Metabolic heterogeneity of gastric cancer cell lines

OBJECTIVE Gastric cancer is one of the most common malignant tumors,and the incidence 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.

Slow-gathering areas of rivers, lakes, and reservoirs: Definition, estimation, and restoration

Recently,humans have been facing serious water pollution problems that aggravate water resources shortages.By 2025,two-thirds of the world's population is likely to live in countries with moderate or severe water shortages(Azizullah et al.,2011).Rivers,lakes,and reservoirs are important sources of water.Globally,there are approximately 8.5 million rivers(Linke et al.,2019),21.15 million natural lakes,and 1.427 million large reservoirs(Messager et al.,2016).Many of them have been contaminated due to urbanization,industrialization,and agricultural activities.In particular,all rivers,lakes,and reservoirs have their own slow-gathering areas(SGAs)where the input contaminants are easily stopped and deposited due to their non-open morphologies and low flow speeds.

无创高频振荡通气对新生儿呼吸衰竭外周血CC16与KL-6表达的影响

目的探究无创高频振荡通气对新生儿呼吸衰竭外周血Clara细胞蛋白16(CC16)与Ⅱ型肺泡表面抗原6(KL-6)表达的影响。方法选取2015年1月—2017年12月于邯郸市第一医院就诊的208例呼吸衰竭新生儿作为研究对象,依照随机数字表法将患儿分成观察组和对照组。观察组采用无创高频振荡通气治疗,对照组采用同步间歇指令通气治疗,比较两组的临床疗效,并检测两组患儿机械通气后0、24和72 h血气指标、治疗前后血清CC16及KL-6的变化和并发症的发生情况。结果观察组总有效率高于对照组(P<0.05),观察组临床疗效优于对照组(P<0.05)。观察组机械通气后的氧合指数、呼吸指数、吸入气氧浓度(FiO2)和动脉血二氧化碳分压(PaCO2)低于对照组(P<0.05),而动脉血氧分压(PaO2)、PaO2/FiO2高于对照组(P<0.05)。观察组与对照组治疗后72 h血清CC16和KL-6均低于治疗前(P<0.05)。观察组治疗后72 h的CC16与KL-6低于对照组(P<0.05)。观察组机械通气时间、住院时间低于对照组(P<0.05)。观察组呼吸机相关性肺炎、并发症总发病率低于对照组(P<0.05)。结论无创高频振荡通气治疗新生儿呼吸衰竭的疗效更佳,能够显著改善患儿血气指标,减轻炎症反应,降低患儿CC16、KL-6水平。

高海拔地区藏族和汉族乳腺浸润性导管癌凋亡相关蛋白及凋亡指数的研究

目的探讨世居青海的藏汉族人乳腺浸润性导管癌组织中Bcl-2、Bax及Caspase-3和凋亡指数(AI)的表达差异。方法采用免疫组织化学法检测藏族和汉族各60例乳腺浸润性导管癌中Bcl-2、Bax及Caspase-3蛋白的表达;采用原位DNA缺口末端标记法检测凋亡指数。结果藏族、汉族乳腺浸润性导管癌组织中Bcl-2的表达率分别为52.2%和38.3%,Bax表达率分别为76.7%和61.7%,Caspase-3表达率分别为60.0%和58.3%,藏族、汉族乳腺浸润性导管癌组织中AI阳性率分别为61.2%和60.0%,差异均无统计学意义(P>0.05)。结论世居青海的藏族和汉族乳腺浸润性导管癌组织中Bcl-2、Bax及Caspase-3的表达无差异,且乳腺浸润性导管癌组织中AI表达比较也无差异。

Anti-esophagus cancer activity and mechanism of DN3,a novel natural diterpenoid derivative,as a dual inhibitor of glycolysis and oxidative phosphorylation

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.

Antitumor activityand mechanism of JD20,a newly synthetic analog of natural ent-kaurane diterpenoid

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.

Immunogenicity noninferiority study of 2 doses and 3 doses of an Escherichia coli-produced HPV bivalent vaccine in girls vs.3 doses in young women

A new HPV-16/18 bivalent vaccine expressed by the Escherichia coli has been proven to be efficacious in adult women.A randomized,immunogenicity noninferiority study of this candidate vaccine was conducted in December 2015 in China.Girls aged 9–14 years were randomized to receive 2 doses at months 0 and 6(n=301)or 3 doses at months 0,1 and 6(n=304).Girls aged 15–17 years(n=149)and women aged 18–26 years(n=225)received 3 doses.The objectives included noninferiority analysis of the IgG geometric mean concentration(GMC)ratio(95%CI,lower bound>0.5)to HPV-16 and HPV-18 at month 7 in girls compared with women.In the per-protocol set,the GMC ratio of IgG was noninferior for girls aged 9–17 years receiving 3 doses compared with women(1.76(95%CI,1.56,1.99)for HPV-16 and 1.93(95%CI,1.69,2.21)for HPV-18)and noninferior for girls aged 9–14 years receiving 2 doses compared with women(1.45(95%CI,1.25,1.62)for HPV-16 and 1.17(95%CI,1.02,1.33)for HPV-18).Noninferiority was also demonstrated for neutralizing antibodies.The immunogenicity of the HPV vaccine in girls receiving 3 or 2 doses was noninferior compared with that in young adult women.

Changing Tendency Analysis of Chinese Normal Male's Semen Quality in Recent 25 Years:Samples from Chinese Documents

Objective To analyze the trend of change in semen quality of Chinese healthy men over recent 25 years. Methods A total of 115 reports were collected on quality inspection of semen of healthy Chinese men between 1985-2009 through literature search, which involved 23 126 persons from 69 counties and cities. Trend of change in semen parameters was analyzed by linear regression model and rank sum test. Results Sperm concentration and non-progressive motility （c%） declined, and the linear equation was of statistics value. Linear equations include y=2.968-0. 004x （P〈0.05） and y=3.204-0.026x （P〈0.05）. As the results of K-S rank sum test showed, the difference of sperm concentration and total sperm count in the three groups, i.e., the group for 1985-1989, the group for 1990-1999 and the group for 2000-2009 was of statistics value. Conclusion The semen concentration decline over recent 25 years （P〈0.05）. of healthy Chinese men appears a possible

Synthesis and antiproliferative activity of novel 4-substituted-phenoxy-benzamide derivatives

A series of novel 4-substituted-phenoxy-benzamide derivatives bearing an aryl cycloaliphatic amine moiety were synthesized and evaluated for antiproliferative activity against SW620, HT29 and MGC803 cancer cell lines in vitro. The pharmacological data demonstrated that the majority of target compounds exhibited moderate efficacy in HT29 and MGC803 cell lines. Compound 10 c showed promising inhibition of hedgehog（Hh） signaling pathway in an Hh-related assay. In addition, the superposition pattern of 10 c showed a good fit for a pharmacophoric model generated by Hh inhibitors and provided a basis for further structural optimization.

Wheat ear growth modeling based on a polygon

Visual inspection of wheat growth has been a useful tool for understanding and implementing agricultural techniques and a way to accurately predict the growth status of wheat yields for economists and policy decision makers.In this paper,we present a polygonal approach for modeling the growth process of wheat ears.The grain,lemma,and palea of wheat ears are represented as editable polygonal models,which can be re-polygonized to detect collision during the growth process.We then rotate and move the colliding grain to resolve the collision problem.A linear interpolation and a spherical interpolation are developed to simulate the growth of wheat grain,performed in the process of heading and growth of wheat grain.Experimental results show that the method has a good modeling effect and can realize the modeling of wheat ears at different growth stages.

基于大肠杆菌表达的重组九价人乳头瘤病毒(HPV 6/11/16/18/31/33/45/52/58型)疫苗免疫原性和安全性的Ⅱ期临床试验

人乳头瘤病毒(HPV)16/18型双价疫苗(大肠埃希菌)已上市并通过世界卫生组织预认证.本研究通过一项随机双盲Ⅱ期临床试验评估其第二代HPV6/11/16/18/31/33/45/52/58型九价疫苗在18~45岁健康女性中的免疫原性和安全性.来自江苏省东台市的627名女性志愿者1:1随机分配至九价疫苗组(313人)或对照组(314人),并按照0、1、6月接种程序分别接种三剂1.0 mL(270μg)九价疫苗或空白佐剂,主要终点为第7月时血清抗体阳转率和抗体水平.结果显示,几乎所有疫苗组符合方案人群在第7月时出现针对9种HPV型别的血清中和抗体阳转,仅2人未发生HPV11型阳转,1人未发生HPV52型阳转.九价疫苗组和对照组的总体不良事件发生率分别为80.8%和72.9%,绝大多数不良事件症状轻微并很快恢复,未发生疫苗相关严重不良事件.该研究证明候选九价疫苗具有良好的安全性和免疫原性,支持在更大人群中进一步进行效力研究.

Unsteady flow structures in centrifugal pump under two types of stall conditions

The stall is an unsteady flow phenomenon that always causes instabilities and low efficiency for pumps. This paper focuses on the unsteady flow structures and evolutions under two types of stall conditions in centrifugal pump impellers. Two centrifugal pump impellers, one with 6 and another with 5 blades, are considered and a developed large-eddy simulation method is adopted. The results show that the alternative stall occurs in the impeller with 6 blades, while, the rotating stall is observed in that with 5 blades. The flow structure and the pressure fluctuation characteristics are further analyzed. For the alternative stall, the stall cells are fixed relative to the impeller, but a large vortex in the stalled passage is always swaying. The outlet vortex is generated from it, and then develops and sheds periodically. For the rotating stall, the stall cells first occur in the suction side of the blade. With the growth of the stall cells, the block area gradually increases until the inlet region is almost blocked, then moves to the pressure side with a continuous decay. When the rotating stall occurs, the amplitude of the pressure fluctuation is much larger than that under the alternative stall condition. The propagation of the stall cells has a significant effect on the pressure fluctuations in the impeller.

Research progress on myocardial regeneration: what is new?

The regeneration capacity of cardiomyocytes(CMs)is retained in neonatal mouse hearts but is limited in adult mouse Myocardial infarction(MI)in adult hearts usually leads to the loss of large amounts of cardiac tissue,and then accelerates the process of cardiac remodeling and heart failure.Therefore,it is necessary to explore the potential mechanisms of CM regeneration in the neonates and develop potential therapies aimed at promoting CM regeneration and cardiac repair in adults.Currently,studies indicate that a number of mechanisms are involved in neonatal endogenous myocardial regeneration,including cell cycle regulators,transcription factors,non-coding RNA,signaling pathways,acute inflammation,hypoxia,protein kinases,and others.Understanding the mechanisms of regeneration in neonatal CMs after MI provides theoretical support for the studies related to the promotion of heart repair after MI in adult mammals.However,several difficulties in the study of CM regeneration still need to be overcome.This article reviews the potential mechanisms of endogenous CM regeneration in neonatal mouse hearts and discusses possible therapeutic targets and future research directions.

Metal-decorated defective BN nanosheets as hydrogen storage materials

Density functional theory computations were performed to investigate hydrogen adsorption in metaldecorated defective BN nanosheets. The binding energies of Ca and Sc on pristine BN nanosheets are much lower than the corresponding cohesive energies of the bulk metals; however, B vacancies in BN nanosheets enhance the binding of Ca and Sc atoms dramatically and avoid the clustering of the metal atoms on the surface of BN nanosheets. Ca and Sc strongly bind to defective BN nanosheets due to charge transfer between metal atoms and BN nanosheets. So-decorated BN nanosheets with B vacancies demonstrate promising hydrogen adsorption performances with a hydrogen adsorption energy of -0.19 - -0.35 eV/H2.