糖原累积病Ⅱ型的临床分析和基因学检测

目的探讨糖原累积病Ⅱ型的酶学诊断方法,并在中国人中进行基因突变分析。方法对1例临床表现为婴儿型糖原累积病Ⅱ型的8个月男性患儿进行研究,采用荧光分光光度计检测外周血白细胞酸性α#葡萄糖苷酶(GAA)活性,并采用16对引物对该家系GAA基因从外显子2到外显子20的编码区进行扩增并进行测序分析。结果该患儿白细胞GAA活性仅为18.72nmol/(mgprotein·hr),正常对照组为(130.5±86.7)nmol/(mgprotein·hr);基因学分析发现该患儿存在2个杂合突变位点,其中之一为1935位点C→A错义突变,是中国人最常见的突变位点,另外一个为988位点T→G错义突变,该位点为新发现突变位点。结论糖原累积病Ⅱ型是由于GAA基因突变引起GAA活性降低所致,酶活性检测和基因检测是目前最有效和可靠的诊断方法。     

1例糖原累积病患儿酸性-α-葡萄糖苷酶基因的新无义突变p.W738X

目的 探讨1例临床疑似糖原累积病Ⅱ型患儿的临床特点和酸性-α-葡萄糖苷酶(GAA)基因突变情况.方法 分析患儿病史,并检测患儿及其父母外周血白细胞酸性-α-葡萄糖苷酶活性,聚合酶链反应(polymerase chain reaction,PCR)扩增GAA编码区,直接测序分析GAA基因突变情况.结果 该患儿生后2个月即出现喂养困难和全身肌无力,4个月发现心脏增大,6个月时死于心肺功能衰竭.患儿白细胞酸性-α-葡萄糖苷酶活性明显降低,仅为正常对照中位数的17.3%.DNA测序分析显示患儿携带一个新无义突变p.W738X和一个已报道的p.E888X突变.患儿及其母亲均携带假性缺陷等位基因c.1726G ＞ A;2065G ＞ A.结论 GAA酶活性测定结合基因诊断明确诊断1例临床疑似糖原累积病(GSD)Ⅱ型,发现1个GAA基因新无义突变p.W738X.根据患儿临床表现可诊断为婴儿型GSD Ⅱ,推测p.W738X突变对GAA酶活性影响严重.由于假性缺陷等位基因c.1726G ＞ A;2065G ＞ A可引起正常人GAA酶活性降低,故GAA基因突变分析对明确GSDⅡ型患者的诊断及其家系的产前诊断有重要意义.     

婴儿型糖原贮积病Ⅱ型5例临床报告并文献复习

2016年11月至2022年3月福建医科大学附属漳州市医院共收治5例婴儿型糖原贮积病Ⅱ型患儿。5例中男3例,女2例,发病年龄均小于1岁,首发症状主要为呼吸困难、心肌肥厚、肝脏增大及骨骼肌无力。4例血酸性ɑ-葡萄糖苷酶浓度明显降低,最低为0.28 mmol/（L·h）。5例均进行了基因检测,其中3例为纯合错义突变,1例为杂合错义突变,1例为复合杂合错义突变,同时存在有致右室心律失常心肌病基因CTNNA3突变C.2122A>G父源。5例均自动出院后死亡。中位生存时间为11个月（6～11个月）。确诊病例中基因突变类型以c.1935C>A最常见,c.2853G>A可能是新的致病突变位点。     

糖原累积病Ⅰa型患儿20例基因突变分析与临床研究

目的 研究糖原累积病Ⅰa型(GSD Ⅰ a)患儿葡萄糖-6-磷酸酶催化亚单位基因(G6PC基因)突变情况,探讨基因型与临床表型之间的关系.方法 依据临床表现、生化检查及饥饿试验、胰高血糖素刺激试验结果,拟诊出48例肝糖原累积病(LGSD)患儿;应用PCR反应直接测序的方法,对疑似LGSD患儿的G6PC基因外显子及其相邻区域进行突变检测.采用50例无血缘关系的健康儿童作为健康对照,以排除基因多态性;使用DNAMAN软件进行多物种序列同源性比较分析,确定其是否具有保守性.结果 48例LGSD患儿中检测到20例患儿存在G6PC基因突变,共检测到8种突变类型,包括1种剪切突变:c.648G＞T,5种错义突变:p.R83H、p.H119L、p.L173P、p.I341N、p.C109Y和2种移码突变:c.262delG、c.260-262delGGinsA.其中c.648G＞T、p.R83H是本组研究最常见的突变,突变频率分别为37.50％、22.50％;p.C109Y、c.260-262delGGinsA为新发突,41.67％(20/48例)拟诊为LGSD的患儿通过G6PC基因分析确诊为GSD Ⅰ a.从临床表现及常规实验室检查分析,20例GSD Ⅰ a患儿均表现为肝大、肝功能异常、高乳酸血症、高三酰甘油血症,88.24％(15/17例)的患儿饥饿试验阳性,82.35％(14/17例)患儿对胰高血糖素刺激试验无反应.结论 c.648G＞T、p.R83H为本组GSD Ⅰ a患儿最常见突变类型,p.C109Y、c.260-262delGGinsA为国内外尚未见报道的新发致病突变.GSDⅠa基因型不同的患儿均有相似的典型LGSD临床表现.     

3例糖原贮积症Ⅱ型患儿的临床特点及GAA基因突变分析

糖原贮积症Ⅱ型(GSDⅡ)是一种主要由酸性α-葡萄糖苷酶(GAA)基因突变引起的常染色体隐性遗传病,主要累及心脏、骨骼肌等脏器。本文报道3例经GAA基因分析确诊的GSDⅡ患儿的临床特点和基因突变结果,1例为婴儿型,年龄为4个月,患儿表现为体重不增,呼吸困难,肌张力低,ALT、CK升高,心脏彩超示肥厚型心肌病。2例为晚发型,年龄分别为8岁、13岁,晚发型患儿均表现为持续肝酶升高,其中1例患儿伴反复呼吸道感染,肺功能示限制性通气障碍;另1例患儿伴肌酶升高明显,而肌电图正常。外周血GAA基因检查结果显示3例患儿中共检测出6种致病突变,其中c.2738CT、c.568CT为未见报道的新突变。外周血GAA基因检测是有效的诊断该疾病的方法。     

糖原累积病Ⅲ型临床和基因研究进展

糖原累积病Ⅲ型(glycogen storage disease typeⅢ,GSD),又称Cori’s或Forbe’s病、界限糊精病。1952年,美国学者Forbe[1]发现患者肝脏、肌肉组织中有短侧链糖原(界限糊精)累积,并报道了本症。1956年Illingworth等[2]明确其病因为糖原脱枝酶(glycogen debrancher deficiency,Amylo"1,6"glu-cosidase,AGL)活性缺乏。据报道,美国GSDⅢ型发病率为1∶100000,北部非洲以色列犹太人发病率为1∶5400,AGL基因突变携带者比例为1∶35,法罗群岛的GSDⅢa型发病率高达1∶3100,中国香港GSDⅢ型的发病率约1∶25650,在中国大陆、西班牙人、因纽特…     

糖原累积病Ⅸa型临床表型基因型及转归分析

目的　总结糖原累积病Ⅸa型（GSD Ⅸa）的临床、实验室检查、基因突变特点及预后,加强对该类疾病的认识。方法　对2016年9月至2020年9月在广州市妇女儿童医疗中心经基因检测确诊的18例GSD Ⅸa型患儿的临床、实验室检查、基因突变及预后资料进行回顾性分析。结果　18例患儿均为男性,发病中位年龄为1岁10月龄。18例均有肝肿大及转氨酶升高;矮小8例（44.44%）;空腹血糖低11例（61.11%）;高甘油三酯血症7例（38.89%）;高胆固醇血症6例（33.33%）;高乳酸血症12例（66.67%）。共检出17种突变,均为半合突变,以错义突变为主,10种为未见报道的突变,其中2种为新发突变。9例进行了肝活检,病理结果均符合典型GSD表现,4例（44.44％）伴有汇管区灶性纤维增生;6例（66.67％）有肝细胞脂肪变性。除1例未治疗外其余患儿均给予改善饮食、口服生玉米淀粉,临床及生化表现均有改善。结论　肝肿大、转氨酶升高与GSD Ⅸa型高度相关,基因检测可明确诊断及分型。该病多数预后好,但部分可能会进展为肝纤维化,需加强随访。     

糖原累积病Ⅰa型1例及其家系的临床和分子遗传分析

目的：研究1例糖原累积病Ⅰa型患者及其家系的基因突变情况。方法：应用聚合酶链反应扩增葡萄糖-6-磷酸酶基因（G6PC基因）全部5个外显子,通过DNA直接测序的方法,对糖原累积病Ⅰa型患者及其家系中父、母、姐姐的G6PC基因进行突变检测。结果：在患者G6PC基因的第5外显子上的第743碱基发生杂和突变,由G转变为A,导致G6Pase蛋白第222位氨基酸由甘氨酸变为精氨酸（G222R）,家系中父亲、姐姐均未发现该突变,而母亲携带与患者相同突变。结论：首次在国内报道G6PC基因的G222R突变,丰富了糖原累积病Ⅰa型在中国人群的突变谱。     

糖原累积病Ⅵ型和Ⅸa型7例病例报告并文献复习

目的总结糖原累积病(GSD)Ⅵ、Ⅸa型的临床、病理和基因突变情况,提高临床对这两型GSD的认识。方法回顾性收集GSD3例Ⅵ型和4例Ⅸa型患儿的临床资料。结果 (1)7例患儿均为男性,确诊年龄2岁3月至5岁。7例均有肝脏肿大和转氨酶升高,身材矮小1例,空腹低血糖、高乳酸血症和高甘油三酯血症各2例,血酮体增高3例,尿有机酸分析结果阳性2例。7例患儿均有肝细胞弥漫性肿大变形和糖原凝聚,4例有肝脏脂肪变性;3例GSDⅥ型有门管区纤维化、肝硬化表现。3例GSDⅥ型检测到6种PYGL基因突变,c.772+1GA、c.244-1GA、c.730CT(p.L244F)、c.2417_2418del TA(p.I806Sfs X9)为新突变,4例GSDⅨa型检测到4种PHKA2基因突变,c.3529CT(p.Q1177X)、c.3574CT(p.Q1196X)为新突变。(2)复习文献共检索到13篇文献,与本文病例合并后共22例Ⅵ型、99例Ⅸa型GSD。肝脏转氨酶增高和肝脏肿大91.9%~100%,有身材矮小18%~23%、空腹低血糖44%~48%、高甘油三酯血症37%~44%、高乳酸血症35%~72%和血酮体增高50%~56%。肝脏活检均可见肝细胞内糖原凝聚,17%有脂肪变性,Ⅵ型25%、Ⅸa型33%检出肝硬化。报道19种PYGL基因突变,多为点突变,剪切位点突变亦较常见,插入突变少见;43种PHKA2基因突变,突变类型多样。结论肝大伴转氨酶升高的患儿需警惕Ⅵ、Ⅸa型GSD;Ⅵ型患儿可早期存在肝硬化,需要进一步随访。     

GBE1基因突变型糖原累积病Ⅳ型1例报告

目的探讨GBE1基因突变的糖原累积病Ⅳ型(GSD Ⅳ)患儿的临床特点及其家系的基因突变情况。方法分析1例GSD Ⅳ患儿的临床表现、肝脏病理结果及其父母的全外显子基因测序情况,并进行文献复习。结果患儿,男,1岁10个月,肝脾肿大6月余伴发热7天;肝脏组织病理示慢性肝损伤,不能除外遗传代谢病。全基因外显子检测示患儿存在2种新的GBE1基因的杂合突变,分别为来自父亲的c.1694GA杂合突变(致病性变异)和来自母亲的c.218AG杂合突变(疑似致病性变异)。结合患儿临床表现、病理及基因检测结果确诊为肝型GSD Ⅳ。结论新发现GEB 1基因c. 1694 GA的致病性杂合突变,丰富了GSD Ⅳ型在中国人群的突变谱。     

Cardiac involvement in glycogen storage disease type III

Twenty patients with enzymatically proven glycogen storage disease type III (GSD III) aged 3–30 years underwent cardiological evaluation. Seventeen showed subclinical evidence of cardiac involvement in form of ventricular hypertrophy on ECG. Of 16 patients in whom an ECG examination was performed, 13 had abnormal echocardiographic features. Only 2 patients had cardiomegaly on X-ray. The cardiac findings in 1 of the patients, a 25-year-old female with clinically evident cardiomyopathy are described in detail. In view of our findings, patients with established GSD III, should not only be investigated regarding their muscular involvement, but should also undergo a detailed evaluation of their cardiac status.Abbreviation GSD glycogen storage disease     

Historical highlights and unsolved problems in glycogen storage disease type 1

Thirty-three years after Von Gierke described the first patient with glycogen storage disease type 1 (GSD1) in 1929, the Coris detected glucose-6-phosphatase (G6Pase) deficiency. The first mutation of this enzyme was identified 41 years later and subsequently the gene was mapped to chromosome 17q21, its enzyme topology defined, a nine transmembrane helical model suggested, an enzyme deficient knockout mouse created and by infusing an adenoviral vector associated murine G6Pase gene, correction of the clinical and laboratory abnormalities was observed. A similar successful gene transfer has been performed in enzyme deficient canine puppies. To explain the function of the G6Pase complex, a multicomponent translocase catalytic model has been proposed in which different transporters shuttle glucose-6-phosphate (G6P), inorganic phosphate (Pi) and glucose across the microsomal membrane. It was suggested that GSD1b patients suffered from a G6P transporter (G6PT) defect and the first mutation in the G6PT gene subsequently recognised. The gene mapped to chromosome 11q23 and its structural organisation was defined which showed a close functional linkage between G6PT and hydrolysis. Nordlie identified the first patient with Pi transport deficiency (GSD1c). However putative GSD1c and 1d patients based on kinetic studies were found to harbour mutations in the G6PT gene so that GSD1 patients are presently divided into 1a and non-1a. G6PT deficient patients suffer from numerical and functional leucocyte defects. A mRNA leucocyte G6PT deficiency has been suggested to account for the glucose phosphorylation and subsequent calcium sequestration defects observed in theses cells. Inflammatory bowel disease which occurs frequently in GSD non-1a patients has been related to their leucocyte abnormalities. Dietary management of GSD1 patients, designed to maintain a normal blood glucose level can be achieved during the night by nocturnal gastric infusions of glucose-containing solution or by the administration of uncooked cornstarch around the clock or by a combination of both. Both therapeutic modalities, if conducted in a meticulous manner, have a major impact on the quality of life, prevention of complications and subsequent prognosis. Open questions relate to the source of endogenous glucose production in GSD1 patients which increases as a function of age from 50% to 100% of normal, concomitant with an improvement in the patients fasting tolerance. Several complications, the nature of which is incompletely understood, tend to occur after the first decade: Liver adenomata with a small risk of transforming into hepatoma, progressive renal disease, which may be related to the hyperlipidaemia observed in this disease, often leading to end stage renal failure, osteopenia apparently based on high bone turnover, growth retardation and delayed puberty.Conclusion: this review highlights the present knowledge of glycogen storage disease type 1 and subtypes, discussing unsolved questions, which reflect the limitation of our knowledge in the understanding of this intriguing group of diseases. Published online: 23 August 2002     

Secondary amyloidosis in glycogen storage disease type Ib

We observed the development of generalized amyloidosis in a girl with glycogen storage disease type Ib (GSD-Ib) who showed neutropenia, neutrophil dysfunction and recurrent infections. Renal and thyroid biopsies showed secondary amyloidosis, characterized by the presence of potassium permanganate sensitive Dylon positive deposits in glomeruli, renal vessels and thyroid interstitium. Immunohistochemistry showed that the deposits were composed of amyloid A (AA) protein. Possibly neutrophil abnormalities are involved in the pathogenesis of amyloidosis.     

Effects of cornstarch treatment in very young children with type I glycogen storage disease

Three children aged 1–2 years with glycogenosis type I were treated with 2 g/kg bodyweight oral cornstarch per meal (4–5 times a day) for a period up to 16 months. In comparison to the previous dietary regimen (day and nocturnal feedings every 3 h) the cornstarch diet stabilised serum glucose profiles and dramatically improved secondary hyperlipoproteinaemia. Mean total triglycerides decreased up to one half, consistent with a fall of very low density lipoprotein-triglycerides up to two thirds. Metabolic acidosis and hyperuricaemia did not occur and normal growth rates (0.7–1 cm/month) were achieved. We conclude that the cornstarch regimen even in the age group up to 2 years can be considered as an efficient alternative in the treatment of glycogenosis type I patients with less frequent feedings and without nocturnal infusion.     

Bone mineralisation in type 1 glycogen storage disease

Radial bone mineral content (BMC) was measured using single photon absorptiometry in 11 prepubertal children, aged 3.4–12.6 years, with glycogen storage disease type 1 (GSD-1), 2 of whom were receiving granulocyte colony stimulating factor (G-CSF) therapy for chronic neutropenia. Patients were short (median height SD score –1.35, range –3.74 to –0.27), and had reduced BMC Z scores (median 1.79, range –6.35 to +0.27) and radial bone width Z scores (median –0.72, range –2.00 to +0.68). Those receiving G-CSF did not differ significantly from the rest of the group. Generally dietary calcium intake was low and urinary calcium excretion increased. Urinary lactate excretion was high but did not correlate with BMC Z scores. Factors regulating bone metabolism (parathyroid hormone and 25-hydroxy vitamin D concentrations) and markers of bone formation (osteocalcin and skeletal alkaline phosphatase) were not increased implying that there was no compensation for increased bone resorption.Conclusion Patients with GSD-1 may be at increased risk of fracture in later life and require close attention to metabolic control and calcium balance.     

糖原贮积症Ⅱ型实验室检测方法的研究进展

糖原贮积症Ⅱ型(glycogen storage disease type2,GSDⅡ)是一种罕见的、进展性的、致死性的溶酶体贮积病.近年来,随着人们对GSDⅡ研究的不断深入,越来越多的实验室检测方法应用于GSDⅡ的诊断.目前,GSDⅡ的实验室诊断方法主要是酶学检测、基因学分析及产前诊断,其中以干血滤纸片法为主的酶学检测和基因学分析在GSDⅡ实验室检测诊断中发挥重要作用.该文对GSDⅡ实验室检测方法作一综述,旨在为GSDⅡ的诊断提供新的理论依据和参考.     

Impaired monocyte function in glycogen storage disease type Ib

Phagocyte (neutrophil and monocyte) function was evaluated in a boy with glycogen storage disease type Ib. Neutrophils were found to be defective in motility and respiratory burst and monocytes showed a defect in respiratory burst but not in motility. These results suggested that the glucose-6-phosphate transport system plays a role in the function of neutrophils and monocytes and that these two phagocytes are different from each other in their energy metabolism for motility.Abbreviations GSD Ia Glycogen storage disease type Ia - GSD Ib glycogen storage disease type Ib - G6P glucose-6-phosphate - G6Pase glucose-6-phosphatase - PHA phytohemagglutinin - CL chemiluminescence - ZAS zymosan-activated serum - FMLP N-formylmethionyl-leucyl-phenylalanine - CSA colony-stimulating activity