Using bioactive compounds 7-hydroxy flavone, salicylaldehyde, cinnamic acid and 4-amino-5- mercapto-1,2,4-triazoles as starting materials, three new types of flavone derivatives containing 1,2,4-triazole structure were synthesized via different step reactions. These new compounds were characterized by 1IHNMR, ESI-MS, IR and elemental analysis. Their scavenging effects on the superoxide radical (O2·-), hydroxyl radical (·OH), DPPH · radical and their total reduction activities were tested. The results showed that all of the compounds possessed some antioxidative activity at the concentration of 0.5 mg · mL(-1), but the scavenging ability of the target compounds was lower than that of the standard compound Vc.

Objective To evaluate the accuracy of stoke volume variation (SVV) determined using FloTrac/Vigileo and Picco-plus technologies in prone position for assessment of the blood volume in the patients undergoing spine surgery,Methods Forty-three ASA physical status Ⅰ-Ⅲ patients of both sexes,aged ＞ 18 yr,weighing 40-100 kg,scheduled for elective posterior approach to lumbar spinal fusion or scoliosis surgery were studied.After induction of anesthesia,a volume expansion was performed in supine and prone positions.Hydroxyethyl starch 130/0.4 sodium chloride injection 5 ml/kg was rapidly infused intravenously over 10 min to carry out the test for fluid responsiveness.Picco-plus and FloTrac/Vigileo systems were simultaneously applied in every subject to measure SVV (SVVP and SVVF).Positive fluid responsiveness was defined as the changing rate of stroke volume index ≥ 10％ as measured by using Piccoplus system.The patients were divided into response group (Rs group) and non-response group (NRs group) according to the changing rate of stroke volume index ≥ 10％ and ＜ 10％.The receiver operating characteristic (ROC) curve for SVV was plotted,and the diagnostic threshold,area under the ROC curve and 95％ confidence interval (CI) were calculated.Results Forty-one patients were included for analysis in this study.In supine position,the area under the ROC curve for SVV in predicting the fluid responsiveness was 0.740 (95％ CI:0.568-0.913),the diagnostic threshold was 12％,and the sensitivity and specificity in determining fluid responsiveness were 86％ and 54％,respectively,for SVVF,and the area under the ROC curve was 0.637 for SVVP.In prone position,the area under the ROC curve was 0.451 for SVVF,and 0.634 for SVVP.Compared with Rs group,the baseline value of SVVFwas significantly lower,and no significant change was found in the other hemodynamic parameters before volume expansion in supine position in NRs group.There was no significant difference in the hemodynamic parameters before volume expansion in prone position between the two groups.Conclusion SVV determined by using FloTrac/Vigileo and Picco-plus systems in prone position can not accurately assess the blood volume in the patients undergoing spine surgery.

Objective To study the protective effect and molecular mechanism of trypsin inhibitor on reperfusion injury after cerebral ischemia. Methods The model of ischemia for 1h and repufusion for 24h of rat cerebrum was set by ligating MCAO described by zea-longa. 24 male rats were divided randomly into the sham operation group, the control group and trypsin inhibitor group. The presence of neurological function deficit was measured by Zea-Longa method, and the immunohistochemical staining and TUNEL were used to detect p53 protein expression and cell apoptosis in the brain tissues respectively. Results The score of neurological function deficit was zero in the sham operation group, 2 8?1 0 in the control group and 1 3?0 7 in trypsin inhibitor group. There were 12 3?2 5 p53 immunostaining positive cells in the control group and 5 5?1 3 in trypsin inhibitor group. The number of apoptotic cells was zero in the sham operation group, 7 6?1 0 in the control group, and 3 5?0 9 in trypsin inhibitor group. There was a significant difference in all above observing indices between the control group and trypsin inhibitor group(P

Objective To compare the advantages and disadvantages of three methods of yeast-like fungi identification of vulvovaginal candidiasis (VVC).Methods The pathogenic candida strains were identified by CHROM agar chromogeinc culture medium,the chromogenic medium of Autobio and VITEK 2 system,and the result of VITEK 2 system was the gold standard.Results All candida strains were appraised.By VITEK2 system,217 candida isolates were identified as candida albicans,and 26 were detected as non-candida albicans.214 isolates were identified as candida albicans and 13 isolates were identified as non-candida albicans by Autobio.214 isolates were appraised as candida albicans,16 isolates were identified as non-candida albicans.The coincidence rate of identification of CHROMagar and the chromogenic medium of Autobio were 94.7％ and 95.06％.Compared with VITEK 2 system,there was no significant difference between these two chromogeinc culture medium identifying candida albicans and non-candida albicans.Conclusion The chromogenic medium of Autobio has higher cost-effectiveness,higher color discrimination and simpler technical operation,and is worthy of promotion in identifying candida species in clinic.

Objective To investigate the change in antimicrobial susceptibility of Enterococcus faecalis (E.faecalis) and Enterococcus faecium (E.faecium) isolated from clinical urine specimens, so as to provide laboratory evidence for clinical anti-infective treatment.Methods Antimicrobial susceptibility of E.faecalis and E.faecium isolated from urine specimens from 20 tertiary hosptials in China between 2004 and 2014 were analyzed, drug-resistant genes of vancomycin-resistant Enterococcus(VRE)were detected with polymerase chain reaction (PCR).Results A total of 788 Enterococcus strains were isolated in 2004-2014, 371 strains were E.faecalis strains, 417 were E.faecium strains.Susceptibility rates of E.faecalis to ampicillin, nitrofurantoin, fosfomycin, vancomycin, and teicoplanin were all>90%, susceptibility rates to rifampin, minocycline, and erythromycin were all<20%, there was significant difference in the susceptibility rate of E.faecalis to fosfomycin betwen July 2011-June 2012 and July 2009-June 2010(P<0.0167).Susceptibility rates of E.faecium to vancomycin and teicoplanin were 96.9% and 97.4% respectively, susceptibility rates to nitrofurantoin, minocycline, and fosfomycin were 41.7%, 51.8%, and 78.2% respectively, susceptibility rates to ampicillin, levofloxacin, rifampicin, and erythromycin were all<10%;susceptibility rates of E.faecium to nitrofurantoin had decreased tendency in different years (any two group comparison, all P<0.0167), susceptibility rates to fosfomycin in July 2011-June 2012 and July 2013-June 2014 both decreased compared with July 2009-June 2010(both P<0.0167),there were no significant changes in antimicrobial usceptibility rates in different years.14 strains of VRE all carried vanA resistance gene.Conclusion E.faecalis strains isolated from urine are susceptible to ampicillin, nitrofurantoin, and fosfomycin, E.faecium are not susceptible to most antimicrobial agents;E.faecalis and E.faecium are both susceptible to vancomycin and teicoplanin, only a few strains are resistant to antimicrobial agents.

Objective To investigate the nature of Th17 cells in murine cytomegalovirus(MCMV)infection during the acute stage,we characterized the frequency of IL-17A-producing CD4 T cells and the level of Th17 cytokines,IL-17A,in MCMV-infected mice.Methods BALB/c mice were randomly divided into two groups.One was infected with MCMV Smith for establishing disseminative infection; the other was sham-inoculated control.On day 3,7,14 and 28 of the experiment,three mice of each group were randomly chosen to be killed separately.Real-time PCR was used to detect MCMV loads in organs of MCMV-infected mice,the pathology of spleen was observed by HE staining.The frequency of CD4+IL-17A+ T cells in total splenocytes of mice was detected by flow cytometry.The level of IL-17A in culture supernatants of splenocytes was measured by double antibody sandwich ELISA.Results MCMV loads in salivary gland reached the peak on day 14 after MCMV infection,the most severe spleen injury was also shown on day 14,the frequencies of CD4+IL-17A+ T cells in total splenocytes increased significantly( all P＜0.01 ) in MCMV-infected mice than those in controls,and reached the peak on day 14 ( 1.14％ ±0.09％ vs 0.19％ ±0.04％,t =17.551,P=0.000).The levels of MCMV-specific IL-17A in culture supernatants were increased dramatically in MCMV-infected mice than those in controls on day 14 [ (81.98± 12.37) pg/ml vs (44.96±8.44)pg/ml,t=4.281,P=0.006].In MCMV-infected mice,correlation was positive between the levels of MCMV-specific IL-17A in culture supernatants and MCMV loads in salivary gland tissues (r=0.54,P＜0.05 ),the levels of IL-17 A in culture supernatants were higher in more severe spleen injury.Conclusion Thl7 cells and IL-17A were involved in the immunity response during acute MCMV infection.They may correlate with the persistence of MCMV and the pathology of spleen in infected mice.

ObjectiveTo observe the changes of AIM2 ( absent in melanoma 2) inflammasome during early murine cytomegalovirus (MCMV) infection.MethodsBALB/c mice were randomly divided into two groups.One was infected with MCMV Smith for establishing disseminated infection,the other was sham-inoculated control.On days 1,3,5 and 7 of the experiment,three mice of each group were randomly chosen to be killed separately.The expression of AIM2,ASC and caspase-1 in splenic macrophages was detected by Western blot,the levels of IL-1β and IL-18 in sera were measured by double antibody sandwich ELISA,and the viral titers in salivary gland tissues were quantified by a standard plaque assay.Results The MCMV titers in salivary gland tissues were gradually increased in MCMV-infected mice on days 3,5 and 7,while the expressions of AIM2 in macrophages were began to increase on day 1 and significantly increased and reached the highest level on day 3 but gradually decreased afterwards.The relative intensity of AIM2 on day 3 differed significantly between the MCMV-infected mice and the controls (1.121±0.243 vs 0.240±0.046,P＜0.01,t test),as did ASC ( 1.318±0.333 vs 0.248±0.090,P＜0.01 ) and caspase-1 ( 1.085±0.243 vs 0.247±0.064,P＜0.01 ).Meanwhile,the levels of IL-1β and IL-18 in MCMV-infected mice were (112.72±5.20) pg/ml and (42.74±4.23) pg/ml,and the levels were significantly higher (P＜0.01 ) than those in controls [ (47.86±4.35) pg/ml and (22.60±2.82) pg/ml].ConclusionThese results demonstrate that AIM2 inflammasome is activated in macrophages during early MCMV infection and could be as a therapeutic target for CMV-induced diseases.

Objective To explore the correlation between the expression of IL-17A and the degree of spleen damage in acute mouse cytomegalovirus(MCMV) disseminated infection in vivo and to understand the mechanism about how IL-17A involved in the pathological damage of the spleen in MCMV infection.Methods An acute disseminated MCMV infection model was established in mice.BALB/c mice were randomly divided into two groups.Mice in group one were infected with MCMV Smith to establish disseminated infection.Mice in another group were sham-infected control.Three mice from each group were randomly chosen to be sacrificed on days 3,7,14 and 28 after the infection.Viral titers in spleen tissues were determined using a standard plaque assay.The expression of IL-17A mRNA and MCMV mRNA in the splenocytes were measured by RT-PCR.The expression of IL-17A in spleen tissues was observed by immunohistochemical staining.The pathology of the infected mice was assessed by histological examination of H&E stained spleen sections.Results Viral titers and MCMV mRNA in the spleen peaked on day 3,but quickly diminished on day 7.Virus was no longer detectable in the spleen on day 14 after the infection.The expression of IL-17A mRNA was significantly increased during the acute infection and reached the highest level on day 14,then decreased on day 28.It is significantly higher than that of the mock infection group.Immunohistochemistry assay also indicated that the expression of IL-17A in spleen tissue gradually increased to climax on day 14,then decreased on day 28.Accordingly,the pathological damages of spleen tissue in the infected mice deteriorated until day 14,then showed signs of recovery on day 28.The most severe pathological injury of spleen tissue and the highest expression of IL-17A appeared in the same period of time.Conclusion Our results showed a close correlation between IL-17A and the pathological damage in spleen.Thus,IL-17A may contribute to the spleen pathological damage during the acute disseminated MCMV infection.

Objective To investigate the influence of murine cytomegalovirus on the expansion of CD+CD25+ Foxp3+ regulatory T cell (Treg) and the activation of CD4+ CD25+Foxp3 - effector T cell (TE) in vivo. Methods Forty-two BALB/c mice were intraperitoneally inoculated with appropriate amount ofMCMV Smith strain for establishing the model of infection, another 42 mice served as mocked-infected con-trois. Day 28 post MCMV infection was determined to be the demarcation point of the acute and chronic in- fection based on the viral load of major visceral organs. On day 1,3, 7, 14, 28, 45 and 60 post infection, splenocytes were prepared by means of routine method. The proportions of CD4+CD25+ Foxp3+Treg and CD4+CD25+Foxp3- activated TE in T lymphocyte were measured by flow cytometry. Results The propor- tion of CD+CD25+ Foxp3+ T cells in T lymphocytes was persistently suppressed since day 7 post infection, and fell to the lowest level on day 28 post infection (P <0.01), then zoomed and reached the peak value on day 60 post infection (P < 0.05). CD4+CD25+ Foxp3 - TE proportion was up to the highest on day 3 post infection(P < 0. 01), then suppressed and in significantly lower level since day 45 post infection (P < 0.05). Treg/CD+TE ratio was in lower level on day 3 to 14 post infection(P <0.05) ; but on day 45 and day 60 post infection Treg/CD+ TE ratio was markedly increased (P < 0.05). Conclusion MCMV infec- tion can increase the CD+CD25+Foxp3+ Treg proportion, and inhibit CD4+T cells activation in chronic in- fection phase, which is likely to suppress the function of antiviral immunity in the infected host to cause a persistent latent infection.

OBJECTIVE To investigate the cytotoxicity of cyflumetofen for SH-SY5Y cells and the mechanism. METHODS SH-SY5Y cells treated with cyflumetofen 0.03, 0.06, 0.125, 0.25, 0.5, 1, 2, 2.6, 4, 6, 8 and 16 mmol·L-1 for 48 h. Cell survival was measured with MTT assay. The reactive oxygen species (ROS) was determined with the DCFH- DA probe, and mitochondrial membrane potential (MMP) was detected by JC-1 staining. The morphological changes in cell nuclei were observed with Hoechst33258 staining. Cell cycle and apoptosis were determined by flow cytometry. The protein levels of phosphorylated Jun Kinase (p-JNK) and p-P38 were measured by Western blotting. RESULTS Compared with solvent (DMSO) control group, cyflumetofen (≥0.06 mmol · L- 1) inhibited the proliferation of SH- SY5Y cells obviously (P<0.05), and the IC50 was 2.6 mmol·L-1. MMP declined and ROS levels increased significantly in cyflumetofen 1, 2, 4 and 6 mmol·L- 1 groups (P<0.01). Cyflumetofen 2, 4 and 6 mmol·L- 1 induced nucleic accumulation, nuclear shrinkage and disintegration in SH-SY5Y cells. Apoptosis rates of cyflu? metofen 2, 4 and 6 mmol·L- 1 groups increased from (0.7±0.1)% in DMSO control group to (6.7±0.1)%, (72.4±8.6)% and (90.7±3.2)% (P<0.01). Cyflumetofen 4 and 6 mmol·L- 1 induced G1 phase cell cycle arrest (P<0.01). In addition, Western blotting showed that cyflumetofen 4 and 6 mmol·L-1 up-regulated the expression of p-JNK (P<0.01), while the level of p-P38 in SH-SY5Y cells was increased in cyflumetofen 6 mmol · L- 1 group (P<0.01). CONCLUSION Cyflumetofen induces cell damage, apoptosis and G1 phase cell cycle arrest in SH- SY5Y cells. The mechanism may be associated with oxidative damage, and activation of P38 and JNK stress-response pathways.