Objective To investigate the clinical manifestation, biochemically detected data, and radiographic features of a pedigree with suspected mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome, and to explore the correlations between the clinical features and the mutant heteroplasmy levels of mitochondrial genome. Methods The personal details, histories of stroke-like episodes and seizures within the proband and 11 members in the maternal lineage of the family were collected. Routine blood examinations and plasma lactate levels before and after movements of these family members were detected, followed by cephalic MRI examinations. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to detect and validate the A3243G point mutation in mitochondrial genome, and real-time PCR were used to quantify the mutation proportion of A3243G. Results Typical symptoms of MELAS such as seizures, stroke-like episodes and hyperlactacidemia and atypical symptoms such as growth failure, exercise intolerance, fevers and migraines were observed on several members in the pedigree. Cephalic MRI findings performed during episode periods were in accord with the typical radiographic features of MELAS and cerebellar atrophy was commonly observed. Family members on the maternal side all harbored the point mutation on 3243 site in mitochondrial genome. Meanwhile, patients with higher heteroplasmy levels relatively manifested more typically and severely according to the clinical observation. Conclusions The pedigree is diagnosed with maternal inheritance of MELAS syndrome. The main cause can be attributed to a mitochonorial A3243G mutation.The mutant heteroplasmy levels of hemocytes in peripheral blood are positively associated with genetic relationship, seizure anticipation, plasma lactate data and other clinical features.

BACKGROUND: Drug treatment has unsatisfactory effect on Alzheimer disease, while many studies have indicated that the transplantation of bone marrow stromal cells (MSCs) is effective on Parkinson disease, cerebral ischemia, etc., but the mechanism is still unclear.OBJECTIVE: To imitate transplantation environment by co-culture of amyloid β1-40 (Aβ1-40) damaged PC12 cells and MSCs, observe the effect of bi-directional information feedback in the microenvironment on the transdifferentiation of MSCs to nerve cells, and observe its protective effect on the apoptosis of damaged PC12 cells.DESTGN: A comparative observation.SETTING: Department of Neurology, China Medical University.MATERIALS: SD rats of 2-3 weeks after birth either male or female were used. PC12 cell lines were purchased from the Institute of Cell Biology, Chinese Academy of Sciences; neuro-specific enolase (1:50, Boster, Wuhan);Methyl-thiazol-tetrazolium (MTT) 15 μL (terminal concentration of 0.5 g/L).METHODS: The experiment was carried out in the Experimental Center (provincial experimental animal center) of China Medical University from June to July in 2004. Bilateral femurs were aseptically removed from 1 SD rat, and MSCs were identified using CD44 antibody immunofiuorescently. PC12 was used to replace nerve cells. The PC12 cells were stimulated by Aβ1-40 then transferred by Transwell. There were 5 groups: Group A: normally cultured PC12 co-cultured with MSCs; Group B: Aβ1-40 stimulated PC12 co-cultured with MSCs; Group C: normal PC12 supernatant+MSCs; Group D: damaged PC12 supernatant+MSCs; Group E: MSCs cultured with common medium 1640.MAIN OUTCOME MEASURES:Routine immunohistochemical staining was performed. NSE positive cells were observed under inverted fluorescence microscope, 10 visual fields (200×) were randomly selected to count the positive cells. MTT metabolic rate was used to detect the proliferation of MSCs in each group. The differences of measurement data were compared using the one-way analysis of variance.fluorescent and bright fields, NSE positive cells appeared as red fluorescence, MSCs were bipolar, multipolar and cone shapes, and appeared as neuron-like forms with dendrite-like structure, and there were extensive connections among some neuron-like cells. The NSE positive rates was obviously higher in group B than groups A, C, D and E (P ＜ 0.01 ).in groups A, C, D and E (F=9.713, P＜ 0.01).

BACKGROUND: The primary pathophysiology of Alzheimer disease (AD) is linked to β-amyloid (Aβ)protein. Neural progenitor cells (NPCs), which have the ability of multipotency, self-renewal and repair,have been detected in the central nerve system (CNS) of adult rat recently. But effective function of these neural progenitor cells are not seen in the AD brain ,which mechanism is unclear.It is unclear if Aβ1-40protein is compromised by the signal pathway of c-Jun N-termial kinase associated with the neurotoxicity to the progenitor cells on the cortex of embryonic rats.OBJECTIVE: To investigate the mechanism of c-Jun N-terminal kinase signal transduction pathway of Aβ1-40 protein, which has neuronal toxicity to progenitor cells(CPC)on the cortex of embryonic rats . To detect the neuroprotective effects of c-Jun N-termial kinase inhibitor (SP600125) against Aβ1-40-induced neuronal toxicity to the cortical progenitor cells on the cortex of embryonic rats.DESIGN: A randomized and controlled trial with cells as objects.SETTING: Department of Neurology, First Hospital Affiliated to China Medical UniversityMATERIALS: This experiment was carried out at the Central Laboratory,China Medical University from May to October 2005. Embryos at age of 14 days from Wistar rats were used in this experiment.METHODS: Cortical progenitor cells harvested from Wistar embryonic rats were cultured in vitro, passaged and identified. Embryonic rat cortical progenitor cells of rats with good growth state were randomly divided into 4groups:Aβ1-40 group (10 nmol/L Aβ1-40 in each well);SP600125+Aβ1-40group (10 μmol/L SP600125 for 30 minutes and then with 10 nmol/L Aβ1-40 in each well); SP600125 group ( 10 μmol/L SP600125 in each well); Normal saline group (same volume of normal saline). The incubated durations were 0,2 hours, 4 hours, 6 hours, 12 hours, 24 hours respectively,8 wells for each time point. The cell survival rate was measured by MTF assay (The concentration of cortical progenitor cells on the cortex was 1×10s L-1 in each group), the apoptosis rate was detected by flow cytometer (The concentration of cortical progenitor cells on the cortex was 1 ×1010 L-1in each group) and the expression of c-Jun N-termial kinase and p-c-Jun N-termial kinase, c-Jun,p-c-Jun were measured by Western Blot(The concentration of cortical progenitor cells on the cortex was 1×1013 L-1 in each group). t test was adopted for the comparison of difference in measurement data.sion of c-Jun N-terminal kinase, p-c-Jun N-termial kinase ,c-Jun and p-c-Jun of embryonic rat CPC .ture time in Aβ group and SP600125 +Aβ group, decreased obviously at 4hours; cellular survival rate in Aβ1-40 group was lower obviously than that in the other 3 groups at 0,2,4,6,12,24 hours (P ＜ 0.01); Cellular survival rate in SP60025 +Aβ1-40 group was lower obviously than that in SP600125 group and normal saline group at 2,4,6,12,24 hours (P ＜ 0.01);Compared with normal saline group, the difference of cell survival rate was not significant without time-dependent manner in SP600125 group (P＞ 0.05).amyloid protein group and SP600125 +Aβ group, increased obviously at 4hours; cell apoptosis rate in Aβ1-40 group was higher obviously than that of the other 3 groups at 0,2,4,6,12,24 hours(P ＜ 0.01); Cellular apoptosis rate in SP60025+Aβ1-40 group was higher obviously than that in the SP600125 group and normal saline group at 2,4,6,12,24 hours (P ＜ 0.01);Compared with normal saline group, the difference of cellular apoptosis rate was not significant without time-dependent manner in SP600125 group 12,24 hours without changes in Aβ1-40 group; the expression of p-c-Jun N-terminal kinase and p-c-Jun in Aβ1-40 group were seen at 0hour ,increased gradually, reached to the peak at hour 4 and decreased gradually.CONCLUSION: Aβ1-40 could inhibit the cell activity of CPC , reduce cellular survival rate and induce cellular apoptosis. c-Jun N-terminal kinase signal transduction pathway may mediate the Aβ1-40 inducd neurnal apoptosis in AD which may be one reason for unseen rescue mechanism in AD. SP600125 (c-Jun N-terminal kinase inhibitor) could inhibit the activation of c-Jun N-terminal kinase and c-Jun and protect the embryonic rats CPC from the Aβ1-40-induced neurotoxicity.

Objective To determine the relationship between serum vitamin B_(12)(VB_(12))level and multiple sclerosis(MS).Methods The pertinent articles of the VB12 and the case control studies of MS were retrieved comprehensively by manual retrieval and computer retrieval.The methodology quality of the retrieved artilces were evaluated,and the arcb'cles were screened.Heterogeneity test was performed.The valid data was extracted and analyzed by Stata 10.1.Results In this study,9 studies were included.A total of 807 subjects were enrolled,including 414 patients and 393 controls.The Meta-analysis showed that the serum level of VB_(12) in patients with MS was lower than that in controls(standardized mean difference-0.24,95% CI-0.39 to-0.10).The sensitivity analysis indicated that the result of meta-analysis was reliable.Conclusion The serum level of VB_(12) might be associated with MS,and the deficiency of VB_(12) may contribute to the development of MS.

Objective To investigate the protective effects of dl-3-n-butylphthalide（NBP）on amyloid β peptide 25-35（Aβ25-35）-induced apoptosis in PC-12 cells.Methods Cultured PC12 cells were divided into 6 groups：normal group,Aβ25-35 treated model group,0.1,1.0,10,100 μmol/L NBP pretreatment groups.MTT assay was employed to analyze the PC12 cell viability.The ultrastructural changes of neuronal mitochondria were viewed under transmission electron microscope.In order to observe the effects of oxidative stress,MDA and SOD activities were detected by spectrophotometry.Results NBP pretreatment could significantly prevent the cell viability induced by Aβ25-35（P 0.05）.Pretreatment with 10 μmol/L NBP could significantly inhibit the viability decrease induced by Aβ25-35（P 0.05）.Compared with the cells in the model group,the number and morphology of neuronal mitochondria changed distinctly in the NBP pretreated cells.The activity of SOD in the NBP pretreated cells was obviously higher than that of the cell in the model group,while MDA activity had opposite result.Conclusion NBP could protect the mitochondria in Aβ25-35 induced apoptosis by inhibiting the MDA activity and activating the SOD activity.

Objective To study the pathologic and molecular genetic characteristics of Chinese patients with oculopharyngeal muscular dystrophy(OPMD).Methods The ultrastructural muscle biopsies in 6 patients were carried out by using transmission electron microscopy.The DNA was obtained through blood samples from patients(n=11) and the at-risk individuals(n=16).Amplification of the PABPN1 gene mutation region was performed by polymerase chain reaction(PCR).The sequences were obtained and compared with the genomic sequence of the human PABPN1 gene.Results Intranuclear inclusions(INIs) were found by electron microscopy in 4 patients,and the rate of appearance was 18%,20%,34% and 40%.Sequence analysis of exon 1 of PABPN1 gene showed abnormal expansions of the GCG-repeat—(GCG)_8 and(GCG)_(10) in 9 patients.Conclusions INIs might be found by electron microscopy in muscle biopsies of OPMD patients.The rate of appearance of INIs should have positive relationship with the amount of the GCG-repeat.PABPN1 gene mutations might be present among Chinese patients with OPMD,and should have a negative relationship with the age of onset.

Objective To establish the co-culture system of marrow stromal cells (MSCs)and A?1-40 injured PC12 in vitro and to evaluate the effect and mechanisms of the system inhibiting apoptosis of PC12 induced by A?1-40.Methods MSCs and PC12 were cultured in vitro and identified by CD44 immunofluorescent staining;PC12 were damaged by A?1-40,and transferred by transwell followed by the classification into 5 groups. PI and Annexin-V co-fluorescent staining was performed,then PC12 apoptosis were detected by flow cytometry and EM;Supernatant was analyzed by enzyme-linked immunoadsordent assay (ELISA) to detected TGF-?,NGF,BDNF,and bFGF. Results About 96% MSCs showed CD44 positive cells. Co-culture group had the lowest rate of PC12 apoptosis(46.17%?8.28%,F=61.637,P0.05). Conclusion The co-culture system of MSCs and A?1-40 injured PC12 in vitro could inhibit apoptosis of PC12 induced by A?1-40. Thus grafted MSCs have the possibility to inhibit neuronal apoptosis by A?in the diseased brain.

Objective To study the clinical,image and pathological features of cerebral gliomatosis.Methods 2 cases with cerebral gliomatosis underwent routine MRI scan, contrast enhance MRI scan and pathologic examination of the lesions. Their clinical manifestations were observed during the past 2 years.Results Main clinic features of the 2 patients were headache, dizziness, nausea, vomit, diplopia, hemiplegia, hemianesthesia and ataxia. Cranial MRI scan showed long T 2 signals in the bitemporal lobes, biparietal lobes, corpus collosum, thalamus, caudate nucleus and putamen. Brain stem and cerebellum were also involved in 1 patient. The borders of the lesions were unclear and no mass-effect phenomenons were found. Contrast enhancement occurred only in 1 patient after Gd-DTPA injection. The biopsies in the 2 patients showed diffuse infiltrative growth of most astroglioma cells. The shape of nucleus was round or ellipse and the staining of nucleus was comparatively deep. Cleavage of nucleus was seldom. The 2 patients died in 4 to 6 months after the onset of the disease.Conclusions MRI scan and pathologic examination are essential diagnostic methods for cerebral gliomatosis. The prognosis of cerebral gliomatosis is poor.

Objective To explore the features of clinical and progression of multiple system atrophy(MSA).Methods The clinic data of 28 subjects diagnosed as probable MSA according to Gilman diagnostic criteria were studied retrospectively.Three aspects of activities of daily living(ADL)(aid-requiring walking,wheelchair-bound state and bedridden state)were used to assess the progression of disease.Kaplan-Meier analysis was also used to estimate the difference between subgroups.Results Three systems were involved in 26 cases(92.9%)and autonomic functional disturbance was common.The calculated median time from onset to evolution to MSA was mean 2 years.The median times from onset to aid-requiring walking,wheelchair requirement and bedridden state were 3,5 and 7 years,respectively.The patients impaired both of motor and autonomic systems progressed fastly within 3 years from onset of the disease(P

Objective To investigate the effects of rhG-CSF on the neuronal cell apoptosis and expression of VEGF after cerebral ischemia in diabetic rats.Methods Wistar diabetic rats were subjected to middle cerebral artery occlusion and randomly devided into control group and rhG-CSF group.The rhG-CSF group received subcutaneous injection of rhG-CSF 50 ?g/(kg?d)for 7 d,14 d and 21 d after cerebral ischemia.Neurological severity scores(NSS),TUNEL,and immunohistological assessments of VEGF were performed to evaluate the rhG-CSF treatment.Results Compared with the control group,the rhG-CSF group showed significantly improved in the NSS,significantly decreased in the TUNEL positive apoptotic neuronal cells and significantly increased in the VEGF positive neuronal cells(all P