小鼠UGT1 A1 mRNA在慢性肝损伤时的表达

目的 探讨慢性肝损伤对小鼠尿苷二磷酸葡萄糖醛酸基转移酶 (UGT ) 1A 1mRNA表达的影响。方法 通过四氯化碳灌胃建立小鼠慢性肝损伤模型 ,30只昆明鼠随机分 3组 :正常对照组 ,实验 1组 (给药 1个月时 ) ,实验 2组 (给药 2个月时 )。检测肝功能 ,并以RT PCR技术检测正常对照组与实验组肝脏UGT 1A 1mRNA表达的差异。结果 各组间UGT 1A 1mRNA表达差异有显著性 (P<0 .0 5 ) ,实验组UGT1A 1mRNA表达比正常组降低 ,且肝损伤程度越严重表达越低。结论 四氯化碳所致慢性肝损伤能降低小鼠肝UGT 1A

Correlation between UGT1A1 Polymorphism and Neonatal Hyperbilirubinemia of Neonates in Wuhan

This study attempts to discuss the correlation between UGT1A1＊28 as uridine diphosphate glucuronosyltransferase gene promoter and coding region Gly71 Arg gene polymorphism with neonatal hyperbilirubinemia of neonates in Wuhan. A total of 168 neonates were divided into the hyperbilirubinemia group（case group, n=108） and healthy neonates group（control group, n=60）. Their DNA was obtained through blood extraction. The gene exon mutation of UGT1A1 was detected by Sanger sequencing, which revealed the relationship between UGT1A1＊28 and Gly71 Arg polymorphism with neonatal hyperbilirubinemia of neonates. The results showed that：（1） The frequency of UGT1A1＊28 allele mutation in the case group and the control group was 9.3% and 10% respectively, with the difference being not significant between the two groups（P〉0.05）.（2） The frequency of Gly71 Arg allele mutation in the case group and the control group was 35.1% and 21.7% respectively, with the difference being significant between the two groups（P〈0.01）.（3） The serum bilirubin level of Gly71 Arg mutant homozygous and heterozygous subgroups（n=66） in the case group was 302.7±31.4 μmol/L, which was significantly higher than 267.3±28.5 μmol/L of the wild subgroup（n=42）（P〈0.01）. It was suggested that the occurrence of neonatal hyperbilirubinemia of neonates in Wuhan was not associated with UGT1A1＊28 gene polymorphism, but closely with the Gly71 Arg gene polymorphism. Meanwhile, the Arg allele mutation was related to the degree of jaundice.

尿苷二磷酸葡萄糖醛酸转移酶1A1与高胆红素血症的关系研究

目的研究尿苷二磷酸葡萄糖醛酸转移酶1A1(UGT1A1酶)在正常人和肝炎患者的水平及治疗前后的变化,同时探讨UGT1A1酶与高胆红素血症的关系。方法 83例受试者分为三组,无肝炎正常人组(A组)19例;无黄疸型肝炎组(B组)23例,血清总胆红素水平小于34μmol/L;黄疸型肝炎组(C组)41例,血清总胆红素水平大于34μmol/L。在入组基线时即抽取受试者外周静脉血并保留血清,检测UGT1 A1酶水平,并记录其肝功检验指标,B组和C组患者在恢复期再次抽取外周血并保留血清,检测UGT1A1酶水平,同时再次记录其肝功检验指标。分别比较A、B、C组受试者在基线时的UGT1A1酶的水平,比较B、C组治疗前后UGT1A1酶的水平;分析UGT1A1酶的水平与胆红素的相关性。结果基线时B、C组受试者的UGT1A1酶的水平较比A组受试者,其水平增加,差异有显著性(P=0.000;P=0.002);B、C组两者酶的水平在基线、恢复期差异无显著性(P>0.05)。B、C组UGT1A1酶在恢复期与基线的差值差异有显著性(P=0.037)。C组UGT1A1酶的水平与总胆红素、直接胆红素呈正相关,差异有显著性(r=0.223,P=0.044;r=0.241,P=0.029)。结论肝脏的炎症反应和高胆红素血症可以诱导UGT1A1酶的水平升高,在高胆红素血症时,UGT1A1酶的水平可能是胆红素代谢的一个关键因素,也是影响疾病预后的一个重要因素。

26例遗传性球形红细胞增多症的临床及基因诊断

目的:遗传性球形红细胞增多症(hereditary spherocytosis,HS)是最常见的遗传性红细胞膜缺陷病,主要表现为贫血、黄疸、脾大。由于部分患者临床表现不典型、家族史阴性,加上传统的实验室检查敏感性和特异性均较低,常导致漏诊、误诊。目前已明确ANK1、SPTB、SPTA1、SLC4A1和EPB42基因突变可引起其对应的编码蛋白质缺失,进而导致红细胞膜缺陷。本研究旨在分析HS基因诊断的可行性和临床应用价值。方法:回顾性收集2018年1月至2021年9月中南大学湘雅二医院血液内科收治的26例中国湖南HS患者的资料,分析其临床表现和实验室检测结果。应用二代测序(next-generation sequencing,NGS)结合Sanger测序,检测HS致病基因突变和胆红素代谢调控关键酶尿苷二磷酸葡萄糖醛酸转移酶1家族多肽A1(uridine diphosphate-glucuronosyl transferase 1 family polypeptide A1,UGT1A1)变异。根据美国医学遗传学与基因组学学会(American College of Medical Genetics and Genomics,ACMG)发布的《序列变异解释的标准和指南》进行致病基因变异判读。分析不同基因变异类型患者的临床特征,并对其临床诊断和基因诊断进行对比分析。结果:在26例HS患者中,贫血23例、黄疸25例、脾大24例、胆石症14例;16例有家族史,10例无家族史;25例HS致病基因突变检测结果为阳性,1例阴性。19个家系共检出18个HS致病基因杂合变异,其中14个为致病性变异,1个可能致病性变异,3个意义未明变异。SPTB突变(12个)和ANK1突变(4个)最多。变异类型以无义突变为主(9个)。SPTB突变组与ANK1突变组相比,外周血红细胞参数及溶血指标的差异均无统计学意义(均P>0.05)。ANK1突变组切脾率高于SPTB突变组,差异有统计学意义(χ^(2)=6.970,P=0.014)。不同突变类型(无义突变、移码突变、剪接位点突变及错义突变)组间外周血红细胞参数及溶血指标差异亦均无统计学意义(均P>0.05)。临床确诊的18例患者中,17例与基因诊断一致;临床疑诊患者8例,均经HS致病基因突变检测确诊。24例HS患者行UGT1A1变异检测,5例患者携带UGT1A1变异导致酶活性降低,19例酶活性正常。酶活性降低组较酶活性正常组的总胆红素(total bilirubin,TBIL)水平高,差异具有统计学意义(U=22,P=0.038)。结论:大多数HS患者有贫血、黄疸和脾大,常合并胆石症。中国湖南HS致病基因突变以SPTB和ANK1突变最常见,基因型与临床表型无明显相关性。基因诊断与临床诊断高度一致。UGT1A1酶活性降低可导致HS患者黄疸程度加重。临床联合基因诊断有利于HS的快速、精准诊断;而UGT1A1酶活性相关基因变异检测对HS黄疸评估有重要意义。

Etiology analysis for term newborns with severe hyperbilirubinemia in eastern Guangdong of China

BACKGROUND Neonatal hyperbilirubinemia is one of the common diseases of newborns that typically presents with yellow staining of skin,resulting in sequelaes such as hearing loss,motor and intellectual development disorders,and even death.The pathogenic factors of neonatal hyperbilirubinemia are complex.Different cases of hyperbilirubinemia may have a single or mixed etiology.AIM To explore the etiological characteristics of severe hyperbilirubinemia in term newborns of eastern Guangdong of China.METHODS Term newborns with severe hyperbilirubinemia in one hospital from January 2012 to December 2021 were retrospectively analyzed.The etiology was determined according to the laboratory results and clinical manifestations.RESULTS Among 1602 term newborns with hyperbilirubinemia in eastern Guangdong of China,32.20%(580/1602)was severe hyperbilirubinemia.Among the causes of severe hyperbilirubinemia,neonatal hemolysis accounted for 15.17%,breast milk jaundice accounted for 12.09%,infection accounted for 10.17%,glucose-6-phosphate dehydrogenase(G6PD)deficiency accounted for 9.14%,and the coexistence of multiple etiologies accounted for 6.55%,unknown etiology accounted for 41.72%.ABO hemolysis and G6PD deficiency were the most common causes in the 20 cases with bilirubin encephalopathy.94 severe hyperbilirubinemia newborns were tested for uridine diphosphate glucuronosyl transferase 1A1(UGT1A1)*6 variant(rs4148323,c.211G>A,p.Arg71Gly),9 cases were 211 G to A homozygous variant,37 cases were 211 G to A heterozygous variant,and 48 cases were wild genotypes.CONCLUSION The main cause for severe hyperbilirubinemia and bilirubin encephalopathy in eastern Guangdong of China were the hemolytic disease of the newborns,G6PD deficiency and infection.UGT1A1 gene variant was also a high-risk factor for neonatal hyperbilirubinemia.Targeted prevention and treatment according to the etiology may reduce the occurrence of bilirubin encephalopathy and kernicterus.