1.Characterization of 5-Enolpyruvylshikimate-3- phosphate Synthase from Sclerotinia sclerotiorum
Hanying YU ; Qian YANG ; Lin LI
Chinese Journal of Biochemistry and Molecular Biology 2006;22(4):301-307
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.
2.The effect of Yili dark bee propolis on oral cariogenic bacteria in vitro
Qian YU ; Upur HAMLAT ; Jing LIN ; Jin ZHAO
Journal of Medical Postgraduates 2014;(12):1269-1272
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.
3.Effects of Yili dark bee propolis on oral cariogenic biofilm in vitro.
Qian YU ; Jing LIN ; Zulkarjan-Ahmat ; Jin ZHAO
West China Journal of Stomatology 2015;33(4):343-346
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.
Actinomyces viscosus ; Animals ; Bees ; Biofilms ; Dental Plaque ; Propolis ; Streptococcus mutans ; Streptococcus sanguis ; Streptococcus sobrinus
4.Effects of repetitive transcranial magnetic stimulation on the lower limbmotor functionof cerebral infarction patients
Yamei LI ; Lin HUANG ; Jing ZHANG ; Jinyan TIAN ; Qian YU
Chinese Journal of Physical Medicine and Rehabilitation 2016;38(11):839-842
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.
5.Abnormality and significance of monocyte subsets in peripheral blood of patients with rheumatoid arthritis
Lei QIAN ; Xin LIN ; Wei CHEN ; Ming LI ; Yuehong YU
Chinese Journal of Immunology 2016;32(10):1519-1523,1531
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.
6.A comparison study on the mechanical strength of two resin cements
Haijun CHEN ; Shuxiang YU ; Lin ZHANG ; Qichun QIAN ; Fuqing NAN ; Junzhou LIN
Chinese Journal of Tissue Engineering Research 2009;13(8):1585-1588
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.
7.Methylation status of promoter of mismatch repair genes hMLH1 and hMSH2 in epithelial ovarian cancer
Shi-Qian ZHANG ; Ai-Feng ZHANG ; Lin-Lin ZHANG ; Le-Le FU ; Hao YU ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
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
8.Enhanced anti-leukemic activity of decitabine to leukemia HL-60 cells by anti-miR-21 oligonucleotide
Yekai WANG ; Qian YU ; Qilong LIN ; Yanzhen YAO ; Peiyu MEI ; Yiwei LI
Chinese Journal of Pathophysiology 2015;(1):109-113
AIM:To investigate the role of anti-miR-21 oligonucleotide ( AMO) in the anti-leukemic activity of decitabine (DCA) in vitro.METHODS:AMO and scramble oligonucleotide (SCR) were constructed and transfected into HL-60 cells.The miR-21 expression was analyzed by real-time PCR to identify the transfection efficiency .The cells were treated with DCA at gradient concentrations (0.5, 2.0 and 4.0 μmol/L) for 48 h.The mRNA expression of human period circadian protein 3 (hPer3) was detected by real-time PCR.The early apoptotic rates were determined by flow cy-tometry with Annexin V/PI staining.Mean fluorescence intensities ( MFI) of CD117 and CD11b were also measured by flow cytometry.RESULTS:The miR-21 relative expression level in AMO group was significantly lower than that in blank group and SCR group (P<0.01).IC50 of DCA in AMO group was significantly lower than that in blank group and SCR group (P<0.01).With the same concentration of DCA, the early apoptotic rate, the mRNA expression of hPer3 and the MFI of CD11b in AMO group were significantly higher than those in blank group and SCR group (P<0.01).The MFI of CD117 in AMO group were significantly lower than those in blank group and SCR group ( P<0.01 ) .CONCLUSION:Activation of hPer3 expression plays an important role in enhanced anti-leukemic activity of decitabine by AMO in vitro.
9.Ultrasonic appearances of intrascrotal adenomatoid tumor
Wenjin LIN ; Ensheng XUE ; Liyun YU ; Shun CHEN ; Rongxi LIANG ; Qingfu QIAN
Chinese Journal of Ultrasonography 2015;24(11):992-995
Objective To explore the sonographic features of intrascrotal adenomatoid tumor.Methods The sonographic features of 21 cases with intrascrotal adenomatoid tumor confirmed by surgery and pathology were retrospectively analyzed.Results Twenty-one cases with intrascrotal adenomatoid tumor could be typed into 3 ultrasonic patterns:(1) The homogeneous echo type:15 cases,their ultrasonic features were homogeneous echo,including 5 cases with low echo,8 case with equal echo,and 2 case with high echo;5 cases with testicular albuginea adenomatous tumors were also homogeneous echo,2 cases of equal echo,2 cases of low echo and 1 case of high echo that was posterior echo attenuation slightly.(2) The edge halo type:4 cases,they showed the epididymis or testis sheath round or oval nodules,of which the middle was low echo and the edge is high echo halo ring.(3) The mixed echo type:2 cases,they showed cystic mixed echo of epididymis nodules,1 case with irregular cystic areas and poor sound transmission,while the other one with good sound transmission cystic areas and a small papillae on the wall.Conclusions Sonography is a valuable method for diagnosis of intrascrotal adenomatoid tumor.The edge halo type has sonographic features.
10.Transcranial magnetic stimulation in the treatment of lateral fissure aphasia
Jing FU ; Qian YU ; Jun XIAO ; Gang YI ; Lin HUANG ; Li XU
Chinese Journal of Physical Medicine and Rehabilitation 2016;38(1):34-38
Objective To use functional magnetic resonance imaging (BOLD-fMRI) to observe the efficacy of low-frequency repetitive transcranial magnetic stimulation (rTMS) combined with speech training in treating those with lateral fissure aphasia.Methods Forty-eight persons with lateral fissure aphasia were divided randomly into a control group and a treatment group,each of 24.The treatment group was treated with repetitive transcranial magnetic stimulation combined with speech training,while the control group was given only speech training.Before the treatment and 2 weeks and 8 weeks after the treatment,BOLD-fMRI images were collected to observe any changes in the language areas of the brain activated.A Chinese aphasia examination (CRRCAE) was used to assess speech function before and after treatment for both groups.Results The average CRRCAE scores of the two groups were not significantly different right before the treatment and at the end of 2 weeks of treatment,but at the end of 8 weeks of treatment the average score of the treatment group was significantly better.The biggest improvement was among those with Broca's aphasia,followed by those with Wernicke aphasia and conduction aphasia.No such differences were observed in the control group.After the 8 weeks of treatment,the average CRRCAE score of the treatment group patients had improved significantly,but in the control group only those with Broca's aphasia had improved significantly.In the treatment group,after 2 weeks and 8 weeks of treatment the activated language areas of the right hemisphere were significantly smaller than before treatment and significantly smaller than the activated areas in the left hemisphere at the same time points.Conclusions Low-frequency,repetitive transcranial magnetic stimulation combined with speech therapy is an early intervention which may promote recovery from lateral fissure aphasia through restoring lingual function after stroke.