Molecular Biology and Omics : Cloning and Characterization of a Novel α-Amylase from a Fecal Microbial Metagenome

( Bo Xu ),( Fu Ya Yang ),( Cai Yun Xiong ),( Jun Jun Li ),( Xiang Hua Tang ),( Jun Pei Zhou ),( Zhen Rong Xie ),( Jun Mei Ding ),( Yun Juan Yang ),( Zun Xi Huang ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.4

To isolate novel and useful microbial enzymes from uncultured gastrointestinal microorganisms, a fecal microbial metagenomic library of the pygmy loris was constructed. The library was screened for amylolytic activity, and 8 of 50,000 recombinant clones showed amylolytic activity. Subcloning and sequence analysis of a positive clone led to the identification a novel gene (amyPL) coding for α-amylase. AmyPL was expressed in Escherichia coli BL21 (DE3) and the purified AmyPL was enzymatically characterized. This study is the first to report the molecular and biochemical characterization of a novel α-amylase from a gastrointestinal metagenomic library.

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>

A Novel Missense Mutation of Keratin 17 Gene in a Chinese Family with Steatocystoma Multiplex

( Wei Wei Ha ),( Jing Wang ),( Wen Wang ),( Hong Yang Fu ),( Hua Yang Tang ),( Xian Fa Tang ),( Jun Zhu ),( Xian Yong Yin ),( Sen Yang ),( Xue Jun Zhang ) 대한피부과학회 2013 Annals of Dermatology Vol.25 No.4

A comparative experimental study on the cross-plane thermal conductivities of nano-constructed Sb2Te3/(Cu, Ag, Au, Pt) thermoelectric multilayer thin films

Yang Gang,Pan Jiahui,Fu Xuecheng,Hu Zhiyu,Wang Ying,Wu Zhimao,Mu Erzhen,Yan Xue-Jun,Lu Ming-Hui 나노기술연구협의회 2018 Nano Convergence Vol.5 No.22

Thermoelectric multilayer thin films used in nanoscale energy conversion have been receiving increasing attention in both academic research and industrial applications. Thermal transport across multilayer interface plays a key role in improving thermoelectric conversion efficiency. In this study, the cross-plane thermal conductivities of nano-constructed Sb2Te3/(Cu, Ag, Au, Pt) thermoelectric multilayer thin films have been measured using time-domain thermoreflectance method. The interface morphology features of multilayer thin film samples were characterized by using scanning and transmission electron microscopes. The effects of interface microstructure on the cross-plane thermal conductivities of the multilayer thin films have been extensively examined and the thermal transfer mechanism has been explored. The results indicated that electron–phonon coupling occurred at the semiconductor/metal interface that strongly affected the cross-plane thermal conductivity. By appropriately optimizing the period thickness of the metal layer, the cross-plane thermal conductivity can be effectively reduced, thereby improving the thermoelectric conversion efficiency. This work presents both experimental and theoretical understanding of the thermal transport properties of Sb2Te3/metal multilayer thin film junctions with important implications for exploring a novel approach to improving the thermoelectric conversion efficiency. Introduction Thermoelectric multilayer thin films used in nanoscale energy conversion have been receiving increasing attention in both academic research and industrial applications. Thermal transport across multilayer interface plays a key role in improving thermoelectric conversion efficiency. In this study, the cross-plane thermal conductivities of nano-constructed Sb2Te3/(Cu, Ag, Au, Pt) thermoelectric multilayer thin films have been measured using time-domain thermoreflectance method. The interface morphology features of multilayer thin film samples were characterized by using scanning and transmission electron microscopes. The effects of interface microstructure on the cross-plane thermal conductivities of the multilayer thin films have been extensively examined and the thermal transfer mechanism has been explored. The results indicated that electron–phonon coupling occurred at the semiconductor/metal interface that strongly affected the cross-plane thermal conductivity. By appropriately optimizing the period thickness of the metal layer, the cross-plane thermal conductivity can be effectively reduced, thereby improving the thermoelectric conversion efficiency. This work presents both experimental and theoretical understanding of the thermal transport properties of Sb2Te3/metal multilayer thin film junctions with important implications for exploring a novel approach to improving the thermoelectric conversion efficiency. Introduction

Effect of Homogenization Temperature on Microstructure and Mechanical Properties of Low-Carbon High-Boron Cast Steel

Fu Hanguang,Song Xuding,Lei Yongping,Jiang Zhiqiang,Xing Jiandong,Yang Jun,Wang Jinhua 대한금속·재료학회 2009 METALS AND MATERIALS International Vol.15 No.3

The effects of quenching treatment on the microstructure, hardness, impact toughness, and wear resistance of low-carbon high-boron cast steel (LCHBS) containing 0.15-0.3 %C, 1.4-1.8 %B, 0.3-0.8 %Si, 0.8-1.2 %Mn, 0.5-0.8%Cr, 0.3-0.6%Ni, and 0.3-0.6%Mo have been investigated by optical microscopy (OM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM), and via an electron probe microanalyzer (EPMA), X-ray diffraction (XRD) analysis, impact tester, hardness tester, and wear tester. The as-cast matrix of LCHBS consists of pearlite and ferrite. There is 8-10 vol.% Fe2(B, C) type borocarbides in the matrix. The micro-hardness of Fe2(B, C) is 1430-1480 Hv. Fe2(B,C) shows no obvious change and the matrix completely transforms into lath martensite upon quenching at 900 °C to 1100 °C. The microhardness of the matrix and the macrohardness of the LCHBS sample show a slight increase with an increase of homogenization temperature. When the homogenization temperature exceeds 1050 °C, no distinct change in the hardness is observed. The change of homogenization temperature has no apparent effect on the impact toughness of LCHBS. The mass losses of LCHBS increase distinctly when the wear load increases. The homogenization temperature is less than 1000 °C and the wear rate of LCHBS decreases with an increase of temperature. The wear rate does not display any obvious change after exceeding a homogenization temperature of 1000 °C. The effects of quenching treatment on the microstructure, hardness, impact toughness, and wear resistance of low-carbon high-boron cast steel (LCHBS) containing 0.15-0.3 %C, 1.4-1.8 %B, 0.3-0.8 %Si, 0.8-1.2 %Mn, 0.5-0.8%Cr, 0.3-0.6%Ni, and 0.3-0.6%Mo have been investigated by optical microscopy (OM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM), and via an electron probe microanalyzer (EPMA), X-ray diffraction (XRD) analysis, impact tester, hardness tester, and wear tester. The as-cast matrix of LCHBS consists of pearlite and ferrite. There is 8-10 vol.% Fe2(B, C) type borocarbides in the matrix. The micro-hardness of Fe2(B, C) is 1430-1480 Hv. Fe2(B,C) shows no obvious change and the matrix completely transforms into lath martensite upon quenching at 900 °C to 1100 °C. The microhardness of the matrix and the macrohardness of the LCHBS sample show a slight increase with an increase of homogenization temperature. When the homogenization temperature exceeds 1050 °C, no distinct change in the hardness is observed. The change of homogenization temperature has no apparent effect on the impact toughness of LCHBS. The mass losses of LCHBS increase distinctly when the wear load increases. The homogenization temperature is less than 1000 °C and the wear rate of LCHBS decreases with an increase of temperature. The wear rate does not display any obvious change after exceeding a homogenization temperature of 1000 °C.

Influence of Li Addition on the Microstructures and Mechanical Properties of Mg–Li Alloys

Jun Zhao,Jie Fu,Bin Jiang,Aitao Tang,Haoran Sheng,Tianhao Yang,Guangsheng Huang,Dingfei Zhang,Fusheng Pan 대한금속·재료학회 2021 METALS AND MATERIALS International Vol.27 No.6

In this study, Mg-xLi (x = 0, 1, 2, 3 and 5 wt%) alloys have been extruded to examine the role of Li content on microstructuresand tensile properties. The results revealed that Li addition increased the grain size and led to the formation of the transversedirection (TD)-split texture. These were mainly attributed to the promoted DRX process and the increased activity of prismatic⟨a⟩ slip during extrusion. Tensile tests revealed that the elongation of Mg-5Li sheet reached ~ 22.4% along the ED. Moreover, it exhibited a higher elongation of ~ 27.3%, three times than pure Mg, along the TD. During tension along the ED,with increasing Li content, more prismatic ⟨a⟩ slip and preferable intergranular strain coordination ability to accommodatethe plastic strain, leading to the enhanced room-temperature ductility. In contrast, more basal ⟨a⟩ slips and extension twinswere also activated along the TD, which further contributed to the enhanced ductility. Therefore, the ductility of Mg sheetsat room temperature gradually improved with Li addition due to the combining effects of basal ⟨a⟩ slip, prismatic ⟨a⟩ slip,extension twin and preferable intergranular strain coordination ability.

Crystal structure of LRG1 and the functional significance of LRG1 glycan for LPHN2 activation

Yang Jimin,Yin Guo Nan,Kim Do-Kyun,Han Ah-reum,Lee Dong Sun,Min Kwang Wook,Fu Yaoyao,Yun Jeongwon,Suh Jun-Kyu,Ryu Ji-Kan,Kim Ho Min 생화학분자생물학회 2023 Experimental and molecular medicine Vol.55 No.-

The serum glycoprotein leucine-rich ɑ-2-glycoprotein 1 (LRG1), primarily produced by hepatocytes and neutrophils, is a multifunctional protein that modulates various signaling cascades, mainly TGFβ signaling. Serum LRG1 and neutrophil-derived LRG1 have different molecular weights due to differences in glycosylation, but the impact of the differential glycan composition in LRG1 on its cellular function is largely unknown. We previously reported that LRG1 can promote both angiogenic and neurotrophic processes under hyperglycemic conditions by interacting with LPHN2. Here, we determined the crystal structure of LRG1, identifying the horseshoe-like solenoid structure of LRG1 and its four N-glycosylation sites. In addition, our biochemical and cell-biological analyses found that the deglycosylation of LRG1, particularly the removal of glycans on N325, is critical for the high-affinity binding of LRG1 to LPHN2 and thus promotes LRG1/LPHN2-mediated angiogenic and neurotrophic processes in mouse tissue explants, even under normal glucose conditions. Moreover, the intracavernous administration of deglycosylated LRG1 in a diabetic mouse model ameliorated vascular and neurological abnormalities and restored erectile function. Collectively, these data indicate a novel role of LRG1 glycans as molecular switches that can tune the range of LRG1’s cellular functions, particularly the LRG1/LPHN2 signaling axis.

Three Component Solvent-free Synthesis of Chroman-2,4-dione-based Heterocyclic Ketene Aminal (HKA) Derivatives by “GAP” Chemistry

Fu-Chao Yu,Xiao-Pan Hao,Xiu-Yang Jiang,Sheng-jiao Yan,Jun Lin 대한화학회 2014 Bulletin of the Korean Chemical Society Vol.35 No.6

A concise and efficient one-pot synthesis of chroman-2,4-dione-based HKA derivatives by three component reaction of HKAs, triethoxymethane and 4-hydroxycoumarin derivatives under solvent-free and catalyst-free conditions is described. This protocol has many advantages, in that the GAP (Group-Assistant-Purification) chemistry process is involved in this method. As a result, the experimenter can avoid cumbersome process steps such as traditional chromatography and recrystallization purifications. The desired products can be easily obtained by washing the crude products with 95% EtOH.

Expression profiles of circular RNAs in sheep skeletal muscle

Cao, Yang,You, Shuang,Yao, Yang,Liu, Zhi-Jin,Hazi, Wureli,Li, Cun-Yuan,Zhang, Xiang-Yu,Hou, Xiao-Xu,Wei, Jun-Chang,Li, Xiao-Yue,Wang, Da-Wei,Chen, Chuang-Fu,Zhang, Yun-Feng,Ni, Wei,Hu, Sheng-Wei Asian Australasian Association of Animal Productio 2018 Animal Bioscience Vol.31 No.10

Objective: Circular RNAs (circRNAs) are a newfound class of non-coding RNA in animals and plants. Recent studies have revealed that circRNAs play important roles in cell proliferation, differentiation, autophagy and apoptosis during development. However, there are few reports about muscle development-related circRNAs in livestock. Methods: RNA sequencing analysis was employed to identify and annotate circRNAs from longissimus dorsi of sheep. Reverse transcription followed by real-time quantitative (q) polymerase chain reaction (PCR) analysis verified the presence of these circRNAs. Targetscan7.0 and miRanda were used to analyse the interaction of circRNA-microRNA (miRNA). To investigate the function of circRNAs, an experiment was conducted to perform enrichment analysis hosting genes of circRNAs using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. Results: About 75.5 million sequences were obtained from RNA libraries of sheep skeletal muscle. These sequences were mapped to 729 genes in the sheep reference genome. We identified 886 circRNAs, including numerous circular intronic RNAs and exonic circRNAs. Reverse transcription PCR (RT-PCR) and DNA sequencing analysis confirmed the presence of several circRNAs. Real-Time RT-PCR analysis exhibited resistance of sheep circRNAs to RNase R digestion. We found that many circRNAs interacted with muscle-specific miRNAs involved in growth and development of muscle, especially circ776. The GO and KEGG enrichment analysis showed that hosting genes of circRNAs was involved in muscle cell development and signaling pathway. Conclusion: The study provides comprehensive expression profiles of circRNAs in sheep skeletal muscle. Our study offers a large number of circRNAs to facilitate a better understanding of their roles in muscle growth. Meanwhile, we suggested that circ776 could be analyzed in future study.

A Clinical Study on Juheli (Recombinant Human Interleukin - 11) in the Second Prevention of Chemotherapy Induced Thrombocytopenia

Xiao, Yang,Liu, Jun,Huang, Xin-En,Guo, Jian-Xiong,Fu, Peng-Chao,Huang, Xiao-Hong,Zhou, Juan,Ye, Ai-Qin Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.2

Objective: to investigate the effect and side effects of recombinant human interleukin - 11 (rhIL - 11, in Chinese Juheli, produced by Qi Lu Biotechnology CO., LTD) in the second prevention of chemotherapy induced thrombocytopenia (CIT). Methods: Cancer patients with CIT were recruited and were treated with rhIL - 11 (treatment phase, TP), and in the following cycle, all these patients administered with rhIL - 11 24 hours immediately after chemotherapy (preventive treatment phase, PTP). Duration and severity of thrombocytopenia between two phases were compared. Results: for patients in TP or PTP, nadir values of platelet were ($29.28{\pm}20.08){\times}10^9/L$ and ($45.24{\pm}19.66){\times}10^9/L$, duration of thrombocytopenia in TP and PTP was ($11.52{\pm}4.33$) and ($8.20{\pm}+2.77$)days, recovery time was ($19.40{\pm}3.89$)and ($13.44{\pm}3.02$)days, duration of rhIL - 11 administration was ($10.68{\pm}2.46$)and ($6.28{\pm}1.77$)days, number of patients needing platelet infusion was 16and4 respectively, all differences were statistically significant (p value were 0.007, 0.002, 0.000, 0.000, 0.034 respectively). For TP and PTP, number of patients with hemorrhage was 8 and 4, duration of bleeding was ($5.00{\pm}0.82$) and ($4.50{\pm}0.71$) days respectively, with no statistically significant difference. Adverse reactions mainly included fever, edema, arrhythmia, joint pain, fatigue, skin rash, headache, dizziness, etc., all were not statistically significant between TP and PTP. Conclusion: rhIL - 11 could be well tolerated and is effective that could reduce the duration, severity of CIT, platelet transfusion, and incidence of bleeding, as well as shorten the recovery time, duration of rhIL - 11 administration. Thus, rhIL - 11 could be commended in the second prevention of CIT for patients with cancer.