南宁市2685例贫血患儿α-地中海贫血基因诊断分析

目的 了解南宁地区贫血患儿α-地中海贫血（地贫）发生情况、基因类型及频率.方法 采用单管多重PCR体系扩增结合琼脂糖凝胶电泳和反向点杂交技术,对诊断为贫血的患儿2685例进行常见α-地贫基因检测,对HbA2和HbF升高者再进行β-地贫基因检测.结果 2685例患儿中,检出α-地贫1243例（46.29%）,22种基因类型,主要为--sea /αα（23.76%）,其次为-α3.7/αα（4.69%）,--sea/-α3.7（3.84%）,--sea/αCSα（3.17%）;包括6种等位基因1563个,主要为--sea（56.62%）,其次为-α3.7（15.74%）和αCSα（14.52%）;α复合β地贫检出率为5.18%.结论 南宁地区贫血患儿α-地贫基因突变率、中间型α-地贫及复合β地贫发生率均较高,应在该类患儿中进行基因诊断,为该类患者的病因诊断和治疗提供依据.     

东莞地区孕龄人群地中海贫血基因检测结果分析

目的了解东莞地区孕龄人群地中海贫血(简称地贫)基因检测结果,为地贫防控工作提供依据。方法以参加东莞市免费婚前健康和孕前优生联合检查的孕龄男女为研究对象,将MCV≤80 fl和或MCH≤27 pg作为血筛查阳性指标,筛查阳性者进行地贫基因检测。结果 74 098例孕龄人群中,地贫初筛阳性11 012例,初筛阳性率为14.86%。检出地贫基因携带者7 214例,检出率9.74%;其中α-地贫基因携带者5 002例,检出率6.75%;β-地贫基因携带者1 973例,检出率2.66%;α-地贫复合β-地贫基因携带者239例,检出率0.32%;发现117对夫妇有妊娠重型地中海贫血胎儿的可能,产前诊断的需求率为65.73%。结论东莞地区育龄人群地贫基因携带高,产前诊断需求率高,医疗卫生部门应重视地贫防控,加强对孕龄人群进行地贫筛查。     

血常规检测对地中海贫血与缺铁性贫血诊断的价值分析

目的：探究与分析血常规检测对地中海贫血与缺铁性贫血诊断的价值分析。方法选取我院抽取的60例缺铁性贫血患者为实验组A,抽取35例轻型地中海贫血的患者为实验组B,并选取55例血常规正常的为对照组。全部的样本都将进行血细胞计数仪血常规测定,血红蛋白电泳仪进行血红蛋白电泳分析,贫血基因检测和血清铁蛋白检测。比较样本的红细胞计数（RBC）,血红蛋白含量（HGB）以及平均红细胞体积（MCV）,红细胞分布宽度（RDW）。结果地中海贫血患者的RBC,HGB比缺铁性贫血患者的高（P＜0.05）,同时缺铁性贫血患者的RDW明显高（ P＜0.05）。结论血常规检测中的检测红细胞体积,血红蛋白电泳,平均红细胞体积,血红蛋白含量对于地中海贫血以及缺铁性贫血的患者有一定的诊断意义,且检测较为简单,方便,快速,并能准确讲贫血类患者与正常人的血常规进行筛查。故值得在临床上大力推广应用。     

新生儿脐血Hb Bart''s水平作为α-地中海贫血携带者早期筛查指标临床应用评价

目的将新生儿脐血Hb Bart'水平作为α-地中海贫血携带者或患儿早期筛查指标进行临床应用评价.方法对1006份新生儿脐血标本同时采取琼脂糖碱性血红蛋白电泳和分子筛查技术分别进行Hb Bart'定量和α-地中海贫血基因分析.结果在上述标本中,检出Hb Bart'阳性样品64份,阳性率为6.36%.此外,还检出异常Hb 6例,在这些阳性样品中,检出2类5种α-地中海贫血基因共64份,其余的906份阴性样品中尚检出α-地中海贫血基因阳性38份,故该市户籍人群中总的α-地中海贫血基因携带率为10.44%.Hb Bart'水平筛查法对正常基因型和α-地中海贫血基因阳性样品的诊断符合率分别为99.8%和62.1%,假阳性率为3.1%;而对静止型α-地中海贫血的漏检率高达36.2%.结论Hb Bart'水平筛查法具有简便、快速且经济,并能对各型α-地中海贫血作出临床早期诊断,但对静止型α-地中海贫血则例外.     

北海地区新生儿脐血血红蛋白电泳实验结果分析

目的了解北海地区新生儿地中海贫血的发病率,为临床诊断及预防提供依据。方法对本院2006年7月～2010年6月共5766份新生儿脐血标本进行琼脂糖电泳,并对Hb区带进行扫描定量分析。结果在5766份脐血标本中,检出Hb Bart阳性（≥1.0%）437例,阳性率为7.579%。根据Hb Bart含量推算,静止型、标准型α-地中海贫血、HbH病、HbBart胎儿水肿综合征的阳性率分别为2.099%、5.376%、0.087%和0.017%。另外发现异常血红蛋白病共17例,发生率为0.295%,其中HbE11例,HbG2例,HbC-S1例,HbJ 2例,HbK1例。结论北海市属于地中海贫血高发区,使用全自动电泳仪对脐血定量检测分析能早期、方便地对新生儿α-地中海贫血进行筛查。     

MCV、MCH及SF在妊娠合并地中海贫血的临床应用

目的：探讨MCV、MCH及血清铁蛋白联合应用在地中海贫血中的诊断意义。方法健康对照组60例,地中海贫血杂合子携带者120例,应用Beckman-Coulter （贝克曼-库尔特） LH750型五分类血细胞分析仪检测红细胞MCV、MCH。用化学发光法测定血清铁蛋白。结果地贫组和IDA组MCV、MCH 均减低,两者与健康对照组比较,差异均有统计学意义（ P＜0.01）。地贫组与IDA组比较,地贫组MCV较IDA组明显减低,两者之间差异有统计学意义（ P＜0.01）。 IDA组SF降低,而地贫组SF不低,两者之间差异有统计学意义（ P＜0.01）。结论（1）MCV、MCH及血清铁蛋白可作为筛查孕妇地贫的有效指标;（2）做好产前地贫的筛查,对预防和降低重型地贫患儿出生具有重要意义。     

微小RNA对β-地中海贫血中γ-珠蛋白基因的调控机制研究进展

β-地中海贫血（β-地贫）是β-珠蛋白基因发生突变导致β-珠蛋白合成减少或缺如,以溶血性贫血为主要临床特征的一类单基因遗传病。目前暂无治疗该病的特效药。研究已证实,一些转录因子通过调控γ-珠蛋白基因增加胎儿血红蛋白表达,从而缓解无效造血,改善β-地贫患者的临床症状,降低对输血依赖性。随着表观遗传学研究的不断深入,发现微小RNA（microRNA,miRNA）不仅在β-地贫患者中存在表达失衡,也参与γ-珠蛋白基因的转录调控。全面了解miRNA对γ-珠蛋白基因的调控作用能够为阐明β-地贫的致病机制并对该病的精准治疗提供新的思路和方向。     

PCR结合寡核苷酸分子杂交技术在基因诊断中的应用——附一例β地贫(IVS—Ⅰ—5G→C)杂合子的报告

本文报告采用~(32)P标记的寡核苷酸探针与聚合酶链式反应(PCR)扩增的β珠蛋白基因的特异DNA片段进行分子杂交,鉴定β地中海贫血(地贫)的基因突变,检出1例病人为地贫基因突变(IVS-1-5G→C)的杂合子,为临床开展其它遗传病及病毒、肿瘤等疾病的基因诊断提供了实用、有效的途径.     

北海市新生儿G6 PD缺乏症与病理性黄疸的相关性调查

目的：了解北海市新生儿葡萄糖6磷酸脱氢酶（ G6PD）缺乏症的发生率及与病理性黄疸的相关性,以便对黄疸进行早期干预,有效减轻G6PD缺乏新生儿溶血的程度和避免发生核黄疸。方法对本院2010年9月-2012年10月共12182份新生儿脐血标本用NADP＋氧化还原酶法定量检测G6PD活性,结合黄疸指数与临床诊断比对分析。结果本市新生儿G6PD缺乏症总发生率4.94％,男婴发生率7.93％,女婴发生率1.50％;G6PD正常新生儿病理性黄疸发生率8.90％, G6PD缺乏症新生儿病理性黄疸发生率34.20％。结论本研究阐明了北海市G6PD缺乏症情况及与新生儿病理性黄疸发生率的相关性,并肯定了新生儿脐血筛查G6PD缺乏、早期诊断、早期防治的重要价值。     

新疆一例汉族家系β地中海贫血ⅣS-Ⅱ-654(C→T)的基因分析

本文报道应用聚合酶链反应(PCR)结合等位基因特异的寡核苷酸(ASO)探针斑点杂交技术,对新疆首例p地中海贫血(p地贫)家系进行基因分析,从12名家系成员中检测出7例p地贫基因突变(ⅣS-Ⅱ-654,C→T)的杂合子。结合家系调查、临床表现及血液学检查结果,证明先证者的致病基因来源于父方。该研究对了解新疆地区p地贫基因突变类型和频率以及开展产前诊断具有重要意义。     

The effects of chemical and radioactive properties of Tl-201 on human erythrocyte glucose 6-phosphate dehydrogenase activity

AimThe inhibitory effects of thallium-201 (²⁰¹Tl) solution on human erythrocyte glucose 6-phosphate dehydrogenase (G6PD) activity were investigated.MethodsFor this purpose, erythrocyte G6PD was initially purified 835-fold at a yield of 41.7% using 2′,5′-Adenosine diphosphate sepharose 4B affinity gel chromatography. The purification was monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which showed a single band for the final enzyme preparation. The in vitro and in vivo effects of the ²⁰¹Tl solution including Tl⁺, Fe⁺³ and Cu⁺² metals and the in vitro effects of the radiation effect of the ²⁰¹Tl solution and non-radioactive Tl⁺, Fe⁺³ and Cu⁺² metals on human erythrocyte G6PD enzyme were studied. Enzyme activity was determined with the Beutler method at 340 nm using a spectrophotometer. All purification procedures were carried out at +4°C.Results²⁰¹Tl solution and radiation exposure had inhibitory effects on the enzyme activity. IC₅₀ value of ²⁰¹Tl solution was 36.86 μl ([Tl⁺]: 0.0036 μM, [Cu⁺²]: 0.0116 μM, [Fe⁺³]: 0.0132 μM), of human erythrocytes G6PD. Seven human patients were also used for in vivo studies of ²⁰¹Tl solution. Furthermore, non-radioactive Tl⁺, Fe⁺³ and Cu⁺² were found not to have influenced the enzyme in vitro.ConclusionHuman erythrocyte G6PD activity was inhibited by exposure for up to 10 minutes to 0.057 mCi/kg ²⁰¹Tl solution. It was detected in in vitro and in vivo studies that the human erythrocyte G6PD enzyme is inhibited due to the radiation effect of ²⁰¹Tl solution.     

Imaging features of thalassemia

Thalassemia is a kind of chronic, inherited, microcytic anemia characterized by defective hemoglobin synthesis and ineffective erythropoiesis. In all thalassemias clinical features that result from anemia, transfusional, and absorptive iron overload are similar but vary in severity. The radiographic features of β-thalassemia are due in large part to marrow hyperplasia. Markedly expanded marrow space lead to various skeletal manifestations including spine, skull, facial bones, and ribs. Extramedullary hematopoiesis (ExmH), hemosiderosis, and cholelithiasis are among the non-skeletal manifestations of thalassemia. The skeletal X-ray findings show characteristics of chronic overactivity of the marrow. In this article both skeletal and non-skeletal manifestations of thalassemia are discussed with an overview of X-ray findings, including MRI and CT findings. Received: 21 August 1999; Revision received: 22 December 1999; Accepted: 15 February 1999     

Degenerative disc disease as a cause of back pain in the thalassaemic population: a case-control study using MRI and plain radiographs

Purpose The aim of this study was to test our observation that back pain in thalassemic patients could be caused by premature and extensive lumbar degenerative disc disease, when compared to non-thalassemic patients with back pain. Methods and materials Sixteen thalassemic patients with their sex- and age-matched controls were recruited into the study, 12 with thalassemia major, and 4 with thalassemia intermedia. Both the thalassemia patients and control subjects suffered from back pain, which was subjective rather than measured/pain scored. All subjects underwent magnetic resonance (MR) imaging of the lumbar spine, and 11 of the cases and 8 controls had lumbar spine radiographs. Each lumbar disc was scored for radiographic appearances and MR features of disc degeneration and disc protrusion. Proportion values for these parameters and median scores were derived at each disc level, and were analyzed and compared. Results There was a statistically-significant difference between proportion values of cases and controls for the MR features (P value=0.01, n=16) and the radiographic features (P value=0.01, n=11 cases, n=8 controls) of disc degeneration. The median disc level scores for the thalassemic group were uniformly high across all lumbar discs, and at all levels except at L 4/5. The control group conversely demonstrated a predilection for disc degeneration at L4/5 level. Conclusion The distribution of lumbar disc degeneration in thalassemic patients with back pain is more extensive, severe and multi-level in nature compared to matched controls, and disc degeneration should be considered as a significant cause of back pain in this population group.     

Exercise-induced oxidative stress in G6PD-deficient individuals

PURPOSE: This study was designed to investigate whether individuals with glucose-6-phosphate dehydrogenase (G6PD) deficiency can exercise without greater perturbations in their redox status compared with non-G6PD-deficient individuals. METHODS: Nine males with established G6PD deficiency and nine males with normal G6PD activity performed two exhaustive treadmill exercise protocols of different duration (the shorter one lasting 12 min and the longer one 50 min). Several hematological parameters, reduced glutathione (GSH), oxidized glutathione (GSSG), thiobarbituric acid reactive substances (TBARS), protein carbonyls, catalase, and total antioxidant capacity (TAC) were measured in the blood before and after each exercise bout. RESULTS: Both GSH and GSSG were significantly higher in the control group compared with the G6PD-deficient group at baseline (0.404 +/- 0.101 vs 0.195 +/- 0.049 mmol.L(-1) for GSH and 0.047 +/- 0.012 vs 0.012 +/- 0.006 mmol.L(-1) for GSSG; P < 0.05); as a result, their ratio was not significantly different between the two groups (P > 0.05). All other oxidative stress indices were not different between groups at rest (P > 0.05). Exercise of both durations affected significantly (P < 0.05) and similarly the levels of all oxidative stress indices either in the G6PD-deficient group or in the control group. Only the long exercise affected GSH status significantly (P < 0.05), whereas both short and long exercise increased the levels of TBARS, protein carbonyls, catalase activity, and TAC to a similar extent (P < 0.05). CONCLUSION: G6PD-deficient individuals are able to exercise until exhaustion without higher oxidative stress compared with non-G6PD-deficient individuals. Exercise duration is an important determinant of the magnitude of exercise-induced changes for GSH, GSSG, and GSH/GSSG, but not for TBARS, protein carbonyls, catalase activity, or TAC.     

MR imaging of the brain: findings in asymptomatic patients with thalassemia intermedia and sickle cell-thalassemia disease

OBJECTIVE: The purpose of this study was to evaluate the spectrum of MR findings of the brain in asymptomatic patients affected with thalassemia intermedia or sickle cell-thalassemia disease to prevent brain damage by identifying patients at risk for stroke so that transfusional or pharmacologic treatment could be implemented. SUBJECTS AND METHODS: Forty-one asymptomatic patients who were younger than 50 years and were affected by minor hemoglobinopathies underwent MR imaging of the brain. Ischemic lesions were classified as small, medium, or large and as single or multifocal. Atrophic changes were graded subjectively as mild, moderate, or severe. A grade of brain damage was assigned to every patient. The frequency and severity of brain damage were correlated with the number of sickle-cell crises per year, hemoglobin level, sickling hemoglobin level, platelet count, sex, and age. RESULTS: Of the patients with thalassemia intermedia, 37.5% showed asymptomatic brain damage, and 52% of those with sickle cell-thalassemia disease showed asymptomatic brain damage. In the thalassemia intermedia group, atrophy was always mild and ischemic lesions were generally small (25%) and single (25%). Among the patients with sickle cell-thalassemia disease, 24% had small, 16% had medium, and 12% had large ischemic lesions. Multifocal lesions were twice as common in the patients with sickle cell-thalassemia disease (20%) as in those with thalassemia intermedia (12.5%). Only in the patients with thalassemia intermedia did the frequency of brain damage increase with age. Moreover, brain damage inversely correlated with hemoglobin level in patients with thalassemia intermedia but not in those with sickle cell-thalassemia disease. Brain damage was more severe in patients with sickle cell-thalassemia disease who had more crises per year. CONCLUSION: This study suggests that patients with thalassemia intermedia and those with sickle cell-thalassemia disease may have asymptomatic brain damage. Our results suggest that MR imaging is useful in identifying patients at risk for stroke so that they can be treated with transfusional or pharmacologic therapy.     

静息态BOLD-fMRI评价鼻咽癌放疗后脑损伤的应用价值

目的 分析静息态血氧水平依赖功能磁共振成像(BOLD -fMRI)对于鼻咽癌放疗后早期脑损伤的诊断价值.方法 收集4组放疗前(G0)、放疗后0~3个月(G1)组、放疗后3~6个月(G2)组及放疗后6~9个月(G3)组鼻咽癌患者进行静息态扫描,通过基于Matlab软件的DPARSF工具进行G1与G0、G2与G0、G3与G0,、G2与G1、G3与G2、G3与G1整理分析及后处理.结果 G1组与G0组相比两侧海马及颞叶的活动降低,一致性减少,而G2组与G0组相比两侧海马及颞叶的活动也降低,但幅度小,较G1组相比有回升,G3组也有回升,但没有恢复到放疗前水平.结论 静息态BOLD-fMRI的异常改变对于早期诊断鼻咽癌患者的放射性脑损伤具有一定临床意义.     

Oxidation of cellular thiols by hydroxyethyldisulphide inhibits DNA double-strand-break rejoining in G6PD deficient mammalian cells

PURPOSE: We investigated the effect of protein- and non protein-thiol oxidation on DNA double-strand-break (DSB) rejoining after irradiation and its relevance in the survival of CHO cells. MATERIALS AND METHODS: We used mutant cells null for glucose 6 phosphate dehydrogenase (G6PD) activity since reducing equivalents, required for reduction of oxidized thiols, are typically generated through G6PD regulated production of NADPH. Cellular thiols were oxidized by pre-incubating the cells with hydroxyethyldisulphide (HEDS), the oxidized form of mercaptoethanol (ME). The concentrations of the intracellular and extracellular non-protein thiols (NPSH), glutathione, cysteine and mercaptoethanol were quantitated by HPLC. Protein thiols (PSH) were estimated using Ellman's reagent. Cell survival was determined by clonogenic assay. The induction and rejoining of DSB in cells was quantitated by Pulse Field Gel Electrophoresis after exposure to ionizing radiation. RESULTS: Much lower bioreduction of HEDS was found in the G6PD deficient mutants (E89) than in the wild-type cells (K1). A 1 h treatment of E89 cells with HEDS produced almost complete depletion of non-protein thiol (NPSH) and a 26% decrease in protein thiols. Only minor changes were found under similar conditions with K1 cells. When exposed to gamma radiation in the presence of HEDS, the G6PD null mutants exhibited a higher cell killing and decreased rate and extent of rejoining of DSB than were observed in K1 cells. Moreover, when the G6PD deficient cells were transfected with the gene encoding wild-type G6PD (A1A), they recovered close to wild-type cellular thiol status, cell survival and DSB rejoining. CONCLUSIONS: These results suggest that a functioning oxidative pentose phosphate pathway is required for DSB rejoining in cells exposed to a mild thiol oxidant.     

Oxidation of cellular thiols by hydroxyethyldisulphide inhibits DNA double-strand-break rejoining in G6PD deficient mammalian cells

Purpose : We investigated the effect of protein- and non proteinthiol oxidation on DNA double-strand-break (DSB) rejoining after irradiation and its relevance in the survival of CHO cells. Materials and methods : We used mutant cells null for glucose 6 phosphate dehydrogenase (G6PD) activity since reducing equivalents, required for reduction of oxidized thiols, are typically generated through G6PD regulated production of NADPH. Cellular thiols were oxidized by pre-incubating the cells with hydroxyethyldisulphide (HEDS), the oxidized form of mercaptoethanol (ME). The concentrations of the intracellular and extracellular non-protein thiols (NPSH), glutathione, cysteine and mercaptoethanol were quantitated by HPLC. Protein thiols (PSH) were estimated using Ellman's reagent. Cell survival was determined by clonogenic assay. The induction and rejoining of DSB in cells was quantitated by Pulse Field Gel Electrophoresis after exposure to ionizing radiation. Results : Much lower bioreduction of HEDS was found in the G6PD deficient mutants (E89) than in the wild-type cells (K1). A 1 h treatment of E89 cells with HEDS produced almost complete depletion of non-protein thiol (NPSH) and a 26% decrease in protein thiols. Only minor changes were found under similar conditions with K1 cells. When exposed to gamma radiation in the presence of HEDS, the G6PD null mutants exhibited a higher cell killing and decreased rate and extent of rejoining of DSB than were observed in K1 cells. Moreover, when the G6PD deficient cells were transfected with the gene encoding wild-type G6PD (A1A), they recovered close to wild-type cellular thiol status, cell survival and DSB rejoining. Conclusions : These results suggest that a functioning oxidative pentose phosphate pathway is required for DSB rejoining in cells exposed to a mild thiol oxidant.