The HLA-DRB1*11 group-specific allele is a predictor for alloantibody production in the transfusion-dependent thalassemia patients

Background and ObjectivesRecently, researchers have shown an increased interest in thalassemia for detecting susceptible factors in alloimmunization development. Alloimmunization, especially against Rh and Kell blood, occurs in 30% of thalassemia dependent transfusion (TDT) patients. The aim of this study is to determine the role of HLA-DRB1*11 and HLA-DRB1*13 group-specific alleles in the production of Rh and Kell alloantibodies.Materials and Methods106 TDT patients were recruited for this study (54 responders and 52 non-responders). Responder patients developed Rh, Kell and/or specificities alloantibodies. HLA genotyping was done with Sequence-Specific Primers (SSP-PCR) and the results were compared between two groups.ResultsA significant association was found between anti-K (P=0.021, OR=2.546, 95%CI) and anti-E (P=0.049, OR=2.304, 95%CI) alloantibodies production with DRB1*11, respectively. Development of Anti-K and Anti-E alloantibodies were associated with DRB1*11 (P = 0.021, OR = 2.546, 95%CI) (P = 0.049, OR = 2.304, 95%CI), respectively. Further analysis showed that DRB1*11 is more frequent in multi responders (responder with both Rh and Kell alloantibodies) than mono-responders, 71% Versus 29%. There was not found any association between the DRB1*13 group-specific allele and the production of alloantibodies (P = 0.584, OR = 0.308, 95%CI).ConclusionsThe evidence from this study suggests that detecting the DRB1*11 group-specific allele before starting transfusion can be useful to identify susceptible patients, increase HSCT transplantation compatibility and blood transfusion management.     

HLA-DRB1＊15与162例儿童急性淋巴细胞白血病发病的关系

为了研究HLA—DRB1＊15与儿童急性淋巴细胞白血病（ALL）的关系,用特异性引物,对162例15岁以下儿童ALL患者进行HLA—DRB1＊15检测,对HLA—DRB5＊进行PCR扩增,并与1000份健康脐血做显著性比较,计算相对危险度。结果表明：儿童ALL患者DRB1＊15的抗原频率为40．12％,等位基因频率为22．62％;对照组抗原频率为30．8％,等位基因频率为16．81％。两组比较,差别有显著的统计学意义（χ^2=5．560,P=0．018,RR=1．506）。结论：儿童ALL患者HLA—DRB1＊15的抗原频率和等位基因频率均显著高于正常对照,此结果对儿童ALL的发病机制研究有重要意义。     

黑龙江地区造血干细胞志愿供者HLA-DRB1基因频率的分布特征

笔者应用PCR-SSP和PCR-SSO方法对黑龙江地区造血干细胞库9548份标本进行HLA-DRB1基因分布调查,了解其多态性。1材料与方法1.1样本选择9548份标本来自2003~2005年哈尔滨、齐齐哈尔、牡丹江、佳木斯、鸡西、大庆等地的造血干细胞志愿捐献者,每份静脉血标本5ml,EDTA抗凝。采用DNA快速提取试剂盒(Gentra公司)提取DNA,每份样本DNA均采用紫外分光光度计(Eppendof公司)测定DNA浓度和纯度,DNA浓度为(30~410)μg/ml,A260/A280为1.6~2.0。1.2试剂与仪器DR SSP UniTray(低分辨分型试剂盒(Pel-Freez公司);Micro SSOtmGeneric HL…     

HLA-DRB1基因全长序列分子克隆及测序方法的建立

目的建立可靠的HLA-DRB1等位基因全长序列的分子克隆和测序方法。方法设计、合成HLA-DRB1基因全长序列PCR引物和探索PCR反应体系,采用长距离PCR技术,对10份经PCR产物直接测序、基因型已知的标本,扩增HLA-DRB1基因5'-UTR区、6个外显子、5个内含子和3'-UTR区,全长约11 kb。PCR产物纯化后作长片段分子克隆,筛选阳性克隆,提取质粒DNA,采用自行设计的测序引物测序。结果 PCR扩增获得了特异性目的片段,克隆测序后获得了全部10种等位基因的全长序列。将HLA-DRB1等位基因全长序列划分为9个亚区,群体遗传学分析发现第2外显子的平均核苷酸变异率(π)8.653%,高于其他外显子,并且非同义突变率(dn)9.029%大于同义突变率(ds)7.846%。第5内含子的平均核苷酸变异率高达13.690%,DRB1*09∶01∶02和DRB1*07∶01∶01∶02这2个等位基因第5内含子的序列与其他等位基因序列差别较大。结论采用HLA-DRB1基因全长序列分子克隆及测序方法获得了HLA-DRB1基因各亚区多态性分布特点等基础资料。     

甘肃汉族HLA-DRB1基因多态性与白血病的相关性研究

为了研究甘肃地区汉族白血病病人易感性与HLA-DRB1基因多态性的相关性研究并寻找白血病的易感基因,采用PCR和特异性寡合苷酸探针杂交（PCR/SSO）法,对74名西北汉族白血病工人和82名健康对照者的HLA-DRB1基因分型进行了研究.结果表明：甘肃地区汉族急性髓性白血病病人HLA-DRB1＊03（x^2=8.125,P=0.004）,HLA-DRB1＊07（x^2=13.526,P=0.000）,HLA-DRB1＊08（x^2=18.855,P=0.000）和HLA-DRB1＊13（x^2=7.039,P=0.008）明显增多;慢性髓性白血病病人HLA-DRB1＊07（x^2=5.689,P=0.017）,HLA-DRB1＊11（x^2=7.73,P=0.005）,HLA-DRB1＊12（x^2=4.234,P=0.040）,HLA-DRB1＊13（x^2=38.333,P=0.000）明显增多.急性淋巴细胞白血病病人HLA-DRB1＊01（x^2=5.294,P=0.021）明显增多.这些数据与对照组比较有差异性.结论：HLA-DRB1＊03,HLA-DRB1＊07,HLA-DRB1＊08,HLA-DRB1＊13与急性髓性白血病正相关.HLA-DRB1＊07,HLA-DRB1＊11,HLA-DRB1＊12,HLA-DRB1＊13与慢性髓性白血病的发病正相关.HLA-DRB1＊01急性淋巴细胞白血病病人的发病有一定联系.可以推测,HLA构象的多态性与白血病发生有相关性.     

HLA-DRB1＊0701/02、DRB1＊1201/02等位基因与新疆维吾尔族白癜风相关性研究

[目的]探讨人类白细胞抗原（HLA-DRB1＊0701/02、DRB1＊1201/02）等位基因与新疆维吾尔族白癜风相关性.[方法]应用聚合酶链反应-序列特异性引物（PCR-SSP）技术检测300例维吾尔族白癜风患者（白癜风组）和300例维吾尔族健康者（对照组）的HLA -DRB1＊0701/02、DRB1＊1201/02等位基因,比较两组基因频率及分布频率.[结果]白癜风患者DRB1＊0701/02、DRB1＊1201/02等位基因频率显著高于对照组（P〈0.05） 儿童型发病、成人型发病、无家族病史型、局限型、泛发型、进展期的白癜风患者的HLA-DRB1＊0701/02和DRB1＊1201/02等位基因频率均显著高于对照组（P〈0.05） 有家族史和稳定期白癜风患者HLA-DRB1＊0701/02等位基因频率与正常对照组相比较有显著性差异（P〈0.05）,有家族史和稳定期白癜风患者DRB1＊1201/02与对照组相比无显著性差异（P＞0.05）.[结论]HLA -DRB1＊0701/02、DRB1＊1201/02 等位基因可能是新疆维吾尔族白癜风的易感基因.     

Unique clinical and pathological features in HLA-DRB1*0401-restricted MBP 111-129-specific humanized TCR transgenic mice

Amino acid residues 111-129 represent an immunodominant epitope of myelin basic protein (MBP) in humans with human leukocyte antigen (HLA)-DRB1*0401 allele(s). The MBP 111-129-specific T cell clone MS2-3C8 was repeatedly isolated from a patient with multiple sclerosis (MS), suggesting an involvement of MS2-3C8 T cells in the pathogenesis. To address the pathogenic potential of the MS2-3C8 T cell clone, we generated transgenic (Tg) mice expressing its T cell receptor and restriction element, HLA-DRB1*0401, to examine the pathogenic characteristics of MS2-3C8 Tg T cells by adoptive transfer into HLA-DRB1*0401 Tg mice. In addition to the ascending paralysis typical of experimental autoimmune encephalomyelitis, mice displayed dysphagia due to restriction in jaw and tongue movements and abnormal gait. In accordance with the clinical phenotype, infiltrates of MS2-3C8 Tg T cells and inflammatory lesions were predominantly located in the brainstem and the cranial nerve roots in addition to the spinal cord and spinal nerve roots. Together, these data suggest a pathogenic role of MBP-specific T cells in inflammatory demyelination within the brainstem and cranial nerve roots during the progression of MS. This notion may help to explain the clinical and pathological heterogeneity of MS.     

3438例山东汉族脐血供者HLA-DRB1等位基因频率的分布特点

为研究山东汉族人群DRB1基因的分布特征，探讨中国广大地区甚至世界其它人种利用山东脐血供者进行造血干细胞移植的可能性，应用PCR-SSP方法对山东地区3438例无血缘关系的汉族健康新生儿脐血进行了HLA-DRB1低分辨等位基因的分布调查。结果显示：在山东汉族人群中前5位高频率等位基因依次为DRB1*15(0.1817)，*07(0.1369)，*09(0.1221)，*04(0.1084)和*12(0.1038)，低频率的等位基因是DRB1*0302／0303(0.0003)，*10(0.0151)，*16(0.0262)和*01(0.0322)。通过山东汉族人群与相关文献中其它人种和不同地域的汉族人群的比较分析显示：没有任何1个DRB1等位基因是某一人种或民族所特有的，但DRB1基因在不同种族的人群中有其各自独特的人群分布特征，并且在不同地域的同一民族人群中亦有其独特的地理分布特征。结论：北方人在山东脐血库中最容易寻找到DR等位基因相合的异基因脐血供者。中国南方人和日本人在山东也完全可能寻找到DR等位基因相合的异基因脐血供者，其机率较北方人稍低。白种人甚至非裔美国人均有可能在山东脐血库中寻找到DR位点低分辨等位基因相匹配的异基因脐血造血干细胞。     

PCR-SBT检测HLA-C座位1-7外显子序列鉴别HLA-C~*07:01:01G和C~*07:02:01G组等位基因

本研究旨在探讨区分人类白细胞抗原(HLA)-C*07:01:01G和HLA-C*07:02:01G组内等位基因,并分析其与HLA-B的连锁情况。通过收集HLA-C*07:01:01G组和HLA-C*07:02:01G组标本,采用聚合酶链反应测序分析方法(PCR-SBT)检测HLA-C座位第1-7外显子编码序列,并采用PCR-SBT方法对标本进行HLA-B基因分型。结果表明,13例HLA-C*07:01:01G组标本中,4例(30.8%)标本为HLA-C*07:01:01,9例(69.2%)标本为HLA-C*07:06;连锁分析显示,HLA-C*07:06与HLA-B*44:03高度连锁。102例HLA-C*07:02:01G组标本全部为HLA-C*07:02:01;连锁分析显示,HLA-C*07:02:01与HLA-B*51:01、B*46:01、B*39:01、B*40:01、B*38:02、B*15:02高度连锁。结论:HLA-C*07:01:01G组中发现存在HLA-C*07:01:01和HLA-C*07:06,而HLA-C*07:02:01G组中以HLA-C*07:02:01为主导。     

实时荧光定量PCR法用于人类SLC01B1和ApoE基因多态性检测的方法学评价

目的：探讨在ISO15189实验室认可体系下,实时荧光定量 PCR法检测人类SLC01B1和ApoE基因多态性的方法学评价。方法本文收集经Sanger测序确定基因型的20份DNA样本。采用实时荧光定量PCR法对其中2份样本的SLCO1B1*1b、SLCO1B1*5、ApoE2和ApoE4共4个多态性位点分别批内重复扩增检测10次,以评价方法的精密度;再使用该方法扩增所有样本的4个多态性位点,每个位点检测一次,并与Sanger测序法结果比对,以评价方法的准确性;使用实时荧光定量PCR法对其中1份经梯度稀释的DNA样本进行扩增,以评价方法的最低检测限。结果2份DNA样本扩增所得Ct值变异系数均小于2．5％（≤1％）;与Sanger测序法结果对比,实时荧光定PCR法结果准确性达100％（20／20）;DNA浓度在0．964 ng／μl时,该方法仍可正确检出其基因型。结论借助经典实时荧光定量PCR法,我们能准确、快速地检测临床样本SLC01B1和ApoE基因多态性,用于指导正确合理使用他汀类药物。     

Irinotecan dose reduction in metastatic colorectal cancer patients with homozygous UGT1A1*28 polymorphism: a single-center case series

ObjectiveThe UGT1A1*28 polymorphism reduces UGT1A1 enzymatic activity, which may increase the risk of severe toxicity in patients who receive standard-dose irinotecan, such as severe neutropenia and diarrhea. This real-world study assessed the optimal irinotecan dose in terms of efficacy and toxicity in metastatic colorectal cancer (mCRC) patients homozygous for the UGT1A1*28 polymorphism and receiving FOLFIRI plus bevacizumab or cetuximab as first-line therapy.MethodsWe analyzed toxicity and treatment outcomes in seven mCRC patients who were homozygous for UGT1A1*28 and received FOLFIRI plus bevacizumab or cetuximab, with an initial irinotecan dose of 120 mg/m².ResultsSix of the seven patients tolerated 120 mg/m² irinotecan without requiring dose reductions in subsequent cycles. The overall response and disease control rates were 43.0% (3/7) and 71.4% (5/7), respectively. The median progression-free survival and overall survival were 11.0 and 33.0 months, respectively. Only one severe adverse event, grade III neutropenia (2.5%), was observed.ConclusionsmCRC patients homozygous for the UGT1A1*28 allele can tolerate irinotecan at an initial dose of 120 mg/m² with favorable oncological outcomes and toxicity profiles. Further prospective studies are warranted to optimize irinotecan-based chemotherapy in these patients.     

补肾健脾中药复方对成骨细胞中ASK1、ERO1、Caspase-12调节作用*

目的：探讨补肾健脾中药复方对成骨细胞中ASK1、ERO1、Caspase-12调节作用机制。方法：将新西兰大白兔9只随机分为中药组、西药组、空白组三组,每组3只。三组分别用中药提取液、雌二醇混悬液按中药剂量和同体积(约10ml)蒸馏水灌胃,1月后抽取含药血浆。选用1日龄SD大鼠获取原代的成骨细胞体外培养,分为中药组、西药组、凋亡空白组、正常空白组。中药组、西药组分别给予对应的含药血浆培养;凋亡空白组和正常空白组均给予常规血清培养。24小时后采用检测ASK1、 ERO1和Caspase-12的含量。结果：在ASK1方面,中药组、西药组与凋亡空白组相比,差异有统计学意义(P<0.05);而且西药组比中药组更能显著降低ASK1 mRNA的表达,其差异有统计学意义(P<0.05)。在ERO1方面,中药组、西药组与凋亡空白组相比,差异有统计学意义(P<0.05),而且西药组比中药组更能显著降低ERO1蛋白的表达,其差异有统计学意义(P<0.05)。在Caspase-12方面,中药组、西药组与凋亡空白组相比,差异无统计学意义(P>0.05)。结论：补肾健脾中药复方通过ASK1、ERO1所介导的方式对成骨细胞的凋亡进行调控。     

补肾健脾中药复方对成骨细胞中ASK1、ERO1、Caspase-12调节作用

目的:探讨补肾健脾中药复方对成骨细胞中ASK1、ERO1、Caspase-12调节作用机制。方法:将新西兰大白兔9只随机分为中药组、西药组、空白组,每组3只。三组分别用中药提取液、雌二醇混悬液按中药剂量和同体积(约10 ml)蒸馏水灌胃,1个月后抽取含药血浆。选用1 d龄SD大鼠获取原代的成骨细胞体外培养,分为中药组、西药组、凋亡空白组、正常空白组。中药组、西药组分别给予对应的含药血浆培养;凋亡空白组和正常空白组均给予常规血清培养。24 h后检测ASK1、ERO1和Caspase-12含量。结果:在ASK1方面,中药组、西药组与凋亡空白组相比,差异有统计学意义(P0.05);且西药组比中药组更能显著降低ASK1m RNA的表达,其差异有统计学意义(P0.05)。在ERO1方面,中药组、西药组与凋亡空白组相比,差异有统计学意义(P0.05),且西药组比中药组更能显著降低ERO1蛋白的表达,其差异有统计学意义(P0.05)。在Caspase-12方面,中药组、西药组与凋亡空白组相比,差异无统计学意义(P0.05)。结论:补肾健脾中药复方通过ASK1、ERO1所介导的方式对成骨细胞的凋亡进行调控。     

中国鲁南地区汉族人群HLA-A,B,DRB1高分辨等位基因多态性分析

本研究旨在分析中国鲁南地区汉族人群HLA-A、B、DRB1等位基因的高分辨多态性分布特征。采用PCR-SBT对中国鲁南地区688名无血缘关系的汉族健康人群进行HLA-A、B、DRB1位点高分辨基因分型,并对分型结果做统计学分析。结果表明：688例样本A、B、DRB1座位分别检出31、63、39个等位基因,其中频率大于0.05的常见等位基因为A＊24：02、A＊30：01、A＊11：01、A＊02：01、A＊33：03、A＊02：06、B＊13：02、B＊44：03、B＊51：01、B＊46：01、B＊15：01、B＊58：01、DRB1＊07：01、DRB1＊15：01、DRB1＊09：01和DRB1＊08：03。上述A、B、DRB1位点常见等位基因累计基因频率分别为0.7223、0.4432、0.5453。最常见的A＊-B＊-DRB1＊单倍型是A＊30：01-B＊13：02-DRB1＊07：01（0.1151）,最常见的两位点连锁单倍型分别是A＊30：01-B＊13：02（0.1303）、A＊30：01-DRB1＊07：01（0.1157）和B＊13：02-DRB1＊07：01（0.1307）。结论：获得了中国鲁南汉族人群HLA-A、B、DRB1高分辨等位基因和单倍型分布特征,为评估该地区患者找到HLA匹配供者的机率,为合理选择移植供者及移植免疫、HLA与疾病相关性研究提供了参考数据。     

血清CEA与CYFRA21-1检测对肺癌诊断价值的探讨

目的测定肺癌患者血清细胞角蛋白19片段(CEFRA21-1)及癌胚抗原(CEA)浓度,探讨其在肺癌诊断中的临床价值。方法对87例确诊肺癌患者和60例肺良性疾病患者测定CYFRA21-1及CEA水平并进行分析。结果肺癌患者血清CEA与CYFRA21-1水平显著高于肺良性疾病患者。CEA在腺癌的敏感性为61.9%,CYFRA21-1在鳞癌的敏感性为79.1%。CEA及CYFRA21-1对非小细胞肺癌的敏感性随临床分期升高而升高。结论CEA及CYFRA21-1是较好的非小细胞肺癌肿瘤标志物,CEA较适用于腺癌,CYFRA21-1较适用于鳞癌,其敏感性是监测肺癌病情的良好指标。     

Prognostic Value of Serum Von Willebrand Factor,but Not Soluble ICAM and VCAM,for Mortality and Cardiovascular Events is Independent of Residual Renal Function in Peritoneal Dialysis Patients

♦ Objective: We explored associations between markers of endothelial dysfunction and outcome events, and whether those associations were independent of residual renal function (RRF) in patients on peritoneal dialysis.♦ Methods: The study enrolled 261 incident patients and 68 healthy control subjects who were followed till death, censoring, or study end. Demographics, biochemistry, markers of inflammation (C-reactive protein) and endothelial dysfunction [soluble intercellular adhesion molecule 1 (sICAM), soluble vascular adhesion molecule 1 (sVCAM), and von Willebrand factor (vWf)] were examined at baseline. Outcome events included all-cause death and fatal and nonfatal cardiovascular (CV) events.♦ Results: Mean levels of vWf, sICAM, and sVCAM were significantly higher in patients than in healthy control subjects. Levels of sICAM and sVCAM, but not vWf, were significantly correlated with RRF. Levels of sICAM and vWf both predicted all-cause mortality and fatal and nonfatal CV events after adjustment for recognizable CV risk factors. The association between sICAM and outcome events disappeared after further adjustment for RRF. However, RRF did not change the predictive role of vWf for outcome events. Compared with the lowest vWf quartile (6.6% - 73.9%), the highest vWf quartile (240.9% - 1161%) predicted the highest risk for fatal and nonfatal CV events (adjusted hazard ratio: 2.05; 95% confidence interval: 1.15 to 3.64; p = 0.014). We observed no associations between sVCAM and RRF, or sVCAM and any outcome event.♦ Conclusions: The prognostic value of vWf, but not sICAM, is independent of RRF in predicting mortality and CV events.     

The use of TP10, soluble complement receptor 1, in cardiopulmonary bypass

Cardiopulmonary bypass (CPB) for cardiac surgery or lung transplantation initiates a systemic inflammatory response characterized by increased vascular permeability, generalized edema, abnormal lung function and oxygenation and impaired ventricular function. This post-CPB syndrome significantly contributes to postoperative morbidity and mortality. Activation of complement during CPB is a key component that initiates and augments this process. TP10, soluble complement receptor 1, is a novel complement inhibitor that is a potent inhibitor of C3 and C5 convertases, blocking activation of the complement cascade at the nexus of all three complement pathways. Recent controlled trials in humans have demonstrated that TP10 effectively inhibits complement activation during CPB. In high-risk adult patients, TP10 decreases the incidence of mortality and myocardial infarction in males but not in females following cardiac surgery. TP10 is also well tolerated and protects vascular function in infants undergoing CPB. In addition, TP10 leads to early extubation in adult lung transplant recipients. TP10 is currently positioned for clinical development in a male-only indication of cardiac surgery on CPB.     

和肽素与大内皮素-1及N末端脑钠肽对心力衰竭的预后价值

目的：通过对心力衰竭患者的随访研究,探讨和肽素与大内皮素-1（Big ET-1）及N末端脑钠肽（NT-proBNP）对心力衰竭患者的预后价值。方法研究159例心力衰竭住院患者,检测入院时血清和肽素,NT-proBNP,肌钙蛋白 I,CKMB和血浆Big ET-1与左室射血分数以及 NYHA分级。并随访观察患者出院后360～490天的心血管事件再发生情况。结果159例心力衰竭患者的中位随访385天,发生心脏事件65例。多元典型相关分析显示心力衰竭患者的年龄越大和NYHA分级越高及 LVEF越低,则心力衰竭患者的和肽素与Big ET-1及 NT-proBNP的浓度越高。Cox比例风险模型分析显示,年龄、和肽素和Big ET-1及NT-proBNP是独立的心脏事件再发生的预后因素,风险比分别为1.215,1.236,4.031和13.052。logistic回归分析显示,和肽素与 Big ET-1及 NT-proBNP是独立的心源性死亡的危险因素,优势比分别为4.003,2.477 和1.235。结论检测和肽素与Big ET-1及 NT-proBNP可对心衰患者进行心脏事件再发生的危险分层和预后分析。