Synaptosomes were isolated from pig brain by homogenization, differential centrifugation and sucrose gradient centrifugation. After synaptosome lysis in hypoosmotic buffer, the plasma membrane vesicles were collected. Following the solubilization of plasma membrane vesicles in Triton X-100, the solubilized protein was applied to calmodulin affinity chromatography colurnn, and the delipidated plasma membrane Ca2 + -ATPase was purified to nearly homogeneity. The novel feature of this purification is the use of large affinity column and heavy washing to facilitate the purified Ca2+ -ATPase with higher activity and protein yield. The specific activity of the purified Ca2+ -ATPase was recovered to a maximum of 3.32 μmol· mg-1· min-1 after incubation with asolectin. Silver staining of SDS-PAGE revealed a single protein band around Mr 140 000, showing the purity was over 90 %. Different Ca2 + concentrations dramatically affect the specific activity of Ca2 + -ATPase.

Objective To identify the factors related to amyotrophic lateral sclerosis (ALS) prognosis in a retrospective study.Methods One hundred and six patients were evaluated with ALS enrolled in our hospital from January 1994 to December 2004.The patients were monitored with a standard evaluation form,which recorded clinical features,function rating scale (FRS) and forced vital capacity (FVC) every 3 or 6 months from visit to death or having tracheostomy.Results Mean age at onset was (52.1±10.5) years.According to revised El Escorial diagnostic criteria (EEDC),52 patients had definite ALS,37 probable ALS,17 possible ALS.The median survival time from symptom onset was 35 months (95% CI 30-40 months).In the univariate analysis,survival was significantly related to lesion site,lag time from onset to diagnosis and predicted FVC at diagnosis (Log rank 6.84,43.30,4.78,all P < 0.05).In the Cox multivariate model,lag time from onset to diagnosis was significantly related to survival (Wald 20.221,hazard ratio 0.351,P<0.05).Age,sex,EEDC classification,FRS at diagnosis were not related to survival.Conclusion Lag time from onset to diagnosis is a strong predictor of survival,which suggests that progression rate is highly related to survival and further study is needed.

Objective To investigate the progression rate and prognosis of spinal decompression surgery after the onset of symptoms of amyotrophic lateral sclerosis(ALS).Methods A total of 329consecutive patients with deftnite or probable ALS were enrolled in the study.The patients were recorded with clinical features,history of spinal surgery,etc. Some of the patients were monitored every 3 months from visit to death or tracheostomy.Results Of 329 typical sporadic ALS,we found cervical spondylosis in 156(47.4%)patients,among whom 19(5.8%)underwent decompressive spinal surgery.No differences were noted regarding age at symptom onset,Sex and ALSFRS-R at time of diagnosis between ALS patients who underwent spinal surgery and other ALS patients.But the time from symptom onset to diagnosis was significantly longer in patients with spinal surgery((23.0±6.5)months vs(13.7±7.9)months,t=4.800,P=0.000).In the folow-up study,there were no differences in the rate of disease progression and survival between 2 groups.Conclusions All patients have inevitably progressed after spinal surgery.Although the surgery does not obviously fasten disease progression rate and shorten survival,it prolongs the time from disease onset to diagnosis,therefore should be handled with caution in patients with concurrent ALS and cervical spondylotic myelopathy.

Objective To identify the gene mutation of Chinese Charcot-Marie-Tooth (CMT) pedigrees and investigate the association of gene mutation to the clinical manifestations and electrophysiology,and the underlying mechanisms.Methods A total of 105 pedigrees with CMT in our hospital were enrolled from January,2007 to December 2013.The clinical features,CMT neuropathy score (CMTNS) and electrophysiological data were collected.Gene mutations were analyzed using multiplex ligation-dependent probe amplification (MLPA) and Sanger gene sequencing.Results We found 31 (29.5％) PMP22 duplication pedigrees,8 (7.6％) GJB1 mutation pedigrees,4 (3.8％) MFN2 mutation pedigrees,4 (3.8％) HSPB1 mutation pedigrees,3 (2.9％) MPZ mutation pedigrees and 1 (1.0％) PMP22 mutation pedigree.In Chinese Han population,the proportion of PMP22 duplication was relatively lower than that in western countries and manifested with classical clinical characteristics of CMT.Subjects with axonal CMT often presented with isolated lower extremity injury and with central nervous system involvement.Hereditary motor neuropathy might be underestimated in clinical setting and should be differentiated from motor neuron disease.Conclusions The gene frequency distribution in patients with CMT in Chinese Han population is different from that in patients from western countries.We should establish our own epidemiological data of CMT in Chinese Han population.

0.05). However, compound muscle action potential (CMAP) amplitude in ims injected group wa s si gnificantly higher as compared with the other two groups[ims injected group (12.50?2.06) mV, iv group (1.50?0.20) mV, control group (10.13?4.04) mV, F=6.347, P=0.033]. The MSC s were able to be observed only in ims injected tissues 3 weeks after implantati on (A large number of small undifferentiated cells were found outside the myofib ers and some were found between the cells.) The atrophy of gastrocnemius in ims injected group was much less severe than that of the other 2 groups. The diameter of muscle fibers was significantly longer on d60 (F=4.537,P=0.021).Conclusion:Intra- muscular injection of MSC was well distributed in denervated muscle, which provides a newway of nerve regeneration in the rat model of sciatic nerve injury.

Objective:To identify the correlation between the progression rate of motor unit number estimate(MUNE) at diagnosis and survival.Methods:We included 129 patents with amyotrophic lateral sclerosis(ALS) enrolled in our hospital from January 2002 to December 2005.We recorded clinical features,ALS functional rating scale(ALSFRS),forced vital capacity(FVC) and electrophysiological data at diagnosis.The patients were monitored every 3 months from visit to death or tracheotomy.Results:Mean age at onset was(52.19?11.00) years.The median survival time from symptom onset was 45.71 months(95% CI = 35 to 51).In univariate analysis of Kaplan-Meier method,outcome was significantly related to progression rate of MUNE(P

Diagnostics probation teaching has important value in the medical teaching for foreign students. The practice experiences of diagnostics probation teaching for foreign students in the last 3 years are reviewed from the management of teaching,practice of teaching,supervision of teaching and examine of teaching in order to improve the effect of diagnostics probation teaching for foreign students.

Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.

To study and optimize the fermentation parameters for expressing human-like collagenⅡduring E. coli high-density fermentation. The effects of pH, temperature, dissolved oxygen and induction instant on the cell growth and human-like collagenⅡproduction were investigated to optimize the fermentation conditions. The results demonstrated that the following conditions were beneficial for cell growth and foreign gene expression, controlling pH in phase induction at 6.8 and initial pH at 6.5, maintaining fermentation temperature and dissolved oxygen concentration was controlled at 34?C and 20% respectively, and implementing induction at the later logarithmic growth phase. Under the optimized condition, the cell density and human-like collagenⅡyield could reach 88.4 g/L and 14.2 g/L, respectively.

This research adopted silt as the sample,and the five highest hydrogen production performing strains contained in the sample were isolated. The strain whose hydrogen production was the highest was identified as Enterobacter cloacae by the analysis of 16S rDNA sequencing and comparison. It is showed by Plackett-Burman Experimental Design that only glucose,citric buffer and reducing agent had significant effects on hydrogen production by Enterobacter cloacae FML-C1. The path of steepest ascent was undertaken to approach the optimal response region of those three factors. Central Composite Design(CCD) and Response Surface Methodology(RSM) were employed to investigate the interaction of the variables and to ascertain the optimal values of the factors,which finally led to the maximum hydrogen production(VH2) . The theoretical optimal medium conditions were:glucose 21.5 g/L,citric buffer 13.6 mL/L,reducing agent10.0 mL/L. The five tentative tests matched this model well. The final VH2 was up to 2347.4 mL/L,which was 127.42% enhanced in comparison to the original. The result shows that PB experiment design and RSM analytical method work well in selecting factors which have significant influences on the hydrogen production and,moreover,achieve the ideal optimal result.