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Article in Chinese | WPRIM | ID: wpr-435968


Clerkship teaching of neurology is one of the important contents in clinical neurological teaching.Application of problem-based learning in combination with heuristic teaching can mobilize the initiative of students,enlighten their thoughts,arouse their flexibility and creativity and deepen the understanding of knowledge so as to achieve the purpose of improving clinical teaching effects.

Article in Chinese | WPRIM | ID: wpr-388299


Objective To study improvement of cognitive function impairment of vascular dementia rats induced by a permanent bilateral ligation of common carotid arteries (2VO) after administration of leptin in brain. Methods Hippocampal neurons was isolated and cultured from SD rats. At oxygen-glucose deprivation condition, protection role of leptin on hippocampal neurons was observed and expression of leptin receptor was detected. Animal model of rats was established by 2VO. Pre-treatment VaD model of leptin was established by administration leptin into hippocampus region. The Morris water test was performed to detect difference in the cognitive function between VaD group and control group. Neuron apoptosis in hippocampus tissue were determined with TUNEL. Results Leptin receptor expression could be seen in hippocampal neurons. After oxygen-glucose deprivation cultured for 12 h , plenty of apoptotic cells were seen in hippocampal neurons, apoptosis rat was up to (72.96 ± 6.25) % , while apoptosis rate was (46.33 ±7.85)% and (23.58 ±5.08)% in 1 ( μg and 5μg leptin treatment group,respectively. Compared leptin treatment group with control group,difference had a statistical significance(P<0.01). Compared with untreated VaD group, latency time was shorter and average velocity was increased in leptin-treat-ment VaD group. Neuron apoptosis in hippocampus tissue of leptin-treated group were different significantly from those of untreated group (P < 0.01). Conclusion Leptin could protect hippocampal neurons from apoptosis in vitro. Cognitive function impairment could be improved by administration of leptin into brain in VaD rats.

Article in Chinese | WPRIM | ID: wpr-411418


Objective  Determination of Wilson disease gene mRNA expression in human fibroblast cell strain (Me32aT22/2L) by reverse transcription-polymerase chain reaction (RT-PCR). Methods Using lipofection reagent, the plasmid vector carrying the Wilson disease gene (pRc/CMV-WD) was transferred into Me32aT22/2L cultured in serum free complement medium. RT-PCR was used to determine WD mRNA expression in Me32aT22/2L. Results  Wilson disease gene expression was detected in Me32aT22/2L, while no specific signals were detected in untransfected fibroblast. Conclusions It demonstrated that Me32aT22/2L strain could express the Wilson disease gene, suggesting that Wilson disease gene transfer might develop a new approach to study Wilson disease.

Article in Chinese | WPRIM | ID: wpr-411416


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.

Article in Chinese | WPRIM | ID: wpr-555385


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.