In order to investigate the effect of malic enzyme on anaerobic mixed acid fermentation,the gene encoding NAD+-dependent malic enzyme was amplified from E.coli DH5? genome by using PCR and cloned into vector pTrc99a to give an expression vector pTrc99a-sfcA.Malic enzyme was overproduced by E.coli FMJ39(ldh,pfl)harboring pTrc99a-sfcA under the condition of IPTG induction.The results of anaerobic fermentation indicated overexpression of malic enzyme would have influence on the pathway of formate,acetate and succinate of FMJ39.The concentration of formate and acetate was 17.58% and 15.27% higher than FMJ39,the succinate concentration was reduced by 26.87%.There was no obvious citrate concentration change.The experimental results also indicated that high concentration of L-Thr and L-Ser would induce the Tdc operon to convert pyruvate into formate and acetate even when pfl gene encoding the pyruvate formate lyase was inactivated.

50 years).The values of ADC,FA,?_1,?_2 and ?_3 in putamen and globus pallidus were calculated after diffusion tensor scanning,and then analyzed the differences between the two tissues.The differences of ADC,FA,?_1,?_2 and ?_3 values among age groups were analyzed by using analysis of variance(ANOVA),and their relationship with age were accessed by using Pearson correlation.Results Between the correspondent age groups,the ADC value was higher in the putamen[(6.68?0.40)?10~(-4),(6.47?0.36)?10~(-4),(6.44?0.34)?10~(-4)mm~2/s]than that in the golbus pallidus[(6.13?0.50)?10~(-4),(6.05?0.33?10~(-4),(6.05?0.52)?10~(-4)mm~2/s],and the value of FA in the putamen(0.20?0.03,0.23?0.03,0.25?0.03)was lower than that in the globus pallidus(0.35?0.03,0.36?0.03,0.37?0.04).ANOVA analysis of the FA value among age groups revealed there were significant differences in the putamen(F=10.082,P=0.000),and the same analysis of eigenvalue ?_3 in the putamen also showed significant differences(F=5.675,P=0.005).The FA value in the putamen was significantly elevated with aging(r=0.555,P

AIM:To observe the effect of spironolactone on myocardial fibrosis of spontaneously hypertensive rats(SHR).METHODS:Sixteen fourteen-week-old male SHRs were randomly assigned to spironolactone and SHR group equivalently(n=8).Rats in each group were given 30 mg ? kg-1 ? d-1 spironolactone and equal sodium chloride respectively for 12 weeks by gavage.Eight fourteen-week-old male SD rats were as control group.Connective tissue growth factor(CTGF),transforming growth factors beta-1(TGF?1),collagenⅠand Ⅲ were measured by qualitative and semiquantitative immunohistochemical staining and semiquantitative reverse transcription polymerase chain reaction.Masson staining was used to determine total collagen in left ventriculum.Alkaline hydrolysis method was used to detect the concentration of hydroxyproline(Hypro)in the myocardium of left ventricle.RESULTS:Left ventricular index(LVI),collagen volume fraction(CVF),Hypro and the expression of TGF?1,CTGF,collagenⅠand Ⅲ in SHR group were significantly higher than those in SD group(P

Objective To understand the basic situation of healthcare-associated infection(HAI)management and prevention and control of multidrug-resistant organism(MDRO)infection in medical institutions in Hubei Province.Methods Questionnaires were used to investigate the basic situation of HAI management and MDRO infection in 47 tertiary and secondary hospitals in Hubei Province.Results 47 hospital were enrolled in this study,HAI management full-time staff was allocated with a median of 0.90/250 beds;in 2015,the isolation rates of methicillin-resistant Staphylococcus aureus,carbapenem-resistant Enterobacteriaceae,carbapenem-resistant Pseudomonas aeruginosa,carbapenem-resistant Acinetobacter baumannii,and vancomycin-resistant Enterococcus were 39.13%,4.13%,19.44%,63.60%,and 2.77%respectively.47 hospitals all carried out surveillance on MDROs,28(59.57%)hospitals were installed HAI monitoring system software,could directly obtain data through collecting with the laboratory system.The diagnosis of MDRO infection was mainly based on the combined diagnosis by HAI full-time personnel and clinicians(26 hospitals,55.32%).44(93.62%)hospitals regularly convened coordination meeting on prevention and control of MDROs,in 2015,hand hygiene compliance rate were 10.0%-89.2%by self-inspection;42(89.36%)hospitals routinely prescribed isolation orders for patients with MDRO infection,33(70.21%)hospitals conducted environmental clean and disinfection twice a day,24(51.06%)hospitals performed personal protection by using complete set of protective equipment.Conclusion This investigation is helpful for understanding the general situation of HAI management in medical institutions,as well as the detection and control of MDROs in this area,and make continual improvement on the problems and weakness found in the investigated project,so as to promote the development of HAI management in this area.

Objective To investigate the effects of methylprednisolone(MP) on the expression of matrix metalloproteinase(MMP)-9 and airway inflammation in murine models of asthma. Methods Thirty female BALB/c mice were randomly divided into asthma group,MP group and control group(n=10).Murine models of acute asthma were established by ovalbumin(OVA) via peritoneal injection and intranasal instillation.The pathological changes of lung tissues were observed with HE staining,and cell quantitation was conducted in bronchalveolar lavage fluid(BALF).The expression of MMP-9 protein was determined by immunohistochemistry and gelatin zymogram,and the expression of MMP-9 mRNA was detected by RT-PCR. Results Compared with control group,there were more significant airway spasm and more infiltration of inflammatory cells in histologic examination,and there was higher eosinophil cell quantitation in BALF in asthma group(P

The GFP(green fluorescence protein)-streptavidin(SA) bi-functional fusion protein was generated and characterized in order to demonstrate novel platform for efficiently and durably modifying the cell surface with SA-tagged bi-functional proteins.The GFP-SA/pET24d construct was generated and expressed in BL21(DE3) host bacteria at the high level.The recombinant protein GFP-SA was purified through the Ni-NTA affinity chromatography,and then refolded.After biotinylation B16 tumor cells were modified with GFP-SA bi-functional fusion protein and then subjected to fluorescent microscopy and FACS analysis.The effect of surface modification on the viability and growth of B16.F10 tumor cells was evaluated by MTT staining.The GFP-SA recombinant fusion protein was expressed in BL21(DE3) at about 20 % of total bacterial proteins.The GFP-SA bi-functional fusion protein exhibited the bi-functionality,i.e.,SA-mediated high-affinity binding to biotinylated cell surfaces and GFP-emitted green fluorescence.The cell surface modification with GFP-SA bi-functional fusion protein did not affect the viability and growth of the modified B16.F10 tumor cells significantly.The GFP-SA bi-functional fusion protein was obtained and could be displayed efficiently on the surface of the biotinylated B16.F10 tumor cells through the specific and tight interaction between streptavidin and biotin,thus can be used as good trace protein and experimental control in the development of surface-modified tumor vaccine.

The chemical consitituents from cytotoxic fraction of the Callicarpa nudiflora extract were isolated and purified by a combination of HP-20 macroporous resin, silica gel and Sephadex LH-20 column chromatographies. The structures were elucidated on the basis of the spectroscopic data and comparison of their spectroscopic data with reported data. The cytotoxicity was evaluated by the MTT assay. The 50% and 70% EtOH elutions of EtOH-extract showed significant cytotoxic activities, leading to the isolation of twelve compounds, which were identified as luteoloside(1), lutedin-4'-O-β-D-glucoside(2), 6-hydroxyluteolin-7-O-β-glucoside(3), lutedin-7-O-neohesperidoside(4), rhoifolin (5), luteolin-7, 4'-di-O-glucoside (6), forsythoside B (7), acteoside (8), alyssonoside (9), catalpol(10), nudifloside(11), and leonuride(12). Compounds 3-6, 10 and 12 were isolated from this genus for the first time, and compound 9 was isolated from this plant for the first time. The cytotoxicity assay demonstrated that flavonoids 1-6, in various concentrations, showed monolithic proliferation inhibitory activities against Hela, A549 and MCF-7 cell lines. Compounds 3, 5 and iridoid glycoside 11 possessed higher cytotoxicacivities. In short, flavonoids are the main components of cytotoxic extract from C. nudiflora, while phenylethanoid glycosides are the predominant ingredient but inactive to cancer cell lines. In addition, the minor iridoid glycoside expressed weak cytotoxic activity.
Callicarpa ; chemistry ; Cell Proliferation ; drug effects ; Cytotoxins ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; MCF-7 Cells ; Molecular Structure

The design principles of a noninvasive ventilator's turbine are studied and discussed in this paper. The design is completed from its several aspects and in combination of related theories, using SolidWorks tools. Abundant experimental results prove that this design's technical specifications meet all the requirements.
Equipment Design ; Respiration, Artificial ; instrumentation ; methods ; Ventilators, Mechanical

Adult ; Delivery of Health Care ; Female ; Health Education ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Occupational Health

OBJECTIVETo exacted analysis each time interval in isovolumic relaxation time (IVRT) of normal subjects through observin the changes of cardiac structure and hemodynamics during the IVRT. Then to provide the evidence of cardiac resynchronization therapy.

METHODSQuantitative analysis was performed for 60 subjects. The dual-channel echocardiography(DCE), pulse wave doppler (PW) and tissue wave dapper (TDI) examination of all the subjects were recorded, and IVRT was divided into two intervals, isovolumic relaxation time of early intervals (IVRTe) and isovolumic relaxation time of late interval (IVRT1). Then measured the time of each interval. Indicators were used including: (1) IVRT; (2) IVRTe; (3) IVRTI; (4) IVRTI/IVRT; calculating the data after heart rate corrected; (5) cIVRT; (6) clVRTe; (7) clVRTI; (8) clVRTI/clVRT; (9) measuring the time difference in mitral blood and tissue (TE-é) of DCE group.

RESULTSThe i-wave within IVRT in PW images was found in 45 subjects, and the i-wave was about 1/2 of IVRT (49.17 +/- 5.37) ms. IVRT was divided into IVRTe and IVRTI by a turning point at descending branch of i-wave as t-point. The j-wave was observed in 84% TDI images, and the j-wave was about 1/2 of IVRT (43.13 +/- 4.83) ms. IVRT was divided into IVRTe and IVRTI by a turning point of the onset of j-wave as t-point. A significant difference was found between PW and TDI with measurement of IVRT, IVRTe, IVRTI (P < 0.05). There were no significant differences between the common group and DCE group (P > 0.05). After heart rate corrected, the data showed no significant difference using pairwise comparisons among the three groups (P > 0.05). The mean and standard deviation of IVRTI/IVRT, cIVRTI/clVRT were (0.50 +/- 0.12) ms. There were little difference of time intervals and good consistenc using DCE measured IVRT with multiple tests confinmed.

CONCLUSIONThe study found that IVRT might be divided into IVRTe and IVRT1 phases. There were i-wave in IVRTe and j-wave in IVRT1. The t-point was nearly midpoint inisovolumic relaxation time.

Adult ; Diastole ; physiology ; Echocardiography ; Female ; Healthy Volunteers ; Humans ; Male ; Middle Aged ; Ventricular Function, Left ; physiology