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 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.

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.

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 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.

AIM:To explore the treatment methods and curative effect of amblyopia in children, and to analyze the effect of compliance on the treatment process.METHODS:The children with amblyopia treatment since January 1st 2010 in General Hospital of Jinan Military Area, were selected and completed follow-up for a total of 129 eyes of 78 patients, including 53 eyes of male in 31 cases and 76 eyes of female in 47 cases, the initial age of 3 to 12 years, the treatment were performed with accurate optometry, reasonable glasses and appropriate to cover the dominant eye based comprehensive therapy and compliance training of the parents and children.The treatment time was 12-36mo, followed up for 3a.The final therapeutic effect was observed and the difference of the stage curative effect between the children with different grade of compliance (good, moderate and poor grade) was compared.RESULTS:In the treatment of 36mo, 106 eyes were cured, 21 eyes were essentially recovered and the total cure rate was 98.4%, which was improved in 2 eyes (1.6%) and was ineffective in 0 eyes (0%);the total effective rate were 100%.The total cure rate was 69% in the treatment of 6mo and there was a significant difference in the cure rate among children with different compliance (overall comparison P<0.001, each comparison P<0.017).In the treatment of 12mo, the total cure rate was 89.1% and the cure rate were significantly different between children with the moderate compliance and the children with good compliance (P<0.05).CONCLUSION:Comprehensive treatment of children with amblyopia can achieve an ideal clinical efficacy.Compliance is an important factor influencing the treatment effect and the treatment process.The parents and children`s compliance training can shorten the course of treatment and improve the curative effect.

Objective To analysis and determine the possibility of the Citellus undulatus infected with Yersinia pestis surviving the winter in an experimental study, and to provide scientific experimental basis for the study on the mechanism of Yersinia pestis preservation. Method In 2006,09 to 2007,04 and 2007,09 to 2008,04 in Xinjiang Wusu-Gurtu natural foci of plague, under natural conditions, the over the winter process of Citellus undulatus carrying the plague bacteria was simulated, and 178 Citellus undulatus were infected with Yersinia pestis (1×107 Bacteria/mouse) using artificial injection method. One hundred seventy-eight Citellus undulatus infected with Yersinia pestis were kept into a construction of the black (1-5 ℃) basement (2 meters under the ground) in the plague focus. In doing so, these Citellus undulatuses almost simultaneously stepped into hibernation. After waking up from hibernation in following year in April, the survived mice carrying the plague bacteria were observed. Results Sixty-eight mice survived among the 178 infected with Yersinia pestis after 6 months of hibernation (through October to the following year in April), and the remaining 110 were all dead without pulling through the hibernation period. The survival rate was 38.2% (68/178). The organ culture of Yersinia pestis of the 110 dead mice(Citellus undnlatus) were tested, 67 were negative(-), 43 positive(+), with a positive rate of 39.1%(43/110). Among the rats with positive plague bacteria, the congestive pulmonary edema and the pathological changes of the hemorrhagic inflammation of the heart, liver, spleen, kidney and injection site could be seen clearly; the plague-free mice were not found to have any pathological changes. The survived 68 mice over the winter were autopsied and observed after being fed up for 20 days. No any pathological changes were found among these mice, and culturing of Yersinia pestis of the heart, liver, spleen, lungs and the tissue of injection site of these mice were all negative (-). Conclusions Citellus undulatus can carry Yersinia pestis during hibernation, but some fail to carry the bacteria through the entire process of hibernation persistently. Yersinia pestis was negative in the survived mice at the end of hibernation. The results showed that Citellus undulatus can not carry Yersinia pestis over the winter.