1.Binding mechanism of rhaponticin and human serum albumin.
Ming GUO ; Xingtao XU ; Zhiwu WU
Acta Pharmaceutica Sinica 2011;46(9):1084-92
The interaction mechanism between rhaponticin (RT) and human serum albumin (HSA) has been studied by fluorescence spectroscopy and absorbance spectra. The mediation effect that the metal ions took part in the interaction has also been discussed in this paper. Based on different theoretical models of fluorescence quenching, the binding constant (K) and binding sites (n) of the interaction were determined and analyzed comparatively. The quenching mechanism of the binding reaction has also been discussed. The binding distance (r) and energy-transfer efficiency (E) between RT/RT-Co(II)/RT-Ni(II) and HSA were also obtained by virtue of the Förster theory of non-radiation energy transfer. The effect of RT acting on the HSA's conformation was analyzed by synchronous fluorescence spectroscopy. The result showed that the result calculated by different theoretical models is generally equivalent and RT bound HSA strongly by forming stable complex, which indicates that HSA under physiological conditions can act as a carrier for RT to be transported to exert effects. The microconformation of HSA changed significantly due to hydrophobicity change in the chemical environment of some fluorescence chromophores in the subdomain IIA and IIB of HSA. Metal ions Co(II) and Ni(II) can mediate RT-HSA interaction, making the binding of the drug to protein stronger, which indicates that Co(II) and Ni(II) can enhance rhaponticin's medical efficacy under physiological conditions.
2.Mechanism of growth hormone inhibiting of alveolar type Ⅱ epithelial cells in rats
Guangfeng MING ; Daomiao XU ; Qulian GUO ; Yuhang AI ; Hong YI
Chinese Journal of Emergency Medicine 2008;17(11):1163-1166
Objective To investigate the mechanism of growth hormone inhibiting IPS-induced apoptosis of alveolar type Ⅱ epithelial cells in rats. Method Isolated and purified AEC Ⅱ cells of SD rats were divided into 5 groups,8 duplicate wells in each group. Group I served as control group; group Ⅱ:LPS 10 ug/ml;group Ⅲ:LPS 10 ug/ml + GH 50 ng/ml;gronp IV :LPS 10 ug/ml + GH 100ng/ml; group V: LPS 10 ug/ml + GH 200 ng/ml. LPS was finally added into wells in group Ⅱ~V . After the cells were incubated for 24 hours, the apoptosis rate and necrosis rate of AEC Ⅱ cells stained with Annexin V/PI were detected by flow cytometry and Fas protein of AEC Ⅱ cells were measured by immunocytochemistry. Results (1) The apoptosis rate and necrosis rate of AECⅡ cells in group Ⅱ,Ⅲ, Ⅳ and V were significantly hitOer than those in group Ⅰ( qapoptosis rate Ⅰ, Ⅱ =12.26,qnecroeis Ⅰ,Ⅱ=18.34, qapoptosisⅠ.Ⅱ=9.63,qnecrosisⅠ,nⅡ=5.75,qapotosisⅠ,Ⅳ= 9.15,qnecrosisⅠ,Ⅳ= 5.39, qapotosisⅠ,Ⅴ = 10.87, qnecrosisⅠ,Ⅴ = 5.91, P 0.05), but lower in group Ⅲ,IV and V than those in group Ⅱ(qapoptosis Ⅱ,Ⅲ= 15.24, qpecrosisⅡ,Ⅲ=16.38, qapoptosisⅡ.Ⅳ = 15.95,qnecrosisⅡ.Ⅳ=16.95, qapoptosis rate Ⅱ,Ⅴ=14.57, qnecrosisⅡ.Ⅴ = 15.61,P<0.05). (2)The positive rate of Fas expression on AEC Ⅱ cells in group Ⅱ,Ⅲ, Ⅳ and V was obviously higher than that in group Ⅰ. ( q Ⅰ.Ⅱ=35.67, qⅠ ,Ⅲ=14.32, qⅠ,Ⅳ = 13.87, qⅠ.Ⅴ=26.16, P<0.05), but lower in gronpⅢ ,Ⅳ and Ⅴ than that in gronp Ⅱ(qⅡ,Ⅲ=12.54, qⅡ,Ⅳ = 13.02, qⅡ,Ⅴ =6.96, P<0.05). Conclusions GH can probably de-crease the apoptosis of AEC Ⅱ cells by inhibiting Fas expression.
3.Evaluation of the immune response to human metapneumovirus multi-epitope antigen in an mouse model
Xiaoyan LI ; Liru GUO ; Mei KONG ; Ming ZOU ; Xu SU
Chinese Journal of Microbiology and Immunology 2015;(9):646-652
Objective To evaluate the immune response triggered by an in-house constructed hu-man metapneumovirus multi-epitope antigen ( MEA) in a mouse model .Methods Female SPF BALB/c mice at age 4-6 weeks were used in the study and divided into 7 groups.Mice in the five groups including MEA+oligodeoxynucleotides containing CpG motifs ( CpG ODN) intraperitoneal injection ( i.p.) treatment group, MEA+Alum i.p.treatment group, MEA+Alum+CpG ODN i.p.treatment group, MEA+CpG ODN intranasal (i.n.) treatment group and MEA+Alum+CpG ODN i.n.treatment group were immunized three times on days 0, 14 and 21, and those in the other experimental group were immunized intramuscularly with MEA+Quickantibody5W on days 0 and 21.A control group without treatment was set up accordingly .All mice were sacrificed two weeks after the last immunization .Antibodies including IgG , IgG1, IgG2a and IgA in serum samples were detected by ELISA .MTS assay was performed to analyze the proliferation of lympho-cytes.The cytotoxicity of cytotoxic T lymphocytes (CTL) was measured by LDH assay.Flow cytometry was used to detect T lymphocyte subsets .The cytokines secreted by T helper cells ( Th1 and Th2) were analyzed with Bio-Rad Liquid Chips.Results High titers of IgG, IgG1 and IgG2a antibodies were produced in MEA treated mice except for those in intranasal treatment groups .Serum samples from three groups including the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups were positive for IgA antibody .The highest titer of IgA antibody was detected in mice from the MEA+Alum+CpG ODN i.p.treatment group, which was 2.15×103.Compared with the control group, significantly enhanced proliferation of lymphocytes was observed in the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups (P<0.05).Enhanced cytotoxic activities of CTL were observed in mice with ip.and i.m.treatments as compared with those in control group (P<0.05).The levels of CD4+/CD8+T cells were slightly increased in mice from the MEA+CpG ODN i.p., MEA+Alum+CpG ODN i.p. and MEA+Quickantibody5W i.m.treatment groups as compared with those in control group (P<0.05).In-creased secretion of IL-2, IFN-γand Th2-type cytokines including IL-4, IL-5 and IL-10 were detected in mice from the MEA+CpG ODN i.p.treatment group.The MEA+Alum i.p.treated mice showed a slightly increased secretion of IFN-γand significantly increased secretions of IL-4, IL-5 and IL-10.Significantly in-creased secretions of IFN-γ, IL-4, IL-5 and IL-10 were detected in mice from the MEA+Alum+CpG ODN i.p.treatment group.Significantly increased secretions of IFN-γ, IL-5, IL-10 and granulocyte-macrophage colony-stimulating factor (GM-CSF) were detected in mice from the MEA+Quickantibody5W i.m.treatment group.Conclusion MEA together with different adjuvants could stimulate high titers of specific antibodies , increase the proliferation of lymphocytes and enhance the cytotoxic activities of CTL .CpG ODN could bal-ance the Th1/Th2-mediated immune responses , and the balance could be enhanced when using CpG ODN in combination with Alum .A similar effect could be achieved by using the commercial adjuvant Quickanti -body5w.This study has paved the way for further investigation on the development of hMPV epitope vaccines and diagnostic reagents for hMPV as well as the epidemiological study of hMPV .
5."Effect of ""brisk walking"" on male elderly with hypertension in community-based health centers"
Enhong DONG ; Ming XU ; Lijun GUO ; Jia BU ; Yong BAO
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(2):230-233
Objective · To explore the effects of brisk walking on blood pressure, body mass index (BMI), and autonomic nerve activity of male elderly with hypertension in community-based health centers in Shanghai. Methods · A total of 630 elderly men received brisk walking treatment (>10000 steps/d) voluntarily, who were selected from six community-based health centers in Pudong New Area of Shanghai. Among them, 210 people stick to training for 12 weeks. According to whether suffering from hypertension, the people were assigned to hypertension intervened group (intervention group, n=110) and normal intervened group (control group 1, n=100). Blood pressure, BMI, heart rate and autonomic nerve activity before and after the training were compared. Results · After 12 weeks of training, systolic blood pressure (SBP), diastolic blood pressure (DBP) and sympathetic nerve activity of intervention group were significantly lower than those before the training (P<0.05). SBP was also significantly lower than control group 2 (P<0.05). However, heart rate, BMI, parasympathetic nerve activity and baroreceptor reflex sensitivity did not change significantly after the training (P>0.05). Conclusion · Brisk walking treatment lasting for 12 weeks can decrease blood pressure and sympathetic nerve activity in elderly men with hypertension. It is worthy to be popularized in the community.
7.Effects of angiotensinⅡtype 1 receptor antagonist in the inhibition of pancreatic cancer in vitro
Hua JIANG ; Zhao-Shen LI ; Guo-Ming XU ;
Chinese Journal of Digestion 2001;0(08):-
Objective To investigate the effects and mechanisms of selective angiotensinⅡtype 1 receptor antagonist ZD7155 on the inhibition of pancreatic cancer in vitro.Methods MTT assays were used to determine the inhibition of pancreatic cancer cell line PaTu8988s by ZD7155 in different concen- trations and at different time.PaTu8988s cell cycle and cell apoptosis were detected by flow cytometry. Transmission electron microscope was used to investigate the apoptosis of PaTu8988s before and after the incubation with ZD7155 under different concentrations.PaTu8988s cell morphology was observed be- fore and after the incubation with ZD7155.Results MTT showed that the increase of inhibition of pan- creatic cancer cell by ZD7155 was in agreement with the increase of the concentrations of ZD7155 and the time of the incubation with ZD7155.The inhibition rates of PaTu8988s cells were 9%,18%,30%, 51%,60% and 78% by ZD7155 with the concentrations of 5?10~(-11),5?10~(-10),5?10~(-9),5?10~(-8),5?10~(-7) and 5?10~(-6) mol/L,respectively.The inhibition rates of PaTu8988s cells were 15%,25%, 36%,51%,67% and 85% by ZD7155 with the same concentration(5.0?10~(-8) mol/L)at 12,24,36, 48,60 and 72 hours,respectively.ZD7155 could also inhibit PaTu8988s cell cycle significantly and was dose-dependent.Cell electron microscopy showed that there were chromatin margination and apoptotic body in the cell nucleus when the cells were incubated with ZD7155,and these changes were increase with the concentrations of ZD7155.The morphology of PaTu8988s cell didn't have any change after in- cubation with ZD7155.Conclusions ZD7155 can inhibit the growth of pancreatic cancer cells in vitro by suppressing the S-phase of cell cycle and induce cell apoptosis without visible cell toxic effects.
8.Significance of Changes of Plasma Levels of Adrenomedullin and Endothelin-1 in Infants with Congenital Heart Disease Complicated with Congestive Heart Failure
ming, LU ; ai-hua, GUO ; yi, WU ; chun-ping, XU
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To investigate the roles of adrenomedullin(ADM) and endothelin-1(ET-1) in left-to-right shunt congenital heart disease(CHD) complicated with congestive heart failure(CHF).Methods Plasma ADM and ET-1 concentrations were measured by radioimmunoassay in heart failure stage and recovery period of 18 infants with left-to-right shunt CHD.Also 20 healthy infants acted as control group were measured.Results Plasma concentrations of ADM and EF-1 were significantly higher in heart failure stage of left-to-right CHD infants than those in their recovery period,and both in their heart failure stage and in their recovery period were higher than that in control group(P
9.Inhibitory effect of angiotensinⅡtype 1 receptor antagonist on pancreatic cancer of nude mice
Hua JIANG ; Zhao-Shen LI ; Guo-Ming XU ;
Academic Journal of Second Military Medical University 1985;0(06):-
Objective:To investigate the inhibitory effect of selective angiotensinⅡtype 1 receptor antagonist ZD7155 on pancreatic cancer xenografts of nude mice.Methods:Sixty nude mice were given subcutaneous injections of PaTu8988s cells to establish the pancreatic cancer xenograft models;then the animal models were evenly randomized into 3 groups:low-dose (10 mg?kg~(-1)?d~(-1))ZD7155,high-dose(20 mg?kg~(-1)?d~(-1))ZD7155 and normal saline groups.Ten mice in each group were sacrificed 10 d after treatment and the tumor sizes and body weights were measured.The microvessel density(MVD)was assessed by immunostaining of endothelial cells for CD31 and the cell apoptoses were observed by transmission electron microscope.Another thirty mice were treated for 30 days;the survival period of mice and toxicity of ZD7155 were observed till the 49th day of treatment.Results:Ten days after treatment,the mean tumor volumes in the control,low-dose and high-dose groups were(35.8?6.7)cm~3,(21.5?6.1)cm~3 and (10.7?4.1)cm~3,respectively(P
10.Effect of lumbar nerve dorsal roots section on the rat bone structural changes of lower limb
Feng XU ; Zhong-Guo FU ; Dian-Ying ZHANG ; Ya-Ming CHU ; Bao-Guo JIANG ;
Chinese Journal of Microsurgery 2000;0(03):-
Objective To investigate the rat bone structural changes of lower limb following lumbar nerve dorsal roots section.Methods Forty-eight mature female Wistar rats were divided into posterior radi- cotomy(PR)and comtrol groups randomly.The bilateral femoral bone mineral density(BMD)and biome- chanics characteristics were analyzed 2,4 and 8 weeks after the radicotomy.The same operation except the radicotomy was done in the sham group.Results In PR group,2,4,and 8 weeks after the radicotomy,the BMD of femur was(0.221?0.008)g/cm~3,(0.213?0.015)g/cm~3 ,and(0.216?0.105)g/cm~3 ,respective- ly;while that was(0.223?0.005)g/cm~3,(0.218?0.014)g/cm~3 ,and(0.208?0.111)g/cm~3 in control group.No significant difference was observed between the two groups(P>0.05).In PR group,2,4,and 8 weeks after the operation,the mean maximum load in three-point bending test of femun midshaft was(93.64?8.76)N,(89.77?11.18)N and(93.21?8.74)N,respectively,and was lower than the values of the con- trol group at the same time point(95.94?6.29)N,(91.63?9.43)N,(95.57?8.64)N,However,there was no significant difference between the two groups(P>0.05).Accordingly,there was no significant difference in the energy absorption in femun midshaft between the two groups(P>0.05).Conclusion The selective rhizotomies of part lumbar never dorsal roots might not cause the loss of the femur BMD and the change of bio- mechanics property significantly in short period.