The 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase activity of Sclerotinia sclerotiorum is one of the multifunctional enzyme AROM activities, which catalyzes a reversible conversion of shikimate 3-phosphate (S3P) and phosphoenolpyruvate (PEP) to EPSP and inorganic phosphate, and is inhibited by the herbicide glyphosate (N-phosphonomethyl glycine). AROM protein has been purified from Sclerotinia sclerotiorum and the EPSP synthase has been analyzed. The results indicated that the optimal pH and temperature of EPSP synthase were 7.2 and 30℃ respectively. The activation energy of the heat-deactivated reaction of the enzyme was found to be 69.62 kJ/mol. Both of the substrates, S3P and PEP, were showed to inhibit the reaction rate when their concentrations exceeded 1 mmol/L and 2 mmol/L respectively. The Km of 140.98 μmol/L for PEP and 139.58 μmol/L for S3P were obtained by Dalziel equation which was a steadystate kinetic equation of the enzymatic reaction with the double substrates. The kinetic pattern of the enzyme was consistent with a sequential mechanism. Inhibition of the EPSP synthase reaction by glyphosate was competitive with respect to PEP, with the Ki 0. 32 μmol/L, and noncompetitive with regard to S3P. Activation by [ K+ ] was observed in the forward reaction. The Km (PEP) was lowered by increasing [ K+ ], while the Km (S3P) changed irregularly and the Ki (PEP) was enhanced.

OBJECTIVETo evaluate the effects of Yili dark bee propolis on the main cariogenic biofilm and mechanisms.

METHODSSusceptibilities to the ethanolic extract of propolis against Streptococcus mutans (S. mutans), Streptococcus sobrinus (S. sobrinus), Streptococcus sanguis (S. sanguis), Actinomyces viscosus (A. viscosus), and Actinomyces naeslundii (A. naeslundii) were analyzed by crystal violet stain method to determine the minimum biofilm eradication concentration (MBEC). The biofilm was initially cultivated for 24 h. Subsequently, the propolis groups with different concentration MBEC and initial pH 7.0 were cultured for 24 h. Moreover, the pH value was measured to evaluate the acid-producing ability of the tested plaque biofilm. The effects of propolis on the insoluble extracellular polysaccharide synthesis of S. mutans biofilm were evaluated by anthrone method.

RESULTSThe MBEC of Yili propolis on S. mutans, S. sobrinus, S. sanguis, A. viscosus, and A. naeslundii were 6.25, 1.56, 3.13, 0.78, and 0.78 mg.mL-1, respectively. Propolis could decrease the ΔpH of the tested plaque biofilm, and the differences between the control and propolis groups were statistically significant (P<0.05). At MBEC, propolis could reduce the ability of S. mutans in synthesizing insoluble extracellular polysaccharides.

CONCLUSIONYili propolis demonstrate remarkable eradicative effects on the cariogenic plaque biofilm, showing inhibition of the synthesis of biofilm-produced acids and insoluble extracellular polysaccharides.

Objective Searching for natural anticariogenic agent has become a focus at home and abroad.The aim of this study was to evaluate the effect of Yili dark bee propolis on the main cariogenic bacteria . Methods ①The minimum inhibitory concentration (MIC) of Yili dark bee propolis on oral common cariogenic bacteria was detected by the liquid diffusion method.②The four different con-centrations lower than MIC were used to confect culture media with pH 7.0 and the normal culture media was used as control group .The pH and the changes of pH (ΔpH) were evaluated to detect the effect of Yili dark bee propolis on acid producing ability of tested bacteria .③The effects of propolis on extracellular polysaccharides synthesis of S.mutans and A.viscosus were evaluated by anthrone method. Results①The MIC of Yili dark bee propolis on S.mutans, S.sobrinus, S.sanguis, L.acidophilus, A.viscosus and A.naeslundii were 0.78, 0.39, 1.56, 0.39, 0.2, 0.2 mg/mL, respectively.The minimum bactericidal concentrations were 1.56, 0.78, 3.125, 0.78, 0.39 and 0.39 mg/mL.②The propolis could decrease theΔpH of tested bacteria, and the differences between propolis groups and control group were statis-tically significant (P<0.05).③The propolis could reduce the ability of S.m and A.viscosus synthtic extracellular polysaccharides at MIC, and the differences were significant (P<0.05). Conclusion Yili propolis could inhibit the growth, acid producing ability and polysac-charides synthesis of the main cariogenic bacteria to some extent, and it could be deemed as a natural anticariogenic agent.

Objective:To explore the role of peripheral blood monocyte subsets in the pathogenesis of rheumatoid arthritis (RA),we therefore decided to compare the percentage of monocyte subpopulations in peripheral blood,as well as cytokines secretion function,to that of healthy controls. Methods:22 patients with RA and 22 cases of healthy controls ( HC) were drew 3 ml fresh venous blood into a tube containing heparin. The percentage of monocyte subsets,expression of Toll-like receptor(TLR)2,HLA-DR,triggering receptor expressed on myeloid cells-1(TREM-1) on intermediate monocyte and mean fluorescence intensity(MFI) of intracellular tumor necrosis factor-α ( TNF-α) were evaluated with the methods of flow cytometry ( FCM ) . The correlation between percentage of monocyte subsets and serum cytokines was explored. Statistical significance between parametric data was determined by the students't-test. Results:Compared to HC controls, the percentages of intermediate monocytes were significant higher in RA patients [ ( 11. 7 ± 1. 6)% vs (4. 6±1. 2)%,P<0. 05],as well as the expression(MFI) of TLR2 (750. 2±110. 3 vs 526. 8±98. 6) and TREM-1 (58. 4± 12. 1 vs 40. 3±10. 2) on intermediate monocytes (P<0. 05). The expression of HLA-DR on intermediate monocytes of RA patients had no difference with HC controls (P>0. 05),while MFI of intracellular TNF-αin intermediate monocytes of RA patients were significant higher than that of HC controls (46. 3±6. 4 vs 36. 7±8. 3,P<0. 05). In addition,RA patients showed a positive correlation between the percentage of CD14highCD16+ monocytes and DAS28 scores(r=0. 538,P=0. 009),as well as the serum levels of TNF-α,IL-17 ( r=0. 471,P=0. 027;r=0. 593,P=0. 003). Conclusion:Monocyte subpopulations from RA patients are abnormally skewed toward to in-termediate monocytes which has high expression of TLR2 , TREM-1 and the function of TNF-α secretion. Therefore, intermediate monocytes may play a role in the pathophysiology of RA. By modulating polarization or blocking monocyte cell surface receptors could be a new treatment of RA.

Objective To explore the effects of repetitive transcranial magnetic stimulation (rTMS) on the motor function of lower limbs of patients with cerebral infarction.Methods Sixty stroke survivors with lower limb dysfunctionwere randomly assigned to an rTMS treatment group or a control group,each of 30.Both groups were given routine medication and rehabilitation treatment,while the treatment group was additionally provided with 4 weeks of rTMS treatment of the contra-lesional M1 at 1 Hz and 90％ motor threshold.The Fugl-Meyer motor assessment (FMA) and maximum walking speed (MWS) were used to assess both groups before and after the treatment and 2 weeks later.Adverse reactions were also recorded.Results Before the intervention,no differences were found between the two groups.After the treatment and two weeks after that,significant improvement was observed in the average FMA and 10 m MWS of both groups,with significantly more improvement in the treatment group than among the controls.No obvious adverse reactions were observed in either group.Conclusions rTMS can improve the motor function of the affected lower limbs of stroke patients safely.

BACKGROUND: There are many species of ceramic bonding systems supplied in market, the mechanical strength is aprerequisite condition for resin cement in clinical use.OBJECTIVE: To study the difference of mechanical strength between Panavia F resin cement and self-made resin cement,DESIGN, TIME AND SETTING: A controlled observational study was performed in the prosthesis laboratory of the FourthMilitary Medical University of Chinese PLA between May 2006 and March 2007.MATERIALS: Panavia F resin cement was sourced from KURARAY MEDICAL INC. (Japan), experimental resin cement wasproduced by College of Stomatological Medicine in the Fourth Military Medical University of Chinese PLA.METHODS: Five cylindrical compressive strength specimens at an 8-mm height and 4-mm diameter were prepared, as well as5 cylindrical diametral tensile strength specimens at a 3-mm height and 6-mm diameter. AGS-500 universal material testingmachine was applied to detect the compressive strength and diametral tensile strength.MAIN OUTCOME MEASURES: The diametral tensile strength test and compressive strength test of the specimens.RESULTS: The compressive strength of Panavia F resin cement was remarkably higher than that of experimental cement[(238.92±24.54), (149.08±12.13) M Pa, P ＜ 0.05]. There were no significances between two resin cements on diametral tensilestrength (P＞ 0.05).CONCLUSION: There were no significance between two resin cements on diametral tensile strength following completecuring. The compressive strength of experimental resin cement can reach the standard of ADA (＞ 70 MPa) although it is lowerthan the compressive strength of Panavia F resin cement.

Objective To explore the methylation status of hMLH1 and hMSH2 promoter region in the epithelial ovarian cancer and its role in oncogenesis.Methods Methylation status of hMLH1 and hMSH2 promoter region was assayed in 20 normal ovarian tissues,25 benign epithelial tumor,56 malignant epithelial tumor and cell lines SKOV3,3AO by methylation-specific PCR (MSP).SKOV3 and 3AO were analyzed before and after 5-aza-2′-deoxycytidine (5-Aza-CdR) treatment.In addition,an alterations of mRNA expression of hMLH1 and hMSH2 was observed by reverse transcription polymerase chain reaction (PT-PCR).Results No methylation of hMLH1 and hMSH2 promoter was found in normal ovarian tissues. CPG islands methylation of hMLH1 and hMSH2 was observed in 4% (1/25),8% (2/25) respectively in benign epithelial tumor,30.4% (17/56),51.8% (29/56) respectively in malignant epithelial tumor. Methylation status in promoter showed obvious correlation with pathological grade and lymph node metastasis (P

Objective To observe the long dwell ultrafiltration volume after using 7.5% icodextrin in different peritoneal transport characteristics of peritoneal dialysis patients. Methods Subgroup analysis of a perspective multicenter randomized double blind and parallel control study was performed. Continuous ambulatory peritoneal dialysis (CAPD) patients were divided into high transport (H) group, high-average transport (HA) group, low-average transport (LA) group and low transport (L) group according to D/Pcr and Twardoski standard. Ultrafiltration volume of night long dwell dialysate was calculated before and after clinic trial for 2 weeks and 4 weeks to evaluate the different effect of transporters on ultrafiltration volume. Results A total of 201 CAPD patients were enrolled in the study, including 98 patients in icodextrin group (ICO group) and 103 patients in glucose group (GLU group). Male and female cases were 96 and 105 respectively. Age was (56.1±13.7) years old (range from 18 to 81). One hundred and ninety-eight patients finished peritoneal equilibrium test (PET), including 24 (12.1%) of H, 72(36.2%)of HA, 81(40.7%)of LA,and 21 (11.0%)of L. After follow-up for four weeks, the ultrafiltration volume was much higher than baseline in H, HA and LA groups. Also ultrafiltration volume in icodextrin group was much higher than that in glucose-based dialysate. Howerve, the increased volume was not significantly difference in L group. Ultrafiltration volume of icodextrin was positively correlated to D/Pcr (R2=0.1681,P＜0.01), while ultratration volume of dextrose was negatively correlated to D/Pcr (R2=0.0949,P＜0.01). Conclusion Compare to glucose-based dialysate (Dineal), 7.5% icodextrin dialysate (Extraneal) improves the ultrafiltration and peritoneal creatinine clearance of long dwell notabily in H, HA and LA group.

Objective To develop an accurate,rapid,high throughout genotyping method based on oligonucleotide microarray for cytochrome P450 gene polymorphisms related to paclitaxel metabolism.Methods The mutant points of 2C8 * 3,3A4 * 18 and 3A5 * 3C from cytochrome P450 gene were regarded as targets.Based on the sequences in the GenBank,the wild-type and mutant-type probes were specially designed for each mutant point.PCR primers were located in the both sides of mutant point,and furthermore the fragments of PCR products were less than 200 bp.Each type of standard plasmids was constructed.Thus,all the olignucleotide probes were modified with 3'amino-group,and the reverse primers were labeled with fluorescein (Cy3).The probes were immobilized onto certain glass slides.The specific fragments of three genes were amplified and then hybridized with oligonucleotide microarray.The results were analyzed by using certain software.Finally this assay was applied to detect 50 clinical blood specimens.Results When PCR products from standard plasmids were hybridized with DNA microarray,the corresponding probes produced positive signals.Meanwhile,the non-specific hybridization signals did not appear.The results of clinical specimens showed that the mutant rate of CYP2C8 * 3 was 2%.The point of CYP3A4 * 18 for all the clinical specimens was wild-type and the mutant rate of CYP3A5 * 3C was 62%. Meanwhile,the results from detecting 50 clinical blood specimens using oligonucleotide microarray were the same as sequencing analysis.Conclusions Oligonucleotide microarray is a reliable and accurate genotyping assay for cytochrome P450 2C8 * 3.3A4 * 18 and 3A5 * 3C polymorphisms related to paclitaxel simultaneously.This genotyping assay is a high-throughout method for guiding personalized therapy and analyzing metabolism of paclitaxel in vivo.

ObjectiveTo evaluate the clinical application of endovascular treatment for severe Takayasu's arteritis (TA).MethodsIn this study,35 target lesions in 32 patients [28 women,mean age (30 ±8) years] with severe Takayasu's arteritis were treated with endovascular merthod.The average length of lesion was 3.1 cm( range 2.7 -5.3).The overall average degree of diameter stenosis was 90％ ± 11％ (range 70- 100)in which 15 lesions were completely occlusive.There were 10 patients whose ESR were higher than 20 mm/h( range 25 -37).Follow-up included physical examination and patency evaluated by color duplex souography/computed tomography angiography/angiography at 6 months and then annually.ResultsRecanalization was unsuccessful in 3 completely occlusive lesions,with a successful rate of 80％(12/15).There was one case in which embolization leading to acute thrombogenesis developed during interventional procedure and resulting in severe stroke.The technical successful rate ( residual stenosis ＜ 50％ ) was 88.6％ ( 31/35 ).The transient cerebral ischemia attack ( TIA ) symptoms disappeared in 31 cases.26 cases were followed up for an average of (19 ± 10) months (range 13 -40).Occipital infarction following severe in-stent restenosis developed 13 months later in one case.Symptomatic in-stent restenosis18monthslaterwasfoundin2cases. Patencyratewas88.5％( 23/26 ).ConclusionsEndovascular treatment is safe and effective for severe TA.Strict indication and accurate targeting the lesions help ensure the success of management.