1.Effect of GLP-1 on high glucose-induced human umbilical vein endothelial cell apoptosis and mechanism
Xiaoyan YUAN ; Ke CHEN ; Honghui HE ; Lilin ZHAO ; Zhaohui MO
Journal of Central South University(Medical Sciences) 2013;38(10):1029-1034
Objective: To investigate the effects of glucagon-like peptide-1(GLP-1)on high glucose-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and the mechanism involved. Methods: HUVECs were cultured under varying conditions for 48 h, and the cell viability was spectrophotometrically measured by MTT assay. Flow cytometry detected the ratio of cell apoptosis. Western blot detected the protein levels of p-Akt and p-eNOS, while NO assay kit detected the NO concentration.
Results: Treatment of high glucose (33 mmol/L) for 48 h signiifcantly decreased the HUVECs viability and induced the apoptosis of HUVECs, concomitant with decreased Akt and eNOS phosphorylation leves and subsequent NO production. Treatment with GLP-1 (3 nmol/L) for 48 h in the high glucose group increased the HUVECs viability (P<0.01), decreased the ratio of HUVECs early apoptosis (P<0.05), ameliorated the reduced protein levels of p-Akt and p-eNOS caused by high glucose, and increased the NO production (P<0.05). The anti-apoptotic effect and the increased NO production of GLP-1were inhibited by PI3K inhibitor wortmannine (100 nmol/L) or eNOS inhibitor L-NAME (100μmol/L). The effect on p-Akt, p-eNOS of GLP-1 was inhibited by wortmannine (100 nmol/L) while L-NAME (100μmol/L) did not have any influence on the expression of p-Akt.
Conclusion: GLP-1 can ameliorate high glucose-induced HUVECs apoptosis, which is probably related to the up-regulation of PI3K/Akt/eNOS pathway.
2.Identification of Volatile Fungistatic Compounds by Spme-GC/MX
Chuan-Kun XU ; Ming-He MO ; Ke-Qin ZHANG ;
Microbiology 1992;0(05):-
The composition of volatile compounds of soils and that of soil bacterial metabolites were identified by using the SPME-GC/MS method. Results showed that some compounds, trimethylamine, 3-methyl-2-pentanoe, dimethyl disulfide, methyl pyrazine, 2,5-dimethyl-pyrazine, benzaldehyde, N,N-dimethyloctylamine and nonadecane, subsisted commonly in soils and soil bacterial metabolites with strong fungistatic activity. These compounds may be the key antifungal factors in soil fungistasis, especially soil volatile fungistasis. Otherwise, the method used in this study was a good tool for further study of soil volatle fungistasis.
3.Influence of psychological nursing on the therapeutic effect of patients with newly diagnosed type 2 diabetes
Qinghua FU ; Huifang YE ; Ke ZHOU ; Guiying MO ; Guangming LAN
Chinese Journal of Practical Nursing 2008;24(18):55-56
Objective To discuss the influence of psychological nursing on the therapeutic effect of patients with newly diagnosed type 2 diabetes. Methods Patients (60 cases) who received psychological nursing were set as the nursing group. Patients (65 cases) who did not received psychological nursing were set as the control group. The number of patients who received early insulin treatment and whose fasting plasma glucose return to normal value on the 7th and 30th days after treatment was observed. Results The number of patients who received early insulin treatment and whose fasting plasma glucose return to normal value on the 7th and 30th days after treatment in the nursing group was higher than that of the control group (P<0.01). Conclusions Psychological nursing of patients with newly diagnosed type 2 diabetes could make patients obey doctors' advice and accept early insulin treatment to control plasma glucose and delete the toxicity of high concentration of plasma glucose as soon as possible.
4.Effect of erythropoietin on the proliferation and apoptosis of neonatal porcine islet cells
Honghui HE ; Tianhui WU ; Jing XIONG ; Ke CHEN ; Zhaohui MO
Journal of Central South University(Medical Sciences) 2010;35(11):1115-1122
Objective To investigate the effect of erythropoietin on the proliferation,differentiation,and apoptosis of the cultured neonatal porcine islet cells in vitro.Methods Neonatal porcine islet cells were separated and pured from neonatal pigs with collagenase digestion and tissue culture,and their viability and purity were tested. The neonatal porcine islet cells were divided into a control group and an experimental group.The experimental group was treated with erythropoietin but not the control group,and the insulin secretion responsiveness induced by low and high glucose stimulation in the islet was tested after 5 days. Cells were counted and the activation of amplification was determined by MTT chromatometry. The rates of cell apoptosis were observed by ethidium bromide/acridine orange (EB/AO) of fluorescent light staining and flow cytometry,and the cell cycle was analyzed by flow cytometry. The expression of bcl-2,bax,caspase-3,glucose transporter 2 (GlUT-2),and pancreatic duodenal homeobox-1 (PDX-1) mRNA was tested by RT-PCR.Results After erythropoietin was treated in the cell culture,the neonatal porcine islet cells had normal morphology,function,and reaction of insulin secretion to the glucose stimulation. Cell count showed more cells in the experimental group than in the control group (P<0.05). MTT chromatometry showed the optical absorbance tended to increase with time,and compared with the control group,the optical absorbance was higher in the experimental group (P<0.05),the expression of PDX-1 mRNA was slightly up-regulated (P<0.05). The expression of GLUT-2 mRNA had no difference in the 2 groups (P=0.34). In the experimental group,the apoptisis rate was lower than that in the control group by flow cytometry and EB/AO fluoscence staining (P<0.01),and the expression of bcl-2 mRNA was higher. Howerer bax mRNA and caspase-3 mRNA were obviously lower than those in the control group (P<0.01).Conclusion Erythropoietin can promote the proliferation but has no effect on the function of neonatal porcine islet cells in vitro. Erythropoietin can protect neonatal porcine islet cells from apoptosis through up-regulating bcl-2 mRNA and downreguling bax and caspase-3 mRNA.
5.Values of the integrated visual and auditory continuous performance test in diagnosing children with attention deficit hyperactivity disorder in clinic and school in China
Wei ZHANG ; Yeping JIANG ; Shuliang MO ; Hongli LIN ; Shanyu KE
Chinese Journal of Behavioral Medicine and Brain Science 2015;24(10):935-938
Objective To explore the psychometric properties of the Chinese version of integrated visual and auditory continuous performance test (IVA-CPT) ,and to assess the diagnosis value of the Chinese version of IVA-CPT in two samples of children with attention deficit hyperactivity disorder (ADHD) in clinic and school.Methods 112 participants were divided into 4 groups:clinic group, clinic control group, school group and control group from school.The participants were measured by IVA-CPT and the results were compared.Results ①The integrated quotients of IVA-CPT had concordance with the dimensions of the DSM-ⅣV and ASRS.Each integrated quotients of IVA-CPT and each dimension of the DSM-ⅣV and ASRS were negatively correlated(school: r=-0.277, P<0.05;clinic: r=-0.423, P<0.05).The score of HI dimension was negatively correlated with integrated attention quotient and integrated inhibition quotient of IVA-CPT (r=-0.480, P<0.01;r=-0.452, P<0.01).② The results of the IVA-CPT and the comprehensive assessment had consistency for diagnosis of children with ADHD from the total subjects,the clinic ,and the school.And there were no difference between the sensitivity, the specificity, the coincidence rate, the missed diagnosis rate and the misdiagnosis rate of IVA-CPT when used to diagnose children with ADHD in clinic and school(x2=3.396, P>0.05).③The IVA-CPT evaluation system had good stability according to the retest of a random sample of 25 subjects(P<0.01).The pretest and posttest results of each full quotients of IVA-CPT were significantly correlated (P<0.01).Conclusion The IVA-CPT not only can be used as auxiliary diagnostic tools to diagnose children with ADHD in clinic,it can be used as auxiliary diagnostic tools to diagnose children with ADHD in the community.
6.Establishment of cell line stably expressing INSIG2 and its effects on fat metabolism
Ke CHEN ; Zhaohui MO ; Xiaowei XING ; Pingan HU ; Yanhong XIE
Basic & Clinical Medicine 2006;0(07):-
Objective To establish the cell line stably expressing INSIG2 and observe its effecet on fat metabolism after overexpression of INSIG2.Methods The eukaryotic expression plasmid pcDNA3.1(+)-INSIG2 was constructed,which was transfected into 3T3-L1 cells.The expression of INSIG2 and related genes were detected by RT-PCR and immunohistochemistry,the contents of FFA in cell culture medium and adipocyte differentiation were detected by ELISA and Oil Red "O"staining respectively.Results After pcDNA3.1(+)-INSIG2 was transfected into the 3T3-L1 cells,the expression of INSIG1 mRNA and FAS mRNA were down-regulated,the content of FFA in the cell culture medium was decreased and adipocyte differentiation was drepressed.Conclusion The cell line stably expressing INSIG2 was successfully established,the transfected INSIG2 may have a drepressant effect on fat metabolism.
7.Effect of taurine on the differentiation of 3T3-L1 preadipocytes
Min LIU ; Pingan HU ; Ke CHEN ; Zhaohui MO
Chinese Journal of Tissue Engineering Research 2007;0(12):-
AIM: Some studies found that the glucose uptake ability of diabetes mellitus mice is stronger after feeding with taurine. Whether the taurine can affect adipocytes or not deserves further studies. This article investigates the effects of taurine on the differentiation of 3T3-L1 preadipocytes and detects the mechanics. METHODS: Experiments were performed in the Laboratory Center of Xiangya Third Hospital of Central South University from July to September 2006. ①3T3L1 preadipocytes were provided by Shanghai Cell Bank of China Academy of Sciences. ②3T3-L1 preadipocytes were cultured in high glucose DMEM medium containing fetal bovine serum of 0.10 volume fraction, 108 u/L benzylpenicillin and 80?1010 U/L streptaquaine. After confluence, cells at 5?107 L-1 were incubated in culture flask, and induced with 0.5 mmol/L IBMX, 0.5 mg/L insulin and 1 ?mol/L dexamethasone. The cells in an experimental group were intervened with taurine, whereas these in a control group were not treated with taurine. Oil O staining was used to observe the development of adipose cells. C3T3-L1 preadipocytes were treated with 10,20 mmol/L taurine respectively for 24 and 48 hours. RNA and protein were extracted in the control group and the experimental group. ③Genes related to adipose cells development was examined by RT-PCR and Western-blot. RESULTS: ①The number of cells stained by oil O was less in the experimental group than the control group in 3T3-L1 preadipocytes treated with 10 mmol/L taurine for 14 days. ②After C3T3-L1 preadipocytes were treated with 10,20 mmol/L taurine respectively for 24 and 48 hours, there were no changes in expression of Insig-2 protein, PPAR, Insing-2, adiponectin, adiponectin receptor, GLUT-4, AP-2 mRNA. CONCLUSION: Taurine inhibits the differentiation of 3T3-L1 preadipocytes, but the mechanics still need to be investigated.
8.Effects of resveratrol on the expression of FAK and the level of phosphorylated FAK in highly metastatic ovarian carcinoma HO-8910PM cell line
Yanmei QIN ; Ke LI ; Taiping HE ; Lier MO ; Nianci LIANG
Chinese Pharmacological Bulletin 1986;0(06):-
Aim To study the effect of resveratrol on the expression of FAK and the level of phosphorylated FAK, and to investigate its possible mechanism of resveratrol on the metastasis-associated ability of human highly metastatic ovarian carcinoma HO-8910PM cells line. Methods MTT assay was used to examine cytotoxicity of resveratrol on HO-8910PM cells after 24 h treatment; The expression of FAK protein and the level of phosphorylated FAK were assessed by Western blotting analysis. Results MTT assay showed that resveratrol had weak cytotoxicity on HO-8910PM cells after 24 h treatment, its IC50 value was 163.40?2.48 ?mol?L-1. The expression of FAK and the level of phosphorylated FAK decreased obviously in HO-8910PM cells treated with 25~100 ?mol?L-1 resveratrol for 24 h. Conclusion The mechanism of resveratrol inhibiting the metastasis-associated ability of human highly metastatic ovarian carcinoma HO-8910PM cells line involves the expression of FAK and the level of phosphorylated FAK.
9.Anti-viral Effects of Lianhua Qingwen Capsule Against Influenza A Virus in Vitro
Hongying MO ; Changwen KE ; Jingping ZHENG ; Nanshan ZHONG
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To study the anti-viral effects of Lianhua Qingwen Capsule against influenza A virus in vitro.Methods Under different administration modes,the inhibitory effect of Lianhua Qingwen Capsule on human influenza A virus(H3N2)in vitro and its time-effect relationship were assayed by crystal violet staining assay with ribavirin as positive reference drug.Results Lianhua Qingwen Capsule showed a multi-access effect on anti-human influenza A virus,such as comprehensive inhibition effect,prevention effect on virus adsorption,inhibition on virus proliferation after adsorption and direct killing the virus effect,and the median effective concentrations(EC50)in above-mentioned effects were 0.042,0.031,0.051 and 0.050 mg/mL respectively.Among them,the prevention effect on virus adsorption was the strongest.With the expanding actuation duration of the drug,the inhibitory effect of Lianhua Qingwen Capsule against influenza A virus weakened at low concentration,but no change at high concentration(≥0.031 g/mL).Meanwhile,Lianhua Qingwen Capsule can remarkably decrease the infectivity of influenza A virus.Conclusion Lianhua Qingwen Capsule has an obvious effect against human influenza A virus in vitro.
10.Effect of dexmedetomidine on myocardial ischemia-reperfusion injury in rats
Xiaofei MO ; Jianjuan KE ; Xiaobo FENG ; Zongze ZHANG ; Yanlin WANG
Chinese Journal of Trauma 2016;32(12):1119-1124
Objective To investigate the effect of different doses of dexmedetomidine (DEX) on myocardial ischemia-reperfusion injury (MIRI) in rats.Methods Seventy-five SD male rats weighing between 250 and 300g were divided into sham group,MIRI group,small-dose DEX group,medium-dose DEX group and high-dose group according to the random number table,with 15 rats per group.Threading the left anterior descending coronary artery was done only in sham group,but the MIRI model was produced in the rest groups by ligation of the artery for 30 minutes followed by 120 minutes of reperfusion.Fifteen minutes before the ligation,small-,medium-and high-dose DEX groups were injected 2.5,5 and 10 μg · kg-1 · h-1 of DEX respectively until the end of reperfusion.Instead,an equal volume of normal saline was given in sham and MIRI groups.At the end of reperfusion,five rats in each group were used to determine the myocardial infarct size,and arterial blood samples and myocardial tissues from ten rats in each group were used to measure serum levels of interleukin-1β (IL-β) and serum tumor necrosis factor-α (TNF-α),expression of myocardial nuclear factor kappa B p65 (NF-κB p65) and change of myocardial pathomorphology.Results Myocardial infract size,degree of myocardial pathomorphology structure damage,serum levels of IL-1 β and TNF-αt and expression of myocardial NF-κB p65 in sham group were significantly lower in sham group than other groups (P < 0.05).Above mentioned parameters in small-,medium-and high-dose DEX groups were all significantly decreased compared to MIRI group (P < 0.05),and the decrease was most significant in medium-dose DEX group (P < 0.05).Conclusions DEX can attenuate the MIRI in rats and the possible mechanism is suppressing the release of NF-κB p65,which can reduce serum pro-inflammatory cytokines like TNF-α and IL-1β.And mediumdose DEX exhibits better protective effect.