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 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.

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 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.

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 effect of sevoflurane postconditioning on mitochondrial connexin 43 (Cx43) during myocardial ischemia-reperfusion (I/R) in isolated rat hearts and the role of mitochondrial ATP-sensitive potassium (mito-KATP) channels in it.Methods Forty-five adult male SpragueDawley rats,weighing 200-250 g,were used in the study.Their hearts were excised and retrogradely perfused with K-H solution in a Langendorff apparatus.The 45 isolated hearts were assigned into 5 groups (n =9 each) using a random number table:control group (group C),I/R group,sevoflurane postconditioning group (group Sev),and sevoflurane postconditioning plus 5-hydroxydecanoate (5-HD,a specific mitoKATp channel blocker) group (group Sev+5-HD) and I/R plus 5-HD group (group I/R+5-HD).The hearts were subjected to 20 main of global ischemia followed by 90 min of reperfusion to establish the model of myocardial I/R injury.From the beginning of reperfusion,the hearts were perfused with K-H solution saturated with 3％ sevoflurane for 15 min in group Sev,with K-H solution saturated with 3％ sevoflurane and containing 100 μ mol/L 5-HD in group Sev+5-HD,and with K-H solution containing 100 μ mol/L 5-HD in group 5-HD.The heart rate (HR),left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and the maximum rate of increase and decrease of ventricular pressure (±dp/dtmax) were recorded at the end of equilibration (T1) and 30 and 60 min of reperfusion (T2,3).At 90 min of reperfusion,the myocardial infarct size was measured by TTC staining,and the expression of total Cx43 (tCx43)and phosphorylated Cx43 (p-Cx43) in mitochondria was determined by Western blot analysis.The percentage of myocardial infarct size and p-Cx43/tCx43 ratio were calculated.Results Compared with group C,the HR,LVDP and ±dp/dtmax were significantly decreased,and the LVEDP was increased at T2,3,and the percentage of myocardial infarct size was increased in the other 4 groups,the expression of mitochondrial tCx43 and p-Cx43 was significantly down-regulated in I/R,Sev+5-HD and 5-HD groups (P＜0.05),and no significant change was found in the expression of mitochondrial tCx43 and p-Cx43 in group Sev (P＞0.05).Compared with group I/R,the HR,LVDP and ±dp/dtmax were significantly increased,and the LVEDP was decreased at T2,3,the percentage of myocardial infarct size was decreased,and the expression of mitochondrial tCx43 and p-Cx43 was up-regulated in group Sev (P＜0.05),and no significant change was found in the parameters mentioned above in Sev+5-HD and 5-HD groups (P＞0.05).Compared with group Sev,the HR,LVDP and ±dp/dt were significantly decreased,and the LVEDP was increased at T2,3,the percentage of myocardial infarct size was increased,and the expression of mitochondrial tCx43 and p-Cx43 was down-regulated in group Sev+5-HD (P＜0.05).There was no significant change in the pCx43/tCx43 ratio between the five groups (P＞0.05).Conclusion The mechanism by which sevoflurane postconditioning attenuates myocardial I/R injury may be related to induction of mito-KATP channel opening and up-regulation of the expression of mitochondrial Cx43 in cardiomyocytes of rats.

Objective The dose and type of light and photosensitizer could seriously affect the curative effect of photodynamic therapy (PDT).The purpose of this study was to observe whether or not PDT with hematoporphyrin monomethyl ether (HMME) can cure laryngocarcinoma in the solid tumor model,and to define the proper laser amount for killing the cancer cells.Methods Forty eight BALB/c mouse models with subcutaneous Hep-2 laryngeal carcinomas were prepared.Mice were divided into six groups depending on the amount of laser received from 30 J/cm2 to 480 J/cm2 including a control group,tumor size in each group was between 8 mm and 10 mm.Tail vein injection were given with HMME prior to applying the laser light,and then illumination was carried out on the tumor at 3 h after HMME administration.Tumor volume,animal weight and histopathologic changes were observed after PDT.Results All mice apparently showed positive results via PDT,and the cancer had been cured in 120 J/cm2 and 480 J/cm2 groups.The laryngeal cancer lesions began to form scab 1 d after PDT and the scab became hard and black after 5 d.The tumor regression began simultaneously and completed around 30 d after PDT.Conclusions PDT may treat laryngeal cancers which sized less than 10 mm in mouse models.The optimum energy to destruct the laryngeal cancer cells may be 120 J/cm2.

Based on photon counting mode,a multi-channel continuous-wave(CW) diffuse optical tomography(DOT) system is designed for galactophore inspection.For the acquisition of dynamic information,the instrument adopts the non-intermittent counters to achieve the function of width-adjustable moving gate.The width of the moving gate can be changed from 50 microseconds to about 10 seconds,and correspondingly the total temporal span of the measurement ranges from several milliseconds to tens seconds,depending on the preset gate number.The data transfer as well as the system control is through the universal serial bus(USB) 2.0 technique,with which the system can readily extended for a variety of applications requiring different spatial resolution.

A patient was treated by multiple dental implants,the implanted 6 was adjucent to impacted 8 .Immediately after implanta-tion,4 month and 3 year after implantation the distace between 8 and 6 implant central line was 4.4,3.2 and 2.5 mm,the angle between 8 long axis and 6 implant central line was 42.3°,45.5°and 50.3°.Then 8 was extracted.