在家系完整的单倍体联合第三方脐血移植中研究供受者HLA单倍型和等位基因错配对移植预后的影响北大核心CSCD

目的在HLA—A、-B、-C、-DRB1、-DQB14高分辨基因分型的基础上,分析供受者单倍型和等位基因错配在单倍体联合第三方脐血移植对预后的影响。方法回顾性分析2012年1月至2014年12月期间行单倍体联合第三方脐血移植的230例恶性血液病患者,采用基因测序和寡核苷酸探针杂交方法进行供受者HLA—A、-B、-C、-DRB1、-DQB1高分辨基因分型。结果供受者根据HLA—A、-B、-C、-DRB1、-DQB1基因分型结果及相合度,分为HLA-5／10、-6／10、-7／10、98／10四组,其3年总生存（OS）率分别为48．7％、59．3％、71．1％、38．3％（P=0．068）。供受者HLA-6／10相合组与HLA-5／10相合组相比,3年OS率差异有统计学意义（P=-0．041）。在供受者HLA-6／10相合组：当HLA—I类位点（HLA-A、-B、-C）相合3年OS率为61．5％,与HLA-5／10相合组相比差异有统计学意义（P=0．017）,且HLA—A位点等位基因相合3年OS及无事件生存（EFS）率分别为90．5％8n84．4％,明显高于HLA-5／10相合组（P=0．013,P=0．013）。供受者与脐血单倍型相合88例,不合51例,两者巨核系重建累积发生率分别为95．3％和86．2％（P=0．007）,粒系重建累积发生率分别为98．8％和96．1％（P=0．022）。结论在单倍体联合第三方脐血移植的供者选择中,HLA相合度及等位基因错配对预后均有意义,选择与供受者单倍型相合的第三方脐血可促进移植后造血重建。     

第一次上讲台

我小时候的梦想是当一名老师.因为我的妈妈曾经是一位老师,妈妈说:老师的职业最高尚,因为他是人类灵魂的工程师.爸爸和妈妈也希望我长大后当一名老师.但事与愿违,参加工作时我成为了一名为病人服务的白衣天使--护士.自从我穿上白大褂的那一刻起,我的梦想就封存在记忆里了.……     

实时定量RT-PCR方法检测初诊白血病患者常见分子标志物

目的 建立实时定量RT-PCR(RQ-RT-PCR)方法,检测初诊白血病患者骨髓细胞中的常见特征性分子生物学标志物的表达水平,评价其在疾病诊断和微量残留病(MRD)监测中的意义.方法 设计TaqMan探针和引物,建立RQ-RT-PCR法对常见融合基因转录本(bcr-abl、AML-ETO、PML-RARα)和abl阳性模板进行扩增,检测202例初诊白血病患者的融合基因转录本含量.结果 初诊白血病患者中的融合基因表达水平以绝对量表示,bcr-abl转录本b3a2或b2a2型拷贝数为47 614.63,bcr-abl转录本e1a2型拷贝数为98 847.53,AML1-ETO转录本拷贝数为300 029.51,PML-RARα转录本拷贝数为25 506.28.以abl校正拷贝数(NCN)表示相对量,bcr-abl转录本b3a2或b2a2型为1.05、bcr-abl转录本e1a2型为0.91、AML1-ETO转录本为5.33、PML-RARα转录本为0.55.结论 以abl校正拷贝数计算融合基因表达水平,结果更准确、直观,优于绝对定量.同时,不同类型融合基因转录本在初诊白血病患者中表达水平不同.     

741对非亲缘关系供、受者HLA高分辨基因分型结果分析

目的 研究造血干细胞捐献者资料库供、受者HLA-A、HLA-B、HLA-Cw、HLA-DRB1、HLA-DQB1位点等位基因高分辨全相合及不相合的概率.方法 采用基因测序分型(sequence based typing,SBT),序列特异性寡核苷酸探针(sequence-specific oligo nucleotide probes,SSOP),聚合酶链反应序列特异性引物(sequence-specific primers,SSP)对741对HLA-A、HLA-B、HLA-DRB1低分辨全相合的无关供、受者进行HLA-A、HLA-B、HLA-Cw、HLA-DRB1、HLA-DQB1基因高分辨检测.结果 HLA-A、HLA-B、HLA-Cw、HLA-DRB1、HLA-DQB1基因位点高分辨分型供、受者全相合的概率为0.1916(142/741);1个等位基因不相合的概率为0.1930(143/741),不相合的位点频率次序为:HLA-A＞-Cw＞-DQB1＞-B＞-DRB1;2个等位基因不相合的概率为0.2362(175/741).其中仅在一个位点(loci)上的概率为0.0175(13/741),位于两个位点上的概率为0.2186(162/741),以A+Cw组合位点不相合的概率最高,其次以B+Cw和DRB1+DQB1组合位点次之.结论 对判断无关供、受者HLA-A、HLA-B、HLA-DRB1低分辨全相合,而HLA-A、HLA-B、HLA-DRB1、HLA-Cw、HLA-DQBI高分辨基因分型全相合或部分相合的概率有重要参考价值,并且不仅能提高非亲缘造血干细胞移植的成功率,还有助于提高造血干细胞捐献者资料库数据的使用率.     

抗HLA和抗MICA抗体的表达对移植肾功能和急性排斥反应的预示作用

Objective To investigate the prediction of anti-human leukocyte antigen antibodies (HLA) and anti-major histocompatibility complex class I-related chain A antibodies (MICA) to the development of acute rejection (AR) and kidney allograft function. Methods Forty-one kidney transplant patients were prospectively tested for anti-HLA and anti-MICA. Thirty-seven patients were screened using Luminex/single-antigen beads to determine the HLA and MICA-specific antibody levels at 0,30,90, 180,360,720 and 1080 days post-transplantation. The patients and donors of HLA and MICA allele typing were determined by PCR-SSOP, and donor specific antibody (DSA) and non-donor specific antibody (NDSA) were identified.Simultaneously,their serum creatinine (SCr) levels and clinical data were analyzed. Results Nine patients (21.95 % ,9/41 ) had pre-existing anti-HLA and(or) anti-MICA, including 6 cases of anti-MICA,2 cases of anti-HLA, and one case of anti-MICA and anti-HLA. Nine patients had pre-existing DSA and NDSA. In the 37 patients, 6 patients (16.2% ) developed de novo anti-HLA, and 3 (8.1%) developed de novo antiMICA. In patients positive for de novo anti-HLA, the titer of antibody was gradually increased during the follow-up of three years. Four patients out of 9 patients with pre-existing antibodies were suffered from AR (44.4%); In 6 patients positive for de novo anti-HLA,three cases (50.0%) were suffered from AR; In three patients positive for de novo anti-MICA,no AR occurred (P＜0.05). In two patients positive for DSA of HLAⅡ antibody detected at the third and seventh day after transplantation, the renal grafts were renovecd due to rejection. The Scr levels in patients positive for pre-existing MICA with AR were higher than in those positive for pre-existing MICA without AR at each scheduled time point during the follow-up period (P＜0.05). The Scr levels in patients negative for antibodies pre-transplantation and having AR were higher than in those having no AR at each scheduled time point during the follow-up period (P＜0. 01 ). The Scr levels in patients positive for de novo HLA and MICA and having AR one month following transplantation were higher than in those negative for antibodies and having no AR (P＜0.01 ). Conclusion Pre-existing and de novo anti-HLA were the irnportant factors for the development of AR, but the mismatch of HLA and MICA alleles in donors and patients was primary causes for generation of de novo antibodies.     

肾移植患者血清中可溶性B7-H3水平的变化及其临床意义

Objective To detect the serum soluble B7-H3 (sB7-H3) in patients before and after renal transplantation, and to investigate the clinical significance. Methods The serum level of sB7-H3 in 34 patients were determined before and 3, 6, 12 months after renal transplantation. Besides, 11 health adults were elected as controls. These 34 patients were divided into two groups according to HLA and/or MICA antibodies using Luminex before operation. After transplantation, all the patients were divided into two groups according to their conditions; group Ⅰ with rejection; group S with stable renal function and no rejection. Results The serum level of sB7-H3 in all 34 patients before operation was significantly higher than health controls [(27. 10 ± 13. 61) μg/L,n = 34 vs (11.61 ±3.77) μg/L,n = 11 ,P ＜0. 01], and significantly higher in patients with antibodies than in those without before operation [(34. 96 ± 17. 37) (μ/L, n = 11 vs (23. 34 ± 9. 75) (μg/L, n = 23, P ＜ 0. 05]. There was no significant difference in the serum level of sB7-H3 between control group and S group after operation (P ＞0. 05). In group Ⅰ, the serum level of sB7-H3 was increased obviously when rejection occurred [(20. 63 ±4. 28) μg/L,n = 12; (18. 95 ±2.98) μg/L,n=6; (28.36 ±19. 83) μg/L,n = 10] , as compared with group S and control group (P＜0. 01), while at the time without rejection, there was no difference among them (P ＞0. 05). Conclusion Monitoring serum sB7-H3 after renal transplantation would be clinically useful in indicating therapeutic effect and outcome of the patients.     

第一次上讲台

我小时候的梦想是当一名老师。因为我的妈妈曾经是一位老师,妈妈说:老师的职业最高尚,因为他是人类灵魂的工程师。爸爸和妈妈也希望我长大后当一名老师。但事与愿违,参加工作时我成为了一名为病人服务的白衣天使——护士。自从我穿上白大褂的那一刻起,我的梦想就封存在记忆里了。我在护理岗位上努力工作,不断提高专科护理水平,全心全意为病人服务。我以为当老师只能是下一辈子的事了,让我没有想到的是,当我在临床第一线工作近二十个春秋时,心中的梦又复苏了——我院成为湖北省三峡大学第一临床医学院,承担着三峡大学护理本科教学任务。在科…     

肾移植术后HLA抗体对移植肾预后的影响

目的：探讨肾移植后产生HLA抗体的影响因素,以及这些抗体对移植肾肾功能的影响。方法：采用Luminex技术检测92例肾移植后患者外周血HLA抗体,并随访1年时间,研究抗体和移植肾预后的关系。结果：HLA抗体的阳性率为58.7%,HLA-Ⅰ类抗体在女性更易产生,而HLA-Ⅱ类抗体的产生和性别无关,但和移植后时间有关,移植后时间越长,HLA抗体阳性率越高;并且发现,移植前诱导治疗采用多克隆抗体者HLA抗体产生的机会较小。在移植肾预后方面,HLA-Ⅱ类抗体和移植肾晚期失功相关,而单独HLA-Ⅰ类抗体的出现和移植肾晚期失功关系不大,HLA-Ⅰ类和HLA-Ⅱ类抗体同时存在对移植肾有协同破坏作用,移植肾生存率最低。多因素分析表明,HLA-Ⅰ类和Ⅱ类抗体同时出现移植肾失功的风险为阴性者的7.9倍（OR=7.9,95%CI：3.2～30.1）,而HLA-Ⅱ类抗体阳性者移植肾失功的风险为阴性者的5.1倍（OR=5.1,95%CI：1.1～24.2）。结论：肾移植术后HLA抗体和移植肾预后密切相关,而HLA-Ⅱ类抗体和移植肾晚期失功相关。