云南苯丙氨酸羟化酶基因内（TCTA）n多态性及其在苯丙酮尿症诊断？…

目的 应用苯丙氨酸羟化酶（ＰＡＨ）基因内（ＴＣＴＡ）ｎ多态性连锁分析进行经典型苯丙酮尿症（ＰＫＵ）的基因诊断和产前诊断。方法 应用聚合酶链反应－扩增片段长度多态性（ＰＣＲ－Ａｍｐ－ＦＬＰ）方法,分析云南省１３个家系苯丙氨酸羟化酶（ＰＡＨ）基因内（ＴＣＴＡ）ｎ多态性。结果 在１３个家系中检测到２２４－２５２ｂｐ的８种等位片段,其ＰＩＣ为０．６９８,杂合频率是５１％。可诊断率为１００％和５０％的家系各     

儿童期脊肌萎缩症的基因诊断及产前基因诊断

为建立脊肌萎缩症（ＳＭＡ）产前诊断方法，应用３个与ＳＭＡ基因紧密连锁的ＣＡ重复序列位点ＪＫ５３ＣＡ１／２、５ＤＳ６３７、ＣＡＴＴ１，对１８个ＳＭＡ家系进行了连锁分析；应用聚合酶链反应－单链构型多态性（ＰＣＲ－ＳＳＣＰ）分析，对３７例ＳＭＡ患儿作了ＳＭＡ基因缺失的检测；并作２例ＳＭＡ产前诊断。结果：１８个家系通过ＪＫ５３ＣＡ１／２位点诊断１２个（可诊断率６６．７％），通过５ＤＳ６３７位点诊断１４个（７．８％），１０个ＳＭＡ家系通过ＣＡＴＴ１位点诊断９．５个（９５．０％）；３７例患儿中３２例（８６．５％）有端粒侧ｅｘｏｎ７的纯合性缺失；２例接受产前诊断者，１例胎儿患病，１例胎儿为正常。结论：应用ＳＭＡ基因旁侧ＣＡ重复序列联合ＰＣＲ－ＳＳＣＰ检测进行ＳＭＡ基因诊断和产前诊断简便、准确、可应用于临床。     

苯丙氨酸羟化酶基因新突变位点与临床的关系

目的 观察新发现的两种苯丙氨酸羟化酶基因突变与临床表现型的关系,对其中一个位点进行基因表达的研究。方法 通过变性梯度凝胶电泳（ＤＧＧＥ）突变检测和固相ＤＮＡ直接序列分析等方法。对５８例苯丙酮尿症（ＰＫＵ）患儿进行突变检测,对新突变进行体外寡革酸定点诱变和在ＣＯＳ细胞中ＰＡＨ活性表达研究,研究 共发现有ＩＶＳ６ｎｔ－１的Ｇ→Ａ突变和Ａｒｇ２５２Ｇｌｎ突变的患儿７例,均有典型的ＰＫＵ临床表现,并有不同     

急性白血病MTS1基因缺失及点突变的研究

目的 探讨多肿瘤抑制基因（ＭＴＳ１）突变与恶性血液病间的关系,方法 应用聚合酶链反应－单链查象多态性（ＰＣＲ－ＳＳＣＰ）分别和ＤＮＡ印迹（ｓｏｕｔｈｅｒｍｂｌｏｔ）方法检测３５例急性白血病患儿ＭＴＳ１基因的改变。结果 急性淋巴细胞白血病（ＡＬ）患儿的ＭＴＳ１基因缺失（包括点突变）为２６％（８／３１）,其中纯合子缺失,Ｂ－ＡＬＬ为１６％（４／２５）,Ｔ－ＡＬＬ为６例中有２例,点突变则两型各占１例。结     

关于一些医学专业术语英文缩略语的使用说明(免疫专业)

为了节省篇幅、方便读者,今后本刊的文章中免疫专业常用术语的英文缩略语不再注中文全称,如族分化抗原（ＣＤ）,Ⅰ类Ｔ辅助细胞（ＴＨ１）,Ⅱ类Ｔ辅助细胞（ＴＨ２）,细胞毒性Ｔ细胞（ＣＴＬ）,自然杀伤细胞（ＮＫ）,外周血单个核细胞（ＰＢＭＣ）,免疫球蛋白（Ｉｇ）,人类白细胞相关抗原（ＨＬＡ）,主要组织相容复合体（ＭＨＣ）,白细胞介素（ＩＬ）,植物血凝素（ＰＨＡ）,脂多糖（ＬＰＳ）,酶联免疫吸附试验（ＥＬＩＳＡ）,单克隆抗体（ｍＡｂ）,聚合酶链反应（ＰＣＲ）,逆转录ＰＣＲ（ＲＴＰＣＲ）,互补ＤＮＡ（ｃＤ…     

儿童再发性腹痛与幽门螺杆菌感染关系探讨

对１２３例非精神心里因素所致的再发性腹痛（ＲＡＰ）患儿进行胃镜检查及幽门螺杆菌（ＨＰ）检测。胃镜所见：正常１７例（１３８％），慢性胃炎（ＣＧ）６６例（５３７％），ＣＧ合并十二指肠球炎１３例（１０６％）、合并食管炎４例（３２％），消化性溃疡（ＰＵ）２２例（１７９％），幽门管狭窄１例（０８％）。采用三种方法检测ＨＰ：ＥＬＩＳＡ方法、ＰＣＲ技术及组织学染色找菌，ＨＰ阳性率５７７％。１２３例中６６例进行了病理学检查，观察到胃炎炎症程度与ＨＰ感染相关，６６例中胃窦粘膜活检正常６例，Ｇｉｅｍｓａ染色找菌ＨＰ全部阴性，轻度ＣＧ３７例Ｇｉｅｍｓａ染色ＨＰ阳性１１例（１１／３７），中重度２２例ＨＰ均为阳性。本组资料显示儿童ＲＡＰ多数存在器质性病变，并与ＨＰ感染有关。对非精神性ＲＡＰ、尤其血清ＨＰＩｇＧ抗体及胃液ＨＰＰＣＲ检测阳性者应进一步做胃镜检查以明确诊断。     

脊肌萎缩症临床及基因诊断研究

３０例脊肌萎缩症（ＳＭＡ），其中Ⅰ型１３例，Ⅱ型１７例。临床表现肢体及躯干进行性对称性肌无力及肌萎缩，合并肋间肌麻痹及肌束震颤。血清肌酸磷酸肌酶（ＣＰＫ）４３～２９０ＩＵ／Ｌ。肌电图（ＥＭＧ）特点：静止状态可见纤颤正向电位，轻收缩时运动单位电位时限增宽，重收缩时为单纯相。肌活检可见萎缩肌纤维及正常肌纤维相间分布，周边可见肥大肌纤维，Ⅰ型病例可见成群或成束状萎缩肌纤维。单链构象多态性（ＳＳＣＰ）检测患儿和父母及无亲缘关系的正常人运动神经元存活基因（ＳＭＮ）外显子７缺失。结果：３０例ＳＭＡ中纯合缺失Ⅰ型１１例，Ⅱ型１３例，总缺失率为８０％。患儿父母及正常人共６８例，其中１例患儿母亲为纯合缺失，其余均无纯合缺失。     

抗T淋巴细胞单克隆抗体治疗再生障碍性贫血患儿近期疗效的观察

抗Ｔ淋巴细胞单克隆抗体（ＭｃＡｂＴ）治疗成人再生障碍性贫血（简称再障,ＡＡ）已有报道［１］,儿童则少见。我们采用ＭｃＡｂＴＳＭＵ３（抗ＣＤ３）和ＳＭＵ８（抗ＣＤ８）治疗儿童ＡＡ２８例,已取得较好疗效,现报告如下。病例选择：ＡＡ的诊断、分型均符合１９８７年全国会议修订的标准［２］。男１７例,女１１例;年龄２～１３岁（中位年龄７岁）。病程最短８．５个月,最长９年（中位数１５个月）。其中急性再障（ＳＡＡⅠ）７例,慢性重型再障（ＳＳＡⅡ）１０例,普通慢性再障（ＣＡＡ）１１例。用ＭｃＡｂＴ治疗前,…     

用PCT—SSCP方法检测中国人Alport综合征COL4A5基因突变

目的 检测中国人Ａｌｐｏｒｔ综合征（ＡＳ）患者编码Ⅳ型胶原α链的ＣＯＬ４Ａ５基因突变。方法 提取１８例中国Ｘ连锁显性遗传型ＡＳ患者基因组ＤＮＡ，据ＣＯＬ４Ａ５基因中与外显子相邻的内含子序列设计引物，分别作ＤＮＡ聚合酶链式反应（ＰＣＲ），扩增了该基因第２０～５１外显子。以琼脂糖凝胶电泳鉴定产物，作单链构象多态性（ＳＳＣＰ）分析检测ＰＣＲ产物，对迁多率异常者进行ＤＮＡ测序。结果 ＰＣＲ－ＳＳＣＰ分析发     

低(无)苯丙氨酸奶粉“华夏2号”治疗苯丙酮尿症的临床观察

苯丙酮尿症（ｐｈｅｎｙｌｋｅｔｏｎｕｒｉａ，ＰＫＵ）是先天性遗传性氨基酸代谢病 ,我国发病率约为１／１１０００。本病患儿因肝中缺乏苯丙氨酸羟化酶 (ＰＨＡ) ,使血苯丙氨酸 (Ｐｈｅ)浓度增高 ,影响了脑发育而导致智能障碍等一系列临床症候群。低 (无 )苯丙氨酸奶粉治疗ＰＫＵ是当今的有效疗法 ,因极大多数ＰＫＵ家庭难以承受昂贵的进口奶粉 ,１９８９年由我们自行设计、研制的国产第一代低苯丙氨酸奶粉问世 ,取得了良好的临床疗效。１９９６年又研制成功第二代奶粉“华夏２号” ,本文总结了３年来“华夏２号”治疗ＰＫＵ患儿的临床疗…     

Prenatal screening for phenylketonuria in 2 families by trophoblast biopsy

Prenatal diagnosis of phenylketonuria (PKU) is now available owing to restriction fragment length polymorphism (RFLP) of the phenylalanine hydroxylase gene. Biopsies of chorionic villi were carried ou at 11 weeks gestation in 2 families who had previously a child with the classical form of PKU and who were considered informative after DNA studies. Fetal DNA was extracted and studied with restriction enzymes selected according to the study of each family. Hybridization studies suggested that one fetus was affected by the disease and that the second was normal. Termination of pregnancy was carried out in the first case; however, study of the fetus could not be performed. In the second family, the pregnancy resulted in the delivery of a normal child, as shown by normal phenylalanine level at birth.     

脊髓性肌萎缩的基因诊断和产前基因诊断研究

目的探讨中国人脊髓性肌萎缩(SMA)基因诊断和产前基因诊断的可行性。方法应用复合聚合酶链反应-限制性片段长度多态(PCR-RFLP)方法对31例SMA患儿进行神经元存活基因(SMN)第7外显子缺失分析,并对2例有SMA阳性家族史的家系进行了产前基因诊断。结果96.8％(30／31)SMA患儿携有SMN基因第7外显子缺失。2例产前基因诊断的病例均无SMN基因第7外显子缺失。结论SMN基因缺失检测技术可用于SMA患儿的基因诊断和产前基因诊断。     

Confidential inquiry into families with two siblings with cystic fibrosis

OBJECTIVE: To audit the care that had been provided to couples before the birth of a child with cystic fibrosis where a sibling had been previously diagnosed. DESIGN: Retrospective review of case notes. SAMPLE: Families where at least one affected child had been born between 1 January 1991 and 30 June 1995 and the diagnosis in the first child was made before the second affected pregnancy reached 20 weeks. The combination of information on these families with data from the prenatal diagnosis register allowed the reconstruction of a cohort of pregnancies in women with a previous affected child. MAIN RESULTS: Forty six eligible families with a second affected child were identified. Details from the paediatrician who had diagnosed the first affected child were obtained in 43 cases: all 43 couples were offered genetic counselling, but where provided by a paediatrician this was difficult to assess as no couple was sent a summary letter. Details were obtained from the obstetrician in the subsequent affected pregnancy in 42 cases: prenatal diagnosis was not offered in 10 (24%), offered and declined in 24 (57%), offered and accepted but termination declined in eight (19%). In the overall cohort of at risk pregnancies, the estimated rate of prenatal diagnosis offer was 97%, prenatal diagnosis uptake 86%, false negative prenatal diagnosis rate 0%, and uptake of termination 95%. CONCLUSIONS: (1) Parental choice was an important determinant of second affected births. (2) Despite widespread availability, prenatal diagnosis was not offered in an estimated 3% of at risk pregnancies. (3) There were shortcomings in counselling documentation, in particular failure to send a summary letter to counselled couples.     

Molecular biology in genetic counseling of Duchenne and Becker myopathy]

From 1985-1991, molecular biology studies were carried out in 115 families affected with X-linked muscular dystrophy (DMD/BMD), including 59 prenatal diagnoses. The approach has changed over the last 6 years when new intragenic markers and cDNA probes became available. The polymerase chain reaction technique allows a rapid detection of dystrophin deletions, but classical Southern blot technique remains useful for restriction length polymorphism analysis. Fifty percent (42/85) of patients with DMD/BMD exhibited deletions of the dystrophin gene. In affected families with a detectable deletion, carrier detection is possible by gene dosage analysis and prenatal diagnosis is reliable. When no deletion is found, carrier detection and prenatal diagnosis depends on linkage analysis using polymorphic probes. Due to the high recombination rate, several markers need to be used. The information provided by linkage analysis must be interpreted given the proper family structure.     

DNA diagnosis of monogene hereditary diseases exemplified by phenylketonuria and mucoviscidosis

In 170 inherited diseases there exists the possibility for diagnosis at the DNA level. Using phenylketonuria (PKU) and cystic fibrosis (CF) as examples we demonstrate the capability of direct and indirect DNA-diagnosis through the use of DNA markers and allelespecific oligonucleotide hybridization respectively. In 88% of our PKU-patients and in 98% of the CF-patients DNA linkage analysis and therefore prenatal diagnosis on the DNA level can be carried out in affected families. The reliability of DNA-diagnosis is 99.0% for PKU and between 96.0-99.99% for CF depending on where the DNA-markers are localized. In contrast to CF, the PKU gene has been isolated and distinct mutations within the phenylalanine hydroxylase gene have been characterized. There is evidence for a correlation between genotype and clinical and biochemical phenotype. Also in CF it is indicated that certain DNA haplotypes correlate with the severity of the disease: less frequent haplotypes seem to be more often associated with a milder course than haplotype "B/B" which represents 85% of the CF chromosomes. Therefore DNA diagnostic methods not only make a major contribution to improved genetic counseling but also offer the possibility for a better future understanding of the heterogeneity of genetic diseases.     

宁夏苯丙酮尿症患儿苯丙氨酸羟化酶基因外显子7突变分析

目的 探讨宁夏地区苯丙酮尿症（PKU）患儿苯丙氨酸羟化酶（PAH）基因外显子7 突变类型及频率,为该地区PKU 的基因诊断和产前诊断提供依据。方法 应用PCR 产物直接测序方法,对宁夏73 例经典型PKU 患儿（回族39 例,汉族34 例）的146 个PAH 等位基因外显子7 及其旁侧内含子区域进行序列分析。结果 共检测出6 种突变基因型,分别是R243Q（14.4%）、R241C（6.8%）、IVS7+2T → A（2.7%）、L255S（0.7%）、G247V（0.7%）和G247R（0.7%）。外显子7 突变基因总频率为26.0%（38/146）,包括错义突变和剪接位点突变两种。回族患儿R241C 等位基因突变检出率高于汉族（10% vs 3%,P<0.05）。结论 宁夏地区PKU 患儿PAH 基因外显子7 突变频率最高的是R243Q,其次为R241C;回族和汉族PKU 患儿R241C 等位基因突变率不同。     

青岛市苯丙氨酸羟化酶缺乏症患儿基因突变分析

目的探讨青岛市苯丙氨酸羟化酶(phenylalanine hydroxylase,PAH)缺乏症患儿的基因突变特点,为青岛市PAH缺乏症的产前诊断、治疗提供科学参考依据。方法对经青岛市新生儿疾病筛查确诊的44例PAH缺乏症患儿,应用第二代高通量测序及多重连接酶探针依赖扩增(multi-ligase probe dependent amplification,MLPA)技术进行基因分析,检测患儿基因突变位点,应用Sanger测序对其父母的PAH基因相应突变位点进行检测并验证。根据患儿血苯丙氨酸浓度,分为经典型苯丙酮尿症、轻度苯丙酮尿症和轻度高苯丙氨酸血症。结果①44例PAH缺乏症患儿PAH基因中均检测到2个突变位点,其中2例为纯合突变,纯合突变的频率为4.6%,所有突变在患儿父母相应突变位点处均能检测到。②44例PAH缺乏症患儿共检测到突变36种,其中c.728G>A突变频率最高(15.9%,14/88),其次是c.1068C>A(10.2%,9/88),再次为c.158G>A(9.1%,8/88)。③21例经典型苯丙酮尿症患儿PAH基因突变19种,其中c.1068C>A突变频率最高(21.4%,9/42),其次是c.728G>A(19.0%,8/42)。10例轻度苯丙酮尿症患儿PAH基因突变14种,其中c.721C>T/722delG突变频率最高(15.0%,3/20),其次为c.1197A>T、c.1301C>A、c.721C>T、c.728G>A(均为10.0%,2/20)。13例轻度高苯丙氨酸血症患儿PAH基因突变17种,其中c.158G>A突变频率最高(26.9%,7/26),其次为c.728G>A(15.4%,4/26)。结论青岛市PAH缺乏症患儿PAH基因突变以复合杂合突变为主,具有明显热点突变(c.728G>A、c.1068C>A、c.158G>A),经典型苯丙酮尿症患儿以c.1068C>A、c.728G>A为主,轻度苯丙酮尿症患儿以c.721C>T/722delG为主,轻度高苯丙氨酸血症患儿以c.158G>A为主。本研究明确了青岛市PAH缺乏症患儿基因的突变类型与特点,为深入开展PAH缺乏症的诊断以及进一步的基因治疗奠定了基础。     

PKU and NON-PKU Hyperphenylalaninemia: Differentiation, Indication for Therapy and Therapeutic Results

The differential diagnosis of phenylalanine hydroxylase deficiency (PAHD) by biochemical methods is difficult. Using standardized oral protein loading or the intravenous deuterated phenylalanine (phe) load in 46 patients with PAHD, three groups could be distinguished: 1) Phenylketonuria (PKU) with plasma phe levels over 20 mg% under the protein load and with residual activities of less than 1% of normal; 2) mild PKU with plasma phe of 10–20 mg% and residual enzyme activities of 1–3%; 3) Non-PKU hyperphenylalaninemia with plasma phe levels of less than 10 mg% and residual enzyme activities of more than 3%. Psychomotor development in 32 untreated patients with PAHD showed that there is a high risk of brain damage for all patients with in vivo residual activities of less than 3% of normal. Restriction fragment length polymorphism (RFLP) haplotypes at the phenylalanine hydroxylase (PAH) locus in 60 German patients with PAHD showed that 90% of the mutant alleles are confined to four distinct haplotypes. Using an oligonucleotide probe for the splicing mutation associated with mutant haplotype 3 a close association between the mutation and the haplotype 3 has been observed. There is also an association between haplotypes 2 and 3 and PKU patients with residual enzyme activities of less than 1%. However, in only 37% of our patients with PAHD could a direct diagnosis of the mutations in the PAH gene be made. More knowledge of other mutations in the PAH gene is necessary to differentiate patients with PAHD on the DNA level.     

Carrier detection by DNA analysis in Duchenne muscular dystrophy families.

We applied DNA analysis techniques to Turkish families whose members were afflicted with Duchenne/Becker muscular dystrophy. The aim of this study was to establish a prenatal diagnosis of this anomaly and to determine the carrier state. All of the techniques used in established diagnosis centers are now applied routinely in our laboratory. Both Southern analysis and polymerase chain reaction (PCR) methods were used for deletion detection in patients and restriction enzyme fragment length polymorphism (RFLP) determination for linkage analysis in women at risk. CA repeated sequence length polymorphism, the most recent technique for linkage analysis, was also applied. About 250 individuals from seventy-nine families were investigated and thirty-six entire families were screened. Twenty-five women were found to be carriers while thirty seven were non-carriers. The carrier state could not be determined in three women.     

Confidential inquiry into families with two siblings with cystic fibrosis

OBJECTIVE—To audit the care that had been provided to couples before the birth of a child with cystic fibrosis where a sibling had been previously diagnosed.
DESIGN—Retrospective review of case notes.
SAMPLE—Families where at least one affected child had been born between 1 January 1991 and 30 June 1995 and the diagnosis in the first child was made before the second affected pregnancy reached 20 weeks. The combination of information on these families with data from the prenatal diagnosis register allowed the reconstruction of a cohort of pregnancies in women with a previous affected child.
MAIN RESULTS—Forty six eligible families with a second affected child were identified. Details from the paediatrician who had diagnosed the first affected child were obtained in 43 cases: all 43 couples were offered genetic counselling, but where provided by a paediatrician this was difficult to assess as no couple was sent a summary letter. Details were obtained from the obstetrician in the subsequent affected pregnancy in 42 cases: prenatal diagnosis was not offered in 10 (24%), offered and declined in 24 (57%), offered and accepted but termination declined in eight (19%). In the overall cohort of at risk pregnancies, the estimated rate of prenatal diagnosis offer was 97%, prenatal diagnosis uptake 86%, false negative prenatal diagnosis rate 0%, and uptake of termination 95%.
CONCLUSIONS—(1) Parental choice was an important determinant of second affected births. (2) Despite widespread availability, prenatal diagnosis was not offered in an estimated 3% of at risk pregnancies. (3) There were shortcomings in counselling documentation, in particular failure to send a summary letter to counselled couples.