Objective:To prepare the recombinant neuron protective protein TAT-Bcl-XL in E.coli expression system and to determine its anti-apoptosis activity.Methods:DNA fragment of TAT-Bcl-XL was obtained by RT-PCR using primers that were specific for Bcl-XL gene. Prokaryotic expression vector(pTBTOPO) was constructed by insertion of TAT-Bcl-XL DNA fragment into pCRT7/CT-TOPO vectors, and the recombinant protein was detected by SDS-PAGE and Western blot analysis using anti-V5 tag epitope antibody as the primary antibody. The recombinant fusion protein was purified by affinity chromatography, and the anti-apoptosis function of the purified protein was determined by flow cytometry.Results:A molecular weight of 30 000 protein was detectable in both SDS-PAGE and Western blot analysis. Immunofluorescent staining showed the fusion protein was distributed in cell plasma after incubation with 200 nmol/L fusion protein. Flow cytometry data indicated that the recombinant protein could enhance cell survival by 40% when 293T cells were treated with 250 ?mol/L zinc chloride as a supplementation in cell culture media.Conclusion:High level expression of the reported neuron protective protein was obtained in E.coli expression system, and the purified recombinant proteins remained its anti-apoptosis activity in our preliminary characterization.

Objective To investigate whether polymorphisms of apolipopretein D gene (APOD) have an effect on the risk for sporadic Alzheimer's disease (SAD).Methods Combination of polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing technique to screen all exons (1-5),along with flanking exon-intron boundaries of the ApoD gene.We investigated the polymorphisms of ApoD in 256 SAD patients and 294 healthy controls from North China by PCR-RFLP technique.Association of every polymorphism with AD was analyzed in this case-control study.Results Two ApoD (rs5952 and rs1568566) polymorphisms were detected and there were significant differences in the genotype or allele frequencies for the 2 ApoD polymorphisms respectively between cases and controls.Logistic analysis showed that rs5952C or rs1568566T allele carrier increased the risk for SAD (rs5952 adjusted OR=1.817,95% CI 1.237--2.669,χ2=9.282,P=0.002 ; rs1568566 adjusted OR=1.563,95% CI 1.060-2.306,χ2=5.072,P=0.024).The APOD polymorphisms showed gender specific associations.The linkage disequilibrium of the 2 single nucleotide polymorphism loci was found between rs5952 and rs1568566 of ApoD.Conclusion Polymorphisms of rs5952 and rs1568566 in APOD might play an important role in modifying risk for SAD.

Objective To study the effects of hypoxia and reoxygenation on expression of a disintegrin and metalloprotease10(ADAM10),?-amyloid precursor protein cleaving enzyme(BACE1) mRNA in SH-SY5Y cells.Methods SH-SY5Y cells were grouped into hypoxic and control groups.The hypoxic cells were incubated in hypoxic condition(95%N_2,5%CO_2) for 12 hours and 24 hours,and then cells were reoxygenated for 0 hours,12 hours and 24 hours.The expression of ADAM10,BACE1 mRNA were tested respectively at different time points after reoxygenation by RT-PCR.Control groups were incubated in normal conditions,seeded and treated at the same time with the hypoxic cells.Results The expression of ADAM10 mRNA was down-regulated by 19.8%,41.4% and 64.6%(P=0.005,0.038,0.001) at different time after reoxygenation with 12 hours hypoxia and down-regulated by 30.1%,75.9% and 86.5%(P=0.009,0.005,0.043)after reoxygenation with 24 hours hypoxia.The expression of BACE1 mRNA was up-regulated by 31.5% and 35.1%(P=0.028,0.005)only at 12 hours and 24 hours points after reoxygenation with 24 hours hypoxia.Conclusion Hypoxia and reoxygenation might alter the expression of ADAM10 and BACE1,which demonstrates that the vascular factors should make the amyloid precursor protein easy to be processed by ?-secrease pathway,thus to involve the pathology of Alzheimer's disease.

A zinc chloride and ethionine resistant mutant 0 5Eth400 5 was o btained from its parent strain Saccharomyces cerevisiae 346 by UV and 60 Co ? ray tr e atment and rational screening The glutathione productivity of the mutant reach e d 165 96mg/L by flask culture, which was 350% higher than the parent strain, an d the glutathione content in the dried cells reached 19 76mg/g, which was 318 6% higher than the parent strain A desecend of only 10 7% in the glutathione yie ld of the mutant was observed after ten times of subculture Therefore, the obtai ned mutant is stable strain that is worthwhile to be studied further

Objective To investigate the correlation between Presenilin 1 gene methylation (PSEN1) and sporadic Alzheimer's disease (SAD).Methods Massarray method was used to perform an analysis of DNA methylation in the promoters of PSEN1 gene in lymphocytes from 35 SAD patients,33 mild cognitive impaired (MCI)patients and 22 age-and gender-matched controls.Reporter gene plasmid of PSEN1 was built by genetic recombination methods,and then transiently transfected into the SH-SY5Y and HeLa cells.After that,cells were treated with several agents including serum deprivation,Aβ25-35,5-aza-2'-deoxycytidine (5-aza Cdr) and S-adenosyl-Lmethionine (SAM).A dual-luciferase reporter assay system was used to calculate the relative luciferase activity (RLA).Results Some CpG sites of PSEN1 (Cytosines at-173,95 and +76) showed hypomethylation patterns in SAD cases as compared with MCI and control group (P＜0.01).A dual-luciferase reporter assay showing significant differences in PSEN1 transcription activities were found in both cell lines under SAM treatment [SY5Y,SAM:(6.40±0.81),control:(9.13±1.67),P=0.000; Hela,SAM:(2.06±0.34),control:(2.99±0.59),P=0.000].Conclusions DNA hypomethylation of PSEN1 gene promoter is significantly associated with SAD susceptibility.PSEN1 may change the gene transcription level by methylation,further affecting the activity of amyloid protein-like γ-secretase,leading to the onset of AD.

This paper reviewed and analyzed the whole 29 years' compiling and revising histo-ry of Neurology the 5-year programming textbook for national undergraduates. The textbook has been becoming a high-quality textbook with novelty, practicability, readability and systematicness, conform-ing to the need of national education reform and reflecting the characteristics of neurology. Through the review of textbook history, we know that material construction can deepen the educational reform, improve the teaching level and discipline development, promote quality education and personnel train-ing.

Stroke is a major cause of death and disability in China.Some strategies to reduce the burden of stroke is to attach importance to thrombolytic therapy within 6 hours,to strengthen public education,to implement systems-level improvements in the prehospital and stroke unit,and to use statins and antiplatelet drugs to prevent and treate stroke.

Objectives: To study the correlation between the dynamic expression of aquaporin-4 (AQP-4) around hematoma and the blood-brain barrier permeability after intracerebral hemorrhage in rats. Methods: Forty-two Sprague-Dawley rats were randomly allocated into sham-operation group, and groups of 6 hours, 12 hours, 1, 3, 7 and 14 days (n=6 in each group) after intracerebral hemorrhage. The intracerebral hemorrhage model in rats was established by injecting autologous arterial blood into caudate nucleus. Immunohistochemical method and Evans blue method were used respectively to detect the dynamic changes of AQP-4 around hematoma and the blood-brain barrier permeability in different time periods after intracerebral hemorrhage in rats. Results: (1) The expression of AQP-4 around hematoma increased gradually 12 hours after intracerebral hemorrhage, it increased significantly at day 1 and reached a maximum at day 3. It was slightly higher than normal level at day 7 and almost returned to normal at day 14 (P

Objective To explore the factors relating to the severity of Guillain-Barr? syndrome (GBS) (i.e. of being the mild form or in need of mechanical ventilation), and to find the predictors in early clinical presentations and laboratory investigations as to guiding the early management of GBS. Methods All 123 cases of patients with GBS admitted to Xuanwu Hospital from 1996-2002 were collected, and divided into two groups based on the international criteria for different severity of GBS. Mildly affected patients (i.e. able to walk unaided during the whole course or the Medical Research Council [MRC] muscle power grading scale≥4 in both lower limbs at nadir) and ventilated patients (i.e. in need of mechanical ventilation) were divided in this study. Characteristics of each group in early clinical presentations and laboratory investigations were analyzed in univariate conditional Logistic model in order to find out the predictors. Results In all 123 patients studied, 32 were having mild GBS; 29 severe GBS (i.e. in need of mechanical ventilation during the course). Men (P=0.047) and patients without bulbar dysfunction (P=0.002) were more likely to become mild GBS. The possibility of being mild form was 3.33∶1 between man and woman, 1∶20.17 between those with and without bulbar dysfunction. Patients with bulbar dysfunction (P

Objective To study the clinical features and pathogenesis of non-traumatic rhabdomyolysis (RM). Methods The causes andclinical features of 11 patients with non-traumatic RM were analyzed retrospectively. Results The causes of non-traumatic RM includeddrugs (3 cases), carbon monoxide poisoning (2 cases), intensive exercise training (2 cases), strenuous farming work (1 cases) and others (3cases). The clinical presentations included muscular weakness, myalgia, muscle swelling, pigmented urine and acute renal failure. The activityof serum creatine kinase increased, electrolyte derangements were common complications. Conclusion Prescribed drugs are the mostcommon cause of non-traumatic RM. The diagnosis of non-traumatic RM is chiefly based on clinical presentations and high levels of serumcreatine kinase. Acute renal failure is the most severe complication and cause of death.