Objective To report clinical and pathological features in a Chinese family with desminopathy caused by a novel mutation in desmin gene. Methods There were 8 patients (7 male and 1 female) in a family manifesting desminopathy in an autosomal dominant inheritance pattern. The onset age ranged from 20--50 years old. Heart disease appeared in 7 cases, diarrhea in 1 case and limb weakness in 3 patients. Muscle biopsies were carried out in proband and his elder brother. The histological, enzymhistochemicai staining, ultrastructural examination and immunohistochemical staining with desmin antibody were performed. The sequence of desmin gene was analyzed in DNA from 4 patients, 5 asymptomatic members and 50 healthy controls. Results Muscle pathology in the proband revealed wide variation in size of fibers. Some fibers had eosinophilic changes with rimmed vacuoles. Muscle pathology in the brother revealed multiple small and round eosinophilic inclusions in the fibers. Desmin staining was positive in the inclusions and in the sarcolemma. Electron microscopy revealed deposits of abnormal granulofilamentous materials. A novel S12F mutation in desmin gene has been identified in 4 affected family individuals, but not in unaffected family members and controls. Conclusions The desminopathy caused by desmiu SI2F mutation dominantly affected male individuals. Most of victims had cardiac abnormalities. The myopatholgical features can vary in the different patients in one family.

Objective To study the single nucleotide polymorphisms (SNP) in 3 sites allele (T189M, R85H, C121W) of SCN1B and the association between gene distribution and epilepsy. Methods All 330 blood samples of refractory (80 cases), non-refractory (100 cases) epilepsy patients and healthy people (150 cases) were collected. Genomic DNA of leucocyte was extracted. SNPs of three sites allele of SCN1B were tested by allele-specific primer-polymerase chain reaction (ASP-PCR).Data were analyzed by SAS 8.1 statistical software. Results Epilepsy group and healthy group had significantly statistical difference in composition of 3 sites allele on single site genotype (x~2=11.19, 11.14 and 6.50, all P < 0.05).There was no statistical significance between refractory and non-refractory epilepsy group. On gene combination, in 27 different combinations of polymorphism, mutation frequency in 3 sites (CT + AG + CG) was highest in epilepsy group (18.40%).The next was one site in CT + GG + CC (16.80%).In healthy group, frequency of non-variant in CC + GG + CC was highest (16.67%), and the next was 2 sites in CT+ AG+CC (13.73%).Thirty-five cases in epilepsy group (28.80%) had 3 sites mutation compared with 10 cases in healthy group (9.71%), and their difference had statistical significance (x~2=12.54, P<0.05).Eighteen cases in refractory epilepsy group (30.51%) had 3 sites mutation compared with 21 cases in non-refractory epilepsy group (28.77%), and the difference had no statistical significance. Fifty cases in epilepsy group (40.00%) had 2 sites mutation compared with 41 cases in healthy group (40.20%), and there was no statistical significance between them; 25 cases in refractory epilepsy group (42.37%) had 2 sites mutation compared with 21 cases in non-refractory epilepsy group (28.71%), and their difference had no statistical significance. Conclusions Mutation, especially multisite mutation of SCN1B is relatively likely to cause epilepsy in human. Gene distribution and combination of three sites allele of SCN1B in refractory epilepsy is close to that in non-refractory epilepsy.

Objective To develop transgenic mice harboring the fusion gene of mutant amyloid precursor protein and two types of fluorescent protein for the future study on Alzheimer's disease.Methods The fusion gene CFP-54 bp-YFP-C99 was introduced into mice by mieroinjection.The presence of CFP-54 bp-YFP-C99 was confirmed by PCR in the founders.Results CFP-54 bp-YFP-C99 gene was injected into pronucleus of 2202 zygotes and 1806 injected eggs were implanted into 56 foster mothers, 13 of which were pregnant.There were 13 foster mothers who borne 52 offspring and 32 of them survived.Recipient mouse pregnancy rate was 23.2% (13/56) and the integration rate was 3.9% (2/52).Conclusion CFP-54 bp-YFP-C99 transgenic mice is obtained, but the transgenic efficiency is low.

Objective To study the clinical characteristics of epidemic Japanese encephalitis in 4 adult patients and to improve the level of diagnosis.Methods The clinical characteristics, laboratory results and MRI features of 4 adult patients with epidemic Japanese encephalitis hospitalized in our hospital between 2007 and 2008 was retrospectively reviewed and analyzed.Results All patients were sporadic.Two were female and others were male with the age from 34 to 68 years old and onset in summer and autumn.One patient had diarrhea at onset while others had fever and headache at onset.All patients had high fever and psychiatric symptoms.Two patients had respiratory failure.Obvious elevation of white cells in cerebrospinal fluid was seen in all patients and Japanese viral antibody of IgM was all positive whereas the image of MRI was abnormal only in one patient.Two patients had neurological sequelae.Conclusions The clinical manifestations of epidemic Japanese encephalitis in adults are severe and untypical and the respiratory failure is likely to occur.

Objective To evaluate the function of motor nerve in patients with diabetic sensory neuropathy (DPN) by estimating the motor units.Methods Fifty-four patients with DPN were divided into two groups according to the nerve conduction abnormality of ulnar nerve.The paresthosia group only included patients of 33 patients (20 male, 13 female) with abnormal sensory conduction.The admixture group included 21 patients (13 male, 8 female) with abnormal motor sensory conduction.The control group included 42 healthy people.A Keypoint4 electromyogram device was used to estimate the motor units of hypothenar muscle, and the fasting blood glucose and the duration of diabetes mellitus were documented.Results The motor units of hypothenar muscle in paresthesia group, admixture group and control group was 97.1±47.6, 63.3±22.3, 173.0±75.5 (F=21.259, P=0.000) respectively.The hypothcnar muscle motor units(84.0±42.8) in 54 patients with DPN had negative correlation (r=-0.414, P=0.002) with the duration of diabetes mellitus (1 month to 26 years) and had no evident correlation with fasting blood glucose (5.0-20.4 mmol/L).Conclusion There is clinical motor nerve damage in patients with diabetic sensory neuropathy, and the loss of motor units is impacted by the duration of diabetes mellitus.

Objective To survey the prevalence, distribution of non-motor symptoms (NMS) in essential tremor (ET) and the relationship with disease severity and duration.Methods Fahn-Tolosa-Matin Tremor Rating Scale (TRS) was used to assess motor symptoms in 62 patients with ET.The Parkinson's disease (PD) NMS Questionnaire and T&T olfacmeter and Mini-mental State Examination (MMSE) were used to explore non-motor symptoms in ET patients.Results In ET, a range of NMS occurred across all disease stages.More than half patients (51.6%, 32/62) had olfactory dysfunction,significantly higher than the healthy control group (30.0%, 18/60, x~2=12.371, P<0.05).A third had hyposmia.16.1% had partial olfactory loss.Each ET patient had 5 different NMS on average.Seven NMS were more common in ET patients than in control, including remembering, olfactory dysfunction, intense vivid dreams, anhedonia, depression, anxiety, sleep disorders.The incidences of remembering, olfactory dysfunction,intense vivid dreams were 58.1% (36/62),51.6% (32/62),48.4% (30/62), ranked top 3 in ET patients.Olfaction had inverse correlation with age, while there was a negative correlation between NMS score and TRS score, gender, disease duration and weather to be treated.Conclusion Besides posture tremor and kinetic tremor,NMS occur in ET,and should be well recognized and treated.

Objective To study the changes of life span and pathology in superoxide dismutase 1 (SOD1)-G93A mice after intracardiac transplantation of human mesenchymal stem cells (hMSCs).Methods hMSCs were isolated from bone marrow cells obtained from healthy donors and cultured.The purity and morphology were assessed by flow cytometry (FCM).hMSCs (3×10~6) resuspended in 0.2 ml DMEM was injected into the heart of 8 week-old SOD1-G93A mice.In non-transplantion control SOD1-G93A mice, only DMEM was injected.The mice were evaluated for signs of motor deficit with 4-point scoring system previously described by Weydt et al.The age of onset and life span in mice were assessed.The pathological change including number of motor neurons was investigated by Nissl staining.Immunofluorescence staining with specific human nuclear antibody was used to confirm the transplant of hMSCs in mice.Results The onset symptoms in untreated SOD1-G93A mice appeared at (156.56±3.60) days of age and the average life span was (188.32±3.51) days.hMSCs transplantation delayed the onset of ALS type symptoms about 16 days (x~2=10.888, P=0.001) and prolonged the life span about 14 days compared to the untreated SOD1-G93A littermates((202.19±4.09) days vs (188.32±3.51) days, x~2=3.917, P=0.04).The loss of motor neurons in untreated mice was earlier and more severe than in hMSCs transplanted mice.At 20 weeks, the number of motor neurons in transplanted mice was significantly higher than those in untreated mice.Human specific nuclear antigen in brain and spinal cord was detected in transplanted SOD1-G93A mice.Conclusion hMSCs can be implanted for a long-term into central nervous system by intracardiac transplantation and the transplantation can prolong life span, and delay the onset of the disease and motor neuron loss in SOD1-G93A mice.

Objective To examine the technique of multiple point stimulation for motor unit number estimation (MUNE) and to establish the normative value range in Chinese.Methods Surface-recorded motor unit action potentials were measured in 80 healthy subjects.The compound muscle action potential (CMAP) amplitude measured by maximum baseline to negative peak was recorded.The stimuli sites included the wrist, 6 cm above the wrist, elbow and 6 cm above the elbow along median nerve and ulnar nerve.Individual motor unit responses were obtained by adjusting location of the stimulate electrode and isolating threshold responses with distinct morphologies.Then, stimulus intensity was increased gradually to detect single motor unit action potentials (SMUPs).SMUPs were recorded three times.Stimulating was increase again to record another SMUPs set.Total of 12 SMUPs were recorded.Repeat the whole procedure for two times.Results MUNE was 230.0±35.7 in abductor pollieis brevis muscle, and 242.5±30.2 in aductor digiti minimi muscle for multiple point stimulation.Test-retest correlation coefficients and coefficients of variation for mean of two MUNE were 0.88-0.91 and 13.20%-15.24%.Conclusions The multiple point stimulation is a useful and replicable method to asses the MUNE.

Objective To evaluate the application of upper trapezins muscle electromyography (EMG) in the diagnosis of lower motor neuron damage in bulbar region in amyotrophic lateral sclerosis (ALS). Methods Standard EMG was recorded over upper trapezius muscle in 100 patients with ALS, 80 patients with cervical spondylotic myelopathy (CSM) and 100 normal controls. In groups of ALS and CSM,EMG was also recorded over sternocleidomastoid, rectus abdominis, first dorsal intercostals muscle and tibialis anterior muscles. Among those CSM patients, 43 patients had operations and the EMG on their trapezius muscle was examined at pre-operation and at 3 months post-operation. The parameters of EMG were analyzed between the groups. Results In ALS patients, spontaneous activity in upper trapezius EMG was detected more frequently in patients with disease duration equal to or less than 8 months than the others (21/30(70%) vs 28/70(40%), X~2=7.56, P=0.004). There was no difference in neurogenic EMG changes including abnormal spontaneous potentials and motor unit action petentials (MUAP) between trapezius and sternocleidomastoid in patients with ALS. Significant differences in MUAP were noted between ALS patients((1086.9±152.6)μV, (17.2±6.5) ms,23.6%±3.4%) and controls ((606.7± 82.7)μV,(11.6±1.8) ms,12. 8%±2.2%;q=9.27, 4.57, 4.12, all P<0.01), and between patients with ALS and patients with CSM ((615.7±90.3) μV,(12.1±2.0) ms,13.5%±2.4%,q=8.32,4. 25, 4. 23, all P < 0. 01). Few spontaneous activities in trapezius EMG were detected in post-operation CSM patients. Conclusion EMG in upper trapezius can assist in assessment of clinical and subclinical involvement of bulbar lower motor neurons in patients with ALS, especially at earlier stage.

Objective To investigate the effect of apolipoprotein E gene knock-out(ApoE KO) and high-fat diet on morphology and the expression of mortalin in hippocampal CA_3 neurons of mice, and to explore the impact of these factors on memory and Alzheimer's disease.Methods Ten wild-type and 10 ApoE KO mice were fed with common chow as the control group and the KO group respectively while 10 ApoE KO mice were fed with high fat diet.Twelve weeks later, the weight and the lid of these mice were measured.The brain tissues were observed using HE staining, nissl staining, protargol staining,immunohistochemistry staining and image analysis by computer.Results In the ApoE KO group, weight,total cholesterol, triglyceride, low-density lipoprotein cholesterol were higher than those in the control group,and these changes were more significant in ApoE KO high-fat diet group.The nissl was higher in the ApoE KO group (0.301±0.031) and in ApoE KO high-fat diet group (0.261±0.020) than those in the control group (0.341±0.035, F=18.068, P<0.05).The mortalin in the ApoE KO group (0.322±0.060) and in ApoE KO high-fat diet group (0.391±0.041) were higher than the control group (0.256±0.061, F=15.230, P < 0.05).Conclusions ApoE KO and high-fat diet can reduce nissl, and improve the expression of mortalin.This protein may be involved in the pathogenesis of Alzheimer's disease.