2.Metabolic heterogeneity of gastric cancer cell lines
Xia-Xia FAN ; Ying-Li ZHU ; Bei-Bei SHAO ; Lu-Lu LI ; Ya-Fei LI ; Ao JIA ; Ai-Feng WANG ; Yong-Cheng MA
Chinese Journal of Pharmacology and Toxicology 2018;32(4):278-279
OBJECTIVE Gastric cancer is one of the most common malignant tumors,and the inci-dence rate is the highest in all kinds of tumors in China. However,it remains unclear that how signifi-cantly gastric cells are dependent on glycolysis,and which type of gastric cells are sensitive to glycolysis inhibition. In this study, several kind of gastric cancer cell lines were used as the research object, and the metabolic characteristics of different cell lines were systematically analyzed to provide theoretical support for the accurate treatment of gastric cancer. METHODS We examined the energy metabolism of four gastric cancer cell lines(MGC-803,SGC-7901,HGC-27 and BGC-823)by using glycolysis inhibitor, 2-deoxy-D-glucose(2-DG)and inhibitor of oxidative phosphorylation,oligomycin.Oxygen consumption rates(OCR)and L-lactate were also measured with an XF96 Analyzer(Seahorse Biosciences)to deter-mine the significance of metabolism of oxidative phosphorylation and aerobic glycolysisin gastric cells. In addition, western blot was used to detect the contribution of AMP-activated protein kinase (AMPK), and anti-apoptotic proteins(Bcl-2 and survivin)to clarify the mechanism of death or survival of gastric cancer cells treated by 2-DG or oligomycin. RESULTS In this study, it was shown that the growth of gastric cell lines were suppressed by 2-DG.However,the sensitivity to 2-DG was quite different among cell lines:IC 50 of 2-DG was from 3.28 mmol·L-1(MGC-803)to 15.57 mmol·L-1(BGC-823).MGC-803 was relatively sensitive to 2-DG (IC 50:3.28 mmol·L-1), consumed more glucose and produced more lactate (waste product of glycolysis) than the three other cell lines. Consequently, MGC-803 could be more dependent on glycolysis than other cell lines, which was further confirmed by the fact that glucose (+)FCS(-)medium showed more growth and survival than glucose(-)FCS(+)medium.Alternatively, BGC-823, most resistant to 2-DG (IC50: 15.57 mmol·L- 1), was most sensitive to oligomycin, and showed more growth and survival in glucose(-)FCS(+)medium than in glucose(+)FCS(-)medium. Thus,we had reasons to think BGC-823 cells depended on oxidative phosphorylation for energy production. In BGC-823,AMPK,which is activated when ATP becomes limiting,was rapidly phosphorylated by 2-DG, and expression of Bcl-2 was augmented,which might result in resistance to 2-DG.Interestingly,AMPK phosphorylation and augmentation of Bcl-2 expression by 2-DG were not observed in MGC-803,which is 2-DG sensitive. CONCLUSION There is a large metabolic difference between gastric cancer cell lines,which will facilitate the future gastric cancer therapy by targeting metabolic pathways.
3.Evaluation of left ventricular function in children with Kawasaki disease by Tei index of adenosine triphosphate-atropine stress echocardiography
Li LIU ; Bei XIA ; Chengrong LI ; Baoming QIU ; Shuna LI ; Xiuming DU
Chinese Journal of Ultrasonography 2009;18(8):672-674
R prognosticate the lesion of coronary artery in KD has influence on left ventricular function.
4.Anti-esophagus cancer activity and mechanism of DN3,a novel natural diterpenoid derivative,as a dual inhibitor of glycolysis and oxidative phosphorylation
Yong-Cheng MA ; Ying-Li ZHU ; Xia-Xia FAN ; Ao JIA ; Ya-Fei LI ; Bei-Bei SHAO ; Ai-Feng WANG
Chinese Journal of Pharmacology and Toxicology 2018;32(4):297-297
OBJECTIVE To probe into the anti-esophagus cancer activity and mechanisms of DN3, a novel natural diterpenoid derivative. METHODS The anti-tumor activity in vitro of DN3 was evaluated by MTT, and by using human esophageal carcinoma cells xenografted into athymic mice model in vivo. The specific mechanisms of DN3, as a dual inhibitor of glycolysis and oxidative phos-phorylation(OXPHOS)were explored through cell and molecular biology techniques.For instance,the manner of cancer cell death induced by DN3 was characterized by hoechst33342, FITC-Annexin V/PI staining and flow cytometric analysis,then these changes of glucose consumption,glucose uptake and lactate production in glycolysis, as well as oxygen consumption rate (OCR) and ATP content in OXPHOS caused by DN3 were performed separately through related kits and SeahorseBioscience XF24 Extra-cellular Flux Analyzer.Furthermore,in order to obtain a clear understanding of the inhibition of DN3 to glycolysis and OXPHOS, these regulatory factors were investigated by Western blot, such as PI3K/AKT, c-Myc and p53 of glycolysis, Bax and HK2 of mitochondrial function. RESULTS DN3 inhibited the growth of esophagus cancer cell EC9706, EC109 and EC1 cells in a dose and time dependent manner,but showed no significant effects on human esophageal epithelial cells(HEECs).DN3 caused significant G2/M arrest of esophagus cancer cell lines and induced apoptosis of these cell lines, which indicated DN3 inhibited the growth of esophagus cancer cell through blocking cell cycle and inducing apoptosis in a dose and time-dependent manner. Importantly, 8 μM DN3 decreased the extracellular acidification rate (ECAR) by 45% in EC109, which indicated glycolysis was inhibited by DN3. Mean-while, DN3 decreased the oxygen consumption rate (OCR) and the OCR linked to intracellular ATP production in EC109 cells,but that was not obvious in HEECs,so which indicated that DN3 could selec-tively block OXPHOS of cancer cells. In addition, the accumulation of reactive oxygen species (ROS) and the drop of mitochondrial membrane potential (MMP) were also observed in EC109 incubated by DN3,which suggested mitochondrial biological function was disturbed.Furthermore,the expression of PI3K/AKT, c-Myc and HK2 related to glycolysis were down-regulated by DN3, but the p53 and Bax were up-regulated in esophageal carcinoma cells. The changes of these enzymes accounted for the decreased glycolysisand OXPHOS in esophageal carcinoma cells treated by DN3. CONCLUSION The new compound DN3 has a strong anti-esophageal carcinoma activity,and it is tolerable that DN3 is seen as a dual inhibitor of glycolysis and oxidative phosphorylation.
5.Antitumor activityand mechanism of JD20,a newly synthetic analog of natural ent-kaurane diterpenoid
Ying-Li ZHU ; Xia-Xia FAN ; Ya-Fei LI ; Ao JIA ; Bei-Bei SHAO ; Ai-Feng WANG ; Yong-Cheng MA
Chinese Journal of Pharmacology and Toxicology 2018;32(4):346-346
OBJECTIVE To study the anti-tumor activity and molecular mechanism of natural diter-pene derivative JD20 in vitro. METHODS Screening the sensitive of gastric carcinoma cell lines to JD20 by cytotoxicity test for 24 h.Cell morphology was evaluated by using DAPI.After staining of can-cer cells with PI or annexin V-FITC/PI respectively,the cell cycle and apoptosis induced by JD20 were detectded by flow cytometry. The change in cell membrane potential was detected by JC-1 test kit. Western blot method was used to detect the apoptosis-related protein. RESULTS The novel natural kaurane diterpene derivative JD20 had a significant inhibitory effect on tumor cells and was particularly active on gastric cancer cell lines HGC-27 (IC50=4.72 ± 1.37 μmol·L- 1) and MGC-803 (IC50=7.36 ± 0.86 μmol·L-1).Further studies found that JD20 resulted in thecell cycle arrest in the G2/M phase,and induced a significant apoptosis in HGC-27. In addition, JD20 also caused the drop of mitochondrial membrane potential of HGC-27 within a short time (3 h). Furthermore, the Western blotting analysis showed that JD20 could induce the up-regulation of p53,Bax and Bim protein expression in gastric can-cer cells,and the releasing of cytochrome c from the mitochondria into the cytoplasm,as well as the ac-tivation of casepase-9/3.CONCLUSION The natural kaurane diterpene derivative JD20 can inhibit the proliferation of various human cancer cells by blocking the cell cycle and inducing apoptosis, and its mechanism of inducing apoptosis may be related to the mitochondria-mediated apoptosis pathway.
6.Effect of Atorvastatin on Plasma MicroRNA-143/145 Expression in Patients With Stable Angina Pectoris
Bei ZHAO ; Lifeng LIU ; Yingqi LIU ; Li LIU ; Jingtao ZHAO ; Li ZHOU ; Huihui XIA ; Shouli WANG
Chinese Circulation Journal 2014;(12):972-975
Objective: To investigate the effect of atorvastatin on plasma microRNA-143/145 expression in patients with stable angina pectoris (SAP).
Methods: A total of 74 SAP patients taken atorvastatin at ifrst time were enrolled in this study, the patients were assigned into 2 groups by the dose of medication:Low dose group, the patients received atorvastatin 20 mg/day, n=36 and Moderate dose group, the patients received atorvastatin 40 mg/day, n=38. Plasma levels of LDL-C and microRNA-143/145 were examined before medication and at 1 month, 12 months after medication respectively. The patients were further divided into another 2 groups by plasma levels of LDL-C:Reach the standard group, plasma LDL-C<2.60 mmol/L and Not reach the standard group, plasma LDL-C≥2.60 mmol/L. Plasma levels of microRNA-143/145 were measured and compared between 2 groups.
Results: ① Compared with baseline condition, plasma levels of microRNA-143/145 were increased in both groups after medication, P<0.001 the levels in Moderate dose group were higher than those in Low dose group at both 1 month and 12 months of time points.②Plasma levels of LDL-C were decreased with medication in both groups, P<0.05. Micro RNA-143/145 expression was similar between Reach the standard group and Not reach the standard group, P>0.05.
Conclusion: Atorvastatin could up-regulate plasma microRNA-143/145 expression, which was not related to lipid-decreasing effect.
7.Studies on Mycoplasma genitalium infection in patients with chronic non-bacterial prostatitis
Shu-Xia ZHONG ; Shan-Shan LI ; Hua ZHENG ; Xiao-Hong LI ; Bei-Lei YUN ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
Objective To study the role of Mycoplasma genitalium(Mg)infection in chronic non- bacterial prostatitis.Methods Culture and two PCR systems were performed on prostatic secretion specimens from 487 patients with chronic non-bacterial prostatitis and 75 health men in Changchun regions. Results positive rate of Mg in the prostatic secretion from the 487 patients was 7.39% (36/487);In the controls,the positive rate of Mg was 1.33%(1/75).The differences between the two groups were statistically significant (X~2=3.88,P
8.Comparison of diffusion tensor imaging and proton MR spectroscopy in the posterior cingulate of patients with Alzheimer disease
Bei DING ; Huawei LING ; Xia LI ; Tao WANG ; Huan ZHANG ; Weimin CHAI ; Kemin CHEN
Chinese Journal of Radiology 2009;43(5):495-499
Objective To compare 1HMRS and DTI findings of Alzheimer disease (AD) patients and normal elderly controls. Methods Fifteen mild AD patients, 20 moderate to severe AD patients and 20 aging controlled normal subjects (CN) were recruited. MRS imaging and DTI were performed on a 1.5 T MRI scanner. A ROI was positioned in the posterior part of the cingulate. MRS data were processed and the metabolite ratios were estimated, including the ratios of NAA/Cr, Cho/Cr, mI/Cr. Comparing with the axial MRS location, we chose the same level to posit the ROIs on both sides of the posterior cingulated fibers on fractional anisotropy map (FA) and mean diffusivity map (MD). Mean spectroscopy data and DTI values for each groups were analysed with Mann-Whitney U non parametric test. Correlations between MRS and DTI values for AD groups were estimated using partial correlations test controlling for the age related bias. Results Compared to normal aging groups, mild AD group showed a significantly lower FA value in the left side of posterior cingulum bundle (0. 549±0. 056 vs 0. 517±0. 058,Z =2. 014,P <0. 05). Whereas, moderate to severe group versus mild AD group revealed significantly elevated MI) value and a decrease in FA value in the right side of posterior cingulate ( FA 0. 517 ± 0. 059 vs 0. 432 ± 0. 073, Z = 3. 216, P < 0. 01 ; MD (0.726±0.041) × 10-3 mm2/s vs (0.761±0.057) × 10-3 mm2/s,Z = 1.970,P <0.05) . Obvious increasing mI/Cr ratio was found in mild AD group ( 0. 61 ± 0. 07 vs 0. 68 ± 0. 12, Z = 2. 911, P < 0. 01 ). NAA/Cr ratio showed gradually decrease in AD groups. Partial correlations analysis revealed a positive correlation between ml/Cr ratio and left posterior cingulated FA value in mild AD group ( r = 0. 586, P < 0. 05) and negative correlation between NAA/Cr and MD value in the right side of posterior cingulated region ( r = - 0. 505, P < 0. 05 ). Conclusions These findings suggested that there were different regional and temporal pattern in different course of AD disease, resulting from axonal loss or gliosis. Combining MRS with DTI alternations could be a better potential indicator and could better explain the pathological changes in AD progression.
9.Comparison of therapeutic effects of implant internal fixation for the treatment of Sanders Ⅱ calcaneal fractures after poking and open reduction
Hao XIONG ; Wei LIU ; Weiwen LIN ; Xiongchao XIA ; Bei LI ; Caiqiong OU ; Maosong LAI ; Ruiliang HUANG
Chinese Journal of Tissue Engineering Research 2013;(26):4919-4925
10.3969/j.issn.2095-4344.2013.26.023
10.Investigation on health policy and policy related behaviors in community nurses
Bei ZHANG ; Yinping ZHANG ; Ruobo LIANG ; Jurong LI ; Ruyi XIA ; Lulu ZHANG ; Xinshuang ZHAO
Chinese Journal of Practical Nursing 2014;30(20):5-8
Objective We aimed to investigate the community nurses' awareness of health policy,related behaviors and paths of learning.Methods A total of 150 community nurses were enrolled in this study by answering a self-completion questionnaire.The survey contents included general conditions of nurses,awareness,related behaviors and paths of learning of community health policy.Results 91.7% of the respondents believed that heahh policy was important or very important,but only 10.4% of the respondents understood health policy fully.71.5% said that they would lecture patients health policy actively and 70.8% would lecture their colleagues actively.51.4% respondents would like to take part in policy-making if they were allowed.Education background,professional title and the awareness of health policy were factors of community nurses' policy related behaviors.They learned health policy by working meeting,training and documents.Conclusions It is necessary to improve their school record and adjust their professional title,to strengthen the publicity of health policy by broaden channels of publicity.By these,community nurses' awareness of health policy would be improved,their policy related behaviors would be promoted.Further,the health policy would be well delivered and implemented.