As an efficient gene transfer vehicle lentiviral vector has been widely used in therapy research. Comparing with other retrovirus vectors, lentiviral vectors have the unique ablility of transfecting nondividing cells and terminal differentiated cells. In addition, lentiviral vectors can accommodate two or more promoters and can carry larger foreign gene insertions. Now the new generation of lentiviral vectors encoding transcriptional control sequence provides effective means for the regulation of foreign gene expression. The development of lentiviral vectors and its application in the gene therapy field were summarized.

512?g/mL; MIC of 16 VIE to vancomycin were 16?g/mL; MIC of 8 VIE to vancomycin were 8?g/mL. It is a risk factor for hospital infection that VRE carriage of inpatients in intestinal tract is high. There is 100% agreement be-tween phenotypes and genotypes in 46 vancomycin resistance enterococci. VRE are multiresistant. Part iso-lates have high homology.

Objective To investigate the expressions of transferrin receptor (TfR) mRNA, ferritin(Fn) mRNA and iron regulatory proteinl (IRP-1) mRNA in the placentas complicated with fetal iron deficiency(ID). Methods Depending on the cord SF,the subjects were divided to fetal ID group and fetal iron sufficient( IS) group. TfR mRNA, Fn mRNA and IRP - 1 mRNA in placentas were measured by RT - PCR. Results 1. The expression of TfR mRNA in ID group was 1.10 ? 0. 26, it was significantly higher than that in IS group (t=0.028 P0.05) ;4. TfR mRNA and Fn mRNA correlated with fetal IS respectively. Conclusion Fetal ID induces highly expressed TfR mRNA and lower level of Fn mRNA to furthest satisfy the iron need for fetus.

Background Cardiomyocytes apoptosis was related to oxidative stress which has been shown to play an important role in ventricular remodeling after myocardial infarction(MI).Probucol is a hypolipidemic and antioxidant agents.Several reports demonstrated its cardioprotective effect after MI.However,the exact effect of probucol on the modification of the apoptosis related gene Bcl-2 and Bax is not clear.Objective To investigate the effects of probucol on mRNA and protein expression of Bcl-2,Bax and oxidative stress in myocardial infarcted rats.Methods Forty-one SD rats that survived 24 h after ligating left anterior descending coronary artery were randomly to receive placebo-saline(5 mL/d,n=20)or probucol(probucol 60 mg/kg?d,n=21).Twelve rats underwent sham operation were served as control(n=12).Six weeks after treatment,hemodynamic pararmeters and left ventricular function were measured with catheterization.Cardiomyocytes apoptosis were determined by TUNEL method.Myocardium mRNA of Bcl-2 and Bax expression levels in the non-infarcted myocardium were as- sessed by RT-PCR.The myocardium protein expression of Bcl-2 and Bax in the non-infarcted myocardium were de- termined by Western blot.Colorimetry was used to determine oxidative metabolism index in myocardium.Results 1)Compared with the sham rats,all MI rats showed marked decrease in Bcl-2 mRNA and protein expression in myo- cardium with increase of Bax mRNA and protein expression and apoptosis index(P

Objective To investigate neural correlates of the generation,execution and inhibition of motor intention. Methods Behavioral performance and event-related potentials (ERP) of 14 participants were recorded when they performed a two-staged Go-Nogo task. At the stage of intention preparation,three kinds of the "what" cues were presented separately to the participants : (1) to press a response key by the left or the right hand (the instructed) ,(2) to decide freely which hand will be used (the free) ,(3) to wait for the second cue (the control). At the stage of intention execution,the "whether" cue suggested the participants execute (Go) or withdraw (Nogo) the previous intention either voluntarily or forcedly. Results The interaction effect between the "what" and the "whether" was significant(F(2,26) = 8.262, P=0.002). The forced responses were faster than those of voluntary action under both the instructed condition ((468.52 ±105.87)ms vs (546.67 ±146.33)ms) and the control condition ((536.78 ± 83.28) ms vs (589.75 ± 140.80) ms). No significant difference was found for the free condition((538.71 ± 127. 39) ms vs (561. 44 ± 146. 51) ms). At the stage of intention preparation,significant ERP effects between the instructed intention and the freely intention were revealed in bilateral frontal regions (150～200 ms) , frontoparietal area (300～700 ms)and right temporal area. At the stage of intention execution,ERP effects between voluntary inhibition and voluntary action were observed in left prefrontal area (160～220 ms) ,fro-nto-cento-parietal area and right frontoparietal area. (300～550 ms). Conclusion The motor intention may be generated in the prefrontal area and maintained in the frontoparietal network. The left prefrontal and fronto-centro-parietal areas inhibit intention in the early stage and the right frontoparietal area involves response inhibition lately.

Objective To investigate neural correlates of the retrieval of temporal order.Methods ERP of 19 channels was recorded from 17 healthy young students while they performed a recency judgment task,a recognition task and a visual discrimination task separately,which were based on Sternberg paradigm.One-way ANOVA of repeated-measurement and post-hoc paired comparisons were carried out for both the task factor(temporal order,recognition and control)and the temporal distance factor correspondingly.The latter contains 3 intervals (1,2 and 3)between the targets.Results The temporal distance effect was significant:the closer targets correspond to longer reaction time((745.5±106.1)ms,(711.7±102.2)ms,(653.7±81.5)ms;F_((1.28,20.48))=43.37,P=0.00]and higher error rate((62.8±7.0)%,(72.5±5.4)%,(84.5±6.4)%;F_((1.32,21.11))=135.99,P=0.00].Significant behavioral effect of the task factor was also found for both reaction time(F_((1.32,21.11))=135.99,P=0.00)and error rate(F_((2,32))=200.31,P=0.00).No significant behavioral difference was found between the temporal order group and the recognition group.The spatiotemporal pattern of statistical parametric mapping(SPM)suggested the ERP effects of temporal order were presented in left temporal region (260～270 ms),bilateral parietal-occipital areas(280～290 ms),and bilateral parietal lobes(300～400 ms)prominently.No ERP effect was found between the temporal order task and the recognition task.Conclusions The neural correlates of the temporal distance effect exist in bilateral parietal regions.Temporal order as well as object information processing involves similar cerebral regions during the retrieval period.

Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Heart ; Heart Diseases ; drug therapy ; Humans ; Medicine, Chinese Traditional

Objective:To remove murine embryonic stem cells(mESC)from the differentiating cell culture and purify the differentiated cells by Magnetic Activated Cell Sorting(MACS).Methods:Neural differentiation of mESC was induced by a 5-stage method.The specific cell surface marker,SSEA-1,was used to identify ES cells in the differentiating cells.The optimal dilutions of mouse anti mouse SSEA-1 IgM primary antibody and FITC conjugated goat anti mouse secondary antibody were determined before the flow cytometry test.The incubation time and incubation temperature of primary antibody were all optimized to make the cytometry test accurate.After the optimization,stage 4 cells were dissociated into single cell suspension,incubated with antibody of SSEA-1 and microbeads conjugated goat anti mouse IgM,and then sorted through the magnetic field.The rate of SSEA-1 positive cells in pre-and post-separation groups was assessed by flow cytometry,and the viability of cells was evaluated by trypan blue staining counting under light microscopy.Results:The proportion of SSEA-1 positive cells in the separated cells can be reduced from(7.19?1.36)% to(1.34?0.80)%.The survival rate of sorted cells was more than 92%,similar to that of pre-separation cells.Conclusions:The MACS system we used can effectively remove mESC from the differentiated cells.The sorted cells will be well provided for the subsequent studies about transplantation therapy.

OTX1 gene is one of the pivotal transcriptional factors involved in the neurogenesis.In order to overexpress the OTX1 gene in distinct cell types and find out its contribution to the proliferation and differentiation of stem cells in vitro,OTX1 cDNA was subcloned into lentiviral vectors.The resulting constructions pDUETOTX1,pDUETGFPOTX1 and pDUETGFP were packaged in 293 cells producing viral particles to transduce 293T cells,SY5Y cells,mouse embryonic stem cells and E15 neural stem cells.It was proved that the transferred OTX1 gene was located in the nuclei of the transduced cells in stead of plasma.Lentivirus is an ideal vector delivering gene to different cells.The overexpression of OTX1 in transduced 293T cells were validated by Western blot and immunofluorescence.