AIM: To investigate whether leukotriene D 4(LTD 4) would stimulates proliferation of cultured human airway smooth muscle (ASMC). METHOD: Human ASMC were isolated and subcultured, varying concentration of LTD 4 were added to the media. Cell counts were obtained, [3H]-thymidine([3H]-TdR) incorporation and inositol 1, 4, 5-trisphosphate (IP 3) accumulation were measured. RESULTS: LTD 4(0.1 nmol?L -1～10 nmol?L -1) increased cell number and also increased incorporation of [3H]-TdR and accumulation of IP 3 in a concentration dependent manner(P

Bronchi ; cytology ; Cell Culture Techniques ; Cells, Cultured ; Humans ; Myocytes, Smooth Muscle ; cytology

Wnt signals widely exist in live creatures,involved in many physical or pathological processes,including the cell prolif-eration, differentiation and apoptosis. Recent studies show that Wnt/β-catenin signaling pathway may play an important regula-tory role in the protective effects of neurovascular unit in ische-mic stroke. Some chemicals,such as sulindac and estradiol,and some compound traditional Chinese medicine which contains gly-coside( e. g. rhodiola rosea ) , volatile oil ( e. g. turmeric ) and compound prescription ( e. g. Dan Long Xing Nao Fang ) can protect the neurovascular unit by regulating the pathway. This aims to review the research progress of the protective effects of Wnt/β-catenin signaling pathway on the neurovascular unit in is-chemic stroke,with combination of chemicals and traditional Chi-nese medicine which can protect the neurovascular unit via Wnt/β-catenin signaling pathway. It aims to provide the methodology basis for elucidating the mechanism of drugs,and also provide a new idea for researching the new drug which can reduce the dis-ability rate caused by sequela of stroke.

Objective To observe the phosphorylation levels of extracellular signal-regulated kinase 5 (ERK5) in acute myocardial infarction (AMI) patients and the effects of ERK5 selective inhibitor XMD8-92 on human platelet activation in vitro,and to explore its mechanism.Methods Western blot was applied to detect the phosphorylation levels of ERK5,Akt473 and Akt308 in AMI patients (n =34) and stable angina patients (n =33,control).The effects of different concentration of XMD8-92 on human platelet aggregation induced by collagen was tested by aggregometer in vitro.The release of ATP was measured simultaneously by luciferase detection.The effects of XMD8-92 on integrin aIIbβ3 were detected by platelet spreading on immobilized fibrinogen and clot retraction.The effects of XMD8-92 on phosphorylation levels of Akt473,Akt308 PTEN370 and ERK5 were detected by Western blot.Results The levels of phosphor-Akt473,Akt308 and phosphor-ERK5 were significantly higher in AMI patients than that in control group (P＜0.05).ERK5 inhibitor XMD8-92 diminished collagen-induced platelet aggregation,ATP secretion,the average area of platelet spreading on immobilized fibrinogen and the clot retraction extent.The levels of phosphor-Akt (Ser-473/Thr-308) and phosphor-PTEN (Ser370) were significantly down-regulated in the presence of XMD8-92.Conclusions ERK5 plays a role in platelet activation in AMI process.It regulates platelet activation by regulating PTEN and Akt phosphorylation.Its specific inhibitor is hoped to be new antithrombotic drug.

Dextroscope workstation is an advanced equipment developed in recent years and isused to make virtual reality model.We not only used it in surgical planning of thyroid disease,but also applied it in clinic teaching.Application of Dextroscope virtual reality system can help to improve students'capabilities of clinical thoughts and clinical techniques,stimulate their learning interest,enlarge the knowledge,shorten learning curve and release the conflict in practice.Dextroscope system will push forward the progress of education reform,so it is worth spreading.

This paper proposes a one-way micro valve with a simple structure and a simulation design for the engineering capsule. We have now got its design parameter selection method and its mechanic characteristic from experiments.
Capsules ; Computer Simulation ; Drug Delivery Systems ; methods ; Equipment Design ; Infusion Pumps, Implantable ; Technology, Pharmaceutical ; instrumentation ; methods

Objective To prepare controlled release microspheres of vascular endothelial growth factor(VEGF)& calcium alginate and observe their effect on proliferation of human umbilical vein endo- thelial cells(HUVEC)in order to provide theoretical basis for controlled release of VEGF facilitating an- giogenesis of tissue engineering bone.Methods VEGF-calcium alginate microspheres were prepared by using the needle extrusion/external gelation method to investigate physicochemical character and in vitro release of VEGF.According to the different ingredients added into the culture medium,the seconda- ry cultured HUVEC were divided into four groups,ie,control group,microsphere group,VEGF group and VEGF-calcium alginate microsphere group,in which the proliferation of the cultured HUVEC was ob- served with cell counting method,MTT method and flow cytometry.Results The calcium alginate mi- crospheres were revealed as spherical shape and evenly distributed,with mean grain diameter of(560?50)?m,carrying capacity of 0.72 ng/mg and the encapsulation efficiency of 54%.Smooth controlled re- lease in VEGF-alginate microspheres lasted for more than 10 days.Proliferation of the cultured HUVEC was accelerated the most in VEGF group at the beginning but in EGF-calcium alginate microsphere group at midanaphase compared with other groups,with statistical difference(P＜0.05).There was no statis- tical difference upon cell counting,cell activity and time point of cell cycle between control group and mi- crosphere group.Conclusion VEGF-sodium alginate microapheres can continue activity of VEGF,re- lease VEGF for over 10 days and promote proliferation cultured HUVEC for a long time.

Objective To construct and express the recombinant plasmid pET28α-Sj26GST-Sj32 of Schistosoma japonicum(Sj) in Escherichia coli BL21 (DE3). Methods Total RNA was extracted from Sj adult worms by ultrasound-breaking, Sj26GST and Sj32 antigen gene was respectively amplified by RT-PCR from the total RNA; Sj26GST-Sj32 fusion gene obtained with gene splicing by overlap extension(SOEing) was cloned into prokaryotic expression plasmid pET28α and transformed into Escherichia coli BL2 (DE3) to construct pET28α-Sj26GST-Sj32;BL21 (pET28α-Sj26GST-Sj32) was induced with isopropyl-β-D-thiogalactopyranosid (IPTG), and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE)and Western blotting. Results The 1991 bp Sj26GST-Sj32 fusion gene was successfully amplified by gene SOEing and cloned into pET28α by restriction analysis and PCR identification, the recombinant plasmid pET28α-Sj26GST-Sj32 was successfully constructed; the relative molecular mass of the expressed recombinant protein was approximately 69 × 103 by SDS-PAGE, and the amount of the expressed protein was 25% of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pET28α-Sj26GST-Sj32 is successfully constructed and highly expressed in Escherichia coli in fused form with His-tag, and the expressed fusion protein shows specific antigenicity.

To study the equipment modular allocation in the emergency medical unit after Beijing emergency public affairs. Using an improved Delphi method, 25 experts from various fields in Beijing were involved into two rounds consultations. The questionnaire of the first round consultation was formed based on analysis of emergency rescue equipment. Based on the result of the first round consultation, the questionnaire of the second round consultation was set up. The two rounds of questionnaires had the responding rates of experts 100% respectively, and the au-thoritative coefficients of the experts were 0.81 and 0.88. An emergency medical rescue equipment solution with three de-grees modular configuration in Beijing was made based on the consultations and experts' advices. The solu-tion develops the emergency medical service ability of mobile medical units and vehicle-mounted mobile hospital. LUO Ying and GAO Hong-wei are the first authors who contributecl equally to the article.

The inhibitory ratio (1%) of artificial musk on cyclooxygenase-2 (COX-2) was determined by enzyme immunoassay (EIA). The dose-effect relationship between concentrations of artificial musk and 1% was established. It was found that artificial musk had obvious inhibitory action on COX-2. The concentration for 50% of maximum inhibitory effect (IC50) was about 2.26 mg x mL(-1). There was a good relationship between the logarithm concentrations of artificial musk and 1% when the concentrations of artificial musk ranged from 0.31-20.0 mg x mL(-1). The results indicated that this EIA method could be applied to evaluate the anti-inflammatory activity of artificial musk quickly, conveniently, sensitively and exactly. This paper provided a novel method and foundational research for the bioassay of artificial musk.