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 explore the association of activated blood platelet and large vacular disease in type 2 diabetes mellitus.Methods A total of 98 cases of type 2 diabetes mellitus patients were divided into two groups(50 cases with large vascular disease and 48 cases,as DM group)and 50 cases healthy subjects served as a control group.From 2002-05 to 2004-11,the doctors of the Department of Endocrinology,Subei Hospital to determined the expression of platelet activation PAC-1、CD62P in large vascular disease by technique of flow cytometry(FCM),and glycosylated hemoglobin(HbA_1c)、C-reactive protein(CRP)、fasting plasama glucose(FBG)、C-peptide(C-P)and blood lipids were measured and analysed for the correlation.Results Activated blood platelet was significantly increased in DM group as compared with that in control group(P

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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.

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.

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.

Objective To observe the effect of Changji'an Prescription(CP) on T lymphocytes count and 5-hydroxytryptamine(5-HT) secretion in rats with diarrhea predominant irritable bowel syndrome(IBS).Methods Forty-six neonatal SD rats were randomized into normal group,model group,low-,middle-and high-dose CP groups(0.94,1.88 and 3.76g?kg-1?d-1 respectively),and Bupi Yifei Pills group(6.76g?kg-1?d-1).Continuous internal rectal stimulation with acetate was used to induce the rat model of intestinal hypersensitive IBS.Flow cytometer was used for the quantitative analysis of peripheral levels of T lymphocytes subsets CD+3,CD+4 and CD+8,and immunohistochemical method was used to observe 5-HT-positive cells number and the immunostaining intensity in the bowels of rats.Results In the model group,the defecation volume and the intestinal sensitivity were increased,CD+4 level and CD+4/CD+8 decreased,CD+8 level increased,5-HT-positive cells number and the immunostaining intensity elevated(P

Objective To explore the relation of fat cells factor and insulin resistance in type 2 diabetes.Methods 60 patients were divided into obesity group, non-obese group and normal control group.To detect the FINs, FPG, TG, TC, LDL,HDL,APN,CTRP3,leptin and TNF-α, according to the formula of HOMA,to analyze the correlation between indicators.Results 1).Compared with normal control group, DM group significantly decrease serum APN, CTRP3 level, compared with non-obesity group, obesity group APN, CTRP3 level significantly decreased(P<0.05 or P<0.01).The leptin level elevated (P<0.05).2)APN concentration and age, BMI, FPG, TG, TC, LDL, FINs, HOMA-IR has negative correlation;CTRP3 andFPG,FINs, HOMA-negative correlation IR(P<0.05), TG was positively correlated (P<0.05);TNF-α and FPG and positively correlated (P< 0.05).Conclusions T2DM patients` serum APN, CTRP3 and high Leptin levels are closely associated with obesity, HOMA-IR, FPG and TG as an independent factor can be used as a new sensitive index of evaluating the degree of insulin resistance.

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.

Objective To investigate the expression level and clinical significance of long non-coding RNA(LncRNA) growth arrest specific gene-antisense 1(GAS8-AS1) in papillary thyroid microcarcinoma(PTMC) patients. Methods We investigated the expression profile of GAS8-AS1 in tissue samples of patients with PTMC as well as nodular goiter(NG) by quantitative real-time polymerase chain reaction(RT-qPCR). Results GAS8-AS1 in cancer tissue was down-regulated in PTMC patients compared with adjacent thyroid tissue and NG samples(P<0.05). Lower level of GAS8-AS1 was also correlated with central cervical lymph node metastasis(CLNM, P<0.05). The area under the ROC curve for GAS8-AS1 was up to 0.717 3 in CLNM prediction(P<0.05). Conclusion GAS8-AS1 may act as a potential biomarker for PTC diagnosis and CLNM prediction.