This study was aimed to analyze the clinical features of paroxysmal kinesigenic dyskinesia (PKD) and extend the understanding of this disease. From August, 2008 to October, 2010, 9 patients were diagnosed with PKD in the Department of Neurology of the First Affiliated Hospital of Zhejiang University, China. The data involving clinical demographic characteristics, somatosensory evoked potentials, results of electromyography, video electroencephalography (EEG), brain magnetic resonance imaging (MRI) and computerized tomography (CT) were collected. All PKD patients exhibited unilateral or bilateral recurrent episodic dyskinetic attacks triggered by sudden voluntary movements. The duration of the attacks ranged from several seconds to one minute. The attack frequency ranged from approximately once in several months to more than 10 times in a day. Patients suffered from no conscious disorders during the attack, and no neurological signs were found during the period between attacks. No abnormal somatosensory evoked potentials were found. Routine EEG, video EEG monitoring or brain imaging showed normal findings. Classical treatment for anti-epilepsy, including carbamazepine and topiramate, was administered to the patients and proved to be effective. It was concluded that PKD is characteristically triggered by sudden voluntary movement; no abnormal electroneurophysiological findings are observed in PKD, and antiepileptic drugs are effective in treating the disorder.

Objective Both QBC Star and Sysmex XP-100 hematology analyzers are convenient to carry,which can be used nor-mally under the condition of the field(emergency).This study would compare their test results and operating performance,so to provide guidance for rational use of the instruments.Methods 100 fresh blood samples of 100 health soldiers anti-coagulated by EDTA-K2 were detected by QBC Star and Sysmex XP-100 haematology analyzers respectively,the results of two analyzers were comparatively analyzed and their test time and operating convenience were analyzed.Results There was no significant difference in the results of hemoglobin concentration (HGB),hematocrit (HCT)tested by the two methods (P >0.05).There were significant difference of the mean corpuscular hemoglobin concentration (MCHC),the sum of lymphocyte percent and middle type cells (LYM%+MID%),neutrophil percentage(NEUT%),white blood cell count(WBC),platelet count(PLT)tested by the two meth-ods(P <0.05).The values of MCHC and LYM%+MID% tested by the QBC Star were significantly lower than that detected by Sysmex XP-100(P <0.05),while the rest indicators tested by the former were higher than that of the latter.It took about 5 minutes to complete a blood sample analysis with QBC Star,while about 1 min was needed for Sysmex XP-100.Conclusion The test results of QBC Star and Sysmex XP-100 hematology analyzers couldn′t exchanged except for that of HCT and HGB.Under the condition of field(emergency),QBC Star hematology analyzer is suitable for individual medical examination,and Sysmex XP-100 hematology an-alyzer can be used for the batch medical examination.

Immobilization conditions of Candida tropicalis to be absorbed in polyurethane foam carrier materials were studied on the xylitol production from corn hemicellulosic hydrolysate. Optimum batch-fermentation conditions were as follows: inoculum amount, 7% (volume ratio); polyurethane foam quantity, 1.0 g/100 mL; 30?C; initial pH, 6.0. Shaking speed was divided into two-phase to accommodate the dissolved oxygen, with 200 r/min at 0~24 h and 150 r/min at 24 h~46 h. The immobilized cells on polyurethane foam carrier have high density and good resistance to inhibitors in the hydrolysates. Average xylitol yield and volumetric productivity of polyurethane foam immobilized fermentation were much higher than the fermentation without immobilization. Corn cob hydrolysates can be directly biotransformed to xylitol without decoloration or ion-exchange treatment. This process can effectively reduce production costs, and it shows broad prospects of applications. Average xylitol yield was 67.6% and xylitol volumetric productivity was 1.92 g/(L?h).

Objective To compare several methods for the rapid detection of methicillin resistant StaphylOCOCCUS aureus(MRSA).Methods Forty-four Staphylococcus aureus isolates from Tianjin Medical University Cancer Institute and Hospital(TMUCIH)were detected by Oxacillin Kirby-Bauer disk diffusion method,Cefoxitin Kirby-Bauer disk diffusion method,Oxacillin micro-broth dilution method,E tests and PBP2a Latex agglutinaltion assay.These above results were compared with PCR analysis.Results PCR analysis showed that 36 MRSA strains containing mecA was identified.Thirty-two MRSA strains were detected by Oxacillin Kirby-Bauer disk diffusion method,Cefoxitin Kirby-Bauer disk diffusion method and Oxacillin micro-broth dilution method.Compared with PCR analysis,the sensitivity and specificity is 88.9%and 100%respectively.Thirty-three MRSA strains were identified by E test,with the sensitivity of 88.9%and specificity of 87.5%.Twenty-nine MRSA strains were identified by PBP2a latex agglutination assay with the sensitivity of 80.5%and specificity of 100%.Conclusions The turnaround time of PBP2a Latex agglutination assay could be reduced 24 h compared with other methods for detection of MRSA.This rapid,convenient and specific method could be applied in clinical laboratories for MRSA detection.

Objective To explore the feasibility of adding a flexible linker between two-pore-domain potassium channel TREK-1 (TWIK related K + channel 1)monomers to construct a tandem-linked dimer.Methods PCR was used to add a flexible linker between the two TREK-1 monomers.The cRNA obtained from in vitro transcription using the above vector was injected into Xenopus oocytes.After 24 -48 h,currents were recorded from these oocytes using a two-electrode voltage clamp.The effects of extracellular Ba2 + and pH on TdTREK-1 were observed and compared with those of native dimeric TREK-1.Results The tandem-linked dimeric TdTREK-1 was highly expressed in Xenopus oocytes.The currents through these channels were inhibited by extracellular Ba2 +and acidification.Furthermore,the responsiveness of the concatenated dimers to these extracellular stimuli was similar to that of native dimers.Conclusion Adding a flexible linker between the two monomers to construct the tandem-linked dimer does not affect the expression and gating properties of TREK-1, suggesting that the method be feasible.Such a method will allow the manipulation of a single subunit,which will help basis study the structure and function of TREK-1.

Objective To explore the interaction of streptococcus suis serotype 2 recombinant suilysin ( SLY ) with platelets, and provide the theoretical basis for clinic treatment of patients infected with S.suis.Methods The nickel column affinity chromatography was used to purify the recombinant SLY.The hemolytic acivity was identified by optical density before the platelets aggregation induced by a SLY was detected by a platelet aggregometer or electron microscope and the effect of aspirin on platelets aggregation was analyzed.The impact of wild type 05ZYH33 and sly-deficient mutant strainΔSLY on platelets of mice was compared to predict the interaction of the SLY with platelets in vivo.Results and Conclusion Hemolytic activity of recombinant SLY was 2000 hemolytic units( HU) and platelets aggregation was induced at 1 μg/ml.The aggregation can be inhibited by aspirin in 5 mmol/L.SLY can also increase the volume and reduce the amount of platelets in mice.

In the present study, packaging system composed of pAAV-CMV-GFP, pAAV-RC and pHelper were transfected into human embryonic kidney 293 cells (HEK293 cells) mediated by polyethyleneimine (PEI) to explore an optimal transfection condition. Different total plasmid DNA dosages (1, 2, 3, 4, 5, 6 μg) and different PEI/Plasmid ratios (1:1, 3:1, 5:1, 7:1) were tested with detection of green fluorescence protein (GFP) with ImagePro Plus6. 0 Software. Then transfection efficiency of the optimized transfection system was further observed for different time periods(12, 24, 36, 48, 60, 72 h). The results showed that total plasmid dosage of 4 μg/well with PEI/plasmid ratio of 3 : 1-5 : 1 was an efficient transfection condition. Transfection efficiency-time curve was an S-shaped curve. Transfection efficiency reached a plateau at 60 h after transfection. The optimized conditions for PEI-mediated transfection at the optimal time result in enhanced transfection efficiency of triple plasmid into HEK293 cells.
Green Fluorescent Proteins ; HEK293 Cells ; Humans ; Plasmids ; Polyethyleneimine ; Transfection ; methods

Objective To investigate the effect of oncolytic adenovirus vector SG600-IL24expressing human melanoma differentiation associated gene-7 (mda-7/IL-24) on hepatocellular carcinoma cell lines with different metastatic potential of HepG2, SMMC7721, MHCC97L and normal liver cell line LO2. Methods The oncolytic adenovirus SG600-IL24 which carrying mda-7/IL-24 gene was transfected into hepatocellular carcinoma cell lines and normal liver cell line. The mRNA and protein expression of mda7/IL-24 in HepG2, SMMC7721, MHCC97L and LO2 cell lines was confirmed by RT-PCR,ELISA assay and Western blot respectively. MTT assay and flow cytometry were used to study tumor cell proliferation and cell cycle in vitro. Hoechst33258 and flow cytometry were studied to indicate the apoptosis effects. Results It was confirmed by RT-PCR, ELISA assay and Western-blot that the exogenous mda-7/IL-24 gene was highly expressed in HepG2, SMMC7721, MHCC97L and LO2 cell lines. MTT and apoptosis detection indicated that MDA-7/IL-24 can induce the growth suppression (the inhibition rate was 75% ±2. 5% ,86% ±3. 5% ,and promotes apoptosis ( the apoptosis rate was 56. 5% ± 4. 0% , 34. 4% ± 2. 0% , 43. 3% ± 2. 5%cell lines at G2/M phase ( the blocking rate was 35. 4% ± 4. 2% , 40. 5% ± 5. 0% , 42. 0% ± 5. 0%metastatic potential hepatocellular carcinoma cell lines but not in normal liver cell line.Conclusions Oncolytic adenovirus vector SG600-IL24 can selectively induce growth suppression, promote apoptosis in hepatocellular carcinoma lines in vitro but not in normal liver cell LO2.

Objective To quantitatively study the distance between the left atrial posterior wall and the descending aorta(DLD)in the normal fetal four-chamber view using fetal echocardiography.Methods The distances between the left atrial posterior wall and the descending aorta in end-systole (DLDs) and in end-diastole(DLDd) were measured in a standard apical four-chamber view using fetal echocardiography in 300 normal fetuses.The gestation ages ranged from 20 to 40 weeks.Fetal non-cardiac biometric parameters included femoral length(FL),biparietal diameter(BPD) and gestation age (GA) based on menstrual age. The correlation between DLDs,DLDd and non-cardiac biometric parameters were analyzed.Results In normal fetuses,the DLDs was (0.32±0.08)cm with a range of 0.19 -0.48 cm,the DLDd was (0.43 ± 0.10)cm with a range of 0.25-0.62 cm.The DLDs and DLDd increased with the growth of GA,and were correlated with GA and BPD and FL significantly (r = 0.545,0.556,0.574,0.530,0.563 and 0.576, respectively,all P <0.001).The linear regression equations between DLDs,DLDd and GA,BPD,FL were:Y =0.009GA+0.041 ,Y =0.038BPD+0.028,Y =0.044 FL+0.073,Y =0.01 1 GA+0.083,Y =0.048 BPD+0.063,Y = 0.054 FL + 0.124,respectively.Conclusions In normal fetuses,the DLDs and DLDd increase with the growth of GA,and have good correlation with GA,BPD,and FL respectively.Normal reference ranges of DLDs and DLDd have been provided.These normative data may be a new tool for assessment of fetal heart,especially has potential applications in screening of total anomalous pulmonary venous connection(TPVAC),absent pulmonary valve syndrome(APVS)or other complex congenital heart defects.

Objective To study changes in synaptic plasticity in the contralesional mirror area of the cortexes of rats with cerebral infarction treated by low-frequency electrical stimulation(LFES)and to explore the therapeutic mechanism of LFES on the molecular level.Methods Forty-eight Sprague-Dawley rats were randomly allocated into a LFES group,a placebo group and a sham-operation group.Following middle cerebral artery occlusion(MCAO),rats in the LFES group were treated with LFES for 7 d(20 min/d),while the ones in placebo group were connected with the same LFES device but without electricity.Rats in the sham-operation group were subjected to a MCAO operation without occlusion and then received no special treatment.Synaptic ultra-structures and the expression levels of glia fibrillary acidic protein(CFAP)and synaptophysin in the contralesional mirror area of the cortexes of the rats in each group were measured with electron-microscopy and Western blotting.Results Compared with the placebo group or the rats before treatment,rats treated with LFES exhibited ultra-structural changes in the form of larger curvature of synaptic interfaces and narrower synaptic clefts.GFAP expression levels did not fluctuate significantly,but the expression of synaptophysin was significantly up-regulated.Conclusion LFES treatment can induce active changes in synaptic plasticity in the contralesional mirror area of the cortex of rats after cerebral infarction.