Correlations among Six Hormone-Induced Transcription Factors and the Alcohol Acyltransferase Gene in Apple

Qing-Qing Li,JIN SHENG,Pengcheng Li,Dapeng Li,Cheng-Chao Zheng,Dequan Li,Huairui Shu 한국식물학회 2012 Journal of Plant Biology Vol.55 No.4

Alcohol acyltransferase (AAT) plays an important role in ester biogenesis in ripening fruit. In apple, MdAAT2 is up-regulated in response to treatment with defense-related hormones. We cloned a novel MdAAT2 promoter via thermal asymmetric interlaced PCR (TAIL-PCR). Sequence analysis indicated the presence of salicylic acid (SA)- and methyl jasmonate (MeJA)-responsive elements in the promoter region. To examine further the operational mechanism for transcription factors (TFs) in regulating expression of the MdAAT2 promoter, we isolated and investigated six potential stress-induced TFs -- MdMYB1, MdMYB2, MdMYB6,MdERF1, MdERF2, and WRKY. These were monitored during various stages of fruit development as well as under several hormonal treatments. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that all of these TF genes, as well as MdAAT2, were detectable in the stigmas. Levels of expression by MdMYB1, MdMYB2, and MdERF1were significantly correlated with that by MdAAT2 during fruit ontogeny. Moreover, transcription of three TFs (MdMYB1,MdMYB6, and MdERF1) was significantly correlated with that of MdAAT2 upon exposure to ethephon, SA, or MeJA. Sequence analysis demonstrated that CAAT, CCAAT elements,and several MYB transcription factor binding sites (CNGTTmotif,AACCA, and CTAACCA elements) existed in the MdAAT2 promoter region. Using that analysis, we performed modeling of MdMYB1 and MdMYB6 based on PROTEINDNA complex1 and complex3 (PDB Accession Numbers 1H88 and 1H8A). There, similarities were found with the protein complex of CAAT or CCAAT consensus sequences. Thus, we propose that two TFs -- MdMYB1 and MdMYB6-- are directly or indirectly involved in the activation of MdAAT2 expression in apple fruit.

Simultaneous Determination of Glutamate, Glycine, and Alanine in Human Plasma Using Precolumn Derivatization with 6-Aminoquinolyl- N-hydroxysuccinimidyl Carbamate and High-Performance Liquid Chromatography

Qing Zhong Li,Qing Xian Huang,Shu Cui Li,Mei Zi Yang,Bin Rao 대한생리학회-대한약리학회 2012 The Korean Journal of Physiology & Pharmacology Vol.15 No.5

A simple, sensitive and reproducible high-performance liquid chromatography (HPLC) method has been validated for determining concentrations of glutamate, glycine, and alanine in human plasma. Proteins in plasma were precipitated with perchloric acid, followed by derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Simultaneous analysis of glutamate, glycine, and alanine is achieved using reversed-phase HPLC conditions and ultraviolet detection. Excellent linearity was observed for these three amino acids over their concentration ranges with correlation coefficients (r)>0.999. The intra- and inter-day precision were below 10%. This method utilizes quality control samples and demonstrates excellent plasma recovery and accuracy. The developed method has been successfully applied to measure plasma glutamate, glycine, and alanine in twenty volunteers.

Simultaneous Determination of Glutamate, Glycine, and Alanine in Human Plasma Using Precolumn Derivatization with 6-Aminoquinolyl-N-hydroxysuccinimidyl Carbamate and High- Performance Liquid Chromatography

Li, Qing Zhong,Huang, Qing Xian,Li, Shu Cui,Yang, Mei Zi,Rao, Bin The Korean Society of Pharmacology 2012 The Korean Journal of Physiology & Pharmacology Vol.16 No.5

A simple, sensitive and reproducible high-performance liquid chromatography (HPLC) method has been validated for determining concentrations of glutamate, glycine, and alanine in human plasma. Proteins in plasma were precipitated with perchloric acid, followed by derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Simultaneous analysis of glutamate, glycine, and alanine is achieved using reversed-phase HPLC conditions and ultraviolet detection. Excellent linearity was observed for these three amino acids over their concentration ranges with correlation coefficients (r)>0.999. The intra- and inter-day precision were below 10%. This method utilizes quality control samples and demonstrates excellent plasma recovery and accuracy. The developed method has been successfully applied to measure plasma glutamate, glycine, and alanine in twenty volunteers.

Study on mechanism of macro failure and micro fracture of local nearly horizontal stratum in super-large section and deep buried tunnel

Li, Shu-cai,Wang, Jian-hua,Chen, Wei-zhong,Li, Li-ping,Zhang, Qian-qing,He, Peng Techno-Press 2016 Geomechanics & engineering Vol.11 No.2

The stability of surrounding rock will be poor when the tunnel is excavated through nearly horizontal stratum. In this paper, the instability mechanism of local nearly horizontal stratum in super-large section and deep buried tunnel is revealed by the analysis of the macro failure and micro fracture. A structural model is proposed to explain the mechanics of surrounding rock collapse under the action of stress redistribution and shed light on the macroscopic analytical approach of the stability of surrounding rock. Then, some highly effective formulas applied in the tunnel engineering are developed according to the theory of mixed-mode micro fracture. And well-documented field case is made to demonstrate the effectiveness and accuracy of the proposed analytical methods of mixed-mode fracture. Meanwhile, in order to make the more accurate judgment about yield failure of rock mass, a series of comprehensive failure criteria are formed. In addition, the relationship between the nonlinear failure criterion and $K_I$ and $K_{II}$ of micro fracture is established to make the surrounding rock failure criterion more comprehensive and accurate. Further, the influence of the parameters related to the tension-shear mixed-mode fracture and compression-shear mixed-mode fracture on the propagation of rock crack is analyzed. Results show that ${\sigma}_3$ changes linearly with the change of ${\sigma}_1$. And the change rate is related to ${\beta}$, angle between the cracks and ${\sigma}_1$. The proposed simple analytical approach is economical and efficient, and suitable for the analysis of local nearly horizontal stratum in super-large section and deep buried tunnel.

Simultaneous Determination of Glutamate, Glycine, and Alanine in Human Plasma Using Precolumn Derivatization with 6-Aminoquinolyl- N-hydroxysuccinimidyl Carbamate and High-Performance Liquid Chromatography

Qing Zhong Li,Qing Xian Huang,Shu Cui Li,Mei Zi Yang,Bin Rao 대한약리학회 2012 The Korean Journal of Physiology & Pharmacology Vol.16 No.5

A simple, sensitive and reproducible high-performance liquid chromatography (HPLC) method has been validated for determining concentrations of glutamate, glycine, and alanine in human plasma. Proteins in plasma were precipitated with perchloric acid, followed by derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Simultaneous analysis of glutamate, glycine, and alanine is achieved using reversed-phase HPLC conditions and ultraviolet detection. Excellent linearity was observed for these three amino acids over their concentration ranges with correlation coefficients (r)>0.999. The intra- and inter-day precision were below 10%. This method utilizes quality control samples and demonstrates excellent plasma recovery and accuracy. The developed method has been successfully applied to measure plasma glutamate, glycine, and alanine in twenty volunteers.

Increased Serotonin Signaling Contributes to the Warburg Effect in Pancreatic Tumor Cells Under Metabolic Stress and Promotes Growth of Pancreatic Tumors in Mice

Jiang, Shu-Heng,Li, Jun,Dong, Fang-Yuan,Yang, Jian-Yu,Liu, De-Jun,Yang, Xiao-Mei,Wang, Ya-Hui,Yang, Min-Wei,Fu, Xue-Liang,Zhang, Xiao-Xin,Li, Qing,Pang, Xiu-Feng,Huo, Yan-Miao,Li, Jiao,Zhang, Jun-Feng Elsevier 2017 Gastroenterology Vol.153 No.1

<P><B>Background & Aims</B></P> <P>Desmoplasia and poor vascularity cause severe metabolic stress in pancreatic ductal adenocarcinomas (PDACs). Serotonin (5-HT) is a neuromodulator with neurotransmitter and neuroendocrine functions that contributes to tumorigenesis. We investigated the role of 5-HT signaling in the growth of pancreatic tumors.</P> <P><B>Methods</B></P> <P>We measured the levels of proteins that regulate 5-HT synthesis, packaging, and degradation in pancreata from Kras<SUP>G12D/+</SUP>/Trp53<SUP>R172H/+</SUP>/Pdx1-Cre (KPC) mice, which develop pancreatic tumors, as well as in PDAC cell lines and a tissue microarray containing 81 human PDAC samples. We also analyzed expression levels of proteins involved in 5-HT synthesis and degradation by immunohistochemical analysis of a tissue microarray containing 311 PDAC specimens, and associated expression levels with patient survival times. 5-HT level in 14 matched PDAC tumor and non-tumor tissues were analyzed by ELISA. PDAC cell lines were incubated with 5-HT and cell survival and apoptosis were measured. We analyzed expression of the 5-HT receptor HTR2B in PDAC cells and effects of receptor agonists and antagonists, as well as HTR2B knockdown with small hairpin RNAs. We determined the effects of 5-HT stimulation on gene expression profiles of BxPC-3 cells. Regulation of glycolysis by 5-HT signaling via HTR2B was assessed by immunofluorescence and immunoprecipitation analyses, as well as by determination of the extracellular acid ratio, glucose consumption, and lactate production. Primary PDACs, with or without exposure to SB204741 (a selective antagonist of HTR2B), were grown as xenograft tumors in mice, and SB204741 was administered to tumor-bearing KPC mice; tumor growth and metabolism were measured by imaging analyses.</P> <P><B>Results</B></P> <P>In immunohistochemical analysis of a tissue microarray of PDAC specimens, increased levels of TPH1 and decreased level of MAOA, which regulate 5-HT synthesis and degradation, correlated with stage and size of PDACs and shorter patient survival time. We found levels of 5-HT to be increased in human PDAC tissues compared with non-tumor pancreatic tissues, and PDAC cell lines compared with non-transformed pancreatic cells. Incubation of PDAC cell lines with 5-HT increased proliferation and prevented apoptosis. Agonists of HTR2B, but not other 5-HT receptors, promoted proliferation and prevented apoptosis of PDAC cells. Knockdown of HTR2B in PDAC cells, or incubation of cells with HTR2B inhibitors, reduced their growth as xenograft tumors in mice. We observed a correlation between 5-HT and glycolytic flux in PDAC cells; levels of metabolic enzymes involved in glycolysis, the phosphate pentose pathway, and hexosamine biosynthesis pathway increased significantly in PDAC cells following 5-HT stimulation. 5-HT stimulation led to formation of the HTR2B–LYN–p85 complex, which increased PI3K–Akt–mTOR signaling and the Warburg effect by increasing protein levels of MYC and HIF1A. Administration of SB204741 to KPC mice slowed growth and metabolism of established pancreatic tumors and prolonged survival of the mice.</P> <P><B>Conclusions</B></P> <P>Human PDACs have increased levels of 5-HT, and PDAC cells increase expression of its receptor, HTR2B. These increases allow for tumor glycolysis under metabolic stress and promote growth of pancreatic tumors and PDAC xenograft tumors in mice.</P>

SPON2 Promotes M1-like Macrophage Recruitment and Inhibits Hepatocellular Carcinoma Metastasis by Distinct Integrin–Rho GTPase–Hippo Pathways

Zhang, Yan-Li,Li, Qing,Yang, Xiao-Mei,Fang, Fang,Li, Jun,Wang, Ya-Hui,Yang, Qin,Zhu, Lei,Nie, Hui-Zhen,Zhang, Xue-Li,Feng, Ming-Xuan,Jiang, Shu-Heng,Tian, Guang-Ang,Hu, Li-Peng,Lee, Ho-Young,Lee, Su-J American Association for Cancer Research 2018 Cancer research Vol.78 No.9

<P>Matricellular protein SPON2 acts as an HCC suppressor and utilizes distinct signaling events to perform dual functions in HCC microenvironment.</P><P>Tumor-associated macrophages (TAM) represent key regulators of the complex interplay between cancer and the immune microenvironment. Matricellular protein SPON2 is essential for recruiting lymphocytes and initiating immune responses. Recent studies have shown that SPON2 has complicated roles in cell migration and tumor progression. Here we report that, in the tumor microenvironment of hepatocellular carcinoma (HCC), SPON2 not only promotes infiltration of M1-like macrophages but also inhibits tumor metastasis. SPON2-α4β1 integrin signaling activated RhoA and Rac1, increased F-actin reorganization, and promoted M1-like macrophage recruitment. F-Actin accumulation also activated the Hippo pathway by suppressing LATS1 phosphorylation, promoting YAP nuclear translocation, and initiating downstream gene expression. However, SPON2-α5β1 integrin signaling inactivated RhoA and prevented F-actin assembly, thereby inhibiting HCC cell migration; the Hippo pathway was not noticeably involved in SPON2-mediated HCC cell migration. In HCC patients, SPON2 levels correlated positively with prognosis. Overall, our findings provide evidence that SPON2 is a critical factor in mediating the immune response against tumor cell growth and migration in HCC.</P><P><B>Significance:</B> Matricellular protein SPON2 acts as an HCC suppressor and utilizes distinct signaling events to perform dual functions in HCC microenvironment.</P><P><B>Graphical Abstract:</B> http://cancerres.aacrjournals.org/content/canres/78/9/2305/F1.large.jpg. <I>Cancer Res; 78(9); 2305–17. ©2018 AACR</I>.</P><P><B>Graphical Abstract</B></P><P> [Figure]</P>

Glucose regulated protein 78 promotes cell invasion via regulation of uPA production and secretion in colon cancer cells

( Zong Wei Li ),( Lic Hao Zhang ),( Han Qing Li ),( Shu Hua Shan ),( Zhuo Yu Li ) 생화학분자생물학회(구 한국생화학분자생물학회) 2014 BMB Reports Vol.47 No.8

Glucose regulated protein 78 (GRP78) is frequently highlyexpressed in tumor cells, contributing to the acquisition of severalphenotypic cancer hallmarks. GRP78 expression is also positivelycorrelated with tumor metastasis, and promotes hepatocellularcarcinoma cell invasion via increasing cell motility, however, other mechanisms involving the prometastatic roles of GRP78remain to be elucidated. Here we report that forced GRP78expression promotes colon cancer cell migration and invasionthrough upregulating MMP-2, MMP-9 and especially uPAproduction. These effects of GRP78 are mediated by enhancingthe activation of β-catenin signaling. Interestingly, we identify thatGRP78 interacts with uPA both in the cells and in the culturemedium, suggesting that GRP78 protein is likely to directlyfacilitate uPA secretion via protein-protein interaction. Takentogether, our findings demonstrate for the first time that besidesstimulation of cell motility, GRP78 can act by increasingproteases production to promote tumor cell invasion.

Risk assessment of water inrush in karst tunnels based on a modified grey evaluation model: Sample as Shangjiawan Tunnel

Yuan, Yong-cai,Li, Shu-cai,Zhang, Qian-qing,Li, Li-ping,Shi, Shao-shuai,Zhou, Zong-qing Techno-Press 2016 Geomechanics & engineering Vol.11 No.4

A modified grey clustering method is presented to systematically evaluate the risk of water inrush in karst tunnels. Based on the center triangle whitenization weight function and upper and lower limit measure whitenization weight function, the modified grey evaluation model doesn't have the crossing properties of grey cluster and meets the standard well. By adsorbing and integrating the previous research results, seven influence factors are selected as evaluation indexes. A couple of evaluation indexes are modified and quantitatively graded according to four risk grades through expert evaluation method. The weights of evaluation indexes are rationally distributed by the comprehensive assignment method. It is integrated by the subjective factors and the objective factors. Subjective weight is given based on analytical hierarchy process, and objective weight obtained from simple dependent function. The modified grey evaluation model is validated by Jigongling Tunnel. Finally, the water inrush risk of Shangjiawan Tunnel is evaluated by using the established model, and the evaluation result obtained from the proposed method is agrees well with practical situation. This risk assessment methodology provides a powerful tool with which planners and engineers can systematically assess the risk of water inrush in karst tunnels.

Multi-Phase MPCA Modeling and Application Based on an Improved Phase Separation Method

Shu Wang,Yu-Qing Chang,Zhen Zhao,Fu-Li Wang 제어·로봇·시스템학회 2012 International Journal of Control, Automation, and Vol.10 No.6

Regarding the multi-phase characteristic of batch process, a new phase separation method is developed in this paper. The method realizes a 3-step sub-phase separation of the batch process using the retained principal components number, loading matrixes and principal component matrixes, which can adequately reflect the features variation of the process. In line with the different features and clas-sification step, automatic identification of ‘burrs’ and transition phases has been expounded. The pro-posed method can directly separate the stable phases and transition phases in the batch process, and deduce high-precision transition phase models. Based on the proposed method, the MPCA modeling and online monitoring is applied in the injection molding process. The experimental results have veri-fied the effectiveness of the proposed method.