NMR分析胶质瘤细胞系CHG5和U87的代谢轮廓

胶质瘤是中枢神经系统最常见的恶性肿瘤,其发病原因和恶性进展机制均不明确,给胶质瘤的临床诊疗带来很大困难.胶质瘤恶性进展伴随着癌细胞代谢改变,但可以反映胶质瘤恶性进展的代谢物信息和分子机制目前仍不清楚.该研究以两种不同恶性程度的胶质瘤细胞系CHG5和U87为研究对象,用基于NMR的代谢组学方法分析这两种胶质瘤细胞系的代谢轮廓,寻找差异性代谢物.结果表明CHG5和U87的代谢轮廓存在明显差异.在高恶性程度的U87细胞中,柠檬酸盐(citrate)等15种代谢物有上升趋势;而乳酸盐(lactate)、牛磺酸(taurine)等6种代谢物有下降趋势.以上结果表明这些差异性代谢物可能与胶质瘤细胞的恶性特性密切相关.这些从胶质瘤细胞系获取的代谢信息将成为临床标本代谢组学研究的重要前提和有益补充.     

肝肿瘤细胞HepG2代谢指纹及代谢足迹的NMR分析

细胞代谢特征的分析是认识细胞生物化学过程物质基础的一个关键点. 该文使用培养72 h的肝肿瘤细胞HepG2为模型,使用一维与二维核磁共振谱学分析方法, 分析了该细胞本身及其培养液中代谢物的组成,确定了50余种覆盖三羧酸循环、糖酵解、氨基酸合成、脂肪酸与胞膜代谢、嘌呤与嘧啶代谢等多个代谢途径的代谢物,发现细胞本身与培养基中代谢物组成能够分别提供“细胞代谢指纹”与“细胞代谢足迹”等互补性信息. 同时发现此方法可用于研究植物次生代谢物槲皮素对肝肿瘤细胞HepG2代谢的影响. 结果表明,核磁共振波谱技术是分析细胞代谢组特征和研究药物对细胞代谢影响规律的有效手段.     

热纤梭菌中心代谢产物的归属及初步代谢组分析

热纤梭菌(Clostridium thermocellum)是一种能够高效利用木质纤维素的嗜热厌氧革兰氏阳性菌,是目前利用整合生物加工技术生产纤维素乙醇的最重要的候选菌株之一.代谢物组可以反映细胞和环境因子相互作用,是系统生物学的重要组成部分.目前,尚无使用核磁共振(NMR)方法对热纤梭菌的代谢组进行研究的报道,而对热纤梭菌代谢物的归属是进行代谢组研究的基础.该文通过1H NMR和2D NMR技术,对热纤梭菌的中心代谢产物进行了归属.在归属过程中,发现了若干特殊的重要代谢产物,包括纤维糊精、磷酸烯醇式丙酮酸、赤藓糖-4-磷酸等.这些代谢物对热纤梭菌的胞内代谢具有特殊的意义.利用已归属的代谢物,对热纤梭菌的野生型和乙醇耐受菌株进行初步的代谢组分析表明,热纤梭菌乙醇耐受性的获得,可能与纤维二糖主动合成纤维糊精途径、非氧化磷酸戊糖途径的增强及糖酵解路径的抑制密切相关.     

NMR 分析胶质瘤细胞系CHG5 和U87 的代谢轮廓

胶质瘤是中枢神经系统最常见的恶性肿瘤，其发病原因和恶性进展机制均不明确，给胶质瘤的临床诊疗带来很大困难．胶质瘤恶性进展伴随着癌细胞代谢改变，但可以反映胶质瘤恶性进展的代谢物信息和分子机制目前仍不清楚．该研究以两种不同恶性程度的胶质瘤细胞系CHG5 和U87 为研究对象，用基于NMR 的代谢组学方法分析这两种胶质瘤细胞系的代谢轮廓，寻找差异性代谢物．结果表明CHG5 和U87 的代谢轮廓存在明显差异．在高恶性程度的U87 细胞中，柠檬酸盐(citrate)等15 种代谢物有上升趋势；而乳酸盐(lactate)、牛磺酸(taurine)等6 种代谢物有下降趋势．以上结果表明这些差异性代谢物可能与胶质瘤细胞的恶性特性密切相关．这些从胶质瘤细胞系获取的代谢信息将成为临床标本代谢组学研究的重要前提和有益补充．     

SIRT7基因低表达胶质瘤细胞株代谢特征的NMR分析

肿瘤是一种代谢疾病，癌基因表达对肿瘤细胞代谢的影响是目前肿瘤研究的热点之一.本文利用基于核磁共振氢谱（¹H NMR）的代谢组学方法对癌基因SIRT7低表达胶质瘤细胞株的代谢特征进行分析，寻找与SIRT7基因表达相关的特征性代谢物和代谢通路.分析结果表明，SIRT7基因低表达组与对照组细胞的代谢轮廓存在显著性差异，其细胞水溶性萃取物中有22种代谢物浓度发生明显变化.与对照组相比，SIRT7基因低表达胶质瘤细胞株中乳酸、甘氨酸、谷氨酸等12种代谢物浓度升高；缬氨酸、亮氨酸、赖氨酸等10种代谢物浓度降低.通路富集分析提示氨酰-tRNA生物合成、氨基酸代谢等代谢通路与SIRT7低表达密切相关.以上结果为进一步阐明癌基因SIRT7调控胶质瘤细胞代谢的作用机制提供了理论依据.     

乌头碱对大鼠尿样代谢产物影响的

将NMR谱和主成分分析(principal component analysis, PCA)相结合用于研究不同剂量下乌头碱（aconitine, AC）急性毒性作用后大鼠尿样的代谢特征. 结果表明灌胃给药后代表能量代谢的柠檬酸（citrate）,2-酮戊二酸（2-oxoglutarate）和顺乌火酸（cis-aconitate）,反应肠道菌群代谢状况的马尿酸盐（hippurate）和苯乙尿酸（phenylacetylglycine）出现异常,并且这些变化在给药后0~8 h达到最大,16 h后基本恢复正常. 且给药剂量越大代谢物异常越明显,需要的恢复时间也越长. 实验中发现2-oxoglutarate的异常增加可能与AC的神经毒性有关,或与2-oxoglutarate的代谢异常有关. 这些信息为进一步阐明AC的毒理学机制和临床毒性监测都会有一定的帮助.     

乌头碱对大鼠尿样代谢产物影响的NMR研究

将NMR谱和主成分分析(principal component analysis,PCA)相结合用于研究不同剂量下乌头碱(aconitine,AC)急性毒性作用后大鼠尿样的代谢特征.结果表明灌胃给药后代表能量代谢的柠檬酸(citrate),2-酮戊二酸(2-oxoglutarate)和顺乌火酸(cis-aconitate),反应肠道菌群代谢状况的马尿酸盐(hippurate)和苯乙尿酸(phenylacetylglycine)出现异常,并且这些变化在给药后0～8 h达到最大,16 h后基本恢复正常.且给药剂量越大代谢物异常越明显,需要的恢复时间也越长.实验中发现2-oxoglutarate的异常增加可能与AC的神经毒性有关,或与2-oxoglutarate的代谢异常有关.这些信息为进一步阐明AC的毒理学机制和临床毒性监测都会有一定的帮助.     

基于NMR的4T1荷瘤小鼠脾脏受不同给药方式影响的代谢组学研究

联合阿霉素和紫杉醇治疗肿瘤的策略在临床上已有广泛应用,为了进一步借助纳米载体对肿瘤组织的靶向优势,以良好生物相容性纳米载体包埋阿霉素和紫杉醇的新型给药方式得以发展.虽然前期研究结果显示纳米给药方式相对传统给药治疗有更好的抑制肿瘤生长的效果,但是它们在分子代谢水平对机体的影响并没有被确切研究.本文采用4T1荷瘤乳腺癌模型小鼠,通过比较荷瘤小鼠分别在不处理、传统给药方式和通过纳米载体给药处理的条件下与健康小鼠脾脏代谢组的差异,发现传统给药处理对小鼠肠道微生物相关的代谢紊乱有较好的恢复效果,而通过纳米载体的给药处理可以更好的调节荷瘤小鼠的葡萄糖和延胡索酸等与能量供应相关的代谢途径.此外,这两种给药方式都不能有效改善荷瘤小鼠显著的脾脏肿大症状,也不能改善脾脏免疫调节功能和供应肿瘤生长而导致的氨基酸、有机酸、核酸和胆碱等一系列代谢物变化.这可能与脾脏对外来肿瘤的免疫反应较为激烈而有效药物治疗时间较短有关.该研究为更全面认识荷瘤小鼠脾脏代谢表型变化和不同给药方式对脾脏代谢组的影响提供了基础数据.     

电离辐射对小鼠肾脏影响的代谢组学研究

电离辐射对生物体影响是目前生物医学领域研究的热点之一.本文利用基于核磁共振氢谱（¹H NMR）的代谢组学技术研究来自暴露于不同的电离辐射的C57BL/6J小鼠的肾脏的水溶性萃取物,并试图寻找电离辐射对小鼠肾脏影响的特异性生物标志物.分析结果表明,辐射组可与相应的对照组明显区分.在所发现的75个代谢物中,有6种代谢物的浓度发生显著性变化,分别为2-氨基丁酸、3-羟基丁酸、高丝氨酸、1,3-丙二胺、β-丙氨酸和抗坏血酸.这些代谢物可能成为高剂量电离辐射对生物体肾脏代谢影响的标志物.     

自旋弛豫时间测定对固体代谢物分子力学性质的研究

自旋弛豫时间测定是研究固体物质分子动力学性质的有效手段,已广泛应用于植物糖、氨基酸、维生素、肾上腺素以及睾丸激素等多类代谢物分子动力学性质的研究中. 近来上述方法也被用于其他代谢物分子动力学性质与其多晶型相变的关系以及对代谢物结构依赖性的研究. 该文将在介绍自旋弛豫时间及其测定方法的基础上,着重对自旋弛豫时间测定研究植物糖、氨基酸及其衍生物分子动力学性质的应用工作进行讨论.     

基于机器学习和可见光光谱的冬小麦叶片氮积累量估算

在拔节期分4次采集了6个施氮水平下的冬小麦冠层图像,同步进行取样并以凯氏定氮法测定叶片含氮量,进而计算叶片氮积累量。利用随机森林算法分割冠层图像之后提取冠层覆盖度、可见光波段(R, G和B)三个分量及其衍生的5个色彩指数。以冠层覆盖度外加色彩指数、色彩分量的两种非线性回归,以及人工神经网络、支持向量回归、随机森林3种机器学习算法建立了冬小麦叶片氮积累量的估算模型。结果表明利用色彩指数的非线性回归模型的估算精度稍低于其他方法,而随机森林算法的拟合精度最高,但存在明显的过拟合现象。其他三种方法,即以冠层覆盖度及色彩分量为输入变量的非线性回归、支持向量回归和人工神经网络方法,均具有较高的拟合精度和泛化性能。     

运用光谱参数冠层覆盖度建立作物长势及氮营养状态模型

为了探索运用数码照片中光谱(红、绿、蓝)的像素计算得到的冠层覆盖度(canopy cover, CC)对玉米长势及氮素营养状态进行非破坏性监测的技术。通过获取玉米冠层的数码照片图像,定量化数码照片色彩参数与作物叶面积指数(leaf area index, LAI)、冠层干重(shoot dry matter weight, DM)、叶片氮素含量(leaf nitrogen content percentage, N%)之间的关系。试验于2012年和2013年在中国农业科学院试验田进行,运用基于Visual Basic Version 6.0研发的玉米冠层图像分析系统,分析了玉米品种中单909在3个氮素水平条件下分别于9叶展时期、抽雄期和灌浆期的CC、11种色彩指数与植株LAI,DM,N%及产量之间的相关性,并对相关性显著的指标进行了拟合与建模。结果表明,CC与LAI(r=0.93, p＜0.01),DM(r=0.94, p＜0.01),N%(r=0.82, p＜0.01)之间均达到了极显著水平;用CC估算LAI,DM和N%的模型均为幂函数,方程式分别是y=3.281 2x0.763 9,y=283.658 1x0.553 6,y=3.064 5x0.932 9;用与建模相独立的数据对模型验证,结果表明,CC估算LAI模型的实测值与模拟值基于1∶1直线的R2,RMSE和RE分别是0.996,0.035和1.46%;CC估算DM模型的R2,RMSE和RE分别是0.978,5.408 g和2.43%;CC估算N%模型的R2,RMSE和RE分别是0.990,0.054和2.62%。综上所述,模型能够较准确的通过CC估算不同氮肥水平条件下玉米9叶展时期、抽雄期和灌浆期的LAI,DM与N%,表明应用数码相机的光谱信息可实现对玉米生长过程中的生长状况及氮素营养状态进行实时无损快速监测与预测。     

Pressure Effects on Spectra of Tunable Laser Crystal GSGG:Cr3+ Ⅳ:Pressure-Induced Shifts of R1 Line, R2 Line, and U Band at 300 K

By means of both a theory for pressure-induced shifts (PS) of energy spectra and a theory for shifts of R2 line, and U band of GSGG:Cr3+ at 300 K have been calculated, respectively. The calculated results are in good agreement with all the experimental data. Their physical origins have also been explained. It is found that the mixingdegree of \t22(3T1)e4T2> and \t322E> base-wavefunctions in the wavefunctions of R1 level of GSGG:Cr3+ at 300 K is remarkable under normal pressure, and the mixing-degree rapidly decreases with increasing pressure. The change of the and the PS of R1 line (or R2 line) due to EPI are quite different. It is the combined effect of them that gives rise to the total PS ofR1 line (or R2 line). In the range of about 15 kbar ～ 45 kbar, the mergence and/or order-reversal between t22(3T1)e4T2 levels and t32 2 T1 levels take place, which cause the fluctuation of the rate of PS for t22(3T1)e4T2 (or t3 2 2T1)with pressure. At 300 K, both the temperature-dependent contribution to R1 line (or R2 line or U band) from EPI and the temperature-independent one are important.     

A comparative study of three cryopreservation protocols for effective storage of in vitro-grown mint (Mentha Spp.)

This study was designed to determine the response of diverse mint genotypes to three commonly used cryopreservation techniques. Four mints [Mentha x piperita nothosubsp. citrata (Ehrh.) Briq.; M. canadensis L.; M. australis R. Br, and M. cunninghamii Benth] were cryopreserved using three protocols: controlled rate cooling (CC), encapsulation dehydration (ED) and PVS2 vitrification (VIT). Regrowth of mint species following controlled rate cooling (93 percent) was significantly (P < 0.0001) better than encapsulation dehydration (71 percent) and vitrification (73 percent). All four genotypes responded well to the controlled rate cooling protocol but there was some variability with the other two protocols. Genotype specific response to the individual protocols showed that there were significant differences in the recovery of Mentha x piperita nothosubsp. citrata and M. australis with CC > VIT > ED. There were also significant differences in the recovery of M. cunninghamii and M. canadensis, with CC and ED significantly better than VIT. Regrowth of the shoot tips of these mints ranged from 60 percent to 95 percent for all but one treatment. The overall results of this study compare favorably to other techniques. These improved results may be due to a combination of favorable growth conditions, cold acclimation and recovery medium. Controlled rate cooling was the most successful technique for the storage of these diverse mint genotypes; however recovery of shoot tips from VIT and ED was high and these techniques could also be used for cryogenic storage of mint germplasm.     

O+NH反应在3A″和1A″势能面上的量子含时波包动力学研究

对于O+NH反应,在3A″和1A″势能面(Guadagnini R,Schatz G C,Walch S P.Global potential energy surface for the lowest 1A′,3A″,and 1A″states of HNO [J].J.Chem.Phys.,1995,10:774)上,我们运用coupled state or centrifugal sudden (CS)近似和close coupling or Coriolis coupled (CC)方法进行了量子动力学计算.通过比较两种方法得到的总的反应几率,我们发现对于两个势能面上的标题反应,CS近似是失效的.我们还讨论了用CS和CC方法得到的速率常数,并进行了结果比较.     

Recovery and characterisation of hybrid firs (Abies alba x A. cephalonica, Abies albax A. numidica) embryogenic tissues after cryopreservation

Embryogenic tissues of hybrid firs were cryopreserved using a slow freezing protocol. The procedure involved preculture of tissues for 24, 48 or 72 h in media with different sorbitol concentrations (0.4 or 0.8 M) and addition of 5% (v/v) DMSO as cryoprotectant. The cell lines tested withstood cryopreservation, even though tissue regrowth after thawing was dependent on treatment and cell line. For cell line AN72, regrowth was 100% for all experimental conditions tested. With cell line AC78, regrowth was 100% except after shorter pretreatment durations, which produced 83% and 86% regrowth for 0.4 M and 0.8 M sorbitol pretreatment, respectively. Cell lines AC1 and AC4 were more sensitive to cryopreservation with 37.5 to 100% regrowth, respectively. Growth parameters evaluated 3 months after cryopreservation showed cell line and treatments effects. In most cases, cryopreservation had no negative effect on growth of tissues. Statistically significant differences in fresh mass accumulation were found for four samples out of 24 investigated, although growth increase of these tissues still reached 79.4-84.6%, compared with non-cryopreserved ones (100% increase). Maturation capacity and genetic fidelity were studied in tissues whose growth was not negatively influenced by cryopreservation. Maturation capacity of embryogenic tissues cryopreserved using the optimal protocol was comparable to that of non-frozen controls. RAPD analysis of 88 genomic regions per cell line did not reveal any changes in genetic fidelity of cryopreserved tissues compared to non-cryopreserved controls.     

Critical current degradation behaviors of ReBCO coated conductor tapes under pressurized liquid nitrogen

In the case of 2G coated conductor (CC) tapes, it has been reported that thin–thick CC tapes with IBAD substrate showed a superior electromechanical property even at smaller bending radius compared with the cases of 1G BSCCO tapes. Considering the application of CC tapes it is significant to evaluate the transport property under operating environment, because CC tapes might experience a change in operating pressure that can affect its current carrying capacity due to temperature variation and deformation. This study was focused on the I_c degradation behavior in bent CC tapes under pressurized liquid nitrogen. Differently processed YBCO and SmBCO CC tapes with IBAD substrate are used as samples. The bending strain characteristics at elevated pressure levels were evaluated by using the ρ-shaped sample holder which can induce different bending strain values at pressured state. Depressurization and thermal cycling were performed to check the reversibility of I_c in CC tapes. Vacuuming tests were also carried out to investigate the characteristics of I_c at different LN₂ temperature levels.     

基于共聚焦显微拉曼光谱的毛竹细胞结构和成分研究

采用共聚焦显微拉曼光谱对毛竹薄壁细胞、薄壁纤维过渡细胞和纤维细胞进行研究。通过构建偏最小二乘(PLS)定量区分模型来对这三种细胞中的差异进行分析,结果表明,该区分模型的建模和交互验证决定系数(R2)分别为0.810和0.800,均方根误差(RMSE)分别为0.323和0.332。根据这一模型的回归系数,发现三种细胞的区别主要体现在1 095,1 319和1 636 cm-1三个波数,这三个波数分别为纤维素、半纤维素和木质素的指纹特征峰。以这三个波数为自变量建立多元线性回归(MLR)模型,该回归模型的建模和交互验证决定系数(R2)分别为0.644和0.643,均方根误差(RMSE)分别为0.442和0.443,表明三种细胞在这三个波数处存在明显的差异。对小波变换基线消除后的拉曼光谱信号进行化学成像分析,结果显示,纤维素微纤维与纤维轴成一个很大的角度,这一结构有利于提高细胞的弹性模量和硬度。半纤维素和纤维素微纤维通过氢键相连,并在范德华力的作用下紧密地结合在一起,因此在拉曼化学成像中可以看到半纤维素和纤维素有相似的分布规律。三种细胞的细胞角和胞间层都高度的木质化,从细胞壁外层到内层木质化程度逐渐降低,表明细胞壁的木质化从细胞角和胞间层开始,且木质化程度并不完全。     

NMR-based metabonomic analyses of the effects of ultrasmall superparamagnetic particles of iron oxide (USPIO) on macrophage metabolism

The metabonomic changes in murine RAW264.7 macrophage-like cell line induced by ultrasmall superparamagnetic particles of iron oxides (USPIO) have been investigated, by analyzing both the cells and culture media, using high-resolution NMR in conjunction with multivariate statistical methods. Upon treatment with USPIO, macrophage cells showed a significant decrease in the levels of triglycerides, essential amino acids such as valine, isoleucine, and choline metabolites together with an increase of glycerophospholipids, tyrosine, phenylalanine, lysine, glycine, and glutamate. Such cellular responses to USPIO were also detectable in compositional changes of cell media, showing an obvious depletion of the primary nutrition molecules, such as glucose and amino acids and the production of end-products of glycolysis, such as pyruvate, acetate, and lactate and intermediates of TCA cycle such as succinate and citrate. At 48 h treatment, there was a differential response to incubation with USPIO in both cell metabonome and medium components, indicating that USPIO are phagocytosed and released by macrophages. Furthermore, information on cell membrane modification can be derived from the changes in choline-like metabolites. These results not only suggest that NMR-based metabonomic methods have sufficient sensitivity to identify the metabolic consequences of murine RAW264.7 macrophage-like cell line response to USPIO in vitro, but also provide useful information on the effects of USPIO on cellular metabolism.     

Combination of sonodynamic with temozolomide inhibits C6 glioma migration and promotes mitochondrial pathway apoptosis via suppressing NHE-1 expression

Temozolomide (TMZ) was used for clinical postoperative or non-surgical chemotherapy patients. However, its effect remains unsatisfactory and gradually discovered that the presence of chemoresistance. To explore more effective therapy using TMZ, we investigate the effects of combination of application of TMZ together with Sonodynamic therapy (SDT), which is based on the ultrasonic activation of a sonosensitizer, with low toxicity, noninvasive, deeper penetrability and a promising approach for treating malignant glioma by inducing apoptosis on glioma cells in vitro. Sodium–hydrogen exchanger isoform 1 (NHE1), which enable glioblastoma cells to escape TMZ-mediated toxicity via increased H⁺ extrusion and affect the apoptosis effect on C6 glioma cells in vitro. The C6 cells survival rate and time point of TMZ resistance were tested by the Cell Counting Kit-8 (CCK8) viability assay. Western blot analysis results showed that the expression of NHE1 and matrix metalloproteinase-2 (MMP-2) protein obviously decreased by TMZ + SDT. Meanwhile, combined treatments enhanced the expression of mitochondrial pathway apoptosis proteins, as well as suppressed MMP-2 to weaken the migration ability in TMZ-resistant C6 cell line. These results provided the first evidence that the sensitivity of TMZ chemotherapy in resistant malignant glioma may be improved by SDT.