Objective:To prepare the recombinant neuron protective protein TAT-Bcl-XL in E.coli expression system and to determine its anti-apoptosis activity.Methods:DNA fragment of TAT-Bcl-XL was obtained by RT-PCR using primers that were specific for Bcl-XL gene. Prokaryotic expression vector(pTBTOPO) was constructed by insertion of TAT-Bcl-XL DNA fragment into pCRT7/CT-TOPO vectors, and the recombinant protein was detected by SDS-PAGE and Western blot analysis using anti-V5 tag epitope antibody as the primary antibody. The recombinant fusion protein was purified by affinity chromatography, and the anti-apoptosis function of the purified protein was determined by flow cytometry.Results:A molecular weight of 30 000 protein was detectable in both SDS-PAGE and Western blot analysis. Immunofluorescent staining showed the fusion protein was distributed in cell plasma after incubation with 200 nmol/L fusion protein. Flow cytometry data indicated that the recombinant protein could enhance cell survival by 40% when 293T cells were treated with 250 ?mol/L zinc chloride as a supplementation in cell culture media.Conclusion:High level expression of the reported neuron protective protein was obtained in E.coli expression system, and the purified recombinant proteins remained its anti-apoptosis activity in our preliminary characterization.

Kawasaki disease,which is an acute multisystem vasculitis of unknown cause,leads to pathognomonic feature of coronary artery lesion.Recent studies show that genetic polymorphism,neutrophils,monocytes,cytokines and other factors are involved in the development of coronary artery disease.Long-term use of anticoagulant drugs may reduce coronary artery damage.

Sepsis is thought to be systemic inflammatory response syndrome that is activated by invasive infection and characterized by the involvement of inflammation,infection,immunological reaction,coagulation and tissue damage.The imbalance between the pro-and anti-inflammatory response mediated by immunologic disorder plays an important part in the progress of sepsis.Therefore,the immunological mechanisms and changes of Toll-like receptors,neutrophils,dendritic cells and regulatory T cells in sepsis are briefly focused on in the article.

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major pathogens of the hospital infection.Its clinical features and drug-resisetance situations have always been concerned.But since the late 1990s,another class of MRSA has become a major concern worldwide as an emerging pathogen in the community.This new class of MRSA is called community-acquired MRSA (CA-MRSA).With the rapid development of the infection of CA-MRSA in 20 years,especially in the latest 3 years,CA-MRSA may be replacing the hospital-acquired MRSA strains(HA-MRSA) as one of the major pathogens in the hospital and the community as well.The characteristics of CA-MRSA are very different from those of HA-MRSA.This review summarizes the current studies of CA-MRSA on the epidemiology and the molecular characteristics.

Most nosocomial bloodstream infections among pediatric patients are related to the usage of an intravascular device.This article reviewed catheter related bloodstream infections from aspects of epidemiology,pathogenesis,diagnosis,prevention and care based on guidelines and new research both in abroad and at home.

Objective To explore the detection meaning of D-dimer(D-D) and fibrinogen(Fg) in patients with coronary disease.Methods 150 patients with coronary disease were randomly divided into UA group,SA group and AMI group,the D-D was detected by ELASA double resistance to clip method and Fg was detected by CLIUSE methods.Results The D-D in SA group,UA group and AMI group showed significant difference between two groups (SA:UA,t =9.340;SA:AMI,t =18.334; UA:AMI,t =10.240.all P ＜0.01 ).The Fg in SA group,UA group and AMI group showed significant difference between each two groups ( SA:UA,t =11.376; SA:AMI,t =11.730; UA:AMI,t =3.121.all P ＜0.01 ).D-D between one coronary artery disease and two coronaries artery disease,three coronary arteries disease showed significant difference( t =7.017,15.603,all P ＜ 0.01 ) ; and between two coronaries artery disease and three coronary arteries disease showed significant difference,too ( t =8.662,P ＜ 0.01 ).The Fg between one coronary artery disease and two coronaries artery disease,three coronary arteries disease,showed significant difference( t =8.687,11.855,all P ＜ 0.01 ) ; and between two coronaries artery disease and three coronary arteries disease showed significant difference,too( t =4.379,P ＜ 0.0 1 ).Conclusion The D-D and Fg showed obvious difference between different types of coronary disease,and the levels increase along with the disease degrees of coronaries

The key point to define the psychological problem in children is the grasp of degree,also,the key point to educate children is the grasp of degree.Are there some principles to clarify the degree? How to quantify the degree? This paper brought forward the gold zone method of psychological guidance in children and adolescents and defined its quantitive expression as golden section.The gold zone method can grasp the scale set according to the goal,form the proper consumption value,control the interpersonal intimacy,cultivate fine thinking and behavioral pattern,and give appropriate guidance to many basic problems about children's growth.

Thiazolidinedione (TZD) is a powerful insulin sensitizer in the treatment of type 2 diabetes. It acts as a ligand to the nuclear receptor PPARγ (peroxisome proliferator-activated receptor-gamma) and induces transcription of PPARγ responsive genes. TZD controls lipid synthesis and storage in adipose tissue, liver and many other tissues through PPARγ. Derivatives of TZD, such as rosiglitazone (Avandia) and pioglitazone (Actos), are more powerful than metformin or berberine in insulin sensitization. Although they have common side effects such as weight gain and edema, these did not influence the side effects in general. However, recent findings of risk for congestive heart failure and bladder cancer have indeed significantly impaired their future in many countries. European countries have prohibited those drugs and in 2011, US will terminate application of rosiglitazone in clinics and hospitals. The multiple country actions may mark the end of TZD era. As a result, there is a strong demand for identification of TZD substitute in the treatment of type 2 diabetes. In this regard, literature about PPARγ ligands and potential TZD substitute are reviewed in this article. Histone deacetylase (HDAC) inhibitor is emphasized as a new class of insulin sensitizer here. Regulators of SIRT1, CREB, NO, p38, ERK and Cdk5 are discussed in the activation of PPARγ.

Large-scale clinical trials have demonstrated that metabolic control achieved early in the course of diabetes can substantially help preventing the onset and slowing down the progression of diabetes and the associated microvascular complications. Various strategies incorporating the use of insulin early in the course of the disease have been developed to meet this goal and to compensate for the progression of β-cell failure. We believe that insulin suppresses inflammation and also reduces glucotoxicity and lipotoxicity, and thus exerts a pivotal and beneficial role. Comprehensive metabolic control, especially when instituted earlier, may alter the natural history of diabetic complications. Therefore, physicians are encouraged to take early and intensive interventions with insulin therapy for patients with type 2 diabetes, especially when attempting to induce "remissions" or in patients suboptimally controlled with oral medications.

Ten-year experience of state-owned public hospitals reform of Shanghai was reviewed and analyzed according to theGuiding Opinion of General Reform of Public Hospitals in Urban areas. The paper focused on administration and operation mechanisms reform,modes transition on service delivery and management with support from information technology were thoroughly introduced,and further working plans were provided based on the analysis of problems and challenges.