免疫吸附治疗在肾移植致敏受者中的应用

目的：探讨蛋白A免疫吸附（immunoads Orption，IA）治疗对清除肾移植致敏受者体内特异性抗HLA抗体的疗效和安全性。方法：10例肾移植致敏（PRA〉50％）受者用彤新蛋白A免疫吸附柱行IA治疗，测定治疗前后血免疫球蛋白及PRA水平。结果：10例患者IA治疗次数为4～15次（中位数9）。所有患者IA治疗后血清总IgG水平都明显下降（P〈0．001），IgA和IgM也较治疗前显著降低（P〈0．01）。PRA8例转阴，1例〈30％，1例仍为100％。结论：对于肾移植致敏受者，IA是一种特异性高、安全有效的治疗措施。     

血液透滤清除血清IL-6在重症急性胰腺炎治疗中的应用研究

目的探讨血液透滤清除IL-6在重症急性胰腺炎治疗中的作用及机理。方法根据诊断标准对2002年7月至2006年6月来我院就诊的178例重症急性胰腺炎患者随机分组：血滤组（HF组）85人,非血滤组（NHF组）93人。比较两组患者局部和全身表现,观察各时相点促炎细胞因子血清IL-6的测定值的差异。结果HF组与NHF组比较：腹痛腹胀持续时间为（18.8±4.2）hvs（89.7±28.1）h（P〈0.05）;治疗后第10天APACHEⅡ积分为（5.5±3.6）分vs（13.8±3.8）分（P〈0.05）;住院天数和医疗费为（28.2±12.4）天vs（42.4±11.2）天和（4.38±2.8）万元vs（7.46±2.2）万元,（P〈0.05）。治疗后各时相点血清IL-6检测结果：HF组较NHF组显著降低（P〈0.05）;MODS发病率和病死率分别为12.47%vs.36.28%和4.28%vs.12.82%,两者差异有统计学显著意义（P〈0.05）。结论早期短时血滤有利于纠正重症急性胰腺炎患者血清促炎细胞因子过度释放,使病情减轻,降低MODS发病率和病死率,提高疗效。     

CD44v6、P27蛋白在乳腺癌组织中的表达及意义

目的 探讨乳腺癌中CD44v6、p27蛋白表达的意义。方法 应用免疫组化S-P法检测65例乳腺癌中CD44v6、p27蛋白表达。结果 65例乳腺癌中CD44v6、p27蛋白表达总阳性率分别为58.4％、38.4％。CD44v6蛋白阳性表达率与乳腺癌组织学类型、腋淋巴结转移呈正相关(P<0.05,P<0.01),与预后呈负相关(P<0.01),与组织学分级无明显相关性(P>0.05);p27蛋白阳性表达与乳腺癌组织学类型、组织学分级及腋淋巴结转移呈负相关(P<0.05),与预后呈正相关(P<0.05)。结论 CD44v6、p27蛋白表达与乳腺癌浸润、转移及预后显著相关,检测乳腺癌中CD44v6、p27蛋白表达水平,对判断乳腺癌恶性程度、复发转移潜能,评估患者预后及术后选择合理治疗方案有一定参考价值。     

抗地高辛人源小分子抗体的获取

目的 从大容量噬菌体抗体库中筛选人源性抗地高辛抗体,并构建双体抗体.方法 以固相化的地高辛对构建的大容量噬菌体抗体库进行筛选,3～4轮后挑取克隆用酶联免疫吸附测定方法鉴定其特异性,对抗地高辛阳性抗体克隆进行DNA指纹分析及测序分析.选取活性好的克隆进行改造,构建双体抗体.结果 在抗体库的筛选过程中可见到明显的富集现象,获得了4株可与地高辛特异性结合的人源抗体,经DNA指纹分析及测序分析证明为不同克隆基因,阳性克隆的可变区基因轻链均属于λ第一亚群,重链分别属于第三和第四亚群.选取4号克隆进行基因改造,构建双体抗体表达载体,获得了活性较好的双体抗体.结论 利用噬菌体抗体技术获得了人源性抗地高辛抗体,并改造成应用前景较好的双体抗体,可能为临床诊断及治疗洋地黄类药物中毒提供具有实用价值的人源抗体.     

精浆抗精子抗体阳性不育患者顶体酶、一氧化氮合酶及超氧化物歧化酶活力变化的研究

目的:研究精浆抗精子抗体(AsAb)阳性对精子顶体酶、精浆一氧化氮合酶(NOS)及超氧化物歧化酶(SOD)活力的影响。方法:精浆AsAb阳性不育者40例,对照组为40例正常生育男性。通过吸光度变化分别计算顶体酶活力(BAEE/ADH联合法)、NOS活力(氧化还原反应)、SOD活力(黄嘌呤氧化酶法)。结果:精浆As-Ab阳性组与正常生育组比较,精子顶体酶活力明显降低(P<0.01),NOS活力明显升高(P<0.01),精浆中SOD活力明显降低(P<0.01)。结论:精浆AsAb阳性引起不育可能与精子顶体酶、精浆中SOD及NOS活力改变有关。     

PCR-SSP法HLA-A、B基因分型与血清学分型的比较

目的比较聚合酶链反应-序列特异性引物(PCR-SSP)进行HLA-Ⅰ类A、B抗原位点分型的准确性,并探讨血清学分型错误发生的原因。方法用PCR-SSP以及单克隆抗体血清学分型技术对HLA-A、B分型并比较。结果34例样本PCR-SSP基因分型无假阳性和假阴性出现。PCR-SSP法与血清学比较,血清学检出错误或漏检率分别为HLA-A位点23.5%,B位点26.5%。血清学发生错误或易混淆的抗原有:A2和A68、A32和A33,B5、B60和61。结论PCR-SSP法进行HLA-A、B抗原等位基因分型具有分辨率高、特异性强、重复性好、实验过程简捷快速、分型结果较血清学更加准确可靠的优点。     

Role of intestinal bifidobacteria in the pathogenesis of gut-derived bacteria/endotoxin translocation and the effect of bifidobacterial supplement on gut barrier function following burns

Early multiple organ dysfunction syndrome appears to be facilitated with bacterial translocation in severely burn injury, yet the mechanisms of bacterial translocation remains in dispute. The aim of this study was to investigate the potential role of intestinal bifidobacteria in the pathogenesis of gut-derived bacteria/endotoxin translocation following burns and the effects of bifidobacterial supplement on gut barrier. Methods: Wistar rats were randomly divided into burn group (Burn, n=60),sham burn group (SB, n=10) in experiment Ⅰ , and burn + saline group (BS, n=30), burn + bifidobacteria group (BB, n=30), and sham-burn + saline group (SS, n= 10) in experiment Ⅱ. Animals in BB group were fed bifidobacterial preparation (5 × 109 CFU/ml) after burns, 1.5ml,twice daily. Animals in BS and SS were fed saline. Samples were taken on days 1, 3, and 5 in burn groups, and on day 3 in sham-burn groups. The incidence of bacteria/endotoxin translocation and counts of Bifidobacterium, Fungi and Escherichia coli in gut mucosa were determined with standard methods. The levels of sIgA in mucus of small intestine were measured by RIA. The positive sIgA expression in lamina propria and ileum mucosal injury was evaluated light microscopically by blinded examiners. Results: Our results showed that the incidence of bacterial translocation was increased after burns, which was accompanied by significant decrease in number of bifidobacteria but significant increase in E. coli and fungi in gut mucosa, and elevation of levels of plasma endotoxin and IL-6 (P<0. 001).The incidence of bacterial translocation was markedly reduced after 3- and 5-day supplementation of bifidobacteria compared with control group (P<0.05). The counts of mucosal bifidobacteria were increased by 4- to 40-fold,while E. coli and fungi were decreased by 2- to 30-fold and 10- to 150-fold, respectively, after bifidobacterial supplementation in contrast to control group. The damage of mucosa tended to be less pronounced after 3-day bifidobacteria-supplemented formula compared with control group [grade 2(0-6) vs. grade 4(3-6), P<0.05]. Moreover, the expression and release of sIgA was markedly augmented after 3-day bifidobacteria-supplementation formula and it returned to normal range on day 5. Conclusion: The decrease in counts and proportion of bifidobacteria in mucous membrane flora may play an important role in the development of bacteria/endotoxin translocation following thermal injury. The supplement of exogenous bifidobacteria could per se improve gut barriers, and attenuate bacteria/endotoxin translocation secondary to major burns.     

Hypertrophy of medial globus pallidus and substantia nigra reticulata in 6‐hydroxydopamine‐lesioned rats treated with L‐DOPA: Implication for L‐DOPA‐induced dyskinesia in Parkinson's disease

The medial globus pallidus plays a crucial role in generation of L‐DOPA‐induced dyskinesia in patients with Parkinson's disease. The 6‐hydroxydopamine‐lesioned rat exhibiting behavioral sensitization to L‐DOPA is one useful animal model for examining L‐DOPA‐induced dyskinesia. To determine neuropathological abnormality responsible for behavioral sensitization, the medial globus pallidus and the substantia nigra reticulata in 6‐hydroxydopamine‐lesioned rats treated with L‐DOPA were examined. Intermittent L‐DOPA treatment induced hypertrophy of the lesioned‐side of medial globus pallidus and substantia nigra reticulata of 6‐hydroxydopamine‐lesioned rats with behavioral sensitization to L‐DOPA. Additionally, coadministration of a 5‐HT_1A receptor agonist, 8‐hydroxy‐2(di‐n‐propylamino)tetralin with L‐DOPA, alleviated the hypertrophy with improvement of the behavioral sensitization. These results suggest that hypertrophy of the medial globus pallidus and substantia nigra reticulata is associated with induction of behavioral sensitization to L‐DOPA in 6‐hydroxydopamine‐lesioned rats. Therefore, neuropathological changes corresponding to hypertrophy might underlie L‐DOPA‐induced dyskinesia in patients with Parkinson's disease.