1.Effect of Elevated O2·-and H2O2 Expression on Abnormal Activation of Platelets in Patients with Type 2 Diabetes Mellitus
Fei CHEN ; Chuanghua LUO ; Hanping LU ; Ti ZHOU ; Zhonghan YANG ; Xia YANG ; Guoquan GAO ; Weiwei QI
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(1):56-62
[Objective]To investigate the molecular mechanism of abnormal platelet activation induced by platelet O2 ·- and H2O2 levels in type 2 diabetes mellitus.[Methods]The platelet parameters in patients with type 2 diabetic patients and normal controls were measured;Immunofluorescence technique was used to observe the platelet morphology changing;Flow cytometry was used to detect platelet intracellular O2 ·- and H2O2 content in two groups,then with platelets in normal controls treated with NADH/PMS system and H2O2 respectively,platelet activation positive percentage was observed. Standard Western blot analysis protocols were used to detect expression difference of Catalase and type 2 super-oxide dismutase(Mn-SOD)in platelets.[Results]The MPV in the group of type 2 diabetic patients was significantly higher than in the normal control group(P < 0.001),but there was no statisticdifference in PLT,PDW,PCT between two groups. Immunofluorescence results showed that morphology of platelets in type 2 diabetic patients changed contrast to normal group. Through flowcytometry detection,the content of mitochondrial O2·-and H2O2 of platelet in type 2 diabetic patients were obviously higher than in normal group(P<0.05),whereas no significant difference in cytoplasmic O2·-. We adopted NADH/PMS system and H2O2 to treat platelets of normal group,heightened activated positive percentage were observed which described O2 ·- and H2O2 can significantly promote platelet activation(P<0.01). Western blot results showed that expression of Catalase in platelet of type 2diabetes patients decreased,while the expression and activity of Mn-SOD had no difference.[Conclusion]It is diabetic platelet Catalase expression decreased that may lead to Diabetic platelet mitochondrial O 2 ·- and H2O2 level increased ,thus regulating aberrant activation of diabetic platelet.
2.Research Progress and Therapeutic Prospect of Angiogenesis Inhibitors in the Microvascular Complication of Diabetic Mellitus
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(2):169-176
Microvascular complications of diabetic mellitus injure entire organs of the body. Diabetic retinopathy,foot ulcer, erectile dysfunction and diabetic nephropathy are the major pathogenesis of blindness,non-traumatic amputation,kidney failure and even death,respectively. Thus,early intervention of microvascular complications is the optimized strategy for improving the quality and prolonging the lifespan. Angiogenesis is the one of most significant pathologic features in microvascular complications of diabetic mellitus.Angiogenesis inhibitors are closely involved in the balance regulation of angiogenesis. Therefore ,this review aims to present the research progress and the therapeutic prospect of angiogenesis inhibitors in the microvascular complication of diabetic mellitus.
3.Quantitative comparison of 68Ga-NGR and 18F-FDG uptake in well-differentiated hepatocellular carcinoma bearing mice
Yongheng GAO ; Zhengjie WANG ; Fei KANG ; Xiaowei MA ; Wenhui MA ; Mingru ZHANG ; Mingxuan ZHAO ; Tianming FU ; Guoquan LI ; Shengjun WANG ; Zhe WANG ; Weidong YANG ; Jing WANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(3):147-152
Objective To quantitatively compare the diagnostic capability of 68Ga-NGR and 18F-FDG in well-differentiated hepatocellular carcinoma (HCC) bearing mice by microPET/CT imaging.Methods The in vitro cellular uptake, in vivo microPET/CT imaging and biodistribution studies of 68Ga-NGR and 18F-FDG were quantitatively compared in SMMC-7721-based well-differentiated HCC.The human fibrosarcoma (HT-1080) and human colorectal adenocarcinoma (HT-29) cells/xenografts were respectively used as positive and negative reference groups for CD13.The expression of CD13 was qualitatively verified by immunohistostaining.The levels of CD13 and glucose-6-phosphatase (G6Pase) were semi-quantitatively analyzed by Western blot test for all 3 types of tumors.Two-sample t test was used for data analysis.Results The in vitro cellular uptake showed that the 68Ga-NGR uptake in SMMC-7721 and HT-1080 cells was higher than that in HT-29 cells, and the 68Ga-NGR uptake was higher than 18F-FDG uptake in SMMC-7721 cells.The in vivo microPET/CT imaging results revealed that the uptake of 68Ga-NGR in SMMC-7721 tumor was (2.17±0.21) %ID/g, remarkably higher compared to (0.73±0.26) %ID/g of 18F-FDG uptake (t=8.826, P<0.01).The tumor/liver ratio of 68Ga-NGR was 2.05±0.16, which was 2.03-fold higher than that of 18F-FDG.In the HT-1080 tumors, the uptakes of 68Ga-NGR and 18F-FDG were both high, and the values were (2.46±0.23) %ID/g, (3.47±0.31) %ID/g.The uptake of 68Ga-NGR was significantly lower than that of 18F-FDG in HT-29 tumors: (0.67±0.20) %ID/g vs (3.17±0.29) %ID/g;t=4.221, P<0.01.Western blot and immunohistostaining results were as follows: HT-1080(CD13+, G6Pase-), SMMC-7721(CD13+, G6Pase+), HT-29(CD13-, G6Pase-).Conclusions The uptake of 68Ga-NGR is higher than 18F-FDG uptake in SMMC-7721 tumor bearing mice, therefore it is worthwhile to consider the feasibility of clinical translation for PET/CT in diagnosis of HCC.Furthermore, because of the difference in 68Ga-NGR and 18F-FDG avidities in tumors with different molecular phenotypes of CD13 and G6Pase, there is an underlying potential for molecular imaging in the determination of molecular phenotypes.
4.Quantitative comparison of 68Ga-NGR and 18F-FDG uptake in well-differentiated hepatocellular carcinoma bearing mice
Yongheng GAO ; Zhengjie WANG ; Fei KANG ; Xiaowei MA ; Wenhui MA ; Mingru ZHANG ; Mingxuan ZHAO ; Tianming FU ; Guoquan LI ; Shengjun WANG ; Zhe WANG ; Weidong YANG ; Jing WANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(3):147-152
Objective To quantitatively compare the diagnostic capability of 68Ga-NGR and 18F-FDG in well-differentiated hepatocellular carcinoma (HCC) bearing mice by microPET/CT imaging.Methods The in vitro cellular uptake,in vivo microPET/CT imaging and biodistribution studies of 68Ga-NGR and 18F-FDG were quantitatively compared in SMMC-7721-based well-differentiated HCC.The human fibrosarcoma (HT-1080) and human colorectal adenocarcinoma (HT-29) cells/xenografts were respectively used as positive and negative reference groups for CD13.The expression of CD13 was qualitatively verified by immunohistostaining.The levels of CD13 and glucose-6-phosphatase (G6Pase) were semi-quantitatively analyzed by Western blot test for all 3 types of tumors.Two-sample t test was used for data analysis.Results The in vitro cellular uptake showed that the 68Ga-NGR uptake in SMMC-7721 and HT-1080 cells was higher than that in HT-29 cells,and the 68Ga-NGR uptake was higher than 18F-FDG uptake in SMMC-7721 cells.The in vivo micro-PET/CT imaging results revealed that the uptake of 68Ga-NGR in SMMC-7721 tumor was (2.17±0.21) %ID/g,remarkably higher compared to (0.73±0.26) %ID/g of 18F-FDG uptake (t =8.826,P<0.01).The tumor/liver ratio of 68Ga-NGR was 2.05±0.16,which was 2.03-fold higher than that of 18F-FDG.In the HT-1080 tumors,the uptakes of 68 Ga-NGR and 18F-FDG were both high,and the values were (2.46±0.23) %ID/g,(3.47±0.31) %ID/g.The uptake of 68Ga-NGR was significantly lower than that of 18F-FDG in HT-29 tumors:(0.67±0.20) %ID/g vs (3.17±0.29) %ID/g;t=4.221,P<0.01.Western blot and immunohistostaining results were as follows:HT-1080(CD13+,G6Pase-),SMMC-7721(CD13+,G6Pase+),HT-29 (CD13-,G6Pase-).Conclusions The uptake of 68Ga-NGR is higher than 18F-FDG uptake in SMMC-7721 tumor bearing mice,therefore it is worthwhile to consider the feasibility of clinical translation for PET/CT in diagnosis of HCC.Furthermore,because of the difference in 68Ga-NGR and 18F-FDG avidities in tumors with different molecular phenotypes of CD13 and G6Pase,there is an underlying potential for molecular imaging in the determination of molecular phenotypes.
5.Effect of neferine on proliferation and invasion of human hepatocellular carcinoma cell line HepG2 and Bel-7402
Yajun ZHOU ; Jing SHI ; Geng TIAN ; Hanxin ZHOU ; Guoquan GAO
Chinese Pharmacological Bulletin 2016;32(11):1539-1542
Aim To investigate the effect of neferine on proliferation and invasion of human hepatocellular car-cinoma. Methods HepG2 and Bel-7402 cells were cultured in vitro with different concentrations of nefer-ine, then cell proliferation was observed by CCK-8 as-say; cell invasion was observed by transwell invasion assay; the protein expression of RhoA, RhoC and ROCK was detected by Western blot. Results CCK-8 results showed that neferine could significantly inhibit cell proliferation in a dose-dependent manner compared with the control group. Transwell invasion assay showed that cell invasion was significantly decreased with neferine 3 μmol · L-1 . Western blot results showed that RhoA, RhoC and ROCK protein expres-sion was decreased when neferine was co-incubated with hepatocellular carcinoma cells. Conclusion Nef-erine can inhibit proliferation and invasion of HepG2 and Bel-7402 cells, which is mediated mainly by the inhibition of RhoA, RhoC and ROCK protein expres-sion.
6.Construction and in vitro characterization of dual-modality single-photon emission computed tomography-MRI nanoprobes targeting HAb18G/CD147 in breast tumors
Xianping LIU ; Mingru ZHANG ; Yao MENG ; Ruirui QIAO ; Xiaowei KANG ; Guoquan LI ; Yu LI ; Jianli JIANG ; Mingyuan GAO ; Hong YIN ; Jing WANG ; Guangquan WEI
Chinese Journal of Radiology 2015;(7):500-506
Objective To prepare dual?modality single?photon emission computed tomography (SPECT)?MRI molecular nanoprobes targeting HAb18G/CD147 expressed on breast cancer cell membranes and investigate the physicochemical and biological properties in vitro. Methods Superparamagnetic iron oxide nanoparticles (SPIOs) were prepared by one?pot reaction method as described. The single?chain antibody fragments HAb18F(ab')2 were conjugated to SPIOs via chemical method and then labeled with 125I using Iodogen method. The final 125I?SPIO?HAbF18(ab')2 nanoprobes were purified. SPIOs or 125I?HAb18F(ab')2 were used as control. We carried preliminary evaluation on their physicochemical properties and biological characteristics in vitro: transmission electron microscope (TEM) and dynamic light scattering (DLS) were used to measure these nanoparticle sizes and the hydrodynamic diameters. The MRI T2 transverse relaxation efficiency of these nanoprobes at different Fe2+concentrations were measured with 1.5 T clinical MR scanner. The 125I?SPIO?HAb18F(ab')2 and 125I?HAb18F(ab')2 radiochemical purity were measured by thin layer chromatography and the radio chemical yield was calculated. We also conducted stability tests in vitro and octanol/water partition coefficient experiments. Two breast tumor cell lines, MDA?MB?231 (HAb18G?overexpressing cells,experimental group) and MDA?MB?468 (control), were used for assessment of cells viability at different Fe2 + concentrations (1, 5, 10, 20, 40 μg/ml) by methyl thiazolyl tetrazolium assay. Specific binding experiments in vitro included two parts:magnetic resonance imaging and radionuclide tests, the above?mentioned breast cancer cell lines were incubated with 125I?SPIO?HAb18F(ab')2 nanoprobes respectively and took MDA?MB?231 cells which were not treated as blank group. First comparing the MR signal intensity differences among experimental group, the control group and blank group, then calculated the rate of MRI signal changes;Two breast tumor cell lines, MDA?MB?231 and MDA?MB?468 were incubated with 125I?SPIO?HAb18F(ab')2 nanoprobes too, then measured radioactivity counting byγcounter at different time and calculated the cell binding rates, and did statistical analysis by using one?way ANOVA. Results The SPIOs were fairly homogeneous with an average core size of (10.32±1.30) nm;the SPIO and 125I?SPIO?HAb18F(ab')2 hydrodynamic diameter of 44.80 and 52.64 nm, and MRI scanning showed that the transverse relaxation efficiency of SPIO and 125I?SPIO?HAb18F(ab')2 were 38.79 and 106.73 mM-1 · s-1, respectively. The radio chemical yield of 125I?SPIO?HAbF18(ab')2 and 125I?HAb18F(ab')2 were 41.90% and 85.50%, respectively. The radio chemical yield of the two groups were >95%, suggesting well stability in vitro. The lipo?hydro partition coefficient values were -0.99 ± 0.03 and-1.49 ± 0.08, respectively, which demonstrated that they were both water?soluble substances. Different Fe2+concentrations (1,5,10,20,40μg/ml) of 125I?SPIO?HAb18F(ab')2 on breast cancer cell lines MDA?MB?231 and MDA?MB?468 showed no significant inhibition of cell proliferation (F values were 0.78, 0.66; P values were 0.58, 0.66). The cell?specific binding experiment showed: MRI signal intensity values on experimental group, the control group and the blank group were (1 670 ± 5), (1 930 ± 8), (2 349 ± 14), respectively, significant differences existed among these groups (F=4 408.48,P=0.000), the rate of signal intensity change of experimental group and the control group were 28.87%,17.78%. SPECT:MDA?MB?231 could uptake 125I?SPIO?HAb18F(ab')2, the cell binding rates were (6.52 ± 0.60)% and (10.52 ± 2.04)% in 20 min and 4 h, respectively.Conclusions Our results suggested that the dual?modality SPECT?MRI nanoprobes 125I?SPIO?HAb18F(ab')2 were prepared successfully with good physicochemical properties and biological characteristics in vitro. These dual?modality molecular imaging nano?probes may have potential to improvearly detection and diagnosis of HAb18G/CD147?expressing cancers and to facilitate the development of HAb18G/CD147?directed interventions.
7.Analysis of the four parameter linear viscoelastic model on prosthetic nucleus and the influence of manufacture on parameters of this model.
Bing JI ; Jin GAO ; Yuanzheng MA ; Zhengqiu GU ; Guoquan LIU ; Haibin XUE
Journal of Biomedical Engineering 2007;24(2):356-359
The purpose of this study is to characterize the stress relax properties of PVA-H prosthetic nucleus via the four-parameter linear viscoelastic model and to analyze the influence of swelling ratio and initial PVA content upon the properties of dissipating compressive stress. The four-parameter linear viscoelastic model can simulate the viscoelastic property of prosthetic nucleus well (corr > 0.99) and be more effective than the three parameter linear viscoelastic model. From the parameters of this model we have obtained the following results: the prosthetic nucleus of higher water content can dissipate compressive stress more quickly than that of lower water content can do, but the quantity of stress relax can not be influenced by water content; the higher the initial PVA content,the slower and smaller the dissipation of compressive stress;the relax time of prosthetic nucleus is similar to that of human nucleus.
Compressive Strength
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Elasticity
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Intervertebral Disc
;
physiology
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Linear Models
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Models, Biological
;
Polyvinyl Alcohol
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chemistry
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Prostheses and Implants
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Viscosity
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Weight-Bearing
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physiology
8.Some Skills of Web-based Courses Design of Biochemistry
Zhihong SONG ; Guoquan GAO ; Jian LU ; Ming RENG
Chinese Journal of Medical Education Research 2006;0(11):-
Aiming at the characteristics of biochemistry and the advantages of web-based courses and combining with teaching practice,a kind of web-based courses has been constructed which practices in the internet.The key of construction will be detailed from general design,page design and some design skills.
9.Swelling characterization of poly (vinyl alcohol) hydrogel for prosthetic intervertebral disc nucleus.
Yong LI ; Jin GAO ; Guoquan LIU ; Zhengqiu GU ; Yuanzheng MA ; Haibin XUE
Journal of Biomedical Engineering 2005;22(5):995-998
In this study the poly(vinyl alcohol)(PVA) hydrogel elastomer was prepared by freezing-thawing method. The influences of percentage of poly (vinyl alcohol) in hydrogel, pH of solution and swelling temperature upon the swelling characteristic of PVA-hydrogel prosthetic nucleus material were studied. Its micropores were observed using SEM, and the swelling dynamics was further discussed. The experimental results showed that the poly (vinyl alcohol) hydrogel was a kind of network with a lot of micropores, the pore size was related with the PVA content. The maximum swelling ratio decreases when the percentage of PVA in hydrogel, the pH of solution and the swelling temperature were enhanced. The swelling process was described by the equation of swelling dynamics equation. The swelling rate was greatly influenced by the PVA content, the pH of solution and the dimension of hydrogel sample.
Biocompatible Materials
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chemistry
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Hydrogels
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Hydrogen-Ion Concentration
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Implants, Experimental
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Intervertebral Disc
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Polyvinyl Alcohol
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chemistry
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Prostheses and Implants
;
standards
10.Establishment and Application of Biochemistry Flash Animation Database
Zhihong SONG ; Xilian WU ; Guoquan GAO ; Sha TAO ; Shihao ZHOU ; Jianquan MA
Chinese Journal of Medical Education Research 2002;0(01):-
Aiming at the characteristics of biochemistry and the special advantage of flash and basing on teaching practice,we set up a set of biochemistry flash animation database,providing a series of immediate flashes and being applied to teaching practice of biochemistry,which shows the great potential of flash in improving biochemistry teaching quality.

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