Objective To construct the short hairpin RNA(shRNA) expression vector of survivin and down-regulate the expression of survivin through RNA interference in neuroblastoma cell line SH-SY5Y. Methods Two pairs of oligonucleotide sequences specific for human survivin mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pBSHH1 plasmid. After being identified by restriction enzyme digestion and sequencing, the recombinant plasmids pBSHH1-S1 and pBSHH1-S2 were transfected into SH-SY5Y cells, respectively. Survivin expression in the transfected cells was assayed by both RT-PCR and western blot. Results Enzyme digestion analysis and DNA sequencing showed that the oligonucleotide fragments were correctly inserted into pBSHH1 plasmid, and survivin expression in the transfected cells was knocked down significantly by pBSHH1-S1 or pBSHH1-S2 at both the protein and mRNA level. Conclusion The shRNA expression vectors of survivin were successfully constructed, and could down-regulate survivin expression in SH-SY5Y cells, which lay a foundation for further research on gene therapy for tumors such as neuroblastoma.

Objective To investigate the clinical characteristics of essential tremor (ET).Methods The clinical materials of 80 patients with ET was analysed.Results In 80 patients with ET, 50 cases for the male, 30 cases for the female,the age at tremor onset was from 3 to 70 years old,the mean age was 34.6?16.3 years old,the course of disease was 6 months to 60 years,the mean 14.2?9.9 years. 45 patients (56 3%) had postive family history,they showed mostly autosomal dominant inheritance.The postural tremor was the main manifestation of essential tremor,tremor affected hand in 92.5% patients,throat in 21.3%,head in 20%,chin in 17.5%. Functional disability and impairment in 17.5% patients because of tremor.91 2% patients had the effect of alcohol occurred in 34 cases who had performed alcohol test.6 3% patients accompanied parkinson’s disease.A small dose of propranolol was effective in 61% patients.Conclusion In the ET group,the patients were mostly young,single model,the number of the male ET cases was more than the female,sole postural tremor,part of patients might accompany PD,a small dose of propranolol was effective.

Objective To explore parkin gene deletion mutations at exons 3 to 7 in Chinese familial patients with Parkinson's disease as well as the association with the clinical features.Methods DNA was extracted from 6 unrelated families of PD patients; the deletion mutations of parkin gene at exons 3 to 7 were identified by PCR amplification, agarose gel electrophoresis. and the clinical data were analyzed together with the above information.Results In the 6 unrelated families of PD patients, 1 case had exon 5 deletion, its hereditary manner was autosomal recessive inheritance,the patient's age at the onset was 60 years old,clinical feature was tremor,rigidity and bradykinesia,but no athetosis.In addition the deletion mutations of parkin gene at exons 3,4,6,7 were not found.Conclusion There are deletion mutations of parkin gene at exons 5 in families PD of Chinese patients.

Objective To explore the clinical characteristics of hereditary spastic paraplegia with thin corpus callosum(HSP TCC).Methods Clinical data of 4 patients with HSP TCC were analysed retrospectively.Results 4 patients were at the onset during youngsters,they revealed mental impairment,walk of spasticity,spasticity of the lower extremities,slowly progressive weakness and hyperreflexia, extensor plantar responses and morbid indication for positive. Sensory impairment was not observed. 2 cases showed ataxia and sphincter disturbance;1 case showed upper limb spasticity and muscular atrophy. Cranial MRI revealed an extremely thin corpus callosum on sagittal image.Conclusion Main clinical characterizations of HSP TCC were slowly progressive spastic paraparesis, mental impairment during youngsters, cranial MRI showed extremely thin corpus callosum.

Objective To investigate the clinical and genetic characteristics of hereditary spastic paraplegia(HSP).Methods The clinical material of 113 patients in 39 families with HSP was analyzed retrospectively.Results The ratio of male to female was 1:1.17.The age at HSP onset was from 2 to 58 years old, the mean age was 21.4 years old, and 81.7% of the patients had HSP before 30. 89.4% of the patients had positive family history and they showed mostly autosomal dominant inheritance. The rate of consanguinity was 28.2%. 24 cases had pure while 89 cases had complicated spastic paraplegia. In the HSP group, we could found the weakness of legs in 65.5% patients, spasticity and hyperreflexia of lower limbs in 96.5%, extensor plantar responses in 68.1%, ataxia in 46.9%, muscular atrophy in 32.7% and dementia in 18.6%.Conclusion In the HSP group, the year of onset was mostly before 30. The female HSP cases were more than the male's, and the complicated cases were more often than the pure. Autosomal dominant was the mostly frequent inheritance, and there were more chances of HSP in the consanguineous families.

Objective To evaluate the clinical and genetic characteristics of Charcot-Marie-Tooth disease (CMT). Methods The clinical materials and hereditary histories of 110 cases in 70 families with CMT were analyzed retrospectively.Results The ratio of male to female was 2.03∶1. The age at onset was from 1 to 61 years old and the mean age was 19.1 years old. 78.2% of the patients had CMT before 30 years old.70 patients (63.6%) had positive family history and they showed mostly autosomal dominant inheritance. The rate of consanguinity was 6.9%. In the CMT group, we could find the amyotrophy of legs in 106 patients (96.4%), distal muscle weakness and atrophy of the upper limbs in 48 patients (43.6%), stork legs in 64 patients (58.2%), pes cavus in 68 patients (61.8%), decreased or diminished tendom reflexes in 108 patients (98.2%). Electromyography examination in 61 patients showed neurogenic damages. Muscle biopsy in 37 patients showed neurogenic amyotrophy. Sural nerve biopsy was performed in 25 patients. 20 patients were charactered by demyelination, Schwann cell proliferation and/or “onion bulbs” change and 5 patients were associated with axis cylinder degeneration, but tomaculous change was not found in all the 25 patients.Conclusions In the CMT group, the onset age was mostly in childhood and adolescence. The male CMT cases were more than the females. Autosomal dominant was the mostly frequent inheretance. Neuroelectrophysiology and pathological examination are important for the diagnosis and type of CMT.

Objective To study the gene mutation and clinical characteristics of hereditary spinocerebellar ataxia type 7 (SCA7).Methods The regions of SCA7 gene containing CAG repeat were amplified by means of polymerase chain reaction (PCR) and polyacrylamide gelelectrophoresis (PAGE) technique in 112 patients with autosomal dominant SCA from 92 families, 16 sporadic SCA patients, 71 family members and 60 healthy controls. The abnormal allele fragments were sequenced by ABI377 DNA sequencing machine. The correlation between clinical manifestations and CAG repeat size in SCA7 gene product was analyzed. Results 2 patients of 6 members in 1 SCA7 family carried a 71-repeat allele. The main clinic features included ataxia, hypopsia, axanthocyanopsia and retinal pigmental degeneration. Alleles from 7 to 9 repeats were seen in the other 4 healthy members. CAG repeats from 6 to 21 were found in other 126 SCA patients, 71 family members and 60 healthy controls. Conclusions Expanded triplet repeats in SCA7 gene contributes to the pathologic phenotype, and molecular genetic analysis is effective for the diagnosis of SCA7. Retinal pigmental degeneration is an important characteristic of SCA7.

Objective To investigate the mutation characteristics of atlastin gene in Chinese patients with hereditary spastic paraplegia(HSP) and establish the base of gene diagnosis of HSP.Methods Mutation analysis of atlastin gene was made by use of polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) combined with DNA direct sequencing in 30 unrelated affected HSP individuals in China in which 20 cases were from autosomal dominant families and ten cases were sporadic HSP patients.Results No abnormal SSCP bands were found in the 30 individuals and the results of DNA direct sequencing were also normal.Conclusion Mutation of atlastin gene may be rare in Chinese HSP patients.

Objective To investigate the mutation of Cx32 gene,clinical and electrophysiological features of Chinese patients with Charcot-Marie-Tooth(CMT) disease.Methods 24 CMT probands were selected for Cx32 mutation screening after the exclusion of the CMT1A 1.5 Mb duplication and male-to-male transmission.The motor and sensory nerve conduction studies were performed in all probands and most of their affected family members to establish the clinical CMT1,CMT2 or CMT intermediate diagnosis.The presence of mutations in the coding region of Cx32 was detected by single-strand conformation polymorphism analysis combined with direct sequencing.Results It was found 7 different point mutations in the coding region of Cx32 in 1 sporadic CMT1 patient and 6 X-linked inherited families,including 4 families with CMT1 diagnosis and 2 families with CMT intermediate diagnosis.There were 20 male CMTX patients,6 female CMTX patients and 12 asymptomatic female carriers among 38 family members bearing Cx32 mutation.All of the 26 patients were mildly to moderately affected clinically.Conclusions Seven different Cx32 point mutations were detected and the percentage of Chinese CMT families with Cx32 mutation is about 10% in our study.The inheritance model of CMT secondary to Cx32 mutation could be X-linked dominant,X-linked recessive or sporadic.Male patients are usually more severly affected than females with slower nerve conduction velocities.Cx32 mutation screening should be firstly performed in those CMT families without male-to-male transmission and CMT1A duplication.

Objective To explore the clinical features of autosomal recessive juvenile Parkinson disease(AR-JP).Methods The clinical materials of 28 patients from 15 families with AR-JP were analyzed retrospectively.Results The onset of all the patients was insidious and the mean age was 26.1 years old.In 23 patients(82.1%),the symptoms began at one limb or one side and progressed bilaterally in a mean time of((4.7?)3.6) years.Bradykinesia(100%),rigidity(100%),resting tremor (85.7%),postural instability(60.7%),hyperreflexia(53.6%),dystonia(32.1%) and diurnal fluctuations with sleep benefit(89.2%) were the cardinal symptoms.The mean improved Webster score was(11.2?)6.1.The mean maintenance dose of DOPA-preparation was((0.40?)0.28) g/d.The mean UPDRS motor score was(27.9?)10.3 before treatment and it decreased to(6.7?)5.4 after therapy(P