1.Study on the relationship between N~( 5,10)-methylenetetrahydrofolate reductase gene polymorphism and the susceptibility to microangiopathy in type 2 diabetes mellitus
Guoqing YANG ; Juming LU ; Changyu PAN
Chinese Journal of Endocrinology and Metabolism 1986;0(04):-
Objective To study the relationship between the polymorphism of N 5,10 methy lenetetrahydrofolate reductase (MTHFR) gene and the susceptibility to diabetic microangiopathy (DMAP) in type 2 diabetes mellitus (DM) of Han nationality in north China. Methods By PCR RFLP, MTHFR C677T mutation was detected in 229 type 2 diabetic patients, including 102 non complication diabetics (NCD group), 67 diabetic nephropathy (DN group) and 60 diabetic retinopathy (DR group), and 62 healthy persons as control (CON group). Results The frequency of homozygous BB genotype and allele B in DN and DR were significantly higher than that in NCD and CON, but NCD and COD showed similar frequency of BB genotype and allele B. Conclusion In type 2 DM of Han nationality in north China, there is a strong correlation between the polymorphism of MTHFR gene and DMAP, and the allele B may be a susceptibile gene of DMAP.
2.Expression and significance of PAI-1 in nasopharyngeal carcinoma (NPC) cell line(CNE-2Z) and its sub-line (CNE-2Z-H5,CNE-2Z-H5-9)
Guoqing PAN ; Hongmei FU ; Li BIAN
China Oncology 2001;0(03):-
0.05) ; whereas it was seen in CNE-2Z-H5-9. Anti-PAI-1 antibody did not inhibit the heterogeneous adhesion and invsion of CNE-2Z-H5-9. Conclusions:PAI-1 maybe inhibit the metastasis and invasion of NPC.
3.Evaluation of efficacy and safety of Gansulin 50R versus Novolin 50R in treatment of diabetic patients:A multicenter,randomized,opened and controlled clinical trial
Guoqing YANG ; Juming LU ; Changyu PAN
Chinese Journal of Diabetes 2008;0(10):-
0.05). Conclusions Gansulin 50R and Novolin 50R have similar efficacy and safty profiles in the treatment of diabetic patients.
4.Study on the role of axon guidance factor Semaphorin 5A in the invasion and metastasis of human gastric cancer
Guoqing PAN ; Xiangling ZHANG ; Junyu REN ; Jianbo LU ; Hongmei FU
Chinese Journal of Digestion 2012;32(2):103-107
Objective To explore the correlation between axon guidance factor Semaphorin 5A and clinicopathological features and its role in the invasion and metastasis of gastric cancer.Methods The expression of Semaphorin 5A in gastric cancer tissues of 171 patients with different gender,age,histological type and TNM stage was detected with immunohistochemistry assay.The expression of Semaphorin 5A was determined by Western blotting assay in gastric cancer cell lines SGC7901 and MKN-45 with metastatic ability and gastric cancer cell lines SNU-1 and AGS without metastatic ability.With RNA interfere technique(RNAi),Semaphorin 5A siRNA expression vector was constructed and transfected into gastric cancer cell line SGC7901.The stable gastric cancer cell line down-expressing Semaphorin 5A was established.The effect of Semaphorin 5A gene silencing on the adhesion,migration and invasion of gastric cancer cell was examined by cell adhesion test,wound healing test and transwell chamber assays.Results The expression level of Semaphorin 5A was correlated with the differentiation degree of gastric cancer(x2 =6.32,P =0.01),lymphnode metastasis(x2 =7.68,P=0.01)and distant metastasis of gastric cancer(x2 =13.67,P =0.00),not correlated with age(x2 =0.21,P=0.79),gender(x2=1.79,P=0.15)and the depth of gastric cancer invasion(x2=1.34,P=0.55).The expression of Semaphorin 5A in cell lines SGC7901 and MKN-45 was significantly higher than that of cell lines SNU-1 and AGS(P<0.01).Semaphorin 5A gene silencing significantly suppressed the adhesion,migration and invasion abilities of gastric cancer cells.Conclusion Semaphorin 5A may play a catalytic role in the invasion and metastasis of gastric cancer through increasing the adhesion,migration and invasion abilities of gastric cancer cell.
5.Effects of PTD-HBcAg induced murine bone marrow-derived dendritic cells maturation on T lymphocyte proliferation in vitro
Xiaohua CHEN ; Qingchun PAN ; Yongsheng YU ; Jinchao HAN ; Guoqing ZANG
Chinese Journal of Infectious Diseases 2009;27(4):198-202
Objective To observe the effects of PTD-hepatitis B core antigen (HBcAg) induced murine bone marrow-derived dendritic cells (DCs) maturation on T lymphocyte proliferation in vitro.Methods Bone marrow derived DCs isolated from BALB/c mice were cultured with recombinant granu|ocyte-macrophage colony-stimulating factor (rGM-CSF) and recombinant interleutin-4 (rIL-4)for 5 days.Tumor necrosis factor (TNF)-a,HBcAg and PTD-HBcAg were added to induce DCs maturation.The distribution and localization of intracellular immunofluorescence were observed by confocal microscopy,and DCs phenotypes were analyzed by flow cytometry.The level of IL-12 p70 in the supernatant was detected by enzyme linked immunosorhent assay (ELISA).The proliferation of T lymphocytes was performed by using cell counting kit-8 (CCK-8).All data were analyzed using t test.Results DCs were cultured and identified successfully.Recombinant PTD-HBcAg could penetrate into DCs cytoplasm while recombinant HBcAg was detected on the surface of cells.DCs surface molecules,such as CD80,CD86 and major histocompability complex (MHC) II were upregulated by PTDHBcAg;IL-12 p70 levels induced by 50 mg/L and 100 mg/L recombinant PTD-HBcAg were (142.50±18.31) ng/L and (124.30±15.12) ng/L,respectively,which were significantly higher than those induced by recombinant HBcAg [(42.31±4.21 ) ng/L,t = 9.234 and 9.045,respectively,P<0.05].The proliferation of T lymphocytes induced by PTD-HBcAg was much higher than that in HBcAg group or positive control TNF-a group.Conclusions PTD-HBcAg could penetrate membrane of DCs and promote the differentiation and maturation of DCs.PTD-HBcAg could up-regulate the expressions of costimulatory molecules on cell surface of DCs,and enhance the ability of DCs on stimulating T lymphocytes proliferation and IL-12 p70 production.
6.Expression of uPA,uPAR,PAI-1 in three nasopharyngeal carcinoma cell(NPC) lines with different ability to metastasize
Guoqing PAN ; Yunhong YAO ; Yuanzhi LU ; Al ET
China Oncology 1998;0(04):-
Purpose:To study the expression of uPA, uPAR ,PAI 1 in 3 nasopharyngeal carcinoma cell lines CEN 2Z,CEN 2Z H5,CEN 2Z H5 9,with different ability to metastasize through lymph.Methods:RT PCR was used to detect expression of uPA,uPAR,PAI 1 at levels of mRNA.Results:mRNA expression of uPA,uPAR was the highest in CNE 2Z H5 9,whereas the lowest in CNE 2Z;mRNA expression of PAI 1was detected in CNE 2Z and CNE 2Z H5, but revealed no differences,but not in CNE 2Z H5 9.Conclusions:uPA?uPAR may promote the metastasis and invasion of NPC,whereas PAI 1 maybe inhibit it
7.Comparison of feasibility of transplanting neural stem cells through three ways to treat newborn piglets with hypoxic ischemic encephalopathy
Guoqing ZHANG ; Jun BU ; Jianhua SUN ; Jun PAN
Chinese Pediatric Emergency Medicine 2010;17(6):529-531,后插2
Objective To compare three ways of transplanting neural stem cells(NSCs) to treat hypoxia-ischemic encephalopathy (HIE) ,such as through axillary vein,internal carotid artery and lumbar puncture. Methods Newborn piglets were divided into three groups randomly,and transplanted NSCs through axillary vein,through internal carotid artery or through lumbar puncture after hypoxic-ischemic damage operations. Each group had five piglets. Two hours after hypoxic ischemic damage,2 × 106 NSCs with green fluorescent protein were injected through axillary vein,internal carotid artery or lumbar puncture. Piglets were sacrificed 24 hours after operations. Slices were gotten at hippocarnpus, anterior horn of lateral ventricle and posterior horn of lateral ventricle. NSCs were counted at four visual fields of each four slices of each lays through fluorescence microscope with 400 amplification factor len. Results The positive cells of axillary vein group,internal carotid artery group and lumbar puncture group were 53. 80 ± 8. 78,69. 80 ± 11.90,265.00 ± 29.65respectively. The positive cells of lumbar puncture group were more than the other groups, and there was statistic significance(P < 0. 01). Conclusion The study proved NSCs injected through lumbar puncture could enter brain tissue. It is feasible to transplant NSCs through lumbar puncture to treat newborn with HIE.
8.Influence of NGF-PEG-PLGA-NPs on neuronal differentiation of neural stem cells and PI3K/Akt signaling pathway
Yan CHEN ; Guoqing BAO ; Feifei LIU ; Jundu ZHANG ; Cuihuan PAN ; Dahong LONG
Chinese Journal of Tissue Engineering Research 2015;(28):4576-4581
BACKGROUND:Our previous studies confirmed that NGF-PEG-PLGA-NPs has good sustained release effect and biological activity in vitro, and can induce the differentiation of PC12 cel s into neuron-like cel s.
OBJECTIVE:To investigate the feasibility of neuronal differentiation of neural stem cel s from septal area of fetal brain induced by NGF-PEG-PLGA-NPs and its influence on PI3K/Akt signaling pathway.
METHODS:According to optimization prescription, NGF-PEG-PLGA-NPs were prepared by multiple emulsion solvent diffusion method. Neural stem cel s were induced to neuronal differentiation in six groups, including control group, NGF group, NGF-PEG-PLGA-NPs group, LY294002 group, LY294002+NGF group and LY294002+NGF-PEG-PLGA-NPs group. Neurons were identified by immunofluorescence, while phosphorylation levels of Akt in PI3K/Akt signaling pathway were detected by western blotting.
RESULTS AND CONCLUSION:The proportions ofβ-Tubulin III-positive neurons in control group, NGF group, NGF-PEG-PLGA-NPs group, LY294002 group, LY294002+NGF group and LY294002+NGF-PEG-PLGA-NPs group were (22.80±2.58)%, (35.80±3.98)%, (35.40±5.77)%, (26.60±3.87)%, (21.20±2.59)%and (25.80±7.22)%, respectively. There were no statistical differences in neuronal differentiation between NGF group and NGF-PEG-PLGA-NPs group (P>0.05), but the ratios of neural differentiation in the two groups were both higher than that in the other four groups (P<0.05). Western blotting results revealed that there were no statistical differences in Akt phosphorylation levels between NGF group and NGF-PEG-PLGA-NPs group (P>0.05), but the phosphorylation levels of Akt were both higher than other four groups (P<0.05). There were also no significant differences between LY294002+NGF and LY294002+NGF-PEG-PLGA-NPs groups and control group (P>0.05), but the phosphorylation levels of Akt were higher than LY294002 group (P<0.05). Results suggest that NGF-PEG-PLGA-NPs promoted neural differentiation of neural stem cel s. The role might be related to increasing phosphorylation levels of Akt in PI3K/Akt signaling pathway.
9.Effects of antioxidants on homocysteine thiolactone-induced apoptosis in human umbilical vein endothelial cells
Weijun GU ; Juming LU ; Guoqing YANG ; Qinghua GUO ; Baoan WANG ; Yiming MU ; Changyu PAN
Journal of Geriatric Cardiology 2006;3(2):107-111
Background and objectives Hyperhomocysteinemia is an independent risk factor for cardiovascular disease. Homocysteine thiolactone (HcyT), one of the homocysteine metabolites in vivo, is toxic both in vivo and in vitro. The aim of this study was to investigate the effect of HcyT on apoptotic damage in human umbilical vein endothelial cells (HUVECs) and the role of antioxidants in the reduction of HcyT-induced apoptosis. Methods HUVECs were cultured in DMEM supplemented with 20% heat inactivated fetal bovine serum cell cultures were maintained in a humidified 5% CO2 atmosphere at 37 ℃. Cytotoxicity was determined by MTT assay,which consists of hypodiploid cells with propidium iodide labeling and intracellular reactive oxygen species levels using 2',7'-dichlorofluorescein diacetate as the probe by flow cytometry. Results HcyT (250-2000μM) induced HUVECs apoptosis in a time- and concentration-dependent manner. Reactive oxygen species levels rose in response to increasing HcyT concentrations at 24-h incubation.The reduction of cell apoptosis by N-acetylcysteine, vitamin E, or pyrrolidine dithiocarbamate, occurred simultaneously with a significant decrease in intracellular reactive oxygen species levels. Conclusion HcyT exerts its cytotoxic effects on endothelial cells through an apoptotic mechanism involving cellular reactive oxygen species production. The capacity of N-acetylcysteine, vitamin E, and pyrrolidine dithiocarbamate to scavenge HcyT-induced cellular reactive oxygen species correlates well with their efficiency to protect against HcyT-promoted apoptotic damage. The protective effect of pyrrolidine dithiocarbamate on cell apoptosis indicates HcyT-generated hydrogen peroxide may provoke cell apoptosis via activating nuclear factor-kappa binding protein.
10.Association of toxic thyroid adenoma with acromegaly: one case report and literature review
Weijun GU ; Jingtao DOU ; Guoqing YANG ; Zhaohui LU ; Jianming BA ; Yiming MU ; Juming LU ; Changyu PAN
Chinese Journal of Endocrinology and Metabolism 2012;(11):901-905
Hyperthyroidism and acromegaly formed an unusual association.An acromegaly patient with a toxic thyroid adenoma was reported here,including clinical features,treatment,and final outcomes.The association of thyroid disease with acromegaly was reviewed.