同步荧光法测定血清过氧化脂质

脂质过氧化损伤在糖尿病、感染性疾病、肿瘤、类风湿性关节炎、动脉粥样硬化等许多疾病的发生与发展中起到了重要作用。丙二醛（ＭＤＡ）是自由基使脂质发生过氧化作用形成的最终产物,通常用ＭＤＡ来反映体内脂质过氧化状态。ＭＤＡ测定大多采用ＴＢＡ比色法,通过ＭＤＡ与硫代巴比妥酸（ＴＢＡ）加热形成红色复合物,用比色或荧光法１测定ＭＤＡ含量,由于荧光法特异性、灵敏度较高,因而得到了广泛地应用。然而,从目前通常使用的荧光法的荧光光谱中可以看出,该法受到瑞利散射光干扰严重,为了降低散射光的干扰,提高测定的特异性和…     

系统性红斑狼疮患者血清中可溶性人类白细胞抗原-G表达水平的研究

目的 探讨血清可溶性人类白细胞抗原(HLA)-G(sHLA-G)在系统性红斑狼疮(SLE)患者中表达水平及与疾病活动度相关性.方法 采用酶联免疫吸附试验(ELISA)方法检测58例SLE患者与60名健康人血清sHLA-G水平.结果 SLE患者血清sHLA-G水平(120±92)ng/ml显著高于健康对照组(35±33)ng/ml(P＜0.01);SLE活动期组血清sHLA-G水平(170±70)ns/mJ显著高于SLE缓解期组血清sHLA-G水平(95±88)ng/ml(P＜0.01);血清sHLA-G水平与SLEDAI呈正相关(r=0.30,P=0.01);与抗双链DNA(dsDNA)、C3、C4及抗核抗体滴度无显著相关性(P＞0.05).结论 SLE患者血清中sHLA-G表达增高,且与疾病活动度呈正相关,sHLA-G在SLE疾病发生、发展中可能发挥了一定的作用.     

年青人脑卒中与抗心磷脂抗体的关系

抗心磷脂抗体(Anticardiolipin Anti-bodies,ACA)是一种自身抗体,常见于系统性红斑狼疮等自身免疫性疾病。近年来,不少学者报道ACA与动脉系统和静脉系统的血栓形成有关。本研究观察年青人脑卒中与ACA之间的关系,现将结果报道如下。资料和方法     

麻风患者外周血中T淋巴细胞亚群的改变

用单克隆抗体检测33例麻风患者外周血中的T淋巴细胞亚群,其中LL15例、BL7例、TT11例;男28例、女5例;年龄23～71岁。另以22名健康检查合格者作对照,其中男13、女9,年龄26～53岁。分离白细胞,配成5×10~6/ml浓度待测。所用抗人淋巴细胞亚群单克隆抗体由武汉生物制品研究所提供,按该所提供的方法测定。结果证实麻风患者外周血中的T淋巴细胞亚群有免疫失衡现象。33例患者的OKT3阳性数都比正常人为低(P<0.01);OKT4阳性与正常人接近(P>0.05).OKT8阳     

高效毛细管电泳技术同时检测随机尿中香草扁桃酸、高香草酸和肌酐含量

Objective To establish a method for detecting urinary vanillylmandelic acid (VMA), homovanillic acid (HVA) and creatinine (Cr) simultaneously by high performance capillary electrophoresis (HPCE). Methods The separations were carried out using a 120 mmol/L phosphate buffer (pH 6.80) in a fused-silica capillary tube of 47 cm×75 μm I.D. by capillary zone electrophoresis (CZE). Injections were made by using the pressure mode for 4 s at 1 p. s. i. after samples were centrifuged and diluted. The detections were monitored by a diode-array detector (DAD) at 200 nm after samples were separated at a voltage of 20 kV. The method developed was validated systematically and applied to urine samples from healthy adults (n = 100) and children (n = 100) for establishing the reference ranges of VMA/Cr and HVA/Cr, respectively. Results Under these conditions, the separations of VMA, HVA and Cr could be completed within 13 min. The linearity ranges of VMA, HVA and Cr were 0-500, 0-500 and 0-4 000 μmol/L, respectively, with the correlation coefficients (r) between 0.997 2 and 0. 999 1 (P < 0.01). The detection limits (S/N= 3) were 1.0 μmol/L for VMA, 1.0 μmol/L for HVA and 50.0 μmol/L for Cr. The mean within-run (n = 10) CVs of migration time for VMA, HVA and Cr in urine were 0.58%, 0.56% and 0.25% respectively, while the mean between-run (n = 10) CVs of migration time were 0.95%, 1.00% and 0.48% respectively. The mean within-run (n = 10) CVs of peak area for VMA, HVA and Cr were 3.78%, 3.97% and 2.76% respectively, while the mean between-rim (n = 10) CVs of peak area were 4.60%, 4.08% and 4.42% respectively. The average recoveries were 98.36% for VMA, 93.56% for HVA and 98.85% for Cr. Other compounds in human urine such as catecholamines, 5-hydroxytryptamine and albumen didn't interfere with the assay. The correlation between CE method and HPLC method was good. And the correlation coefficients (r) of VMA and HVA were 0.954 9(P <0.01) and 0.945 1 (P < 0.01), respectively. Skewness distributions were presented for VMA/Cr and HVA/Cr in random urine from both adults and children, and the 95% reference ranges were established by the percentile method. For adults, the reference ranges of VMA/Cr and HVA/Cr were 0-4. 26 and 0-1.69 (μmol/mmol), respectively. For children, the reference ranges of VMA/Cr and HVA/Cr were 0-10.39 and 0-4.31 (μmol/mmol), respectively. Conclusions The CE method devised here for direct measurement of urinary VMA, HVA and Cr is simple, fast,precise and automatic with good repeatability. It is an ideal method for routine detection and mass screening of pheochromocytoma and neuroblastoms.     

尿蛋白超薄琼脂糖凝胶电泳及初步临床应用

报道尿蛋白超薄琼脂糖凝胶电泳，采用考马斯亮蓝染色，对人血清白蛋白（HSA）灵敏度为3μg／ml，对尿本─周氏蛋白灵敏度为5μg／ml．通过改进凝胶制板工艺和琼脂糖板的贮存方法，使每次可以大量制板，保存期可在三个月以上。该法具有快速、简便、不需浓缩标本、便于临床常规开展的优点。对52例临床尿标本的电泳分析表明，该法对肾脏损害程度的鉴别诊断及指导临床用药方面有主要实用价值。     

慢性乙型肝炎患者外周血单个核细胞内与血清HBV DNA载量的相关性及超微结构研究

目的研究慢性乙型肝炎患者外周血单个核细胞(PBMCs)与血清中HBV DNA载量之间的相关性及PBMCs超微结构的变化。方法应用荧光定量聚合酶链反应(FQ-PCR)检测60例慢性乙型肝炎患者PBMCs与血清中HBV DNA的载量;并采用透射电镜分析观察其中20例PBMCs的超微结构所见。结果血清中HBV DNA载量为(5.25±2.11)log10copies/ml,PBMCs中HBV DNA载量为(3.74±1.08)log10copies/ml,两者的HBV DNA表达呈正相关(r=0.84,P<0.0001)。电镜观察到血清HBVDNA载量高(≥105copies/ml)的患者PBMCs胞浆中易见到HBV颗粒,线粒体膨胀、嵴消失,内质网也可见膨胀。病程5年以上或血清HBV DNA载量<103copies/ml的患者,虽然PBMCs中病毒颗粒不多见,但细胞器多有改变。结论慢性乙型肝炎患者PBMCs与血清中HBV DNA载量呈正相关,可作为疗效观察指标。随着病程延长,尽管血清HBV DNA载量明显减少,但PBMCs细胞器损伤明显,提示机体免疫功能受损。     

血、尿β痕迹蛋白检测对2型糖尿病肾损伤早期诊断价值研究

目的探讨血、尿β痕迹蛋白(βTP)检测在2型糖尿病肾损伤中的诊断价值。方法明确诊断为2型糖尿病患者178例,按照尿Alb/Cr比值将其分为三组:Alb/Cr正常组60例,微量白蛋白尿组65例,大量白蛋白尿组53例,另设健康对照组80名。采用胶乳增强散射免疫比浊法原理检测血、尿βTP,比较各组患者血、尿βTP水平,并与Scr、eGFR以及实验及临床相关指标进行相关性分析。结果尿Alb/Cr正常的2型糖尿病患者血清βTP为(0.70±0.36)mg/dl,较健康对照组[(0.59±0.12)mg/dl]显著升高(P<0.01)。微量白蛋白尿组血清βTP(0.82±0.30)mg/dl,较Alb/Cr正常组显著升高(P<0.05);大量白蛋白尿组血清βTP(1.79±1.19)mg/dl,与其他两组比较显著升高(P<0.01);三组2型糖尿病患者血清βTP阳性率分别为28.3%、60.0%和87.8%;尿液βTP/Cr阳性率分别为20%、84.6%和90.6%。相关分析结果表明,血清βTP与SC呈显著正相关,r为0.705 0(P<0.01),eGFR值与血β-TP呈显著负相关(r=-0.805 1,P<0.01)。尿βTP与Scr、HbA1c、收缩压、病程呈正相关(r分别为0.387 9、0.240 2、0.164 3、0.241 5,均P<0.05)。结论血清βTP对2型糖尿病患者早期肾损伤具有重要的诊断价值,可以早期反映肾小球滤过功能,而尿βTP/Cr可以作为评价2型糖尿病患者肾小管损伤的有效指标。     

探讨血清25羟基维生素D水平在糖尿病肾损伤中的作用

目的探讨血清25羟基维生素D[25-hydroxy vitamin D,25-（OH）D]水平与2型糖尿病肾损伤的关系。方法选择2012年12月至2013年5月我院内分泌科住院的2型糖尿病患者148例,选择同期健康体检者50例作为对照组。依据尿白蛋白／肌酐（albumin／creatinine,Alb／Cr）水平将糖尿病患者分为正常蛋白尿组（Alb／Cr〈30mg／gCr）、微量蛋白尿组（Alb／Cr30～299mg／gCr）和大量蛋白尿组（Alb／Cr≥300mg／gCr）。检测血清中25-（OH）D、空腹血糖（fasting plasma glucose,FPG）、糖化血红蛋白（hemoglobin Alc,HbAlc）、Cr、尿酸（uric acid,UA）、总胆固醇（total cholesterol,TCH）、甘油三酯（triglyceride,TG）、高密度脂蛋白（highdensity lipoprotein,HDL）、低密度脂蛋白（low density lipoprotein,LDL）。并对所得数据进行统计学分析。结果糖尿病患者的蛋白尿水平随着病程的延长而升高。三组糖尿病患者的病程差异有统计学意义（P〈0．01）。三组患者的收缩压、Alb／Cr、FPG、HbAlc、TCH、TG、LDL、UA和血Cr水平均高于对照组,而肾小球滤过率（glomerular filtration rate,eGFR）和HDL低于对照组,差异均有统计学意义（P均〈0．05）。大量蛋白尿组和微量蛋白尿组血清25-（OH）D水平较正常蛋白尿组和对照组明显降低,差异均有统计学意义（P均〈0．01）,而正常蛋白尿组与对照组比较差异无统计学意义。病程〉10年的患者25-（OH）D水平显著低于病程5—10年及〈5年的患者,差异均具有统计学意义（P均〈0．05）。Pearson相关分析结果表明,血清25-（0H）D与尿Alb／Cr、血Cr和UA均呈负相关,而与eGFR呈正相关（P均〈0．05）。多元logistic回归分析结果表明,25-（0H）D与DN呈独立相关（β=-0．39,P〈0．01）,且OR为2．98。结论血清25-（0H）D是糖尿病肾脏损伤的独立危险因素,其水平降低可能导致糖尿病肾病的发生。