小儿常染色体隐性遗传多囊肾病临床分析

目的探讨小儿常染色体隐性遗传多囊肾病（autosomal recessive polycystic kidney disease，ARPKD）的临床特点。方法回顾我院1995年1月～2006年12月收治的16例小儿ARPKD的临床资料。结果16例ARPKD中，男11例，女5例，影像学诊断14例，病理学诊断2例，肾脏影像学显示囊肿广泛分布于皮质和髓质。起病时以泌尿系症状就诊仅7例（43．75％），以肾脏外症状就诊3例（18．75％），其他就诊原因包括早产1例，出生时重度窒息1例，血尿素氮-肌酐（Bun—Cr）升高2例，贫血1例，外伤后肾囊肿破裂性腹痛1例。确诊时有12例（75％）肾衰竭，8例（50％）生长迟缓，10例（62．5％）合并肾脏外病变。随诊8例，4例死亡（分别死于先天性肺发育不良、重度窒息后多器官功能衰竭、进行性肾衰竭以及终末期肾病合并胆管细胞癌肾脏转移），1例透析，1例肾移植，1例肾功能正常，1例肝硬化。结论ARPKD为先天遗传性、进行性的肾脏和肝脏损害，小儿期临床表现形式多样，产前诊断对减少ARPKD畸形儿的出生有重要意义。     

常染色体显性遗传性多囊肾病临床及基因突变分析

目的 分析常染色体显性遗传性多囊肾病（ADPKD）患者临床特征及基因突变特点。方法 入选ADPKD患者23例,收集临床数据,并进行家系调查;抽取外周血经高通量测序方法进行多囊肾基因检测。结果 23例ADPKD患者主要临床表现为腰腹痛、血尿、感染,肾功能不全;与女性患者相比,男性ADPKD患者血尿酸水平明显增高;基因检测PKD1基因突变19例;PKD2基因突变4例。同处于慢性肾脏病（CKD）5期的ADPKD患者,PKD1基因突变患者血红蛋白明显低于PKD2基因突变患者（65.89±13.59 vs 97.5±17.02,P<0.01）。结论 ADPKD可进展至肾功能衰竭,基因检测有助于早期诊断和预后评估,终末期ADPKD患者,PKD1基因突变患者预后更差。     

两个中国常染色体显性遗传性多囊肾病家系的表型与基因型分析

目的研究中国人多囊肾病基因1（polycystic kidney disease 1 gene，PKD1）突变的特点，检测基因突变位点。方法25例多囊肾患者，正常对照16名，扩增PKD1基因的第44、45外显子的基因片段，变性梯度凝胶电泳突变检测系统进行初筛，然后测序。结果发现1个移码突变（12431delCT）、1个无义突变（C12217T）、1个多态性（A50747C），突变检测率为8％（2／25）。结论检测到2个新的可能的致病突变：1个移码突变（12431delCT）、1个无义突变（C12217T）。     

成人型多囊肾病一家系6例报告

成人型多囊肾病(aduit polycystic kidney disease；APKD)是一种常染色体显性遗传病，常合并多囊肝，多囊脾、多囊胰等，把这两个或两个以上脏器同时罹患囊肿病，称为内脏多囊病。现将我们遇到的一家系报告如下。     

一个可能与PKD2基因连锁的常染色体显性多囊肾病家系

目的 研究常染色体显性多囊肾病（autosomal dominant polycystic kidney disease,ADPKD）在中国人中的遗传异质性。方法 采用聚合酶链反应 （polymerase chain reaction, PCR）、非变性聚丙烯酰胺凝胶电泳,检测了1个ADPKD家系各成员中与PKD1基因连锁的4种和与PKD2连锁的4种微卫星标记的基因分型。然后以软件辅助构建单倍型,并推测疾病单倍型。结果 发现该ADPKD家系中,与PKD1紧密连锁的4个微卫星KG8、SM6、CW4和CW2是有信息的;与PKD2基因紧密连锁的3种微卫星DNAIMS1563、D4S414和D4S423是有信息的。推定的单倍型提示,在这个家系中疾病可能与PKD2连锁,而不与PKD1连锁。结论 在此家系中,受累成员间存在表型异质性,并且有一个早发的儿童患者。与PKD2连锁的家系较少,这个家系的报道表明中国人中存在ADPKD的遗传异质性,PKD2的异常也可能会引起中国人ADPKD的发生。另外,发现有遗传早现现象存在,且疾病通过母亲传递。这提示在与PKD1不连锁的家系中后代可能早发病。     

多囊蛋白-1的研究进展

多囊蛋白-1（polycystin-1,PC-1）是由多囊肾病基因-1（PKD1）编码的跨膜蛋白,属多囊蛋白家族成员,主要在上皮细胞、内分泌细胞、心肌细胞和骨骼肌细胞上表达。PC-1可与多囊蛋白-2（polycystin-2,PC-2）、踝蛋白、纽蛋白等蛋白结合,参与细胞-细胞、细胞-基质黏附和细胞间信号转导。PC-1可调节肾小管上皮细胞的增殖和分化、介导细胞间的黏附,在肾脏的正常发育和常染色体显性遗传性多囊肾病（ADPKD）发病起重要作用。     

多囊肾病基因1和多囊肾病基因2在不同肾组织和肾细胞株中的表达及意义

目的检测多囊肾病基因1（PKD1）和多囊肾病基因2（PKD2）在不同肾组织和肾细胞株中的表达,探讨其表达产物的正常生理功能、相互作用关系及病理意义。方法用免疫沉淀、Westernblotting、原位杂交和免疫组织化学、免疫细胞化学方法检测PKD1和PKD2在不同肾组织和肾细胞株中的表达水平。结果PKD1和PKD2在正常肾组织和常染色体显性多囊肾病（ADPKD1）肾囊肿组织中都有表达。PKD1 mRNA和PKD2mRNA及其表达产物有着共同的组织分布,且表达水平差异无统计学意义。在PKD1突变的多囊肾组织中,二者在囊肿衬里上皮细胞和远端肾小管上皮细胞中都有高水平表达,且明显强于他们在正常肾小管中的表达（P〈0．01）,差异有统计学意义。但在多囊肾临近囊肿壁的小管细胞中,二者表达量明显减低。此外,二者在ADPKD肾囊肿衬里上皮细胞系和猪近端肾小管上皮细胞株（LLC-PK1）中也有表达。多囊蛋白-1（PC-1）的表达位于胞膜和胞质,多囊蛋白-2（PC-2）的表达则局限于胞质。结论PKD1和PKD2几乎共同的表达分布模式及其在正常肾组织和多囊肾组织的表达分布差异提示PC-1和PC-2在功能上的关联可能是以他们分子结构的相互作用为基础的,多囊蛋白可能具有维持肾小管结构稳定性的功能。     

抗多囊蛋白1氨基端单克隆抗体的制备和鉴定

目的 :用杂交瘤技术制备抗多囊蛋白 1胞外区氨基端的单克隆抗体 (mAb) ,并对其特异性进行鉴定。方法 :以肾组织总RNA为模板 ,用RT PCR扩增多囊蛋白 1胞外区氨基端的编码基因PKD1cDNA。将该基因克隆到融合蛋白表达载体pQE3 0中 ,转染大肠杆菌M15。以异丙基硫代半乳糖苷 (IPTG)诱导表达多囊蛋白 1胞外区氨基端的组氨酸融合蛋白 (PC1 e2 ) ,用亲和层析法纯化。以纯化的融合蛋白作为抗原免疫BALB/c小鼠 ,取小鼠脾细胞与小鼠骨髓瘤细胞株Sp2 / 0进行细胞融合 ,间接ELISA筛选阳性克隆 ,有限稀释法进行单克隆化。mAb的特异性用间接ELISA和Westernblot鉴定。结果 :克隆到两个编码多囊蛋白 1氨基端的cDNA片段 (50 2bp和 471bp)。构建的表达质粒经酶切和DNA测序证实 ,为所需要的质粒。表达出相对分子质量 (Mr)分别为 1980 0和 1890 0的融合蛋白 ,经Westernblot鉴定 ,均为多囊蛋白 1的融合蛋白。用Mr 为 1890 0的融合蛋白免疫小鼠 ,得到杂交瘤细胞株 7B1。Westernblot分析表明 ,该细胞株分泌的mAb能特异地与多囊蛋白 1氨基端结合。结论 :本实验表达了PKD1多拷贝区所编码的PC1 e2 ,成功地制备了抗多囊蛋白 1胞外区N端的mAb。     

家族性多囊肾病五例

先证者　男 ,72岁 ,因右足第一跖趾关节部红肿疼痛 ,皮肤溃破流出白色粉笔沫样物 7天就医。患者多囊肾伴高血压病史3 0余年 ,5年来右拇跖趾关节多次红肿疼痛发作 ,经休息及抗感染治疗好转。查体 :轻度贫血貌 ,血压 170 /10 0 mm Hg,心界轻度向左下扩大 ,A2 >P2 。肺及腹部未见异常。右足第一跖趾关节部皮肤溃破 ,有白色结晶样物暴露。血常规 RBC 3 .8× 10 9/L ,HGB 93 g/L ,余正常。肾功能检查 :血尿素氮 2 0 .4mmol/L ,血肌酐 3 17μmol/L,血尿酸 (uric acid,UA) 890 μmol/L。尿常规 :蛋白 +红细胞 +。ECG示慢性冠状动脉供血不足…     

Autosomal recessive polycystic kidney disease

Summary Autosomal recessive polycystic kidney disease is a rare inherited disorder which usually becomes clinically manifest in early childhood, whereas autosomal dominant polycystic kidney disease usually is a disorder of adult onset. With increasing knowledge and improving diagnostic techniques, it becomes evident that the spectrum of both entities is much more variable than generally known. The presentation of autosomal recessive polycystic kidney disease at later ages and survival into adulthood have been reported. The diagnostic criteria, clinical course, genetics and differential diagnosis of autosomal recessive polycystic kidney disease will be presented.Abbreviations ADPKD, ARPKD autosomal dominant/recessive polycystic kidney disease - CHF congenital hepatic fibrosis     

Genetics and phenotypic characteristics of autosomal dominant polycystic kidney disease in Finns

Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited kidney disease, leading to renal insufficiency and renal transplantation. Mutation screening in the major gene for ADPKD, the polycystic kidney disease type 1 (PKD1) gene, has often been incomplete because of multiple homologous copies of this gene elsewhere on chromosome 16. Furthermore, there are only a few studies investigating genotype–phenotype correlations in patients with ADPKD. In this study, we screened the entire coding region of the PKD1 and PKD2 genes in 17 Finnish families with ADPKD via long-range polymerase chain reaction, single-strand conformation polymorphism analysis, and direct sequencing. We were able to identify mutations co-segregating with ADPKD in all 16 families linked to PKD1 by haplotype analysis. Of these mutations, six were insertions/deletions, five nonsense mutations, and five missense mutations. In the only PKD2-linked family, we found a missense mutation, R322Q. With the exception of one mutation (L845S in PKD1), all mutations were novel. Mutations and their location did not have a strong correlation with the phenotype with the exception of subarachnoidal hemorrhage or brain aneurysm, where mutations were located more often at the 5 end of the PKD1 gene than at the 3 end of the PKD1 gene.Electronic Supplementary Material Supplementary material is available for this article at .     

Autosomal recessive polycystic kidney disease

 Autosomal recessive polycystic kidney disease (ARPKD) is a rare inherited disorder which usually becomes clinically manifest in early childhood, although the spectrum of ARPKD is much more variable than generally known. Presentation of ARPKD at later ages and survival into adulthood have been observed in many cases. The responsible gene has been mapped to chromosome 6p. Thus there is no evidence of genetic heterogeneity. The most important indication for DNA diagnosis is the prenatal diagnosis in families with at least one affected child. The critical region has been narrowed with the use of recombinant families of about 4 cM. Several possible candidate genes have been excluded. Received: 23 April 1997 / Accepted: 12 August 1997     

PGD for autosomal dominant polycystic kidney disease type 1

Autosomal dominant polycystic kidney disease (ADPKD) is primarily characterized by renal cysts and progression to renal failure. It is a genetically heterogeneous disease, with mutations in the PKD1 gene accounting for the majority of cases. Direct mutation detection for PKD1-linked ADPKD or type 1 is complicated by the large size and complex genomic structure of PKD1. This paper describes a microsatellite marker-based assay for PGD in couples at risk of transmitting ADPKD type 1. During PGD, genetic analysis is carried out on single blastomeres biopsied from preimplantation embryos obtained after IVF, and only embryos unaffected by the disease under investigation are selected for transfer. Single-cell genetic analysis relied on a fluorescent duplex-PCR of linked polymorphic markers followed by fragment length determination on an automated sequencer. The co-amplification of the intragenic KG8 and the extragenic D16S291 marker at the single-cell level was evaluated in pre-clinical tests on lymphoblasts and research blastomeres. The developed assay proved to be efficient (96.1% amplification) and accurate (1.4% allele drop-out and 4.3% contamination), and can be applied in all informative ADPKD type 1 couples. From five clinical cycles carried out for three couples, two pregnancies ensued, resulting in the birth of two healthy children.     

Hepatorenal findings in obligate heterozygotes for autosomal recessive polycystic kidney disease

Autosomal recessive polycystic kidney disease (ARPKD), characterized by progressive cystic degeneration of the kidneys and congenital hepatic fibrosis (CHF), is the most common childhood onset ciliopathy, with an estimated frequency of 1 in 20,000 births. It is caused by mutations in PKHD1. The carrier frequency for ARPKD in the general population is estimated at 1 in 70. Given the recessive inheritance pattern, individuals who are heterozygous for PKHD1 mutations are not expected to have clinical findings. We performed ultrasound (USG) evaluations on 110 parents from 64 independent ARPKD families and identified increased medullary echogenicity in 6 (5.5%) and multiple small liver cysts in 10 parents (9%). All ARPKD parents with these abnormal imaging findings were asymptomatic; kidney and liver function tests were unremarkable. Complete sequencing of PKHD1 in the 16 ARPKD parents with abnormal imaging confirmed the mutation transmitted to the proband, but did not reveal any other pathogenic variants. Our data suggest that carrier status for ARPKD is a predisposition to polycystic liver disease and renal involvement associated with increased medullary echogenicity on USG. Whether some of these individuals become symptomatic as they age remains to be determined.     

一个成人多囊肾疾病家系研究分析

目的通过一个家系分析,探讨常染色体显性遗传性多囊肾疾病的病因学、病理学、遗传学因素。方法通过对一个多囊肾家系三代20例家庭成员进行调查,并对六例患者进行B超、肝肾功能检查。结果患者主要表现为双侧肾脏多发性囊肿,部分患者合并肝脏多发性囊肿,多于35岁以后起病,伴有腹胀,高血压,血尿等症状,部分患者行囊肿去顶开窗手术,有一例患者合并多囊脾。家系遗传分析表明该疾病属于常染色体显性遗传性多囊肾疾病,其中多囊肝为肾外表现。结论本病发病机制与基因缺陷相关,尤其是PKD1、多囊蛋白-1、PKD2、多囊蛋白-2、PKD3等,家系中如果亲代发病,子代中亦会有发病,年龄在35岁以后为多。     

5例成人型多囊肾家系调查

目的 了解5例成人型多囊肾先证者家族中的患病情况及患病规律。方法 对76例家族成员进行B型超声波检查．在肾脏中发现多个囊肿为主要诊断依据。结果 家族中患病率43．6％，男42．8％，女44．1％，男女患病率无明显差异，符合常染色体显性遗传患病规律。结论 家系调查对开展遗传咨询、优生指导有重要意义。     

Mutational analysis of PKD1 gene in a Chinese family with autosomal dominant polycystic kidney disease

Autosomal dominant polycystic kidney disease (ADPKD) is a hereditary disease and common renal disease. Mutations of PKD genes are responsible for this disease. We analyzed a large Chinese family with ADPKD using Sanger sequencing to identify the mutation responsible for this disease. The family comprised 27 individuals including 10 ADPKD patients. These ADPKD patients had severe renal disease and most of them died very young. We analyzed 6 survival patients gene and found they all had C10529T mutation in exon 35 of PKD1 gene. We did not found gene mutation in any unaffected relatives or 300 unrelated controls. These findings suggested that the C10529T mutation in PKD1 gene might be the pathogenic mutation responsible for the disease in this family.