5例;肾小球肾炎和淋巴瘤各2例.101例M蛋白病人中初次诊断怀疑为M蛋白病的有54例;贫血查因的有9例;肾损伤和蛋白尿查因的有23例;骨折的有4例;其它,如肺部感染、心功能不全、消化道疾病、神经系统疾病等有11例.结论 用免疫固定电泳法检测尿液本-周蛋白具有特异性和高灵敏度,对M蛋白病的诊断有着重大意义.