The aim of this study is to clone the mouse T-bet promoter and enhancer, construct and identify the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS for T-bet transcription regulation study and its function in signaling of multiple sclerosis. The promoter and CNS of T-bet were predicted by bioinformatics assay. The predicted fragment of mouse T-bet promoter plus CNS was amplified by PCR and cloned into pGL4.10. The recombinant plasmid pGL4.10-TBX21pr-CNS was transferred into Escherichia coli DH5α. The positive clone was identified by double digestion with Kpn I and Sfi I and DNA sequencing. Finally, pGL4.10-TBX21pr-CNS was cotransfected with pRL-TK into 293T cells and Jurkat cells, pRL-TK and pGL4.10 as a control. The luciferase activity in 293T cells (P = 0.012 2) and Jurkat cells (P = 0.002 2) was higher than that of the control group. A fragment of 1 028 bp mouse T-bet promoter plus 1 308 bp CNS was successfully cloned and the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS was constructed. In 293T cells and Jurkat cells, pGL4.10-TBX21pr-CNS has the promoter functions. This work offers a basic material for the research of T-bet transcription.
Animals ; Gene Expression Regulation ; Genes, Reporter ; Genetic Vectors ; Luciferases ; Mice ; Multiple Sclerosis ; Plasmids ; Promoter Regions, Genetic ; T-Box Domain Proteins ; genetics

Objective To observe the impacts of tirofiban on plasma levels of sCD40L and HIF-1α, and iNOS activity in patients undergoing emergency PCI. Methods A total of 80 patients with acute STEMI received primary PCI were randomly divided into a tirofiban group and a control group. The tirofiban group received tirofiban of 10 μg/kg, followed by an infusion of tirofiban of 0.15 μg/(kg·min) for 24 ~ 36 hours; while the control group directly received stent implantation after balloon angioplasty. Plasma levels of sCD40L , HIF-1α, and iNOS were detected in baselines and 12 hours after the procedrue. Results 12 hours after the procedure, serum levels of sCD40L, HIF-1α, and iNOS were higher than the baseline levels (P < 0.05). As compared with the control group, levels of sCD40L, HIF-1α, and iNOS were significantly lower (P < 0.05). Conclusions This study preliminarily sugguests tirofiban has a role in anti-inflammation and protection of vascular endothelium besides anti-platelet effects.

Objective To establish a cell line with repressed expression of p53 by transfecting a plasmid construct expressing short hairpin RNA(shRNA)targeting p53 into human skin fibroblasts(HSFs),and to evaluate the effect of repression of p53 expression on the senescence in HSFs.Methods The eukaryotic expressing plasmid pGCsi-p53 containing shRNA targeting p53 gene was transfected into HSFs with lipofectamine.Subsequently,the cells were selected by G418,and resistant cell clones were chosen and expanded.Reverse transcription-PCR and real time fluorescence-based quanitative PCR were performed to determine the expression of p53 gene,and Western blot to detect the expression of p53 protein in HSFs.The senescence in HSFs was evaluated by SA β-gal staining,and cell proliferation by methyl thiazolyl tetrazolium(MTT)assay.Results A HSF clone with repressed expression of p53 was established successfully.The expressions of p53 mRNA and protein were downregulated in transfected HSFs compared with untransfected HSFs(0.09 ± 0.03 vs.0.32 ± 0.04,0.11 ± 0.04 vs.0.84 ± 0.05,both P ＜ 0.01).The percentage of senescent cells was 13.47％ ± 1.01％ in the transfected HSFs,significantly lower than that in untransfected HSFs(18.10％ ± 0.66％,P ＜ 0.05).As MTT assay showed,the proliferation was accelerated in transfected HSFs compared with untransfected HSFs(P ＜ 0.05).Conclusions The repression of p53 expression decelerates the senescence in HSFs,but promotes the proliferation of HSFs.

Objective To explore the stress perception among acute myocardial infarction (AMI) patients during acute stage.Methods Phenomenological method of qualitative study was adopted.18 AMI patients during the acute stage of 3～4 days after the onset of the disease were interviewed.Results Stress made AMI patients hard to accept the fact of illness,worry about the treatment of illness and the impact on future life,be difficult to alter living habits,be hard to relieve negative emotion.Conclusions AMI patients during acute stage will experience different levels of psychological stress.Stress management should be fabricated and effective measures should be adopted to relieve their negative emotions in terms of inducing factors.

Objective To discuss the efficacy of edaravone combined with batroxobin in the treatment of patients with acute progressive infarction,and its influence on neurological deficit and coagulation.Methods 64 patients with acute and progressive infarction were selected,they were randomly divided into the control group and the observation group,32 cases in each group.The patients of the observation group were given edaravone combined with batroxobin,while the patients of the control group were given batroxobin only.Results The total effective rate of the observation group was 91.63%,which was obviously higher than 78.13% of the control group(χ2 =11.274,P <0.05).The NDS of the observation group was (14.8 ±2.9 )points,which was obviously lower than (17.9 ± 3.3)points of the control group(t =9.98,P <0.05).The level of Fib of the observation group was (2.9 ±0.3)g/L, which was obviously lower than (3.2 ±0.4)g/L of the control group(t =9.34,P <0.05).Conclusion Edaravone combined with batroxobin in the treatment of patients with acute and progressive infarction has significant effect and deserve promotion.

Traditional sample entropy fails to quantify inherent long-range dependencies among real data. Multiscale sample entropy (MSE) can detect intrinsic correlations in data, but it is usually used in univariate data. To generalize this method for multichannel data, we introduced multivariate multiscale entropy into multiscale signals as a reflection of the nonlinear dynamic correlation. But traditional multivariate multiscale entropy has a large quantity of computation and costs a large period of time and space for more channel system, so that it can not reflect the correlation between variables timely and accurately. In this paper, therefore, an improved multivariate multiscale entropy embeds on all variables at the same time, instead of embedding on a single variable as in the traditional methods, to solve the memory overflow while the number of channels rise, and it is more suitable for the actual multivariate signal analysis. The method was tested in simulation data and Bonn epilepsy dataset. The simulation results showed that the proposed method had a good performance to distinguish correlation data. Bonn epilepsy dataset experiment also showed that the method had a better classification accuracy among the five data set, especially with an accuracy of 100% for data collection of Z and S.
Algorithms ; Electroencephalography ; Entropy ; Epilepsy ; diagnosis ; Humans ; Multivariate Analysis ; Nonlinear Dynamics

Objective To investigate the neural mechanism of inhibitory control in sensation seeking by using the event-related potential(ERP) technique. Methods High and low sensation seekers( 16 people in each group ), who were selected according to their sensation seeking scores, performed a Go/Nogo task in which the stimuli possessed two levels of difficulty. Electro- encephalogram(EEG) signals were recorded continuously by a set of 32 Ag/AgCI electrodes. Results For two types of stimuli ( Congruent, Incongruent) :( 1 )The amplitude(FCZ) of Nogo N2 and Nogo P3 were( (1.61 ±4.25)μV,(-2.32±4.55)μV)and((16.44±5.74)μV,(17.00±5.71)μV). (2)There was no significant main effects of group for the Nogo N2 amplitude( F (1.30) =0.31, P=0. 59,η2=0. 01;F(1.30) =0.07,P=0.80,η2=0.002) ,the N2d amplitude( F(1.30) =1.18,P=0.29,η2=0.04;F(1.30) =0.004, P=0.95, η2 ＜ 0.001 ) ,the Nogo P3 amplitude( F (1.30) =0.13, P=0.72, η2 =0.004;F(1.30)=0.28, P=0.60, η2 =0.009) and the P3d amplitude( F(1.30) =0.08, P=0.50, η2 =0.02; F (1.30) =0.56,P=0.46, η2 =0.02). (3)Neither of main effects for the N2 and P3 latency was significant (P＞0.05). Conclusion The inhibitory control is similar across high and low sensation seeking groups,indicating that there is no relationship between the sensation seeking behaviors and the individual inhibitory control.

Abstract: Neurotensin receptor-1 (NTR1), which can stimulate the intracellular cascade signal pathway, belongs to the large superfamily of G-protein coupled receptors. NTR1 is related to the occurrence and development of several kinds of diseases. In order to screen the inhibitors for the cancers associated with NTR1 protein, we established a CHO (Chinese hamster ovary) cell line in which human neurotensin receptor-1 was highly expressed. The method is to construct the recombinant plasmid which was lysed with the hNTR1 gene and transfect it into CHO cells. After selected with G418, the cell line was evaluated by Western blotting analysis and calcium flux assays. Through the calcium flux assays on FlexStation 3, we got the EC50 value of neurotensin peptide which is the natural NTR1 agonist, and the IC 50 value of SR48692 which is the known NTR1 antagonist. The established human CHO/NTR1 cell line can be used to study the profile of NTR1 biological activity and further screen of NTR1 antagonists and agonists.

Early use of statins after onset of acute ischemic stroke can improve the functional prognosis of patients and reduce in-hospital mortality.This article reviews the preclinical and clinical trials of statins for the treatment of ischemic stroke.

Objective To investigate the clinical value of radiotherapy combined with xeloda monotherapy in the treatment of elderly patients with rectal cancer.Methods 80 elderly patients with rectal cancer were retrospectively collected.The patients were assigned into study group (n=43) or control group (n=37) according to the treatment way.The study group adopted radiotherapy combined with xeloda,while the control group only treated with radiotherapy.The main indicators included major clinical outcomes (3-year survival rate,recurrence rate,progression free survival),and postoperative health related quality of life and major complications of the two groups were observed.Results Compared with the control group,the 3-year recurrence rate of the study group decreased significantly (25.58% vs.48.65%,x2=4.579,P=0.032);the progression free survival was significantly prolonged [(42.58±7.63)months vs.(34.95±6.30)months,t=7.495,P=0.000];the quality of health related life after 1 year increased significantly [(75.50±8.11) vs.(69.76±9.58),t=3.295,P=0.002].There were no significant differences in postoperative major complications (granulocyte reduction,diarrhea,nausea,vomiting,hand foot syndrome and sensory neuropathy) and 3-year survival rate between the two groups (all P>0.05).ConclusionRadiotherapy combined with xeloda chemotherapy helps improve progression free survival and recurrence rate in elderly patients with rectal cancer.