Aim:To develop a practical chiral CE method for the quantitative determination of the unwanted enantiomer[( R )-enantiomer]presented in zolmitriptan. Methods:The background electrolyte was 20 mmol/L sodium dihydrogenphosphate solution with 1% S-β-CD,adjusted to pH 3.50 with phosphoric acid. A fused-silica capillary(60 cm×50 μm ID,effective length 51.5 cm)was used at 20 ℃ for the separation. The applied voltage was -30 kV. The samples were loaded by hydrodynamic injection(50 mbar pressure,6 s). UV was measured at 220 nm. Results:Zolmitriptan and its chiral impurity were baseline resolved ( R s=6.66). The linearity was good over the concentration range from 4 to 80 μg/mL( r =0.999 8) of ( R )-enantiomer. The injection precision (expressed as CV%) was 2.83%. The average recovery was 99.97%( n =9). The limit of detection was 1.5 μg/mL. The host-guest complex binding constants were 964 and 905 mol-1 for ( R )-enantiomer and zolmitriptan,respectively. Conclusion:The method is suitable for the determination of ( R )-enantiomer in zolmitriptan and binding constants of zolmitriptan enantiomers to S-β-CD.

ObjectiveTo investigate left ventricular (LV) radial dyssynchrony and LV rotational change of isolated complete left bundle branch block (CLBBB) and CLBBB with heart failure (HF).Methods Twenty-six isolated CLBBB patients,twenty-four CLBBB patients with HF and twenty healthy controls were studied.Two-dimensional speckle tracking imaging (2D-STI) was applied to short-axis views to calculate LV radial dyssynchrony and LV rotational and twist degree and time to peak rotational and twist degree.ResultsLV radial dyssynchrony and LV rotation and twist degree were intermediately damaged in isolated CLBBB patients and was significantly damaged in CLBBB patients with HF,compared with healthy controls.ConclusionsFor isolated CLBBB patients,even if LV ejection fraction was in the normal range,another parameters of cardiac mechanic had changed and 2DSTI could be used to evaluate these alterations.

Animals ; Carcinoma, Neuroendocrine ; metabolism ; pathology ; physiopathology ; Carcinoma, Small Cell ; metabolism ; pathology ; physiopathology ; Cell Differentiation ; Chromogranin A ; metabolism ; Humans ; Male ; Neuroendocrine Cells ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; physiopathology

Objective To evaluate the patterns of intracardiac flow of left ventricle (LV) in patients with premature ventricular complexes (PVCs) from the right ventricular outflow tract (RVOT) by analyzing the vortex during isovolumetric contraction phase and the distribution rules of flow-time curves in each layer of LV.Methods Twenty-seven patients with frequent isolated PVCs from RVOT were involved and 25 healthy subjects as control.The color Doppler image of LV at apical four-chamber view was acquired.Vector flow mapping (VFM) was performed to assess the parameters of vortex during isovolumic contraction phase, including diameter (transverse and vertical diameter), velocity (maximal positive and negative velocity) and the number of vortex rings.Positive flow during systole and negative flow during diastole of LV in each layer were measured by flow-time curve.All values of patients with PVCs were recorded during sinus beats (PVC-S) and PVC (PVC-V) respectively.Results Significant differences were demonstrated in all parameters of vortex between the PVC-V and control subjects.And the flow-time curves disarrayed in PVC-V.The velocity of vortex in PVC-S was lower than that in control subjects.And the distribution pattern of flow-time curves in LV of PVC-S differed from that of control subjects.Conclusions Alternation of intracardiac fluid pattern in LV was demonstrated in patients with PVCs from RVOT during both sinus beats and PVC.VFM can be used to analyze the intracardiac flow field in normal and pathological electrical activation.

AIM: To investigate the expression of microRNA-375 (miR-375) in hepatocellular carcinoma (HCC) and to analyze the target genes and signaling pathways regulated by miR-375.METHODS: The expression of miR-375 was examined at tissue microarray of HCC by in situ hybridization.The whole human genome chip and bioinforma-tics analysis were applied to screen out the differential expression genes and signaling pathways in 4 HCC cell lines trans-fected with miR-375 mimic.RESULTS:In situ hybridization showed the expression of miR-375 in HCC tissues were obvi-ously higher than that in tumor-adjacent tissues (P<0.05).There were 20 co-upregulated genes and 17 co-downregulated genes in all 4 cell lines.Bioinformatic analysis showed that there were 54 signaling pathways related to up-regulated genes and 48 signaling pathways related to down-regulated genes in all 4 cell lines.CONCLUSION: miR-375 may play a key role in the pathological process of HCC.The bioinformatic analysis is able to screen the target genes and signaling pathways regulated by miR-375 and to provide an explicit direction for further mechanism research on HCC.

Objective Our purpose was to evaluate the teaching satisfaction of clinical epi-demiology among medical postgraduate and to come up with measures for further improvement of teaching quality. Methods A self-administered questionnalre was given to all the medical postgradu-ates and doctoral students of Grade 2013 by cluster sampling when they finished the course of clinical epidemiology. A total of 559 graduate students, including 324 graduate students (58%), 235 doctoral students (42%), recruitment graduate students 350 (62.6%), on-the-job graduate student 209 (37.4%).The contents of the questionnalre included many aspects such as the investigation object in general, teaching materials evaluation satisfaction, curriculum and teacher satisfaction evaluation. Respondents ' self-administered manner was adopted. Parallel input was done by using Epidata software; data were analyzed with SPSS 19.0 software, continuous variable was made by x±s, categorical variables was expressed by n(%). Continuous variables were compared among groups by t test and analysis of vari-ance. Classification grouping variable was compared by chi-square test, and P<0.05 for the difference was statistically significant . Results Overall evaluation of theory was below that of the internship teaching materials. The satisfaction rate of practicability, meeting the learning needs and difficulty degree for theory and practice teaching material were 83.7% (468/559), 87.5% (489/559), 67.1% (375/559) and 92.7%(518/559), 89.6%(501/559), 83.0%(464/559) respectively. 41.6%(87/209) of on-the-job students and 36.7% (119/324) of postgraduates considered theory teaching material was difficult for them. The satisfaction rate for faculty teaching attitude (99.5%, 556/559), faculty teaching method (98.6%,551/559), the theories combining with practice teaching method (97.5%, 545/559) were high, but the satisfaction rate for teaching hours was low (67.4%, 377/559), with one third students consider more hours for the course. Conclusion The Teaching effect of clinical epidemiology is falrly good in our school. The postgraduates are satisfied with the teaching materials, curriculum setting, and teach-ing faculty. But there are also some shortcomings. In the future teaching work, we should appropriately increase the hours, and set corresponding teaching contents and methods according to the different learning characteristics of the on-the-job graduate students and recruitment graduate students, to improve the teaching effect and teaching quality.

BACKGROUND: The content of type Ⅰ and Ⅲ collagen and the ratio of both are crucial factors to promote heart geometric morphology change,and ventricular systolic and diastolic myocardial performance.Matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase 1 (TIMP1) are primary regulatory substances of collagen metabolism.After myocardial infarction,chronic heart failure rats were subjected to bone marrow meseuchymal stem cell transplantation.What changes in MMP-2 and TIMPI would occur?OBJECTIVE: To observe the content of type Ⅰ and Ⅲcollagen and the ratio between both,as well as expression levels of MMP-2 and TIMP1 in the left ventricular tissue of rats with myocardial infarction-caused chronic heart failure following bone marrow mesenchymal stem cell (MSC) transplantation.DESIGN: A randomized controlled experiment.SETTING: Department of Senile Angiocardiopathy,General Hospital of Chinese PLA & Department of Biochemistry,Peking University Health Science Center.MATERIALS: This study was performed at the laboratory of Department of Biochemistry,Peking University Health Science Center between July 2004 and December 2005.Male Sprage Dawley rats of clean grade,aged 4 weeks old,were provided by the Laboratory Animal Center,Beijing Medical University and used for preparation of MSCs.Fourteen female rats,weighing 200-250g,were developed into models of heart failure-caused by myocardial infarction.METHODS: MSCs were isolated and,purified by gradient centrifugation and adherent cells were allowed to proliferate.Female rats underwent coronary artery ligation to induce chronic ischemic heart failure.Four weeks later,the rats were randomly divided into 2 groups: (1) experimental group (n=7),rats received transplantation of MSCs harvested from male rats [5×106 in 50 μL phosphate buffered saline(PBS)]by injection into the ischemic myocardium; (2) control group (n=7),rats received the same volume of PBS.MAIN OUTCOME MEASURES: Twenty-one days after therapy,(1) left ventricular fusion was tested by hematoxylin-eosinstaining and Masson staining; (2) Expression of MMP-2 and 1TMPI as well as contents of type I and llI collagen was analyzed by immunohistochemistry; (3) MMP-2 and TIMP1 expression levels were examined by Western blot.RESULTS: Fourteen rats were included in the final analysis.Type Ⅰ collagen expression in the scar area was much higher in theexperimental group than in the control group,while type Ⅲ collagen expression was much lower in the experimental group.MMP-2 expression was reduced and TIMPI expression was increased in the experimental group compared with the control group.Together,ventricular wall was thickened,ventricular chamber was reduced,and heart function was strengthened in the experimental group compared with the control group.CONCLUSION: MSC transplantation alleviated left ventricular remodeling in chronic ischemie heart failure,which results from dynamic regulation of MMP-2/TIMP1.

BACKGROUND: There are still few effective methods to repair injured myocardium after myocardial failure and pathologically rebuild reverral myocardium. As a new therapy, normal myocytes and therapeutic gene to interfere injured myocardium have advantageous effects in improving heart function.OBJECTIVE: To observe the efficiency and stability of adenovirus-medicated gene transferred into different passages of bone marrow mesenchymal stem cell (MSC) and investigate the effect of MSC-based sarcoplasmic reticulum Ca2+ ATPase gene (SERCA2a) gene therapy for rats with chronic heart failure. To compare the effects of gene therapy, cell transplantation and MSC-based SERCA2a gene therapy for chronic heart failure. DESIGN: Randomized controlled study.SETTING: Department of Senile Angiocardiopathy, General Hospital of Chinese PLA; Department of Biochemistry, Beijing Medical University. MATERIALS: Male Sprague-Dawley (SD) rats with 4 weeks old, clean grade and weighing 45-50 g provided by the Animal Experimental Center, Peking Medical University were used as donators of bone marrow. Other female SD rats of 12 weeks old, clean grade and weighing 200-250 g were used as receptors of cell transplantation and gene therapy. Sry gene of Y chromosome in male rats was used to evaluate whether transplanted cells of donators lived in myocardium of receptor rats. Ad-SERCa2a and Ad-EGFP were constructed by Doctor Lu Xiao-chun; MSC in the 3rd and 8th generations was isolating cultured on its own. METHODS: The experiment was carried out in the Zhou CY Laboratory (BSL-2), Department of Biochemistry, Beijing Medical University from July 2004 to December 2005. Thirty female SD rats received ligation at the left coronary artery to make models with chronic cardiac failure following acute myocardial infarction. And then, 29 rats were randomly divided into four groups, including gene therapy group (n=7), MSC group (n=7), gene-modified MSC group (n=8) and control group (n=7). Rats in the four groups were given the interventions of SERCA2a gene, MSC transplantation, MSC+Ad/SERCa2a and empty adenoviral vector, respectively. MSCs were separated and cultured, and then Ad-SERCA2a-GFP was used to transfer MSC in the 3rd and 8th generations.MAIN OUTCOME MEASURES: Ad-SERCA2a-GFP transfection rate of MSC was measured by using flow cytometer. Before and at 14 and 21 days after treatment, cardiac function was evaluated by ultrasonic echocardiogram. Expression of cytokine Ⅷ was tested by immunohistochemical staining. SERCA2a gene and protein expression were evaluated by RT-PCR and Western blot respectively, as well as SERCA2a enzyme activity. RESULTS: ① Transfection rate: The infection efficiency of adenovirus-medicated gene into different passages of MSC was over 80%, and there was no difference between passage three (P3) MSC and P8 MSC (P > 0.05). ② Heart function: Left ventricle wall was thickened obviously in group MSC and group MSC+Ad/SERCa2a on the 21st day after treatment, while volume was shortened and gradually rounded. Compared to control group, ejection fraction (EF) and shortening fraction (FS) of group Ad-SERCa2a, group MSC and group MSC+Ad/SERCa2a were elevated significantly on the 14th day after therapy (P < 0.01). While the elevation values of EF and FS began to reduce in group Ad-SERCa2a on 14th day after therapy, it continued to increase in both group MSC and group MSC+Ad/SERCa2a (P < 0.01). Improvement rate of EF at 21 days after therapy (EF D21) increased in group MSC and group MSC+Ad/SERCa2a respectively, but decreased in group Ad-SERCa2a. Compared to group Ad-SERCa2a, peak systolic flow velocity of anterior wall and interventricular septum in group MSC+Ad/SERCa2a increased significantly on the 21st day after therapy, and peak diastolic flow velocity of anterior wall and interventricular septum elevated in group MSC+Ad/SERCa2a, too (P < 0.01). ③ SERCA2a gene, protein expression and enzyme activity in group MSC+Ad/SERCa2a were significantly stronger in group MSC and control group. Parts of MSC transplanted into scar zone expressed Ⅷ.CONCLUSION: ① MSC is an effective platform for the targeted delivery of therapeutic gene. It suggests that different passages of MSC from P3 MSC to P8 MSC are regarded as high-effectively gene vehicles. MSC-based SERCA2a gene therapy showed much strong and lasting beneficial effect on exhausted myocardium. ② Effect of MSC transplantation on improving heart function may be related to promoting vascular neogenesis.

Objective To assess alternations in left ventricular volume and systolic synchrony in patients with frequent premature ventricular complexes(PVCs) from the right ventricular outflow tract(RVOT).Methods Twenty-nine patients with frequent isolated PVCs from RVOT were included and 30 healthy subjects as control.Instantaneous full-volume imaging(IFI) was performed to evaluate left ventricle volumetric parameters,including end-systolic volume (ESV),end-diastolic volume (EDV),stroke volume (SV),ejection fraction (EF),and systolic synchrony parameters,including systolic dyssynchrony index (SDI),dispersion end-systole (DISPES),mean end-systolic time (MES),pre-contraction time volume (PreContr) and post-contraction time volume (PostContr).Contraction front mapping was performed to visualize volumetric contraction sequence.All values of patients with PVCs were recorded during sinus beats (PVC-S) and premature ventricular beats (PVC-V) respectively.Results Significant differences were observed in left ventricular systolic volumetric and synchrony parameters between PVC-V and control subjects (P＜0.01),as well as in MES and PreContr between PVC-S and control subjects (P＜0.01).Conclusions Left ventricular systolic dysynchrony was demonstrated in patients with PVCs from RVOT.IFI was a novel tool to analyze left ventricular global and regional volumetric alternations.

Objective To compare the effect of autologous bone marrow stem cells transplantation on liver function between first and second transplantation in decompensated liver cirrhosis patients.MethodsA total of 45 decompensated liver cirrhosis patients were enrolled, and 23patients in first transplantation group were transplanted with autologous bone marrow stem cells through femoral artery when condition was stable after medical treatment.In second transplantation group, 22 patients were accepted second transplantation in 4-12 month after the first transplantation.All the patients undergone routine blood test, congulation test and liver function examination at the fourth week and eighth week after transplantation.ResultsEight weeks after transplantation, the liver function was improved obviously in both first and second autologous bone marrow stem cells transplantation.The level of albumin in patients of second transplantation group increased from (37.26± 5.90) g/L before transplantation to (42.49 ± 4.80) g/L (P＜0.01), alanine aminotransferase (ALT) decreased from (57.05±45.51) U/L to (44.86±29.19) U/L (P＜0.05),aspartate aminotransferase (AST) decreased from (39.14-±-15.07) U/L to (53.73 ± 24.98) U/L(P＞0.05).Congulation parameters were also improved, prothrombin time (PT) decreased from (16.15±3.01) s to (14.63±2.32) s (P＜0.01), fibrinofen (Fib) increased from (2.44±0.61) g/L to (3.00±0.81) g/L (P＜0.01).Compared with first transplantation group, the albumin level was higher in second autologous bone marrow stem cells transplantation group, which increased from (38.00±6.33) g/L to (42.49±4.80) g/L (P＜0.05), AST and ALT also improved obviously, and there was significant difference between two groups.Meanwhile, Child-Pugh scores decreased from (7.22±0.67) to (6.67±[0.71) (P＜0.05).But there was no significant difference in bilirubin, FIB and PT.ConclusionThe second transplantation of autologous bone marrow stem cells could further improve liver function and maintain symptoms remission of liver cirrhosis.