2型DR患者同型半胱氨酸和胱抑素C水平的变化及相关性研究

目的:探讨同型半胱氨酸(Hcy)与胱抑素C(Cystatin C)及超敏C反应蛋白(hs-CRP)在2型糖尿病视网膜病变(DR)发病机制中的改变及其相关性。方法:选择经诊断为2型糖尿病患者无视网膜病变组45例;背景期DR组42例;增殖期DR组46例。同时选择40例健康正常人做对照组。Hcy,CysC采用酶联免疫分析法、hs-CRP采用颗粒增强免疫比浊法进行检测。双变量相关分析采用直线相关分析。结果:患者各组胱抑素C及同型半胱氨酸、超敏C反应蛋白含量明显高于对照组P<0.01,组间有非常显著差异P<0.01,且随着病程的进展升高明显P<0.01。Hcy与Cystatin C水平均有一定增长趋势,且两指标呈正相关(无视网膜病变组r=0.675,背景期DR组r=0.823,增殖期DR组r=0.756,P<0.01)。结论:2型糖尿病视网膜病变患者血浆胱抑素C、同型半胱氨酸、超敏C反应蛋白含量远高于正常对照,表明检测各指标在2型糖尿病视网膜病变的早期诊断、治疗等方面有很重要的临床意义,联合检测可推断糖尿病视网膜病变的进程,更全面的评价早期糖尿病视网膜病变,为临床早期干预血同型半胱氨酸及胱抑素C等提供理论基础。     

Complement activation and depletion during LDL-apheresis by heparin-induced extracorporeal LDL-precipitation (HELP)

The heparin-induced extracorporeal elimination of low density lipoproteins (LDL) is a well-established clinical procedure to markedly reduce cholesterol levels. The biocompatibility of this artificial filter system (HELP) was investigated by quantitation of representative complement proteins within the extracorporeal circuit using established ELISA procedures, based on monoclonal antibodies recognizing exclusively either native (C6, C7) or activated proteins (act.C3, C5a, TCC). HELP was found to be a self-limiting extracorporeal system with respect to complement activation, since act.C3 and TCC, generated mainly at the plasma filter, were partially adsorbed to the following HELP specific filters to concentrations which were lower than those obtained before the plasma filter. C5a, which increased 14.5-fold at the plasma filter was not eliminated by the following filters; however, elevated levels were not found in the patients at the end of apheresis and no leucocytopenia was observed.     

结直肠癌血清P53抗体和组织P53蛋白的临床意义

目的 :检测结直肠癌患者血清p5 3抗体 (p5 3Ab)的敏感性和特异性 ,并了解其与肿瘤组织p5 3的相关性和临床意义。方法 :检测 2 0 0 2年 7月至 2 0 0 3年 12月的结直肠癌病例 178例。对照组为 4 0例正常健康志愿者。血清p5 3Ab检验采用酶联免疫法 :肿瘤组织p5 3蛋白表达测定应用免疫组化SP染色法。结果 :178例结直肠癌中33例血清p5 3Ab阳性 ,阳性率 18.5 4 % ;4 0例对照组检测阴性 ,特异性 10 0 %。血清p5 3Ab与患者的年龄、性别和肿瘤发生的部位无关 (P >0 .0 5 ) ;血清p5 3Ab阳性为Dukes’分期B期 (16 / 71,2 2 .5 4 % )、C期 (12 / 5 5 ,2 1.82 % )和D期(5 / 30 ,16 .6 2 % )。 10 3例肿瘤组织p5 3蛋白阳性 (5 7.87% ) ,75例阴性 (4 2 .13% ) ;血清p5 3Ab阳性者 6 9.7%组织表达阳性 ,30 .3%组织表达阴性 (P <0 .0 5 )。随访 6～ 2 4个月 ,肿瘤复发率在血清p5 3Ab阳性和阴性患者间无显著性差异 ;平均存活时间亦无统计学意义。结论 :研究表明血清p5 3Ab的产生伴有肿瘤组织p5 3蛋白的积聚 ;因为抗体检测的低敏感性 ,血清p5 3Ab不能作为术前的肿瘤指标 ;但由于它的高特异性 ,对今后的随访可能具有一定临床意义     

“桥抗体”对酶免疫分析的影响

在对 T-2毒素检测的 ELISA 研究中,T2-HS(T2-琥珀酸半酯)与 BSA 或 PLL联接,前者(T2-C_4-BSA)为免疫原,后者(T2-C_4-PLL)为检测抗原。在进行 ELISA 时,将 T2-C_4-PLL 包被于微板,并与兔抗 T-2抗体和标准 T-2样品一起保温。T-2抗体结合量用酶(HRP)标羊抗兔 IgG 与酶底物反应进行测定。在此情况下,我们不仅观察到抗 T-2抗体及抗 BSA 抗体,而且还有抗“-C_4-”抗体(即“桥抗体”)存在,同时发现“桥抗体”对T-2检测的灵敏度有明显干扰,甚至使 ELISA 不能进行。如以 T2-C_2-TT 代替 T2-C_4-PLL为检测抗原,由于“-C_4-”桥的排除,“桥抗体”的干扰也明显减弱。     

Induction of specific anti-actin antibodies and their measurement by ELISA technique.

Using the ELISA technique we have been able to quantify antibodies directed against actin and to follow the kinetics of antibody production. Specific anti-actin antisera have been raised in rabbits by immunization with chemically modified white muscle rabbit actin. Two or three dinitrophenyl groups linked per actin molecule were sufficient to break natural tolerance, while linkage of three phosphorylcholine groups to actin was not.     

~(60)Co照射及低温保存对转基因细胞影响的分析

目的　观察经放射前后的转基因细胞Tca8113/TNF α的TNF α的分泌情况 ,为转基因细胞经致死剂量的放射线灭活作为“瘤苗”应用于临床前TNF α基因的有效表达提供依据。方法　利用基因转染技术 ,用逆转录病毒载体将肿瘤坏死因子基因 (TNF α基因 )转导入人舌癌Tca8113细胞 ,并获得表达 ;在此基础上 ,通过ELISA法分析转基因细胞Tca8113/TNF α在60 Co放射组及未放射组的TNF α分泌量的变化以及经液氮储存 4 8h、一周复苏后的TNF α分泌量的变化。结果　 (1) 60 Co放射组转基因细胞Tca8113/TNF α的TNF α的分泌量 (x±s) (pg/ml/ 10 6细胞 /2 4h)平均高达 (110± 5 6 2 ) ,而未转基因的Tca8113细胞的上清液中未检测到TNF α ,统计学上有明显的差异性 (P <0 0 1) ;(2 )经频率为 4 39 5cGy/min ,总剂量为 10 0 0 0cGy的60 Co放射后TNF α分泌量降低 ,但在 4～ 6d时有一分泌高峰 ;(3)转基因细胞Tca8113/TNF α经60 Co放射后在经 4 8h及一周的液氮内储存后TNF α能够分泌 ,其分泌量的变化趋势同储存前。 4 8h组和一周组之间无明显的统计学差异性。结论　转基因细胞Tca8113/TNF α作为“瘤苗”应用时 ,目的基因的表达可初步评价为有效     

人醛缩酶A单抗ELISA对原发性肝细胞癌诊断价值的探讨

从人骨骼肌纯化醛缩酶（ＡＬＤ）Ａ，制备抗人ＡＬＤ－Ａ单克隆抗体，首次建立了测定血清ＡＬＤ－Ａ的竞争性ＥＬＩＳＡ，最小检出量０．１ｎｇ／ｍｌ，对ＡＬＤ－Ｂ和Ｃ无交叉反应。６０例正常人血清含量１３１±２４ｎｇ／ｍｌ，上限１７９ｎｇ／ｍｌ，均未超过此限；原发性肝细胞癌４５例５５４±１９３ｎｇ／ｍｌ，阳性率９５．５％（４４／４５），两组差异有非常显著意义（Ｐ＜０．００１）。对肝癌诊断有实用价值。单抗可大量制备，能使ＡＬＤ－Ａ检测标准化。     

包虫病人唾液抗体的三种免疫学检测方法比较

本用间接法、点免疫法、ＡＢＣ－ＥＬＩＳＡ法三种免疫学试验检测４６例包虫病人唾液抗体，并做了比较，结果表明：点免疫法简便、敏感、准确，综合评估成绩最高，为流行病学调查提供了有力的手段；间接法特异性好，ＡＢＣ－ＥＬＩＳＡ法敏感度高，二组合使用，可提高免疫诊断价值，适于科学研究和临床分析。     

Neurocysticercosis immunodiagnosis using Taenia solium cysticerci crude soluble extract, excretory secretory and lower molecular mass antigens in serum and urine samples of Indian children

Neurocysticercosis (NCC), the most common neurological disorder of parasite etiology, results from lodgment of Taenia solium cysticerci in the central nervous system and is now increasingly being recognized in children. The confirmed diagnosis is based collectively on radiological findings and serodiagnostic techniques. The serodiagnostic techniques have variable sensitivity and specificity depending upon the technique, antigens used, location and number of cysts. Crude soluble extract (CSE), excretory secretory (ES) and lower molecular mass (LMM) (10-30 kDa) antigenic fraction of T. solium cysticerci were evaluated for antibody detection in serum and urine samples by ELISA. Serum and urine samples were collected each from 125 clinically suspected and radiologically proven NCC (111 with single Computed Tomography (CT) lesions and 14 with multiple CT lesions) and 125 control subjects (60 with neurological disorders other than NCC, 40 with other parasitic diseases and 25 apparently healthy subjects). The sensitivity of the ELISA with the use of CSE, ES and LMM antigenic fractions was 38.4%, 63.2% and 30.4% with serum (cut off dilution 400), 46.4%, 44% and 47.2% with neat urine and the specificity was 88%, 76.8% and 85.6% with serum (cut off dilution 400), 66.4%, 65.2% and 58.4% with neat urine samples, respectively. The study suggests that detection of antibody to ES antigen in serum samples may serve useful purpose for the serodiagnosis of human NCC.     

ELISA法诊断实验性大鼠肝毛细线虫病

目的：实验研究肝毛细线虫对大鼠的血清抗体水平的影响，为人体肝毛细线虫病的免疫诊断提供科学依据。方法：捕获阳性鼠分离肝毛细线虫虫卵，将虫卵培养至感染期，感染大白鼠。制作虫卵可溶性抗原，用ELISA法对大鼠体内血清抗体作动态检测，并选用5种对照血清作比较，以观察该法的特异性。结果：大鼠在感染4周后可全部阳性检出，该法对大鼠旋毛虫、日本血吸虫、犬弓蛔虫、广州管圆线虫和肺吸虫抗体阳性血清分别有2／10、2／10、2／10、1／10和0的假阳性。结论：ELISA法具有较高的敏感性和特异性，适用于检测肝毛细线虫感染。