Objective To Construct fusion gene DNA vaccine pcDNA3/STxB-VP1 and fusion gene DNA vaccine pcDNA3/MDC-VP1,then the two vaccines were inoculated to mice and stuy their immunological effects.Methods B subunit of Shiga Toxin(STxB)gene fragment were amplified by PCR from the DNA of Shigella dysenteriae and Macrophage-derived chemokine(MDC)gene fragment was amplified by RT-PCR from the total RNA of a mouse spleen cells.The DNA fragments of STxB and MDC waere respectively linked to coxsackievirus B3(CVB3)VP1 by a DNA sequence which encode a flexible polypeptide(15 amino acids)to construct eukaryotic expression plasmids pcDNA3/STxB-VP1 and pcDNA3/MDC-VP1.BALB/c mice were randomized to 6 groups which were respectively immunized pcDNA3,pcDNA3/STxB,pcDNA3/MDC,pcDNA3/VP1,pcDNA3/STxB-VP1 and pcDNA3/MDC-VP1 for 3 times at 3-week intervals,intramuscularly(i.m.)in tibialis anterior muscle.The levels of the serum neutralizing antibodies were detected 20d after each inoculation.The mice were challenged with 7 LD50 CVB3 3 weeks after the last immunization.Results The fusion gene vaccines pcDNA3/STxB-VP1 and pcDNA3/MDC-VP1 were constructed successfully.The survival rates of each group were 10%,10%,15%,40%,20% and 75%,respectively,and the levels of neutralizing antibody titers,virused titers,were all consistent with those survive rates in each group.Conclusion Comparing with pcDNA3/VP1,pcDNA3/MDC-VP1 vaccine can induce a high level of neutralizing antibody titers and result in a higher survival rate in mice,while pcDNA3/STxB-VP1 induce a low level of neutralizing antibody titers and result in a lower survival rate in mice.

Objective To investigate the efficacy of milrinone micropump in treatment of chronic heart failure(CHF),and the influence to the level of amino-terminal B-type natriuretic peptide precursor(NT-proBNP) in serum.Methods 120 cases with CHF were selected,they were randomly divided into the observation group and the control group according to a random number table.The patients of the control group were treated with standardized anti -conventional treatment of heart failure according to their heart function,the patients of the observation group were given milrinone micropump on the basis of the standardized anti-conventional treatment of heart failure.The efficacy of clinical conditions 1 week after treatment and NT-proBNP levels in serum before and after treatment were compared between two groups.Results The clinical efficacy of the observation group was 95.00％,which was higher than 76.67％ of the control group,the difference was statistically significant(x2 =8.67,P ＜ 0.05).The serum level of NT-proBNP of the observation group was (38.9 ± 9.2) ng/L,which was significantly lower than (86.3 ± 32.4) ng/L of the control group,the difference was statistically significant (t =9.86,P ＜ 0.05).Conclusion Milrinone micro pump used in the treatment of CHF can significantly improve the clinical efficacy,and lower the serum level of NT-proBNP,improve heart function,it is safe and reliable and worth to application in the clinical.

Objective: To determine 17 types of phthalic acid esters (PAEs) in drinking water using solid-phase extraction coupled with gas chromatography-mass spectrometry (GC-MS) in multiple reaction monitoring (MRM) mode. Methods: One litre of commercially available bottled water was purified using an HLB solid-phase extraction column, and was eluted with ethyl acetate, dichloromethane and average methanol. Seventeen types of PAEs were detected using a triple quadrupole gas chromatography-mass spectrometer in MRM mode. By optimizing the temperature programming and adjusting the mass spectrometry collision energy, collection efficiency was improved and matrix interference was reduced. The precision and accuracy of this method were assessed by determining the standard curves, detection limits, quantification limits, relative standard deviations (RSD) and average spiked recovery rates for the 17 types of PAEs. Results: The 17 types of PAEs showed good linear relationships between mass concentration and chromatographic peak areas in the range of 0.02 to 1.0 mg/L, with correlation coefficients all greater than 0.999 1. The detection limits ranged from 0.002 9 to 0.009 7 mg/kg, the quantification limits ranged from 0.008 7 to 0.029 1 mg/kg, the RSD ranged from 0.8% to 3.0%, and the average spiked recovery rates ranged from 88.8% to 111.8%. Conclusion Solid-phase extraction coupled with MRM of GC-MS can better determine low concentrations of PAEs in drinking water.

Objective To explore the preoperative assessment of airway morphology affecting the early-phase prognosis by observing congenital tracheal stenosis survival after surgical treatment.Methods Totally 52 cases of CTS surgically treated in the Shanghai children's Medical Center,from April 2007 to June 2013,were retrospectively analyzed to study their clinical characteristics,survival condition and related factors influencing the early-phase prognosis.Using log-rank test and Cox multiple factors analysis for statistical analysis.Results Overall postoperative survival rate of 3 months was 73.1％.There were 14 deaths among 48 patients.Single factor and multiple factors analysis showed that the cross-sectional area(CSA) of stenosis and Anton-Pacheco classification were both associated with early-phase prognosis as independent factors(P ＜0.05).Long CPB duration was a prognostic factor for the outcome of surgical treatment for CTS(P ＜ 0.05).CPB time was significantly longer in death cases than in surviving cases.Conclusion CSA and Anton-Pacheco classification were significant prognostic factors for CTS.The proper utilization of radiologic imaging allow for improved patient care.

Objective:Adjuvant arthritis(AA)rats interfered with recombinant hIL-10,methotrexate(MTX)separately,we detected the changes of T cell subsets in rat′s blood and IL-17 A in rat′s serum.Methods:AA rats interfered with recombinant hIL-10, MTX and IL-10 plus MTX.The changes of the CD4+and CD8+T cell subsets were detected by flow cytometry;the Levels of IL-17A in serum of rats were measured by ELISA method.Results:Compare to normal control group ,the CD4+T cell total quantity and CD4+/CD8+T cell Proportion was significantly decreased , there were significant differences between the treatment group and the untreated group(P<0.05 ), the IL-17A of treatment group was significantly decreased , there was a significant differences ( P<0.05 ).The combination group had significant difference compared with IL-10 group.Conclusion:Recombinant hIL-10 can down-regulate the blood of AA rats in the number of CD4 +T cells and CD4+/CD8+ratio,reduce serum levels of IL-17A.The combination group compared with the IL-10 group effect on serum IL-17A more significantly,the results displayed recombinant hIL-10 plus MTX in the treatment of rheumatoid arthritis can better play the role.

AIM: To investigate the role of post-hemorrhagic shock mesenteric lymph (PHSML) in the enhancementof vascular permeability .METHODS: Eighteen Wistar rats were randomized into sham group , shock group,and shock plus mesenteric lymph drainage (shock +drainage) group.The rats in shock group and shock +drainagegroup were routinely subjected to hemorrhagic shock and hypotension [(40 ±2) mmHg] was maintained for 90 min, andthen the fluid resuscitation was performed.Mesenteric lymph was drained in the rats in shock +drainage group from resuscitationfinished to 6 h, for the observation of PHSML drainage on the vascular permeability in multiple tissues of hemorrhagicshock rats.Afterwards, human umbilical vein endothelial cells (HUVECs) were incubated with the PHSML in vitro to observethe effects of PHSML on the morphology and permeability of HUVECs .RESULTS: The degree of blue color and concentrationsof Evens blue in the lung, myocardium, kidney, liver, spleen and small intestine were significantly increased inthe shocked rats than that in sham group, while the ratios of the dry weight to the wet weight were decreased .The mesentericlymph drainage reversed these changes .Meanwhile, 4% and 10% of PHSML at 0 ~3 h and 3 ~6 h after resuscitation,and lipopolysaccharide (10 mg/L) all caused the damage of HUVECs, decreased the viability and trans-endothelial electricalresistance of HUVECs, and increased the permeability of HUVECs to fluorescein isothiocyanate -labeled albumin. CONCLUSION: PHSML is a vital factor in the enhancement of vascular permeability .

Objective&Methods The mutagenicity of natural Borneol and synthetic Borneol was studied by using Ames test.Results Natural Borneol and Synthetic Borneol in the dose range of 0.4～250.0 ?g/plate had no effect in reducing the reversion frequencies of Salmonella Typhimurium histidine auxotrophic mutant(TA97,TA98,TA100,TA102)whether if metabolic activation system(S9mix)existed or not.Moreover,the backward mutant colony had normal background.Conclusion Natural Borneol and Synthetic Borneol do not possess evident mutagenicity under experimental condition.

ObjectiveTo explore the effect of the intestinal lymph drainage on lung tissue cell apoptosis in rats with hemorrhagic shock after resuscitation,rich ALI intestinal lymphatic pathway theory.MethodsTUNEL method was used to determine the apoptosis of lung tissue cells,the brown nuclei were apoptotic cells.SABC was used to determine Bcl-2 and Bax protein expression.ResultsThe shock group and shock + drainage group lung tissue cell apoptosis rate were significantly higher than those in the sham operation group,but the shock + drainage group,the apoptosis of lung tissue cells was significantly lower than the shock group.Sham operation group showed Bcl-2,Bax protein of expression; In shock group,lung tissue cell Bcl-2 expression was significantly lower than the sham operation group,the Bax expression was significantly higher than that in the sham operation group; In shock + drainage group,lung tissue cells shored enhanced expression of Bcl-2,Bax expression was reduced,and the shock + drainage group lung tissue cell Bcl-2 expression was significantly higher than that in the shock group,the expression of Bax was significantly lower than the shock group.ConclusionThe excessive apoptosis of lung tissue cells was one of the mechanisms of lung injury after shock.Intestinal lymph drainage could reduce lung tissue cell apoptosis,the mechanism invdved the regulation of Bcl-2/Bax protein expression.

IFIT family genes are a kind of interferon stimulated genes (ISGs), and play important roles in antiviral sector and immunity regulation. To study the regulatory effect of IFIT family genes during influenza A virus (IAV) infection, we used RNA-sequencing analysis (RNA-Seq) technique and found that when 293T cells were infected by A/WSN/33 (WSN), the concentration of IFIT family genes were increased. Further study reveals that overexpression of IFIT2 or IFIT3 could inhibit IAV replication and transcription, and cause the dose-dependent inhibition of polymerase activity of vRNP. In addition, IFIT2 and IFIT3 encoding protein could colocalize with NS1 in 293T cells infected by WSN, indicating that they might interact with each other. The results suggest that IFIT family genes can inhibit the replication and transcription of IAV, which contributes to our understanding of the regulatory effect of host factors during influenza virus infection.
HEK293 Cells ; Humans ; Influenza A virus ; physiology ; Influenza, Human ; genetics ; Intracellular Signaling Peptides and Proteins ; genetics ; Proteins ; genetics ; Virus Replication

Objective To evaluate the imaging features of pneumonia caused by A (H1N1) influenza virus. Methods Imaging data of 51 patients with pneumonia caused by A H1N1 influenza were retrospectively reviewed. All patients underwent mobile chest radiographs and 44 patients underwent CT as well. On the basis of the lesion degree in the lung, the patients were classified into mild, moderate and serious types. Results Mild type showed patchy consolidation at chest imaging in 4 patients. Moderate type demonstrated consolidation and(or) ground -glass opacities more than 2 lung fields in 33 patients, including 30 bilateral and 3 unilateral. Serious type displayed diffuse consolidation and ground-glass opacities, probably accompanying with interstitial lesions in the lungs in 14 patients, including 6 patients with ARDS, 2 with infection and 1 with cutaneous emphysema. Conclusion The imaging features of pneumonia caused by A H1N1 influenza mainly manifest as consolidation and ground-glass opacities, probably accompanying with interstitial changes. The imaging findings show various in patients with infection. Some serious patients even develope to ARDS.