Aims To construct the N-terminal Strep-tagged ( NS-tagged) fusion protein expression vector, and to apply the vector to express NS-tagged fusion proteins of Chlamydia RNA polymerase subunit. Meth-ods By using PCR method, NS fusion protein tag and a new multiple cloning sites (MCS) were inserted into pET21c-DH plasmid by primers to replace the original T7 protein tag and MCS. The newly introduced Not I cutting site was chosen for self-ligation of PCR prod-uct. Then, the cyclized PCR product was transformed into DH-5α competent cells. The positive clones were selected by PCR and sequencing. To get NS-tagged fu-sion proteins of chlamydial RNA polymerase subunits, the α, β and β′ subunits were inserted between BamH I and Sal I cutting sites of the newly constructed ex-pression vector. Then, the NS-α, NS-β and NS-β′ ex-pression vectors were transformed into Arctic Express expression cells. The fusion protein expression statuses of transformed cells were identified by Commassie blue staining and Western blot. Results The NS-tagged fusion protein expression vector pET21c-NS-MCS was successfully constructed, and NS-α, NS-β and NS-β′fusion proteins were obtained by using this newly con-structed expression vector. Conclusions In this pro-ject, we constructed an NS-tagged fusion protein ex-pression vector and applied it to express NS-α, NS-βand NS-β′ fusion proteins. Our study can lay a solid foundation for the study of transcriptional regulation of Chlamydia genes.

Objective Bone cement joint prosthetic aseptic loosening always occurs in the bone cement-bone interface, while the mechanical strength of the interface mainly depends on the microstructure and adhesive strength.The aim of this study was to ex-plore the bone microstructure and the cement bone interfacial biomechanics of osteoarthritis(OA) and rheumatoid arthritis(RA), and also discuss the correlation. Methods Twenty trabecular bone specimens of tibial plateaus were taken from OA and RA patients who underwent total knee arthroplasty( TKA) .The microstructural parameters of the trabecular bone specimens were measured by Skyscan1176 microcomputed tomography.The bone specimens were made into cementbone models, which were tested by INSTRON strength tester.The association of bone microstructure and interfacial shear stress was analyzed subsequently. Results ①Bone volume fraction ( BV/TV) ( r=-0.313) , trabecular thickness ( Tb.Th) ( r=-0.340) ,trabecular spacing ( Tb.Sp) ( r=0.345) of OA had obvious correlation to shear strength (P<0.05).The cement-bone interfacial strength of medial tibial plateaus[(87.45±52.50)N] was lower than lateral tibial plat-eaus[(177.25±71.11)N] of OA (P<0.05).②Bone volume fraction (BV/TV)(r=0.343), trabecular number (Tb.N)(r=0.391) of RA had obvious correlation to shear strength (P<0.001).The cement-bone interfacial strength of lateral tibial plateaus[(62.23±46.22) N] was lower than medial tibial plateaus[(79.20±56.37N)] of RA (P<0.05).③The interfacial strength of OA[(132.35±76.64)N] was higher than RA[(71.05±51.55)N] (P<0.05). Conclusion There are differences of microstructure between OA and RA, which lead to the distinction of strength of cement -bone interface.And it has a certain guiding role of analyzing the biomechanics in TKA.

Objective To investigate the clinical effect of milrinone in the treatment of children with congenital heart disease complicated by severe pneumonia and heart failure. Methods 94 patients with congenital heart disease complicated with severe pneumonia and heart failure in our hospital from March 2014 to February 2016 were selected,and randomly divided into experimental group and routine group,47 cases in each groups. The routine group were treated with conventional treatment, the experimental group in the conventional group received routine treatment based on the same given milrinone treatment. The clinical efficacy, symptom disappearance time and adverse reactions were compared between the two groups. Results The treatment in the experimental group the total efficiency (93.62%) was significantly higher than that of the conventional treatment group with the total effective rate (78.72%), significant difference, statistically significant (P<0.05); the experimental group in children with fever, cough, rales and other symptoms disappeared time is lower than the conventional group, the data had significant difference, has statistical significance (P<0.05); two groups of patients during treatment were not any serious adverse reactions, the adverse reaction condition no difference does not have statistical significance.Conclusion The clinical effect of milrinone in the treatment of children with congenital heart disease complicated by severe pneumonia and heart failure significantly, clinical efficacy, low incidence of adverse reactions, worthy of promotion.

ObjectiveTo understand the relationship between mycoplasm load in bronchoalveolar lavage lfuid (BALF) with the status of Th1/Th2 immune response in children withMycoplasma pneumoniae pneumonia (MPP).MethodsThe levels of IL-4, IFN, IL-8 , TNF-α in BLAF and total IgE, ECP in serum from 90 children with MPP were measured by ELISA.MP DNA in BALF was detected quantitatively by lfuorescent real-time PCR. Children with MPP (n=90) were divided into two groups of low MP-DNA load (n=24) and high MP-DNA load (n=26) according to the copies of MP DNA in BALF. The cytokines in BALF, and total IgE and ECP in serum were compared between the two groups. The relationship between the levels of cytokines in BLAF and the copies was evaluated.ResultsThe levels of IL-4 and the IL-4/IFN ratio in BALF from the high DNA-load group were signiifcantly higher than that of the low group (t=4.280, 2.076, allP<0.05). The level of IL-4 was signiifcantly correlated with the copies of MP-DNA in BALF from children with MPP (r=0.509,P<0.05). The percentage of total IgE and ECP positive result in serum from the high DNA-load group is higher than that of the low group. (χ2=24.638, 6.392,allP<0.05).Conclusion Infection with high-load MP in children may cause the imbalance of Th1/Th2. And the Th2 cytokines response seems predomi-nant.

Objective To explore the competency of community health administrators in a survey of such administrators at large in Shanghai. Methods McClelland's Competency Dictionary was used to set two benchmark positions, while the behavioral event interviewing and questionnaires were made to survey 19 community health service centers in Shanghai. Results The competency model benchmarking on the head of a medical branch consists of 6 character clusters, 11 characteristic items, 27 evaluation dimensions and 27 typical behavior descriptions. The model benchmarking on the community team leader consists of 5 character clusters, 8 characteristic items, 21 evaluation dimensions and 21 typical behavior descriptions. Conclusion Findings of the survey provide objective criteria for competence appraisal of community health administrators, and serve as reference for formulating their development plans as well.

AIM:To investigate the effect of pretreatment of stem cell factor(SCF) and granulocyte colony-stimulating factor(G-CSF) on the proliferation and the differentiation of mesenchymal stem cells(MSCs) into cardiomyogenic cells.METHODS:The MSCs,isolated primarily from bone marrow,and purified by passage culture,were obtained from the adult rats of four groups:the rats were pretreated by 5 daily injections of SCF;the rats were pretreated with G-CSF;the rats were pretreated with SCF and G-CSF;the rats were treated without any intervention.The 4th passage of MSCs was labeled by DAPI and cellular cycle analysis was conducted by flow cytometry before co-culture.The neonatal rat cardiomyocytes cultured for 3 days were co-cultured with DAPI-MSCs.The percentage of the differentiation of MSCs into cardiomyogenic cells during the five co-culture days was analyzed.The morphologic changes of MSCs and the proteins expression of cardiac myosin heavy chain(MHC) and troponin T(TnT) were recorded respectively with digital microscope camera system and immunofluorescence technique.The percentage of the differentiation of MSCs into cardiomyogenic cells was also calculated.RESULTS:The percentage of MSCs in G0/G1 phase in SCF/G-CSF group was significantly lower than that in SCF group,G-CSF group and the control group.The percentage of MHC protein-positive MSCs in SCF/G-CSF group was markedly higher than that in SCF group,G-CSF group and the control group,and that in SCF group and G-CSF group was significantly higher than control group.The percentage of TnT protein-positive MSCs in SCF/G-CSF group,SCF group and G-CSF group was significantly higher than that in control group.CONCLUSION:SCF and G-CSF show the ability to stimulate the proliferation of MSCs and induce MSCs to differentiate into cardiomyocytes.The combination of using SCF and G-CSF is more effective than using only SCF or G-CSF.

Objective To evaluate the value of double stapling technique with curved cutter stapler in colorectal anastomosis,especially in low colorectal anastomosis. Methods The clinical data of 168 cases of rectal carcinomas treated with double stapling technique with curved cutter stapler were retrospectively reviewed.The intraoperative condition,postoperative complications and findings during follow up were analysed. Results During the operations,the processes of closure and anastomosis of all the patients were satisfactory,and no operative death occurred.After the operations,4 cases(2.4%) had anastomotic leakage,3 cases(1.8%) had anastomotic bleeding,and 2 cases(1.2%) had rectovaginal fistula.All the complications were cured.There was no anastomotic stricture. Conclusion Double stapling technique with curved cutter stapler may help to accomplish low colorectal anastomosis which is a difficult task for handed suture.

For treatment of pediatric inguinal hernia, we fabricated a device, i.e. so called "filling type pediatric hernia sac", which treats the problem from the abdominal cavity, through the abdominal and is a self-adaptive closer, using synthetic material. The device includes filling rack, self-adaptive umbrella support bar, bottom piece, outside pulling line and device fixing lines. The filling rack is composed of 2 concentric circles of 3.0 cm diameter with peripherally fixed together and can be pulled into the shapes of a ball or an olive. The supporting bar is structured of 3 pieces with 0.5 cm wide, 4.0 cm long, cross-fixed on top of the filling rack. The bottom piece is in a circular structure with a diameter of 3.0 cm, and it is connected to the filling rack bottom. Adjust positioning stay outside the fixed on the top of the device are connected at one end, and the other end free through filling the top frame connected with the bottom slice of central fixation. By using this device, we treated 37 pediatric inguinal hernia cases with 38 side-inguinal hernia successfully. The mean duration of post-operation follow-ups was 14.6 ± 5.89 months, without hernia recurrence, obvious scar and hard sections of inguinal region. This device could provide a convenient, safe and effective plugging technology for children's pediatric hernia.
Child ; Hernia ; therapy ; Herniorrhaphy ; instrumentation ; Humans

Objective: To investigate the effects of taraxerol and taraxeryl acetate on cell cycle and apoptosis of human gastric epithelial cell line AGS cells. Methods: The inhibitory effects of taraxerol and taraxeryl acetate at different concentrations on AGS cell growth were measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) method and the concentrations of taraxerol and taraxeryl acetate to be used in following experiments were decided. Then, cell cycle analysis was performed by FACScan flow cytometry after culture with taraxerol or taraxeryl acetate. Annexin V-fluorescein isothiocyanate/propidium iodide staining was used to measure cell apoptosis. Results: Taraxerol significantly inhibited AGS cell proliferation in a dose- and time-dependent manner. Taraxerol arrested the AGS cells at G(2)/M stage. 110 μmol/L taraxerol elevated the population of AGS cells arrested in G(2)/M phase compared with solvent (P<0.05). Taraxerol also promoted early cell apoptosis in AGS cells. 110 μmol/L taraxerol increased the early cell apoptosis rate from 4.45% to 10.29%, which was 1.31 times higher than that of the untreated cells. However, taraxeryl acetate had a lower inhibitory effect than taraxerol, and it showed a tendency of G(2)/M arrest and apoptosis promotion but with no statistical significance (P>0.05). Conclusion: Taraxerol has inhibitory effects on AGS cell growth through inducing G(2)/M arrest and promotion of cell apoptosis. Taraxeryl acetate has less effect on cell cycle arrest and apoptosis of AGS cells than taraxerol.

BACKGROUND:Adherent migration of vascular smooth muscle cells and proliferated remodeling of vessel walls following vascular endothelial injury play a key role in onset of restenosis after percutaneous transluminal coronary angioplasty,while expression and phosphorylation activation of focal adhesion kinase are attacked during this period.OBJECTIVE:To observe the interventional effect of angiotensin receptor antagonist,telmisartan,on the expression and activation of focal adhesion kinase during vascular-injured remodeling.DESIGN:Randomized controlled animal study.SETTING:Shenzhen Center for Chronic Disease Control and Prevention;Department of Cardiology,Affiliated Hospital of Xianning Medical College;Department of Cardiology,Tongji Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology.MATERIALS:The experiment was carded out in the Cardiovascular Laboratory,Wuhan Tongji Hospital from March to May 2005.A total of 36 male Wistar rats weighing 300-360 g were randomly divided into control group,model group and treatment group with 12 in each group.METHODS:Aortal restenosis models were established after endothelial denudation.Foley's tube technique was used to strip vascular endothetium of rats in the model group and the treatment group;while,rats in the control group and the model group were fed and drunk normally; in addition, rats in the treatment group were also given 5 mg/(kg·d) telmisartan solution.Thirty clays after successive administration,vessels at injured sites were collected to observe the morphological changes and extract RNA and protein.Reverse transcriptase polymerase chain reaction (RT-PCR) was used to measure mRNA expression of focal adhesion kinase and Westem blot technique was USod to measure total protein and phosphorylated protein of focal adhesion kinase.MAIN OUTCOME MEASURES: Proliferation of vessel wall at injured sites; mRNA expression, total protein and phosphorylated protein of focal adhesion kinase at injured sites at 30 days after administration.RESULTS:A total of 36 rats Were involved in the final analysis.Thirty days after operation, aortic tunica intima was thickened in the model group, mRNA expression of focal adhesion kinase was increased, and total protein and phosphorylated protein were higher in the model group than those in the control group;however,proliferation of tunica 5ntima vasorum was lightened in the treatment group, activity of mRNA expression of focal adhesion kinase was decreased, and total protein and phosphorylated protein were lower in the treatment group than those in the model group.CONCLUSION: Telmisartan can remarkably relieve proliferation of tunica intima after denudation and inhibit expression and activation of focal adhesion kinase after vascular injury.Effect of telmisartan on vascular-injured remodeling may be related to inhibiting expression and activation of focal adhesion kinase.