Objective Study on the inhibitory effect of gallnut extract extract on MRSA β-lactamase. Methods Determination of inhibitory effect of gallnut extract on MRSA3002 by TTC method. β-lactamase was repeated by freezing and thawing method . Synergistic effect of gallnut extract and gentamicin was detected by TTC. Results The MIC and MBC of MRSA3002 by gallnut extract were 8mg/mL and 32mg/mL.Gallnut extract can reduce strains of β-lactamase activity,the MRSA300224h 1/2MIC after the effect of gallnut extract, beta lactam enzyme activity inhibition compared with the control group there were significant differences (P<0.01),compared with the positive control group, the difference was not significant. Synergistic effect of gallnut extract and gentamicin can significantly reduce the MIC of MRSA3002. Conclusion Gallnut extract can reduce β-lactamase activity recovery sensitivity of drug-resistant bacteria.

Objective To investigate the inhibition mechanism of rohdea roth on hyphae formation by Candida albicans.Methods MTT assay was used to detect the minimum inhibitory concentration(MIC)and minimum fungicidal concentration(MFC)of C.albicans.The inhibitory effect of fungicidal adherence was detected by MTT assay.Inverted fluorescence microscope was used to observe effect on hyphae formation.The influence on Efg1 and Hwp1gene expression were detected by RT-PCR method.ResultsMIC and MFC of C.albicans were 16 mg/mL and 32 mg/mL, respectively.The inhibitory effects of rohdea roth on C.albicans adherence and hyphae formation were significantly inhibited,and the concentration was dose-dependent.After the concentration of 16 mg/mL acted on C.albicans for 6 h, hyphae disappeared completely.The results of RT-PCR showed that the gene expression of Efg1 and Hwp1 could be inhibited by rohdea roth.Compared with the control group,the expression of Efg1 and Hwp1in the experimental groupwere reduced by 84.18% and 59.57%(P<0.01).Conclusion The inhibitory effect of rohdea roth on the adherence and hyphae formation of C.albicans is mainly through inhibiting the expression of Efg1 and Hwp1genes.

Baicalein (BAI) is an effective bactericide. The antibacterial activity and mechanism experiments were carried out by determining conductivity and content of macromolecules of membrane penetrability, the oxidative respiratory metabolism and protein synthesis changes and the inhibition of DNA topoisomerase activities. Electrical conductivity and the number of large molecules of BAI increased 2.48% and 1.8%, respectively, than that of the control. However, the membrane integrity did not destroyed by BAI directly. With BAI treatment, inhibition rates of activities for SDH and MDH were 56.2% and 57.4%, respectively, demonstrating that BAI could inhibit cell respiratory. After treated with BAI for 20 h, the total soluble content of proteins decreased by 42.83%. Moreover, the activities of DNA topoisomerase I and II were inhibited completely by 0.2 mmol x L(-1) BAI. These results indicated that BAI had obvious antibacterial activity on Staphylococcus aureus. The mechanism is that it could affect bacterial membrane penetrability, inhibit protein synthesis and influence SDH, MDH and DNA topoisomerase I and II activities to exert its antibacterial functions.

Objective To investigate effect of emodin on mice immune function and its hemolysis toxicity.Methods The mouse specific immune cells of T, B lymphocytes and nonspecific immune cell of macrophages and NK cells were prepared and incubated in vitro.The different immune cells were treated by emodin with different concentrations of 5,10,15 and 20μM, and DMSO as control group.The effect of emodin on immune cells function was detected by neutral red assay and MTT assay.The hemolysis test in vitro was conducted by emodin with different concentrations of 20, 40, 60 and 80μM, physiological saline as blank control group and sterile distilled water as positive control group, then the hemolysis toxicity of emodin was observed. Results There were no significant difference of T and B lymphocyte proliferation among control group, 5, 10, 15 and 20 μM group(F=0.009,P=1.000;F=0.003,P=1.000), the phagocytic ability of macrophages enhanced in each dose group and was concentration dependent(F=665.525,P=0.000), the proliferation rate of macrophages enhanced and was concentration dependent(F=134.812,P=0.000), the activity of NK cells enhanced and was concentration dependent(F=200.190,P=0.000).Hemolysis test results showed the hemolysis rate was less than 5% in the range of 20 to 80μM emodin.Conclusion Emodin could significantly promote the nonspecific immune cells activity.Within the concentration of experiment, emodin has no hemolysis toxicity.

Objective To compare the anti-tumor activity and antioxidant activity of ethanol extracts of F.veluties and A.auricula.Methods Three human carcinoma cell lines, including MCF-7, HeLa and A375 were assessed by MTT assay to measure cell viability.The antioxidant activity was detected with a DPPH and RFAP assays.Results Two domestic fungus have different anti-tumor activity on the inhibition of MCF-7, HeLa and A375 cells at 25-400μg/mL concentration rage.F.velutipes was better than the A.auricula.The difference was statistically significant ( P<0.05 ).Compared with the control group, the inhibition of F.velutipes were 48.20%, 52.61%and 50.58%at the concentration of 400μg/mL, respectively (P<0.01).At the same concentration, the inhibition of A.auricula were 37.62%、50.21%and 41.59%, respectively (P<0.01).The ethanol extracts of two domestic fungus have significant antioxidant activity.F.velutipes was better than the A.auricula.The difference was statistically significant ( P<0.01 ).Compared with the control group, the scavenging radical capacities on DPPH of ethanol extracts of F.velutipes and A.auricula were 60.30%and 40.43%at the concentration of 1.6 mg/mL, respectively (P<0.01).At the same concentration FRAP value were 9.5 and 7.0 mmol/mL(P<0.01).Conclusion The F.velutipes and A.auricula both have strong anti-tumor and antioxidant activities, F.velutipes is better than A.auricula.

Objective To investigate the antimicrobial and antioxidant activity of the water extract and ethanol extract of the powder of G.lucidum.Analysis and comparison of two kinds of antibacterial and antioxidative effects of the extracts.Methods G.lucidum powder was prepared by using a pulverizer and extracted with water and ethanol respectively.The inhibitory effects of water extract and alcohol extract of G.lucidum on the tested strains were determined by TTC method.The antioxidant activities of aqueous extract and ethanol extract of G.lucidum were determined by DPPH and FRAP.Results The minimum inhibitory concentration ( MIC) of the aqueous extracts and ethanol extracts of G.lucidum were 32 mg/mL and 16 mg/mL, respectively.The MIC for S.Aureus respectively was 64 mg/mL and 32 mg/mL, the MIC for M.luteus respectively was 32 mg/mL and 8 mg/mL, MIC for B.terom respectively was 64 mg/mL and 8 mg/mL, The MIC of B.subtilis respectively was 32 mg/mL and 8 mg/mL.The antioxidant activity of G.lucidum powder extract and ethanol extract showed good antioxidant activity, the scavenging radical capacities on DPPH of the water extract and ethanol extract were 56.22%and 20.67% at a concentration of 2 mg/mL ( P<0.5 ) , the FRAP value respectively were 3.52 and 3.77 mmol/mL. Conclusion The powder of G.lucidum has the effective antimicrobial activity and antioxidant activity.The two kinds of extracts extract of G.lucidum had good antibacterial activity and antioxidant activity.The ethanol extract of G.lucidum powder antimicrobial effect and total antioxidative ability in water extracts, aqueous extract of Ganoderma lucidum granules on DPPH free radical scavenging rate better than that of ethanol extract.

Objective To investigate inhibitory effects of Sanguisorba officinalis L.on biofilms of MRSA41577.Methods Congo red agar method and crystal violet semi-quantitative method were used to detect the biofilms-forming ability of tested strains; TTC assay was used to detect inhibitory effects of Sanguisorba officinalis L.on biofilms formation and mature biofilms of MRSA41577,as well as effects of Sanguisorba officinalis L.in combination with vancomycin on mature biofilms of MRSA41577.Results Sanguisorba officinalis L.showed significant inhibitory both on biofilms formation and mature biofilms, minimum inhibitory concentration(MIC) and minimal bactericidal concentration(MBC) of biofilms formation were 1 mg/mL and 8 mg/mL,MIC of mature biofilms was 4 mg/mL.The sensitivity of mature biofilms to vancomycin was greatly increased when Sanguisorba officinalis L.was combined with vancomycin with subinhibitory concentrations.Sanguisorba officinalis L.at 1/4 MIC can inhibit mature biofilms when combined with vancomycin at 4 μg/mL, while vancomycin didn't show inhibitory effects on mature biofilms when concentrations were below 64 μg/mL. Conclusion Sanguisorba officinalis L.has significant inhibitory on biofilms formation, the mechanism may be related to Sanguisorba officinalis L.destroyed biofilms and make vancomycin penetrate into the biofilms to finish the bactericidal activity.

Objective To study the inhibitory effects of andrographis paniculata and silybum marianum on the efflux system of MRSA 41577. Methods Inhibitory effects of andrographis paniculata and silybum marianum on efflux system of MRSA 41577 was evaluated using fluorescence spectrophotometry.PCR was applied to detect the norA efflux gene.By RT-PCR method for detection of andrographis paniculata and silybum marianum influence of the expression of norA efflux gene.Results Andrographis paniculata and silybum marianum significantly increased the accumulation of ciprofloxacin in MRSA 41577 in a time-dependent manner.At 12 minute, andrographis paniculata and silybum marianum respectively increased ciprofloxacin in MRSA41577 by 49% and 76%( P <0.05 ) , which is superior to that of reserpine. Further mechanism studies indicated that andrographis paniculata and silybum marianumcould reduce the expression of norA in MRSA 41577.After incubated with andrographis paniculata and silybum marianum for 16 h, the relative expression of norA of MRSA41577 was respectively reduced by 35% and 42% ( P <0.05 ). Conclusion Andrographis paniculata and silybum marianumcould inhibit MRSA efflux system through reducing pathogen ’s expression of norA and NorA protein.

Objective To investigate the effect of Omithogalum caudatum Ait(OCA)on apoptosis of Candida albicans,illustrated the antifungal mechanism of OCA.Methods Annexin Ⅴ-FITC/PI double stainingwas used to detect the effect of OCA on the apoptosis of C.albicans;JC-1 and DCFH-DA staining were used to detectthe effect of OCA on mitochondrial membrane potential(MTP)and reactive oxygen species(ROS)of C.albicans.Results OCA had a good antifungal activity,the minimum inhibitory concentration(MIC)and the minimum fungicidal concentration(MFC)were 8mg/mL and 32mg/mL respectively.OCA could induceapoptosis of C.albicans,promote the reduction of MTP and increase of ROS.Conclusion OCA induced cell apoptosismainly through disrupting mitochondrial function.

Objective To investigate the inhibition mechanism of gallnut on biofilm formation by MRSA 41577.Methods TTC assay was used to detect inhibitory effects of biofilms formation and mature biofilms.The of PIA on biofilm formation was studied using Congo red agar method.Micro-Ultraviolet Spectrophotometer was used to detect inhibitory effects of the release of eDNA.The influence for Baicalein on icaA and cidA gene expression were detected by RT-PCR method.Results The inhibitory concentration (MIC) and minimum bactericidal concentration (MIC) of MRSA 41577 BF were 0.5 mg/mL and 1 mg/mL, respectively.The inhibitory effect of galla on MRSA 41577BF formation and mature BF was significantly inhibited.Inhibition of MRSA 41577,the MIC and MBC of mature BF were 4 mg/mL and 16 mg/mL.Congo red test results show that Galla can inhibit the synthesis of MRSA 41577 PIA, and the concentration was dose-dependent.The results showed that gallnut could inhibit the release of MRSA 41577 eDNA, and the release amount of eDNA was 3.61μg/OD595 and 11.91μg/OD595 , respectively, when the concentration of gall was 1/2MIC.The release of eDNA was reduced by 69.7% (P<0.01).The expression of icaA and cidA genes in the control group was 9.7% and 6.67%, respectively.The expression of icaA and cidA in the control group was significantly lower than that in the control group ( icaA and cidA, and cidA gene expression were 100%, the expression of icaA and cidA genes were reduced by 90.3%and 93.3%, respectively (P<0.01).Conclusion The inhibitory effect of gallnut on the biofilm of MRSA 41577 is mainly through inhibiting the expression of icaA and cidA genes, and then affecting the synthesis of PIA and the secretion of eDNA .