p-mTOR、p-4EBP1及p-S6K1在结直肠腺癌组织中的表达及临床意义

目的 研究p-mTOR、p-4EBP1及p-S6K1在结直肠腺癌发生、发展过程中可能的作用及临床意义。 方法 应用免疫组化检测60例结直肠腺癌组织及40例癌旁正常组织中p-mTOR、p-4EBP1及p-S6K1的表达情况,比较各组差异,分析其与相关临床病理特征的关系,阳性率间相关性分析采用Spearman相关分析检验,检验水准α=0.05。 结果 p-mTOR、p-4EBP1及p-S6K1在结直肠腺癌组织中表达水平均高于癌旁组织,差异有统计学意义（P<0.05）。p-mTOR的表达与患者的淋巴结转移、TNM分期及分化程度有关（P<0.05）。p-S6K1的表达与患者的TNM分期及分化程度有关（P<0.05）。p-mTOR与p-4EBP1在结直肠腺癌组织中的表达有一定的正相关性（P<0.05）。p-mTOR与p-S6K1在结直肠腺癌组织中的表达有明显的正相关性（P<0.05）。 结论 p-mTOR、p-4EBP1及p-S6K1在结直肠腺癌粘膜组织中过度表达可能是结直肠腺癌发生的重要机制之一,此三种磷酸化蛋白与结直肠腺癌的发生发展存在某种内在关联。     

结直肠腺癌中PTEN和NDRG1的表达及其与预后的关系北大核心CSCD

目的探讨PTEN和分化相关基因（NDRG1）在结直肠腺癌组织中的表达及其临床病理意义。方法收集91例结直肠腺癌、30例结直肠腺瘤和21名正常肠黏膜组织标本及其临床资料,采用免疫组织化学法检测PTEN和NDRG1蛋白的表达,并结合临床资料分析其临床意义。结果PTEN和NDRG1在结直肠腺癌中的阳性表达率分别为55．0％（50／91）和76．9％（70／91）,与正常肠黏膜组和结直肠腺瘤组比较,差异有统计学意义（P〈0．05）。PTEN表达下调和NDRG1过度表达与淋巴结转移明显相关（P〈0．05）。结直肠腺癌组织中PTEN和NDRG1的表达存在负相关关系（rs＇=0．251,P=0．016）。PTEN蛋白阴性表达组的结直肠腺癌患者无病生存时间和总生存时间明显低于PTEN蛋白阳性表达组,差异有统计学意义（P〈0．05）。结论PTEN失表达有可能成为结直肠腺癌癌变早期的分子生物学标志物之一,并可作为评估结直肠腺癌患者预后的指标之一。NDRG1与结直肠腺癌的发生发展有关,但不能作为临床评估结直肠腺癌患者预后的指标。联合检测PTEN和NDRG1,可作为疑难病例的辅助诊断指标。     

血清CYFRA21—1和NSE联检对肺癌诊断的价值

本文分析了１１２例肺癌患者的血清ＣＹＦＲＡ２１ -１和ＮＳＥ的检测结果 ,并与肺良性疾病组的结果进行对比 ,认为血清ＣＹＦＲＡ２１ -１、ＮＳＥ对肺癌的诊断有较好的临床应用价值 ,现报告如下。材料和方法一、病人资料 :１１２例（男７４ ,女３８）肺癌患者为我院１９９７年５月～１９９８年１０月的住院病人 ,均经病理组织学证实。年龄３６～７６岁 ,平均５４岁。组织学分类为鳞癌４６例、腺癌３７例（包括鳞腺癌）、大细胞癌１４例、小细胞癌１５例。肿瘤分期按ＴＮＭ法 ,肺良性疾病组３２例（肺炎、慢支、肺结核、炎性假瘤等）平均年…     

缺氧诱导因子-1α基因G1790A多态性与慢性牙周炎易感性分析

目的 了解深圳地区不同分度慢性牙周炎（CP）患者缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子（VEGF）、25-羟基维生素D[25-(OH)VitD]水平及HIF-1α基因（HIF-1α）G1790A位点多态性，并探讨其与牙周临床指标的相关性。方法 选择2020年3月至2022年5月CP确诊者137例（CP组），其中男性81例，年龄16～58岁，平均年龄31.89岁；女性56例，年龄17～56岁，平均年龄32.04岁；疾病程度，轻度79例，中度31例，重度27例。同期非CP健康体检人群115例（对照组），其中男性69例，年龄15～57岁，平均年龄30.28岁；女性46例，年龄16～59岁，平均年龄32.86岁。采用酶联免疫吸附分析法检测HIF-1α和VEGF水平，并运用C8000化学发光分析仪检测25-(OH)VitD水平，同时采用限制性片段长度多态性-聚合酶链式反应分析HIF-1α G1790A位点多态性。结果 CP组HIF-1α、VEGF水平（31.37 pg/mL±7.56 pg/mL,102.35 pg/mL±29.68 pg/mL）明显高于对照组（5.62 pg/mL...     

COPD肺心病患者血浆ET—1含量变化的临床观察

本文对30例慢性阻塞性肺疾病（COPD）患者急性发作期和缓解期（稳定期）、30例肺心病患者、30例健康体检者的血浆ET-1含量进行了检测,现将结果报告如下。 对象和方法 一、对象:本院住院COPD患者30例（男20,女10）。年龄47～73岁,平均60.5岁,全部患者均分别在急性发作期和缓解期抽血测定ET-1。30例肺心病患者（男23,女7）,年龄50～80岁,平均67岁。其中8例有右心衰竭合并Ⅱ型呼衰。对照组30例均为健康体检者。     

结直肠腺癌患者外周血CNRIP1基因启动子甲基化的临床病理学分析

目的 分析结直肠腺癌患者外周血大麻素受体互作蛋白1(cannabinoid receptor interacting protein 1,CNRIP1)基因启动子甲基化程度与临床病理学参数的相关性,探讨外周血CNRIP1基因启动子甲基化检测在结直肠腺癌早期筛查及病程评估中的价值.方法 采用定量甲基化特异性PCR技术(quantitative methylation-specific PCR,qMSP)检测结直肠腺癌患者(50例)、结直肠腺瘤患者(20例)、健康志愿者(20例)外周血CNRIP1基因启动子CpG岛甲基化程度,并通过焦磷酸测序技术确定该基因启动子甲基化位点.登记结直肠腺癌患者临床病理参数.采用Fisher检验分析CNRIP1基因启动子CpG岛甲基化与结直肠腺癌临床病理参数的相关性.结果 结直肠腺癌、结直肠腺瘤患者及健康志愿者外周血CNRIP1基因启动子CpG岛的2246位点甲基化程度分别为(85.50±8.24)％、(81.33±5.81)％、(63.66％±3.61)％(P＜0.01);结直肠腺癌CNRIP基因启动子CpG岛2246位点甲基化程度分别与肿瘤直径、原发灶浸润深度、TNM分期具有显著相关性(P＜0.01),与腺癌分化程度、淋巴结转移具有一定相关性(P＜0.05).结论 CNRIP1基因启动子CpG岛2246位点甲基化程度与结直肠腺癌病程进展相对一致,可作为结直肠腺癌早期筛查及病程评估的标志物.     

钠/氢交换调节因子1是结直肠腺癌临床预后的预测因子

目的寻找与结直肠腺癌侵袭和迁移相关的因子。方法收集TCGA及GEO数据库中结直肠腺癌相关数据进行SAM差异表达基因分析;将得到的差异基因通过DAVID进行聚类分析;寻找与侵袭迁移相关的基因;并对TCGA数据库中的患者临床信息进行回顾性分析及生存预后分析;同时收集50例具有详细临床及预后信息的结直肠腺癌组织及配对癌旁组织进行免疫组织化学染色来验证上述结果,并绘制ROC线进行评估。结果 DAVID聚类分析发现6个与迁移相关的基因集(P0.05),其中下调的基因有DLC1、EPB41L3、KIT、PARVA、SLC9A3R1和TPM1。而SLC9A3R1编码的钠/氢交换调节因子1(NHERF1),是一种能与细胞骨架及多种信号蛋白相互作用的多功能连接蛋白。TCGA数据分析显示,结直肠腺癌中NHERF1的mRNA水平较正常结直肠组织降低。独立临床队列数据的免疫组化结果显示,在高级别及发生淋巴结转移的组织中NHERF1表达降低。结论 NHERF1表达降低是影响结直肠腺癌患者生存期的一个独立影响因子,可能与其促进肿瘤转移有关。NHERF1表达降低是一个潜在的诊断和预测结直肠腺癌不良预后的指标。     

Expression and clinical significance of CD151 and integrin α3β1 in colorectal tumors

目的:探讨CD151和整合素α3β1在直肠腺瘤及结直肠腺癌组织中的表达及其与临床各个病理因素之间的关系,并且研究两者的相关性.方法:应用免疫组织化学双染方法对正常结直肠黏膜、结直肠腺瘤及结直肠腺癌组织各120例进行CD151和整合素α3β1检测,并进行Kaplan-Meier生存分析.采用Spearman等级相关分析CD151和整合素αβ1之间的相关性.结果:CD151结直肠正常黏膜、腺瘤、腺癌组织的阳性率分别为21.7％、52.5％、72％,腺瘤和腺癌组织分别与正常黏膜比较均具有统计学意义.整合素α3β1在结直肠正常黏膜、腺瘤、腺癌组织的阳性率分别为34.2％、55％、70％,腺瘤和腺癌组织分别与正常黏膜比较均具有统计学意义.在结直肠腺癌中,CD151和整合素α3β1的表达与患者年龄、性别和肿瘤的部位、大小无相关性,与肿瘤的分化程度、浸润深度、淋巴结转移及Duke's分期有关.CD151和整合素α3β1在大肠正常黏膜、腺瘤及腺癌组织中的表达经双变量相关分析,表达呈正相关.从图的Kaplan-Meier生存曲线及Log-Rank检验可知,CD151+、α3β1+、CD151+α3β1+与大肠癌患者5年生存期密切相关,是影响大肠癌预后的因素.结论:CD151和整合素α3β1在大肠癌的表达密切相关,提示CD151与整合素α3β1复合物存在于大肠癌,其表达对预后产生明显的影响.CD151与整合素α3β1联合表达是临床预后判断的可靠指标.     

肺鳞癌和肺腺癌中Pin1与cyclin D1的表达及意义

目的探讨Pin1（the peptidy-proly1 isomerase）在肺鳞癌和肺腺癌组织中的表达及其与组织类型、肿瘤分化、分期及淋巴结转移关系，并分析其与细胞周期素（cycinD1）是否存在相关性。方法采用SP免疫组化方法检测69例肺鳞癌和肺腺癌组织中Pin1和cyclin D1的表达。结果免疫组化结果显示：Pin1与cyclinD1在肺鳞癌、肺腺癌中的阳性表达率分别为78．3％（54／69）、52．2％（36／69）。Pin1与cyclinD1在肺癌组织中的表达水平均高于正常肺组织。Pinl的表达与患者的性别、年龄、组织类型、分化、淋巴结转移与否等临床病理学参数无相关。cyclin D1的表达与分化程度负相关（P=0．0274），而与患者的性别、年龄、组织类型、淋巴结转移与否等临床病理学参数无相关。Pin1和cyclin D1二者的表达呈正相关（P=0．0048）。结论在肺癌中，Pin1的表达高于正常肺组织，但与临床病理学参数无关，而与cyclin D1正相关。其对cyclin D1的影响可能是Pin1发挥作用的主要机制之一。     

Wnt信号通路成员蛋白NKD1、β-catenin和Cyclin D1在结直肠腺癌中的表达及意义

目的观察NKD1、β-catenin和Cyclin D1蛋白在结直肠正常黏膜、腺瘤和腺癌组织中的表达,探讨NKD1、β-catenin和Cyclin D1在结直肠正常黏膜-腺瘤-腺癌这一癌变过程中的表达变化及临床意义。方法应用免疫组化SP法检测38例结直肠正常黏膜、70例腺瘤、128例腺癌组织中NKD1、β-catenin和Cyclin D1蛋白的表达。结果 NKD1在结直肠正常黏膜、腺瘤、腺癌组织中的阳性率依次降低(P0.05),其在轻度异型增生腺瘤组织中的阳性率高于重度异型增生腺瘤组织(P0.05);β-catenin异位表达率与Cyclin D1阳性率在结直肠正常黏膜、腺瘤、腺癌组织中均依次升高(P0.01),且Cyclin D1在重度异型增生腺瘤组织中的阳性率高于中度及轻度异型增生腺瘤(P0.05)。NKD1、β-catenin、Cyclin D1在结直肠腺癌组织中的表达均与癌分化程度、Duke分期、淋巴结转移有关(P0.05)。在结直肠腺癌组织中NKD1的阳性率和β-catenin异位表达率呈负相关(r_s=-0.645,P0.01)。在结直肠腺癌组织中β-catenin异位表达率和Cyclin D1阳性率呈正相关(r_s=0.618,P0.01)。结论 NKD1、β-catenin、Cyclin D1三者可能参与结直肠肿瘤的发生、发展,有望成为评价结直肠腺癌恶性程度及预后的指标。     

扩散峰度成像在预测直肠癌KRAS基因突变中的应用价值

目的 探讨扩散峰度成像(DKI)定量参数在预测直肠癌KRAS基因突变中的应用价值。方法 回顾性分析山西省肿瘤医院2016年11月-2018年6月经病理证实的152例直肠腺癌患者的资料,其中男92例,女60例;年龄33～86岁,平均61岁。患者均于术前行常规MRI和功能DKI序列检查,由2名放射科医师双盲勾画感兴趣区,应用MatLab软件计算两组患者DKI定量参数平均表观扩散系数(MD)、平均峰度(MK)以及表观扩散系数(ADC),并采用组内相关系数进行一致性分析。术后均进行KRAS基因检测,依据检测结果将患者分为野生组和突变组两组,采用独立样本t检验对比两组患者的MD、MK、ADC。绘制DKI定量参数诊断KRAS基因突变的受试者工作特征曲线(ROC) ,根据约登指数确定各定量参数的最佳诊断阈值,以及相应的灵敏度、特异度,并采用DeLong检验比较各定量参数的ROC的曲线下面积(AUC)。结果 152例直肠癌患者中,KRAS基因突变组74例,野生组78例,基因突变率为48.6%(74/152)。突变组患者的ADC、MD、MK值分别为(1.18±0.18)×10^-3 mm²/s、(1.28±0.18)×10^-3 mm²/s、0.97±0.11,野生组分别为(1.33±0.18)×10^-3 mm²/s、(1.42±0.17)×10^-3 mm²/s、0.82±0.09;突变组ADC、MD值均小于野生组患者,而MK值则大于野生组患者,差异均有统计学意义(t=5.424、4.882、-8.809, P值均＜0.01)。ROC曲线显示,ADC、MD、和MK值预测KRAS基因的AUC分别为0.758、0.740、0.845,灵敏度分别为75.7%、82.4%、77.0%,特异度分别为68.0%、57.8%、84.6%。DeLong检验结果显示,MK值的AUC明显高于ADC和MD值(P值均＜0.01),而ADC值和MD值间AUC差异无统计学意义(P＞0.05)。结论 DKI定量参数MD、MK和ADC值,在预测直肠癌的KRAS基因突变方面均有一定的价值,其中MK值有更高的AUC和特异度,有更高的诊断价值。     

弥散峰度成像与直肠癌临床病理预后因素的相关性研究

目的 探讨弥散峰度成像(DKI)定量参数和表观扩散系数(ADC)与直肠癌临床病理预后因素之间的潜在关系,为临床预测评估直肠癌的恶性程度提供一定的参考依据。方法 对2016年11月—2017年4月山西省肿瘤医院122例术前行MRI检查的直肠腺癌患者的影像和临床资料进行回顾性分析,其中女48例(39.3%)、男74例 (60.7%),年龄42~81岁。利用相关软件测得平均表观扩散系数(MD)、平均峰度(MK)和ADC值,通过独立样本t检验或Mann-Whitney U检验、ROC曲线和Spearman相关性分析进行统计学分析。结果 直肠癌高T分期及淋巴结转移与低MD值(r=-0.367、-0.240)和低ADC值(r=-0.391、-0.254)相关,不同分组间差异均有统计学意义(P值均＜0.05);同样,不同组织病理学分级的MD值之间差异亦有统计学意义(P＜0.05)且呈负相关(r=-0.210, P＜0.05);随着组织病理学分级的升高、淋巴结受累、瘤周血管浸润(LVI)或神经侵犯及环周切缘(CRM)受侵,MK值相应增大,差异均有统计学意义(P值均＜0.05)呈正相关(r=0.478、0.206、0.237、0.228, P值均＜0.05)。MD、MK和ADC值均与淋巴结转移有相关(P值均＜0.01)性;ROC曲线显示MK值较其他参数在诊断淋巴结转移与否的曲线下面积高为0.784(95%CI 0.703~0.865),当其阈值取0.984时,具有较高的敏感度及特异度,分别为65.9%和88.7%。结论直肠癌DKI定量参数值和ADC值,尤其MK值,与其重要的临床病理预后因素均有明显相关性,对于预测直肠癌患者预后有一定意义。     

UGT1A1 基因多态性与转移性结直肠癌伊立替康化疗毒性及疗效的关系

目的: 探讨 UGT1A1 ^*28和 UGT1A1 ^*6基因多态性与伊立替康治疗转移性结直肠癌患者的不良反应和疗效之间的关系。方法: 外周血中抽提基因组DNA,采用PCR扩增目的基因片段,直接测序法分析2010年4月至2012年3月在我院做基因检测的207例消化道肿瘤患者 UGT1A1 ^*28和 UGT1A1 ^*6基因多态性的分布情况,并对其中56例采用含伊立替康方案化疗的转移性结直肠癌患者出现的不良反应情况、肿瘤进展时间及化疗疗效进行观察并记录,比较不同基因型患者之间的差异。结果: 207例消化道肿瘤患者中, UGT1A1 ^*28位点野生型TA6/6有164例(79.2%),杂合突变型TA6/7有41例(19.8%),纯合突变型TA7/7有2例(1.0%); UGT1A1 ^*6位点野生型G/G有154例(74.4%),杂合突变型G/A有51例(24.6%),纯合突变型A/A有2例(1.0%)。在56例转移性结直肠癌患者中,^*6位点突变型(G/A和A/A)可以增加发生3级以上腹泻(38.9% vs 7.9%,P<0.05)和中性粒细胞减少(61.1% vs 29.0%,P<0.05)的风险;^*28位点突变型(6/7和7/7)可以增加发生3级以上血小板减少(33.3% vs 2.1%,P<0.05)的风险;肿瘤进展时间和化疗疗效在^*28和^*6位点各基因型之间差异无统计学意义。结论: 在采用含伊立替康方案化疗的转移性结直肠癌患者中, UGT1A1 ^*6位点突变型增加发生3级以上腹泻和中性粒细胞减少的风险; UGT1A1 ^*28位点突变型增加发生3级以上血小板减少的风险。     

Distribution of selected gene polymorphisms of UGT1A1 in a Saudi population

Introduction

Glucuronidation is an important phase II pathway responsible for the metabolism of many endogenous substances and drugs to less toxic metabolites, which undergo renal excretion. The aim of the current work was to evaluate genotype and allele frequencies of certain UDP-glucuronosyltransferase 1A1 (UGT1A1) variants in an Arab population.

Material and methods

Genomic DNA was isolated from 192 healthy unrelated Saudi males of various geographic regions and genotyping of UGT1A1*6, *27, *36, *28, *37, and *60 was carried out using polymerase chain reaction (PCR) amplification followed by direct sequencing.

Results

The most common allele for (TA) repeats was the wild type (TA)6 with a frequency of 74.3% followed by the mutant (TA)7 (i.e., UGT1A1*28) with a frequency of 25.7%. The distribution of UGT1A1*60 allele was 62.4% among subjects with the homozygous mutant genotype of 35.4%, while the wild type variant represents 10.6% only. Both UGT1A1*6 and *27 were not detected as all screened subjects showed a homozygous wild type pattern. Similarly, UGT1A1*36* and *37 were either not present or rarely found, respectively. In comparison to other populations, the frequency of UGT1A1*60 and *28 in the studied population was less than that of African Americans but higher than Asians. The geographical origin of the study subjects also implied some differences in genotype distribution of (TA) repeats and UGT1A1*60.

Conclusions

Our data indicate that Saudis harbor some important UGT1A1 mutations known to affect enzyme activity. Additional studies are warranted to assess the clinical implications of these gene polymorphisms in this ethnic group.     

弥散峰度成像对局部进展期直肠癌新辅助放化疗疗效的评估价值

目的 探讨弥散峰度成像 (DKI) 定量参数和表观扩散系数(ADC)对局部进展期直肠癌患者新辅助放化疗疗效的应用价值。方法 回顾性分析。纳入2016年12月—2018年5月山西省肿瘤医院直肠癌患者108例,其中女42例、男86例,年龄42~81岁,均行新辅助放化疗,并于新辅助放化疗后4~8周接受手术治疗。根据病理结果按肿瘤退缩评级系统(TRG)进行评分分级,TRG1~2级为新辅助放化疗疗效好,TRG 3~5级为新辅助放化疗疗效差。然后利用Matlab软件测得两组治疗前后的平均弥散系数(MD)、平均峰度系数(MK)和ADC,并通过配对样本t检验、独立样本t检验、受试者操作特征(ROC)曲线和组内相关系数进行统计分析。结果 108例患者术后评估疗效好者27例(25%),其中肿瘤退缩分级评分(TRG)1级5例,TRG 2级22例;疗效差者81例(75%),其中TRG 3级41例,TRG 4级26例,TRG 5级12例。新辅助放化疗前,疗效好者ADC与MD值均低于疗效差者,MK值高于疗效差者,但差异均无统计学意义 (P值均＞0.05)。新辅助放化疗后,疗效好患者的MK值低于疗效差者,ADC与MD值显著高于疗效差者,差异均有统计学意义(P值均＜0.05),并且两组间的MD、ADC和MK比值比较差异均有统计学意义(P值均＜0.05)。ROC曲线结果提示,新辅助放化疗后的治疗后MK值较其他参数在评估新辅助放化疗疗效好的曲线下面积大(0.823),诊断效能最高,当其最佳阈值为0.635时,敏感度为76.5%,特异度为63.0%。结论 直肠癌DKI的MK、MD定量参数值和ADC值,尤其治疗后MK值,对于预测局部进展期直肠癌患者新辅助放化疗疗效有一定意义。     

Validation of rapid polymerase chain reaction-based detection of all length polymorphisms in the UGT 1A1 gene promoter.

UDP glucuronosyltransferase (UGT) 1A1 gene promoter polymorphism can affect the expression level of the UGT 1A1 enzyme. The polymorphism consists of an insertion of a TA nucleotide sequence into a (TA)6TAA sequence in the gene promoter resulting in (TA)7TAA (UGT1A1*28). This results in a reduced UGT 1A1 expression with 70% less glucuronidation capacity for bilirubin and other UGT1A1 substrates. Other polymorphisms include (TA)8TAA (UGT1A1*37) and (TA)5TAA (UGT1A1*36). The longer the TA repeats the lower the enzyme expression level. The anticancer agent irinotecan is metabolized to the active SN-38, which is further glucuronidated and detoxified by UGT 1A1. Decreased glucuronidation leads to SN-38 accumulation with severe neutropenia and diarrhea. We have developed a rapid polymerase chain reaction (PCR)-based detection of all length polymorphisms in the UGT 1A1 gene promoter. It uses PCR and DNA fragment analysis using an ABI Genetic Analyzer. Thirty-two blood samples were analyzed for UGT 1A1 promoter polymorphism. We found 2 (TA)(5)TAA/(TA)(5)TAA, 4 (TA)(5)TAA/(TA)(6)TAA, 2 (TA)(5)TAA/(TA)(7)TAA, 9 (TA)(6)TAA/(TA)(6)TAA, 11 (TA)(6)TAA/(TA)(7)TAA, 2 (TA)(7)TAA/(TA)(7)TAA, and 2 (TA)(7)TAA/(TA)(8)TAA in our sample group. To confirm the results, 6 samples with different repeats were also analyzed by DNA sequencing method. This is a rapid and reliable method for analysis of the promoter length polymorphisms of UGT 1A1 gene.     

Establishment of multiplex hybri-probe method--analysis using the polymorphism of UGT1A1 gene

There are a number of methods for gene analysis of a point mutation and deletion/insertion of several nucleotides. In 2011, we reported an improved hybridization probe methods (Hybri-Probe method) that are highly sensitive and accurate, and excellent in cost and time effectiveness. Here, we have developed the Multiplex Hybri Probe method for several types of mutations or polymorphisms including the microsatellite polymorphisms, especially of palindromic sequence such as (TA)n and (GC)n. In addition, several mutations are analyzed at a time. In this research we focused on the three types of polymorphism on the Uridine diphosphate glucuronyltransferase (UGT) gene. Design of the probes for the detection of UGT1A1*6(211 G --> A G71R) and UGT1A1*27 (686 C --> A P229Q) was not difficult because the mutations were a single base substitution. However, UGT1A1*28 (A (TA)6TAA --> A (TA)7TAA) has tandem and palindromic sequence. Since the probes to detect the mutation in such sequence resulted in failure, we made several mismatches, i.e., TATATATATATA --> TATGTGTATATA. As a result, the probes designed for the three polymorphisms above did not overlap in the Tm and were separated by approximately 10 degree intervals between 63.2 degrees C and 37.5 degrees C. In this Multiplex Hybri-Probe method, the three kinds of probe are added tube into the PCR product in the same tube and followed by the measurement of the Tm using the LightCycler. Thus, it is very simple, and performed by one step for any mutation including the microsatellite polymorphisms, and it also has good cost performance and favorable time efficiency. It takes two hours including the running time of PCR for completion of the analysis. It may also be available to the detection of other gene abnormalities.     

Prevalence of uridine glucuronosyl transferase 1A1 (UGT1A1) mutations in Malay neonates with severe jaundice

A number of genetic risk factors have been implicated in the development of neonatal severe hyperbilirubinaemia. This includes mutations in the uridine glucoronosyl transferase 1A1 (UGT1A1) gene which is responsible for unconjugated hyperbilirubinemia in Gilbert's Syndrome. We studied the prevalence of UGT1A1 gene mutations in a group of Malay neonates to determine whether they are risk factors to severe neonatal jaundice. One hundred and twenty-five Malay neonates with severe hyperbilirubinemia were studied. Ninety-eight infants without severe hyperbilirubinaemia were randomly selected from healthy Malay term infants (controls). DNA from EDTA cord blood samples were examined for UGT1A1 mutations nt211G > A and nt247T > C using established Taqman SNP genotyping assays and the UGT1A1*28 variant was detected by the Agilent 2100 bioanalyzer. All samples were also screened for common Malay G6PD variants using established techniques. The frequency of UGT1A1 211G > A mutation is significantly higher in the severely hyperbilirubinemic group (13%) than the control group (4%; p = 0.015) and all the positive cases were heterozygous for the mutation. There was no significant difference in the frequency of UGT1A1*28 mutation between the severely hyperbilirubinemic (3.5%) and the control group (0.01%; p = 0.09). None of the neonates in both groups carried the nt247 T > C mutation. The prevalence of G6PD mutation was significantly higher in the severely jaundiced group than control (9% vs 4%; p = 0.04). In conclusion, nt 211 G > A alleles constitute at least 12% of UGT1A1 mutations underlying unconjugated hyperbilirubinemia and appears to be a significant independent risk factor associated with severe neonatal hyperbilirubinemia in the Malay newborns.     

A case of concomitant Gilbert's syndrome and hereditary spherocytosis

We describe moderate hyperbilirubinemia in a 28-year-old man who suffered from gallstones and splenomegaly, with combined disorders of hereditary spherocytosis (HS) and Gilbert''s syndrome (GS). Since it is difficult to diagnose HS in the absence of signs of anemia, we evaluated both the genetic mutation in the UGT1A1 gene and abnormalities in the erythrocyte membrane protein; the former was heterozygous for a UGT1A1 allele with three mutations and the latter was partially deficient in ankyrin expression. This is the first report of the concomitance of HS and GS with three heterozygous mutations [T-3279G, A (TA)7TAA, and G211A] in the UGT1A1 gene.     

Fast diffusion kurtosis imaging (DKI) with Inherent COrrelation‐based Normalization (ICON) enhances automatic segmentation of heterogeneous diffusion MRI lesion in acute stroke

Diffusion kurtosis imaging (DKI) has been shown to augment diffusion‐weighted imaging (DWI) for the definition of irreversible ischemic injury. However, the complexity of cerebral structure/composition makes the kurtosis map heterogeneous, limiting the specificity of kurtosis hyperintensity to acute ischemia. We propose an Inherent COrrelation‐based Normalization (ICON) analysis to suppress the intrinsic kurtosis heterogeneity for improved characterization of heterogeneous ischemic tissue injury. Fast DKI and relaxation measurements were performed on normal (n = 10) and stroke rats following middle cerebral artery occlusion (MCAO) (n = 20). We evaluated the correlations between mean kurtosis (MK), mean diffusivity (MD) and fractional anisotropy (FA) derived from the fast DKI sequence and relaxation rates R₁ and R₂, and found a highly significant correlation between MK and R₁ (p < 0.001). We showed that ICON analysis suppressed the intrinsic kurtosis heterogeneity in normal cerebral tissue, enabling automated tissue segmentation in an animal stroke model. We found significantly different kurtosis and diffusivity lesion volumes: 147 ± 59 and 180 ± 66 mm³, respectively (p = 0.003, paired t‐test). The ratio of kurtosis to diffusivity lesion volume was 84% ± 19% (p < 0.001, one‐sample t‐test). We found that relaxation‐normalized MK (RNMK), but not MD, values were significantly different between kurtosis and diffusivity lesions (p < 0.001, analysis of variance). Our study showed that fast DKI with ICON analysis provides a promising means of demarcation of heterogeneous DWI stroke lesions.