加热抗原修复对乳腺癌组织中内源性抗生物素蛋白结合物的影响及其对策

目的:探讨加热抗原修复对甲醛固定石蜡包埋乳腺癌组织中内源性抗生物素蛋白结合物(EABA)的影响,以及消除EABA对免疫组织化学染色干扰的对策。方法:采用微波加热抗原修复法,免疫组织化学SP染色法,蛋清液封闭法,抗生物素蛋白-生物素封闭法及Envision二步法对10例甲醛固定石蜡包埋乳腺癌标本内的EABA进行检测。结果:1经甲醛固定石蜡包埋的乳腺癌组织中的EABA被封闭;2加热抗原修复可造成EABA暴露,且EABA仅存在于细胞质中;3蛋清液封闭法及抗生物素蛋白-生物素封闭法可封闭暴露的EABA。采用非生物素检测系统也可消除EABA对正常免疫组化染色的干扰。结论:微波加热抗原修复可使甲醛固定石蜡包埋乳腺癌组织中的EABA重新暴露。因而对生物素-抗生物素蛋白-辣根过氧化物酶检测系统的免疫组织化学染色造成干扰;暴露的EABA可通过蛋清液封闭法、抗生物素蛋白—生物素封闭法封闭,或采用非生物素检测系统,达到避免EABA对正常免疫组化染色干扰的目的。     

免疫组织化学中肝组织EABA的封闭

目的:初步探讨蛋清液在封闭肝组织EABA中的作用;方法:采用免疫组化SP法及蛋清液封闭法,对40例肝细胞肝癌组织进行EABA检查;结果:20 %蛋清液可封闭肝组织中的EABA;结论:以蛋清液封闭肝组织中的EABA的方法经济便捷,效果良好.     

免疫组织化学中肝组织EABA的封闭

目的 :初步探讨蛋清液在封闭肝组织EABA中的作用 ;方法 :采用免疫组化SP法及蛋清液封闭法 ,对 4 0例肝细胞肝癌组织进行EABA检查 ;结果 :2 0 %蛋清液可封闭肝组织中的EABA ;结论 :以蛋清液封闭肝组织中的EABA的方法经济便捷 ,效果良好     

高压锅和微波抗原修复法的对比分析

免疫组织化学技术是用已知抗体或抗原检测组织细胞中的未知抗原或抗体的技术 ,具有操作简单、敏感性和特异性强等优点 ,已广泛用于病理诊断。但在甲醛固定、石蜡包埋后的组织有部分抗原决定簇与核酸或其他蛋白发生交联 ,形成网络 ,固定液中的醛基在使蛋白质变性的同时 ,自身也会交联形成网络结构 ,使组织抗原决定簇封闭。导致表达不佳或假阴性。因此 ,充分的抗原修复是关系免疫组化结果的重要因素 ,抗原修复技术对免疫组织化学方法的敏感性起着决定性的作用 [1 ] 。微波炉和高压锅加热是目前常采用的抗原修复方法 [2 ] ,本文比较了这两种方…     

微波技术在免疫组化中的应用

免疫组织化学 (简称免疫组化 )自 1 974年用于病理诊断以来 ,已成为病理科一种常用手段。但目前进行的常规石蜡切片标本 ,一般用甲醛固定 ,其免疫组化标记敏感性明显降低 ,造成显色困难。因此在染色时 ,有些抗原须先进行修复或暴露。笔者采用本科总结的微波修复技术先对病变组织进行修复。然后再观察病变组织中激素受体的表达。并与对照组进行比较。1　材料与方法1 .1　材料　 40例乳腺浸润性导管癌病变组织均为我院 1 997年 4月～ 1 998年 1 0月乳腺手术切除标本。所有标本均为 1 0 %福尔马林固定 ,常规石蜡包埋 ,HE染色。每例取病变最为…     

应用不同免疫组化抗原修复技术的体会

免疫组化成败的关键因素之一是抗原决定簇是否充分暴露,经甲醛固定的组织常引起蛋白质空间结构的改变而导致抗原决定簇的封闭,抗原修复技术在石蜡切片免疫组织化学染色中是一种简单有效的增强方法,但无论是高压热修复还是微波热修复均会出现不同程度的脱片现象,给技术人员带来烦恼,并加大了工作量.     

高压锅和微波炉加热抗原修复方法的比较

免疫组化技术广泛应用于肿瘤病理诊断,其抗原的妥善保存及其恢复是免疫组化染色成败的关键.由于日常工作中所使用的10%福尔马林固定的石蜡包埋组织常产生蛋白交联反应,引起许多抗原决定簇被封闭,因此导致免疫组化染色呈假阴性或表达微弱的结果.要充分暴露抗原决定簇,采用微波炉和高压锅抗原修复是当前常用的方法[1,2].我们比较了两种抗原修复的方法对20种抗体免疫组化标记结果的影响,现总结报告如下.     

免疫组织化学染色中消除内源性生物素的方法研究

内源性生物素(endogenous avidin-bindingacticity,EABA)指得是体内能够与亲和素结合的分子或基因.虽然早已被发现,但是由于对它的研究或观察的样本较少,所以未引起人们的高度重视.随着免疫组织化学技术的发展,在应用单克隆抗体检测石蜡包埋组织,加热抗原修复在增强抗原决定簇表达的同时,也增强了组织中内源性生物素的反应.常用的SP法属于亲和素一生物素法之一,如果待检组织和细胞中富含EABA,则可出现假阳性,导致假阳性结果.     

家用微波炉及金属溶液在恢复陈旧石蜡包埋组织抗原性中的应用

应用家用微波炉及饱和硫氰酸铅溶液对３５倒不同陈旧性甲醛固定石蜡包埋组织切片进行免疫组化染色，其中间叶性恶性肿瘤组织５例，流行性出血热肺、肾上腺、甲状腺组织各１０例，分别使用Ｖｉｍｅｎｔｉｎ、Ｓｅｒｏｔｏｎｉｎ、Ｃ１同相应组织中的抗原反应，同时与常规免疫组化进行对照。结果表明：改良的免疫组化法与常规免疫组化法相比，不需要酶的消化处理，阳性染色强度大，范围广。提示，应用微波炉及金属槽液能促使陈旧性石蜡包埋组织中抗原的恢复，其染色效果明显优于常规酶液处理之方法。     

利用微波促进组织抗原恢复在石蜡切片免疫组化染色中的应用

将石蜡切片浸泡于抗原恢复液，利用微波进行高强度辐射，以恢复组织中因甲醛固定而被封闭的抗原，提高抗原的检出率和染色强度。结果表明，抗原被封闭的组织经抗原恢复处理后。免疫组化染色强度有不同程度的增加，获得了满意的染色效果。一些常规免疫组化方法呈细胞角蛋白，波形蛋白和结蛋白染色阴性或弱阳性的组织，特别是经甲醛长时间固定的组织因抗原的恢复而呈阳性或强阳性反应。这对改善免疫组化染色效果，提高外科病理对肿瘤的鉴别诊断水平，对经甲醛长时间固定的尸解组织进行回顾性免疫组化研究有一定的意义     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.