微波-EDTA脱钙技术在免疫组织化学染色中的应用研究

目的探讨微波(MW)-乙烯二胺四乙酸钠(EDTA)快速脱钙技术对骨及软组织抗原活性的影响.方法根据实验条件将待脱钙组织分为4组,即MW-EDTA(15℃),Jenkins脱钙-固定液(20℃),10%EDTA(20℃)和10%硝酸(20℃),其中MW-EDTA方法中EDTA的浓度分别是4%、7%、10%、13%和16%.应用免疫组织化学LSAB染色,光镜观察免疫组织化学染色结果.结果MW-EDTA脱钙的免疫组织化学染色阳性细胞显著高于Jenkins脱钙-固定液(20℃)、10%ED-TA(20℃)及10%硝酸脱钙(20℃)3种脱钙方法,其时间明显缩短.其中用10%和13%浓度的EDTA脱钙的组织,其免疫组织化学染色阳性细胞又明显多于EDTA浓度为4%、7%和16%者.结论以MW-10%EDTA技术进行组织脱钙,其脱钙时间短、免疫组织化学染色效果好.     

光波-微波-硝酸快速脱钙技术对骨组织免疫组织化学染色的影响

目的:探讨光波微波硝酸快速脱钙技术对骨及软组织抗原活性的影响。方法:用8%硝酸分别在单纯微波、单纯光波、光波微波组合Ⅰ和光波微波组合Ⅱ条件下脱钙,同时以室温脱钙作为对照;应用标记的链霉菌素卵白素生物素(LSAB)免疫组织化学(免疫组化)染色技术检测不同脱钙条件下心钠素(ANP)、5羟色胺(5HT)、S100、神经微丝蛋白(NF)、神经胶质酸性蛋白(GFAP)和波形蛋白(Vimentin)的表达,并对其染色结果进行图像分析和统计学处理。结果:8%硝酸室温脱钙时间最长,光波微波组合脱钙时间最短;微波组、光波组、光波微波组合Ⅰ组、Ⅱ组的免疫组化染色均明显比对照组好(P<0.01);光波微波组合Ⅰ组、Ⅱ组均明显好于单纯微波组(P<0.01)或单纯光波组(P<0.01);光波微波组合Ⅰ组与Ⅱ组,单纯光波组与单纯微波组之间的免疫组化染色无显著差异(P>0.05)。结论:光波微波硝酸脱钙所用时间比室温常规脱钙、单纯光波、微波脱钙时间明显缩短,免疫组化染色效果最好。     

微波技术在免疫组织化学染色中的作用

本研究在免疫组织化学染色体应用微波炉技术，建立快速有效的免疫组织化学染色方法，结果表明，采用微波技术可以将传统的ＡＢＣ免疫组织化学染色时间由２４ｈ缩短为１ｈ用１２种抗体对染色表明效果稳定，与传统染色效果相比一部分呈色明显增强，起到了常规方法中起不利的良好的抗原修复作用，且脱片率降低，非特异性背景染色减少，认为可以在免疫染色体甚至在其它病理中推广应用。     

光波-微波-硝酸快速脱钙技术对骨组织免疫组织化学染色的影响

目的:探讨光波-微波-硝酸快速脱钙技术对骨及软组织抗原活性的影响.方法:用8%硝酸分别在单纯微波、单纯光波、光波-微波组合Ⅰ和光波-微波组合Ⅱ条件下脱钙,同时以室温脱钙作为对照;应用标记的链霉菌素卵白素-生物素(LSAB)免疫组织化学(免疫组化)染色技术检测不同脱钙条件下心钠素(ANP)、5-羟色胺(5-HT)、S-100、神经微丝蛋白(NF)、神经胶质酸性蛋白(GFAP)和波形蛋白(Vimentin)的表达,并对其染色结果进行图像分析和统计学处理.结果:8%硝酸室温脱钙时间最长,光波-微波组合脱钙时间最短;微波组、光波组、光波-微波组合Ⅰ组、Ⅱ组的免疫组化染色均明显比对照组好(P＜0.01);光波-微波组合Ⅰ组、Ⅱ组均明显好于单纯微波组(P＜0.01)或单纯光波组(P＜0.01);光波-微波组合Ⅰ组与Ⅱ组,单纯光波组与单纯微波组之间的免疫组化染色无显著差异(P＞0.05).结论:光波-微波-硝酸脱钙所用时间比室温常规脱钙、单纯光波、微波脱钙时间明显缩短,免疫组化染色效果最好.     

微波低温改良脱钙在钙化软组织免疫组化染色中的应用

目的 探讨运用微波低温改良脱钙法对提高钙化软组织在免疫组织化学染色中的效果.方法 选取本院送检伴有钙化灶状的甲状腺组织标本62例,每例采用室温和微波低温进行脱钙,免疫组化染色光镜观察效果.结果 运用微波低温改良脱钙在免疫组化染色中明显比室温下的10％硝酸脱钙定位准、染色阳性细胞显示多、假阴性减少,背景清晰.结论 微波低温改良脱钙法能更好的运用于钙化软组织的免疫组化染色.     

大体骨组织免疫组织化学染色的比较

骨组织因其十分坚硬,脱钙时间长,且在常规HE及免疫组织化学染色中均极易脱片,不能观察完整的骨组织结构。常规骨组织采用混合酸即45％盐酸、55％甲酸和EDIA脱钙。20例兔股骨头10例采用混合酸液(福尔马林100ml,生理盐水800ml,甲酸70ml,盐酸80ml,三氯化铝结晶60g,冰醋酸25ml)脱钙,10例采用45％盐酸、55％甲酸脱钙,均用10％中性福尔马林     

免疫组织化学染色在病理诊断中的应用

人类的历史有多长,医学的历史也就有多长,这是因为疾病与生命相伴而至的,医学的任何一个进步都是建立在当时科学与技术发展的相应水平上,因此,了解医学的历史也就是了解科学技术发展史,医学的目的是解除人体的病痛,其核心是疾病的诊断和治疗两个五一节,而这两个五一节的关系可以明确为：任何疾病的有效治疗均来源于正确的诊断,在当今医学发展的现状下,虽然疾病的诊断手段有几十种之多,但就其准确性来说首推病理诊断准确性高,即病理组织学诊断要比化验、放射、ＣＴ、核磁共振、核素等任何一种诊断方法更准确、更可信、更有权威性。而在病理组织学诊断中,免疫组织化学（以下简称免疫组化）已成为其中的一种常用手段。     

Von Ebner氏脱钙液利用超声微波技术在骨组织脱钙中的研究与分析

用氯化钠75g加入500ml蒸馏水中,再加入浓盐酸15ml,最后加蒸馏水至1000ml配制成Von Ebner脱钙液,此脱钙液与超声微波技术相结合运用到骨组织脱钙中,此方法操作简单易行,脱钙时间缩短,脱钙程度很容易掌控,脱钙效果良好,可以制出优质切片,不易落片,不褪色,组织结构清晰,染色鲜明,为病理诊断提供优质切片。此方法容易被广大病理工作者所掌握,同时所需试剂及设备易得,经济方便,特别适用于基层医院的推广。     

免疫组织化学染色技术的应用体会

随着免疫组织化学染色技术的广泛应用,病理学研究产生了划时代的飞跃。根据标记物的不同,免疫组织化学技术可分为免疫荧光细胞化学技术、免疫酶细胞化学技术、免疫铁蛋白技术、免疫金-银细胞化学技术、亲和免疫细胞化学技术、免疫电子显微镜技术等。不同的免疫细胞化学技术各具有独特的试剂和方法,但其基本技术方法是相似的,都包括抗体的制备,组织材料的处理。免疫染色,对照试验,显微镜观察等步骤。下面以美国ZYMED公司SP试剂盒免疫组化染色步骤为例谈几点体会。     

原位杂交与免疫组织化学双标记技术的应用价值

目的 探讨cRNA探针原位杂交结合免疫组织化学技术的临床应用价值.方法 在15例痤疮及毛囊炎患者的石蜡切片上进行原位杂交和免疫组织化学标记,观察双标信号的共存状态.结果 15例痤疮患者皮肤石蜡切片的毛囊上皮、皮脂腺及导管部位获得蓝紫原位杂交阳性信号,同时在表皮及皮脂腺处可见明确的棕黄色树突状阳性信号.结论cRNA探针原位杂交结合免疫组织化学双重标记的方法能够在皮肤石蜡切片上对微生物及表皮细胞成分进行准确定位.     

Pharmacokinetics of ethylene in man; body burden with ethylene oxide and hydroxyethylation of hemoglobin due to endogenous and environmental ethylene

The inhalation pharmacokinetics and the endogenous production of ethylene has been determined in healthy volunteers with respect to the formation of the carcinogen ethylene oxide. Ethylene showed a low degree of accumulation in the body determined in six subjects, the thermodynamic partition coefficient body/air being 0.53±0.23 (mean ± SD) and the accumulation factor body/air at steady-state being 0.33±0.13 (mean ± SD). The rate of metabolism was directly proportional to the exposure concentration. Only 2% of ethylene inhaled was metabolized to ethylene oxide, whereas 98% of ethylene was exhaled unchanged. The rate of the endogenous production of ethylene was 32±12 nmol/h (mean ± SD), as calculated from exhalation data from 14 subjects. The resulting body burden was 0.44±0.19 nmol/kg (mean ± SD). By analyzing published data on ethylene oxide in man its half-life was estimated to be 42 min. Using the pharmacokinetic parameters of ethylene and ethylene oxide, the body burden of ethylene oxide due to the sum of the exposure to environmental ethylene of about 15 ppb and to endogenous ethylene exposure of 0.44 nmol/kg was predicted to be 0.25 nmol/kg. In the blood of five nonsmokers and one smoker the hemoglobin adduct resulting from the reaction of ethylene oxide with the N-terminal valine, N-(2-hydroxyethyl)valine, was quantified by gas chromatography/mass spectrometry. The value of 20±5 pmol/g Hb (mean ± SD) found in the non-smokers corroborated the steady-state level of 18±3 pmol/g Hb (mean ± SD) calculated from the pharmacokinetic approach.     

Extension of a PBPK model for ethylene glycol and glycolic acid to include the competitive formation and clearance of metabolites associated with kidney toxicity in rats and humans

A previously developed PBPK model for ethylene glycol and glycolic acid was extended to include glyoxylic acid, oxalic acid, and the precipitation of calcium oxalate that is associated with kidney toxicity in rats and humans. The development and evaluation of the PBPK model was based upon previously published pharmacokinetic studies coupled with measured blood and tissue partition coefficients and rates of in vitro metabolism of glyoxylic acid to oxalic acid, glycine and other metabolites using primary hepatocytes isolated from male Wistar rats and humans. Precipitation of oxalic acid with calcium in the kidneys was assumed to occur only at concentrations exceeding the thermodynamic solubility product for calcium oxalate. This solubility product can be affected by local concentrations of calcium and other ions that are expressed in the model using an ion activity product estimated from toxicity studies such that calcium oxalate precipitation would be minimal at dietary exposures below the NOAEL for kidney toxicity in the sensitive male Wistar rat. The resulting integrated PBPK predicts that bolus oral or dietary exposures to ethylene glycol would result in typically 1.4-1.6-fold higher peak oxalate levels and 1.6-2-fold higher AUC's for calcium oxalate in kidneys of humans as compared with comparably exposed male Wistar rats over a dose range of 1-1000 mg/kg. The converse (male Wistar rats predicted to have greater oxalate levels in the kidneys than humans) was found for inhalation exposures although no accumulation of calcium oxalate is predicted to occur until exposures are well in excess of the theoretical saturated vapor concentration of 200 mg/m³. While the current model is capable of such cross-species, dose, and route-of-exposure comparisons, it also highlights several areas of potential research that will improve confidence in such predictions, especially at low doses relevant for most human exposures.     

免疫组织化学法和流式细胞术检测非小细胞肺癌骨髓微转移

目的 流式细胞和免疫组织化学技术检测非小细胞肺癌骨髓微转移状况的差异比较.方法 选取2004年3月至2007年5月我院手术治疗的非小细胞肺癌160例,术前未经化疗及放疗,术中抽取肋骨骨髓,分别应用流式细胞技术和免疫组化法检测骨髓中阳性细胞表达率.结果 160例非小细胞肺癌患者免疫组化检测骨髓转移阳性率为30.0％(48例),流式细胞术检测骨髓转移阳性率为34.4％(55例).112例免疫组化检查阴性的患者中,流式细胞术检测发现12例患者骨髓中存在微转移;而48例免疫组化检查阳性的患者中,有5例流式细胞术检测为阴性.结论 流式细胞术和免疫组化技术都可检测非小细胞肺癌骨髓微转移,流式细胞技术有着更高的敏感性和检测效率.     

甲酸脱钙液的浓度对骨组织脱钙后切片染色效果的研究

目的探讨骨组织经不同浓度的甲酸脱钙液24小时脱钙后切片染色效果,选择最佳甲酸脱钙工作液。方法350例骨组织标本每例取材3份,分别用40％、30％、20％的甲酸脱钙液常温下脱钙24小时,观察三种不同浓度的甲酸脱钙液脱钙后骨组织的切片和HE染色情况。结果骨组织经40％甲酸脱钙液脱钙后,切片完整,但HE染色核质偏浅,胞浆偏红;30％甲酸脱钙液脱钙后,切片完整,HE染色核质清晰,胞浆鲜艳;20％甲酸脱钙液脱钙后,切片不够完整,HE染色核质偏浅,胞浆偏浅。结论脱钙液脱钙后应保持组织切片完整,染色效果良好,且30％的甲酸脱钙液是较理想的工作液。     

Dosimetry considerations in the enhanced sensitivity of male Wistar rats to chronic ethylene glycol-induced nephrotoxicity

The developmental consequences of exposure to the polychlorinated biphenyls (PCBs) have been widely studied, making PCBs a unique model to understand issues related to environmental mixture of persistent chemicals. PCB exposure in humans adversely affects neurocognitive development, causes psychomotor difficulties, and contributes to attention deficits in children, all of which seem to be associated with altered patterns of neuronal connectivity. In the present study, we examined gene expression profiles in the rat nervous system following PCB developmental exposure. Pregnant rats (Long-Evans) were dosed perinatally with 0 or 6 mg/kg/day of Aroclor 1254 from gestation day 6 through postnatal day (PND) 21. Gene expression in cerebellum and hippocampus from PND7 and PND14 animals was analyzed with an emphasis on developmental aspects. Changes in gene expression (> or =1.5 fold) in control animals identified normal developmental changes. These basal levels of expression were compared to data from Aroclor 1254-treated animals to determine the impact of gestational PCB exposure on developmental parameters. The results indicate that the expression of a number of developmental genes related to cell cycle, synaptic function, cell maintenance, and neurogenesis is significantly altered from PND7 to PND14. Aroclor 1254 treatment appears to dampen the overall growth-related gene expression levels in both regions with the effect being more pronounced in the cerebellum. Functional analysis suggests that Aroclor 1254 delays maturation of the developing nervous system, with the consequences dependent on the ontological state of the brain area and the functional role of the individual gene. Such changes may underlie learning and memory deficits observed in PCB exposed animals and humans.     

TCT结合DNA定量及Ki67 p16免疫组化联合对宫颈癌及其癌前病早期筛查的应用研究

目的 评估液基细胞学(TCT)结合DNA定量分析及免疫组化(Ki67、P16)对宫颈上皮内瘤病变筛查的应用价值.方法 对参预宫颈癌前病变筛查的妇女,同时进行液基细胞,DNA定量分析及免筛组化(Ki67、P16)方法学评估及临床应用价值探讨.结果 液基细胞学筛查阳性率为11.63％,敏感性与特异性分别为81.28％、27.50％.DNA定量分析筛查阳性率为11.87％,敏感性及特异性分别为:89.83％、19.51％; TCT与DNA联合筛查阳性率为13.54％,敏感性及特异性分别为94.11％、15.36％.免疫组化(Ki67、P16)的阳性表达率分别为84％、56％,两者之间差异有统计学意义(P＜0.05).结论 液基细胞学结合DNA定量分析,再辅以免疫组化(Ki67、P16),不但可以提高筛查的阳性率、敏感性、特异性,而且可以了解宫颈癌及癌前病变各阶段的预后与转归,病变程度.更有利于早期诊断及早期治疗.     

两点法腰麻—硬膜外联合麻醉在剖宫产术中的应用

目的:观察两点法腰麻-硬膜外联合麻醉CCSEA应用于剖宫产术的临床效果和并发症,探讨其最优化的穿刺方式。方法:80例剖宫产患者实施腰麻—硬膜外联合麻醉,随机分成两组,单点法穿刺组(SST组)和两点法穿刺组(DST组),比较两组的血流动力学、穿刺操作时间、硬膜外腔用药量、穿刺及置管困难发生率、麻醉失败率、腰背痛、头痛等情况。结果:硬膜外腔用药量和腰麻平面不对称发生率SST组高于DST组(P<0.05),其他方面两组无差异(P>0.05)。结论:两点穿刺法应用于腰麻—硬膜外联合麻醉,比单点穿刺法更合理安全,值得推荐。