PCR－SSP快速HLA－DR52组基因分型与临床应用

采用顺序特异引物聚合酶链反应技术（ＰＣＲ－ＳＳＰ）对６５例肾移植供受者ＨＬＡ－ＤＲ５２组基因分型，同时与血清学方法对比研究。显示血清学方法错误率高达３６％；而ＰＣＲ－ＳＳＰ分型全部成功，无一例假阳性和假阴性；重复性１００％，总耗时５小时。分型结果经标准细胞株、限制性内切酶分析和寡核苷酸探针杂交等确证，特异性达１００％。结果表明：ＰＣＲ－ＳＳＰ用于ＤＲ５２组基因分型，快速、特异，结果可靠，适合于临床应用。     

高致敏肾移植供受者的HLA配型研究

目的　探讨人类白细胞抗原 (HLA)配型在高致敏受者肾脏移植中的临床意义。　方法　对 18例高致敏受者采用酶联免疫吸附法 (ELISA)检测体内预存的群体反应性抗体 (PRA IgG)水平及其特异性 ;采用补体依赖性细胞毒试验 (CDC)和微量序列特异性引物聚合酶链反应 (Micro PCR SSP)技术进行HLA I类和II类分型。　结果　 18例高度致敏受者的PRA IgG水平为 40 %～ 96 % ,平均 5 6 % ;供受者之间按传统的HLA A、B、DR抗原错配 (MM )原则 ,0～ 1MM者 5例 (2 8% ) ,2～3MM者 13例 (72 % ) ,而按交叉反应组 (CREGs)错配原则 ,0～ 1MM者 11例 (6 1% ) ,增加了 33 % ,而2～ 3MM者仅 7例 (39% ) ;肾移植术后仅 4例发生急性排斥反应 ,排斥发生率为 2 2 % ,经OKT3 治疗后逆转。　结论　CREGs配型可显著提高供受者的HLA配合率 ,良好的HLA配型对减少高致敏受者肾移植的排斥反应、提高移植物存活率具有重要临床意义     

PCR－SSP快速HLA－DR2、DR7、DR9基因配型研究

采用顺序特异性引物聚合酶链式反应（ＰＣＲ－ＳＳＰ）ＤＮＡ分型技术，首次对３５例肾移植供受者和４份标准ＤＮＡ进行ＨＬＡ－ＤＲ２、ＤＲ７、ＤＲ９基因配型。ＰＣＲ扩增条件为９４℃变性３０秒，６０℃退火５０秒，７２℃延伸４０秒，５个循环后降低退火温度至５８℃共３４个循环，即可在凝胶电泳上出现清晰可辨的特异性产物和阳性对照条带。经分子量标志初步鉴定，标准ＤＮＡ和地高辛标记的特异性探针Ｓｏｕｔｈｅｒｎ杂交确定性鉴定证实。实验结果稳定可靠，特异性和重复性达１００％，无假阳性和假阴性，耗时５小时。显示ＰＣＲ－ＳＳＰ基因配型具有高分辨度、高特异性和简捷快速的特点，适合于临床应用．     

肾移植供受者HLA－DR基因配型结果分析（附276例报告）

为了进一步提高肾移植的水平，采用顺序特异引物聚合酶链反应（ＰＣＲ－ＳＳＰ）方法用于２７６例肾移植供受者ＨＬＡ－ＤＲ基因配型，以探讨其临床可行性和实用性．结果显示：２７６例供受者３０８份样本，ＰＣＲ－ＳＳＰ分型均获成功，检测时间５小时内。供受者ＤＲ等位基因频率分布大致相同，无显著性差异。已行移植的１３５例受者分为配型选择组５１例和随机选择组８４例。配型组０～１个位点错配率５４．９％，明显高于随机组的２８．６％，而２个位点错配率为４５．１％，明显低于随机组的７１．４％，具有非常显著性差异（Ｐ＜０．０１）。研究表明，ＰＣＲ－ＳＳＰ快速ＨＬＡ－ＤＲ基因分型技术选择供受者适合于临床应用并具有很强的实用性。     

应用荧光偏振免疫法监测肾移植患者全血环孢素A浓度的临床评价

应用荧光偏振免疫法(FPIA)对274例肾移植术后患者行2312人次全血环孢素A(CsA)浓度测定。结果分析表明:(1)CsA在三联免疫抑制疗法中的理想治疗窗浓度范围是:术后第1、2、3和4～12个月全血CsA浓度分别为300～430、280～350、180～280和110～250ng/ml;(2)FPIA法具有快速、准确、操作简便,适于批量测定等优点。讨论了肝肾中毒、排斥反应、高危人群及合并用药与全血CsA浓度的关系。     

1例肾癌异基因造血干细胞移植病人的护理

造血干细胞移植是利用移植时诱导移植物抗肿瘤 (ＧＶＴ)效应 ,杀伤体内转移癌细胞 ,达到治疗肿瘤的目的[1] 。 2 0 0 1年 4月我科对 1例肾癌病人行造血干细胞移植 ,护理体会报告如下。1　病例简介　　男 ,38岁。于 2 0 0 0年 4月无明显诱因出现肉眼血尿 ,ＣＴ示右肾占位性病变 ,行右肾癌根治性切除术。同年 8月胸片示两肺肿块进行性增大 ,行健择 +氟尿嘧啶 +卡铂及紫杉醇 +氟尿嘧啶 +卡铂各两周期化疗和长期生物治疗 (罗扰素 6 0 0ＩＵ肌内注射 )无效。 2 0 0 1年 4月在我科采用异基因造血干细胞移植治疗 ,供者为其胞妹 ,ＨＬＡ抗原全相合 ,…     

肾移植后特定尿微量蛋白分析的临床意义

为探讨尿微量排泄蛋白与肾移植的关系，应用免疫散射测浊技术检测了３０例肾移植患者四种特定尿微量蛋白：尿微量白蛋白（ＵＡＬＢ）、尿转铁蛋白（ＵＴｆ）、尿α１微球蛋白（Ｕα１Ｍ）、尿免疫球蛋白Ｇ（ＵＩｇＧ）。结果显示，除ＵＩｇＧ外，ＵＡＬＢ、Ｕα１Ｍ与ＵＴｆ均与血清肌酐（Ｓｃｒ）呈显著正相关。动态观察发现，Ｓｃｒ升高前１周左右，在常规蛋白尿尚不明显时，特定尿微量蛋白已开始出现异常，并于Ｓｃｒ增高期间达到高峰。肾移植急性排斥以ＵＡＬＢ增幅最大（２０７５±１１２７ｍｇ／Ｌ），而慢性ＣｓＡ肾中毒则以Ｕα１Ｍ增高为主（９５０±３４１ｍｇ／Ｌ）。结果认为：特定尿微量蛋白变化与移植肾功能密切相关，对移植后并发症的鉴别有一定帮助     

PCR－SSP快速HLA－DR基因分型与临床应用

应用分子生物学技术，建立顺序特异引物聚合酶链反应方法（ＰＣＲ－ＳＳＰ），首次对１１２例肾移植供受者和９份标准ＤＮＡ进行ＨＬＡ－ＤＲ基因分型。每份样本同时２０管ＰＣＲ反应，扩增条件为９４℃３０秒，６０℃５０秒，７２℃４０秒，共３４个循环。即可对ＤＲ各等位基因作出准确分型，总耗时５小时。分型结果经标准ＤＮＡ，限制性内切酶分析和特异性探针杂交等予以确证，特异性和重复性１００％，无一例假阳性和假阴性。显示本方法快速、精确，适合于临床应用     

单基因肾钙质沉着症的研究进展

肾钙质沉着症常与肾结石病伴发,近年来越来越多的肾钙质沉着症被发现是由单基因病所致,其发病机制尚未完全阐明。随着分子遗传学的发展,已发现30多种基因是肾钙质沉着症的致病基因。同时,随着基因检测技术的广泛开展,更多的患者得到了早期诊断和及时干预。现就单基因肾钙质沉着症的临床和基础研究进展进行综述。     

Cerebral vasospasm diagnosis by means of angiography and blood velocity measurements

Summary We investigated 76 patients with known subarachnoid haemorrhage (SAH) in order to compare the results of angiography and non-invasive Doppler recordings of cerebral artery blood velocity in the diagnosis of cerebral vasospasm. One radiologist and one neurovascular surgeon assessed angiographic spasm visually on a four-level scale. The radiologist's ratings were the term of reference for the study.When there was angiographic spasm of the middle cerebral artery (MCA), the MCA blood velocity was higher and the blood velocity in the distal extracranical internal carotid artery (ICA) was lower than when MCA spasm was scored as absent. Analysis by Kappa statistics, a measure for the agreement between two independent judges with correction for random coincidence, revealed moderate agreement between angiographic spasm and the absolute MCA blood velocity (Kappa=0.47). However, there was substantial agreement (Kappa=0.64) between angiographic spasm and the index calculated from dividing the blood velocity in the MCA (V_MCA) by the blood velocity in the ipsilaterial ICA (V_ICA). The results indicate that this V_MCA/V_ICA index gives more appropriate information on MCA spasm.Congenitally asymmetric circles of Willis with one wide dominant ACA showed normal blood velocities. In asymmetry induced by vasospasm, the diameter of the major ACA was normal or even reduced, and the blood velocity was significantly elevated. Hence, the agreement between blood velocity recordings and angiographic findings was substantial (Kappa=0.64) when considering together the findings from both sides of the anterior circle of Willis. The agreement between angiographic spasm of the posterior cerebral artery (PCA) and the PCA blood velocity was substantial (Kappa=0.68).The two observers' rating of angiographic spasm agreed substantially on the MCA (Kappa=0.67) and the ACA (Kappa=0.71). Moderate agreement (Kappa=0.59) was obtained for the PCA. These findings show that assessing cerebral vasospasm angiographically is difficult. The present study therefore indicates that with regard to diagnosing cerebral vasospasm, the reliability of blood velocity measurements match the reliability of judging angiograms visually.This work was supported by The Norwegian Council on Cardiovascular Diseases.     

Difficulty in blood typing after infusion of plasma substitutes

Recently we experienced one incident that the patient with A blood type reacted as AB type on macroscopic observation when the blood type was tested immediatly after the infusion with Dextran₇₀ in saline. The patient was confirmed as having the A type by reexamination later. Subsequently 90 cases were subjected to blood typing following infusion with one of the seven plasma substitutes such as dextran, MF-gelatin and HE-starch. One patient demonstrated an abnormal reaction. She had marked acceleration of erythrocyte sedimentation rate (E.S.R.) and positive serological reactions for syphilis before the infusion with Dextran₇₀. It was thought that the rapid increase in high molecular colloids in circulation accelerated rouleaux formation and caused the change in agglutination thus resulted in misjudgement of blood typing.