BACKGROUND: It is of great importance to study the genotype distribution of hereditary ataxia in understanding its epidemiologic rule and pathogenetic pathway.OBJECTIVE: To analyze the distribution of different genotype of hereditary ataxia in south China.DESIGN: A case-control observation.SETTING: Department of Neurology, the First Affiliated Hospital of Sun Yat-sen University.PARTICIPANTS: Forty-three patients (26 males and 17 females) with hereditary ataxia from 36 families and 38 patients with sporadic hereditary ataxia (24 males and 14 females) were selected from the Outpatient Clinic of Neurogenetics, Department of Neurology, the First Affiliated Hospital of Sun Yat-sen University between September 1998 and September 2002. At the same time, 60 healthy individuals from the patients' families and 44randomly-selected healthy physical examinees were taken as controls. All the participants were enrolled voluntarily.METHODS: The fragments of trinucleotide repeats at different sites of mutant genes were amplified with polymerase chain reaction (PCR), and then the lengths were calculated with polyacrylamide gel electrophoresis and imaging analytical software. The repeated numbers of trinucleotide repeats in all the normal and abnormal amplified alleles were calculated respectively.MAIN OUTCOME MEASURES: Different genotype distribution in patients with hereditary ataxia.RESULTS: All the subjects were involved in the analysis of results. Of the detected patients with hereditary ataxia, the Machado-Joseph disease/spinocerebellar ataxia (SCA) 3 was the most common type of autosomal dominant SCA in South China, which was 42.0%, and was followed by SCA2 (7.4%), SCA1 (4.9%), SCA7 (3.7%), SCA6 (2.5%), SCA12 (1.2%).No patient was detected to have SCA8 SCA 10, SCA 17 dentatorubropallidoluysian atrophy (DRPLA) and Friedreich ataxia (FRDA).CONCLUSION: Autosomal dominant SCA3 is the most familiar genotype in South China. Clinical detection of hereditary ataxia should be done firstly aiming at the SCA3 genotype.

Objective 　To screen for gene mutation of exon 18 in Chinese patients with Wilson disease. Methods　PCR-SSCP was used to screen exon 18 in 45 Wilson disease patients among 39 Chinese families and 10 normal controls. Those with abnormality were further analyzed by necleotide sequence analysis. Results　There were 16 mobility shift with two different styles in exon 18. All abnormal mobility shifts were sequence analysed. No gene mutation was found. Conclusions　Our result suggest that, contrary to findings in Caucasians, exon 18 is not a frequent mutation point in Chinese patients with Wilson disease.

0 05) Conclusions Arg778Leu mutation was related to the onset age in patients with WD, but not to patients′ sex, first symptoms and copper metabolic disturbance Arg778Leu mutation is suggested to postpone the onset age of WD patients

AIM: To construct the liver-specific transgene vectors encoding wild-type as well as most common disease mutant STBXATP7B cDNA under the control of mouse albumin promoter and to explore their expression. METHODS: Two Kbpala/Alb-STBXATP7B mutants containing the Arg778Leu and His1069Gln mutations were constructed using site-directed mutagenesis system plus site-subcloning technique. The vectors expressed wild-type and mutants of human STBXATP7B in mouse liver cells were obtained and transiently transfected into BRL and BHK cell lines. Western blotting analysis was utilized to detect the expression of human STBXATP7B. RESULTS: Enzyme analysis and sequencing analysis confirmed that the target genes were STBXATP7B and in right position. The results of Western blotting showed that the gene products were expressed specifically in liver cells. CONCLUSION: The Kbpala/Alb-STBXATP7B vectors were constructed successfully and the liver-specific expression of human STBXATP7B proteins were conformed.

Objective To study the molecular genetic diagnosis and clinical characteristics of spinocerebellar ataxia type 6 (SCA6).Methods 43 patients with autosomal dominant SCA from 36 families and 38 sporadic SCA patients were enrolled in the study. SCA6 (CAG)n dynamic mutations were detected by polymerase chain reaction (PCR). Abnormal allele fragments were sequenced and repeated numbers were calculated. The clinical data of two cases with SCA6 were analyzed.Results CAG repeat of normal SCA6 allele ranged from 10 to 13. CAG repeat of abnormal SCA6 allele expanded to 25 in one familial patient and 24 in one sporadic patient in our study. The basic characteristics of these SCA6 patients were slowly progressive cerebellar ataxia, nystagmus and dysarthria.Conclusion Diagnosis of SCA6 can be confirmed by detection of abnormal CAG repeat expansion. There is no obvious difference of clinical features between SCA6 and other SCA subtypes.

Objective To investigate the change and role of nitric oxide (NO), nitric oxide synthase (NOS) in serum of children with Epilepsy.Methods The serum NO and NOS levels of 58 epileptic children and 23 healthy children were measured with enzyme linked immunosorbent assay (ELISA). Results The concentrations of the serum NO [( 5.86? 1.21)?mol/ml] and NOS [( 28.26? 8.49)U/ml] in patient group were significantly higher than those in control group [( 3.78? 0.74)?mol/ml,( 17.86? 4.58)U/ml)]( P

0.9 mg/L in 6 patients.Kayser-Fleischer ring were found in 85.5 % of all the patients. The abnormal hepatic function in the liver type HLD was more common than that of in the brain type. The liver injury was detectable by B mote ultrasonic wave in different type HLD.MRI examination was taken in 79 patients, 65 of them had showed the symmetry abnormal signs in basal ganglia. Conclusions CP has independent diagnostic values when its content ≤0.08 g/L. Kayser-Fleischer ring is an excellent discriminatory test for the diagnosis of HLD patients with neurological or psychiatric symptom. 24 h urine copper is the best single screening test because it increases over 100 ?g/24 h in all patients who were taken the test. The B mote ultrasonic wave test for the liver and MRI for brain were helpful in detecting the damage in the liver and brain.

Objective To study the gene mutation and clinical characteristic of hereditary spinocerebellar ataxia type 7 (SCA7).Methods The SCA7 (CAG) trinucleotide repeat mutations were detected by polymerase chain reaction(PCR) and polyacrylamide gel electrophoresis technique in 24 patients with autosomal dominant SCA from 15 families, 20 sporadic SCA patients and 41 normal persons from the same family and 30 healthy persons from different family,the abnormal allele fragments were sequenced by ABI 373 DNA sequencing machine.Results 24 patients with SCA had CAG repeat numbers of SCA 7 allele from 9~18.Normal alleles of SCA 7 had CAG repeat number from 9 to 19. One sporadic SCA patient had one abnormal SCA 7 allele with the CAG repeat expanded to 63 repeats, being confirmed by DNA sequencing.Conclusion CAG expansions were pathogenic cause of SCA 7. The technique of gene mutation detection could provide an effective way for the prediction of asymptomatic and genetic counseling,which was a basis for gene typing.

Objective To screen polymorphisms and mutations in the promoter region of WD gene.Methods DNA from peripheral blood was obtained from 71 subjects of 36 family (48 WD patients,23 patients first-degree relatives) and 20 healthy people from Feb.2001 to Feb.2004.DNA sequence of the genes was analyzed by PCR amplification and direct genomic sequencing.Results There were three polymorphisms at positions-190,-78,+260(transcription start site as +1) of the promoter region of WD gene.Normal controls,WD patients and patients’ first-degree relatives all showed the polymorphisms;three of 48 WD patients presented C→T base substitution mutations at the same position -183:two were homozygous mutation,while the other was heterozygous.Normal control subjects and patients' relatives didn’t show this kind of mutation.Conclusion It suggests that the mutation of the promoter region is one of WD pathogenesis.

Objective 　To study the sequence and structure of intron 8 in WD gene in order to further understand the relationship between intron 8 and WD. Methods　We utilized polymerase chain reaction (PCR) to the amplification of exon 8-intron 8-exon 9 which were then sequenced by a dideoxy chain termination methon in 10 normal controls and 32 members of 11 families(20 WD patients and 10 of their relative). The results were analyzed by the computer. Results　The sequence of intron 8 was 703 bp with the G　+　C content of 42.7%. There were one short tandom repeats, 7 direct and inverted repeats in it. An open reading frame coded with 82aa was found at 323 base pairs of downstream of a TATAbox. There were two DNA polymorphisms at 408 and 487 nucleotides. The sequence analysis showed that the 5end has the sequence of 5-GTAAC, 3end has the sequence of CCTAG-3, and branchpoint of 5-TTTCGA-3.Conclusions　The sequences and structures of intron 8 in WD familiess members are not different from normal controls. Our data suggest that the WD gene intron 8 might not play an important role in the pathogenesis of WD.