Molecular and Biochemical Characterization of a Novel Intracellular Low-Temperature-Active Xylanase

( Jun Pei Zhou,),( Yan Yan Dong ),( Xiang Hua Tang ),( Jun Jun Li ),( Bo Xu ),( Qian Wu ),( Ya Jie Gao ),( Lu Pan ),( Zun Xi Huang ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.4

A 990 bp full-length gene (xynAHJ2) encoding a 329- residue polypeptide (XynAHJ2) with a calculated mass of 38.4 kDa was cloned from Bacillus sp. HJ2 harbored in a saline soil. XynAHJ2 showed no signal peptide, distinct amino acid stretches of glycoside hydrolase (GH) family 10 intracellular endoxylanases, and the highest amino acid sequence identity of 65.3% with the identified GH 10 intracellular mesophilic endoxylanase iM-KRICT PX1-Ps from Paenibacillus sp. HPL-001 (ACJ06666). The recombinant enzyme (rXynAHJ2) was expressed in Escherichia coli and displayed the typical characteristics of low-temperatureactive enzyme (exhibiting optimum activity at 35 o C, 62% at 20 o C, and 38% at 10 o C; thermolability at ≥45 o C). Compared with the reported GH 10 low-temperature-active endoxylanases, which are all extracellular, rXynAHJ2 showed low amino acid sequence identities (<45%), low homology (different phylogenetic cluster), and difference of structure (decreased amount of total accessible surface area and exposed nonpolar accessible surface area). Compared with the reported GH 10 intracellular endoxylanases, which are all mesophilic and thermophilic, rXynAHJ2 has decreased numbers of arginine residues and salt bridges, and showed resistance to Ni 2+ , Ca 2+ , or EDTA at 10 mM final concentration. The above mechanism of structural adaptation for low-temperature activity of intracellular endoxylanase rXynAHJ2 is different from that of GH 10 extracellular low-temperature-active endoxylanases. This is the first report of the molecular and biochemical characterizations of a novel intracellular low-temperatureactive xylanase.

The Study of Influencing Factors of Mobile E-commerce Software Use Intention-based on TAM and TRA

Yan Zhao,Ya-Ru Xue,Yan-Fei Jin,Jun-Wei Li,Yi He 보안공학연구지원센터 2016 International Journal of Grid and Distributed Comp Vol.9 No.8

With the wide popularity of mobile devices, e-commerce software in the mobile client forms a larger consumer market, which makes the volume of e-commerce transactions having a wide range of growth. As the consumers have a frequent contact with mobile devices, which makes consumers pay more attention to security and entertainment of the e-commerce software. Past mobile e-business research typically focus on the related factors about transaction process, like interface design, services and user experience, but seldom study the factors about consumers. On the basis of the TAM, this paper not only added perceived security and perceived entertainment, but also the consumers’ personality factors like subjective norm in TRA, into the model. The study shows that PU, PEU, PS and PE all will have direct or indirect effects on use attitude and use intention, and subjective norms and use attitude will directly affect the consumers' use intention. Through the empirical analysis we found that e-commerce operators not only make the consumers experience more of usefulness and ease of use, security, and entertainment in the technology and services, but also improve the popularity with the aid of the mainstream media propaganda and star power to enhance the public's normative beliefs, which will ultimately affect the user behavior of the consumers.

Synthesis, Crystal Structure and Characterization of Cu(II) and Cd(II) Coordination Compounds Based on Ligand 2-(3-(Pyridin-2-yl)-1H-pyrazol-1-yl)acetic Acid

Ya-Jun Zhang,Cui-Juan Wang,Kai-Li Mao,Xiao-Lei Liu,Shuai Huang,Yan Tong,Xian-Li Zhou 대한화학회 2014 Bulletin of the Korean Chemical Society Vol.35 No.7

Two novel coordination compounds [Cu2(pypya)3(H2O)2]·Cl·(H2O)5 (1) and {[Cd(pypya)(ta)1/2]·H2O}n (2) (Hpypya=2-(3-(pyridin-2-yl)-1H-pyrazol-1-yl)acetic acid, H2ta=terephthalic acid) were synthesized and characterized by single X-ray diffraction. Structure determination reveals that complex 1 and complex 2 crystallize in the triclinic system, with the P-1 space group. The asymmetric unit of 1 contains two Cu(II) ions, and their coordination modes are different. These units of complex 1 are linked together via hydrogen bonds and π-π interactions, and the 3D structure of complex 1 was formed. Complex 2, a mononuclear Cd(II) coordination compound, has a 2D structure which was constructed via coordination bonds. TGA and fluorescence spectra analysis of complex 1 and complex 2 have also been studied. In addition, the geometry parameters of complex 1 have been optimized with the B3LYP method of density functional theory (DFT) to explain its coordination behavior. The electronic properties of the complex 1 and ligand Hpypya have been investigated based on the nature bond orbital (NBO) analysis at the B3LYP level of theory. The result verifies that the synergistic effect have occurred in the compound.

Molecular characterization of the sans fille gene in Antheraea pernyi: A highly conserved gene during the evolution of animals

Ya-Jie Li,Rui Mi,Nan Meng,Zhi-Xin Wen,Xue-Jun Li,Mo Chen,Yan-Qun Liu,Shu-Ying Li 한국응용곤충학회 2013 Journal of Asia-Pacific Entomology Vol.16 No.3

Sans-fille (SNF) is the Drosophila homologue ofmammalian general splicing factors U1A and U2B″, and plays an important role in sex determination in Drosophilamelanogaster. In this study, the snf gene fromAntheraea pernyi (Lepidoptera: Saturniidae), an economically important insect, was isolated and characterized. The obtained 925 bp cDNA sequence contains an open reading frame of 669 bp encoding a polypeptide of 222 amino acids,showing 78% sequence identity to that from D. melanogaster. A database search revealed that SNF protein homologs are present in many animals, including invertebrates and vertebrates, with more than 70% amino acid sequence identities, suggesting that they were highly conserved during the evolution of animals. Phylogenetic analysis revealed that A. pernyi SNF was closely related to Bombyx mori SNF. Quantitative real-time PCR (qRT-PCR) analysis showed that the A. pernyi snf gene was transcribed during five larval developmental stages,and in six tested tissues (ovaries, testes, silk glands, fat body, integument, and hemolymph),with the most abundance determined in the gonads (ovaries or testes). Investigation of expression changes throughout embryonic development indicated that A. pernyi snfmRNAwas expressed at a lowlevel fromdays 0 to 4, and reached amaximum level at day 10, but decreased to a low level before hatching. These results suggest that the product of the snf gene may play important roles in the development of A. pernyi.

Neuron Discharge and c-Fos Expression in the Nucleus of the Solitary Tract Following Electroacupuncture at Acupoints of the Yangming Stomach Meridian of Foot

Jun-Feng He,Jie Yan,Jiang-Shan Li,Jian-Hua Liu,Chao Wang,Xiao-Rong Chang,Ya-Ting Qu 사단법인약침학회 2013 Journal of Acupuncture & Meridian Studies Vol.6 No.2

The nucleus of the solitary tract (nucleus tractus solitarii; NTS) is a primary center for both visceral afferents and somatic afferents. Previous experiments have demonstrated that the NTS is closely connected to the stomach and acupoints in the Yangming Stomach Meridian of Foot (ST Meridian). In this study, extracellular recording and immunochemistry methods were used to analyze the discharge of neurons and c-Fos protein expression in the NTS following acupuncture at different acupoints and a nonacupoint. A total of 104 discharging neurons were detected in the NTS of 52 rats, of which 86 provided complete data. After acupuncture at Sibai (ST 2), Zusanli (ST 36), Neiting (ST 44), Quanliao (SI 18), and the nonacupoint, the neuron response rate in the NTS was 65.12%, 51.16%, 46.51%, 34.88% and 31.40% respectively. For neuron response rate, there was a significant difference among Sibai (ST 2), Zusanli (ST 36), Neiting (ST 44), Quanliao (SI 18), and the nonacupoint (p < 0.01 or p < 0.05). In the other 48 rats, the number of c-Fos immunoreactive neurons in the NTS by electroacupuncture (EA) at Sibai (ST 2) group was significantly higher than that EA at other acupoints and the nonacupoint (p < 0.05 or p < 0.01). EA at both Zusanli (ST 36) and Neiting (ST 44) increased c-Fos immunoreactive neurons significantly over EA at Quanliao (SI 18) and the nonacupoint (p < 0.05 or p < 0.01), while there was no difference between EA at Quanliao (SI 18) and the nonacupoint group (p > 0.05). The experiments demonstrated that the afferent convergence in NTS are different by body surface points stimulus, which suggests that the NTS might be a primary center in the central nervous system receiving acupoints stimulus from the ST Meridian.

Nematicidal Activity of Paecilomyces spp. and Isolation of a Novel Active Compound

Ya-Jun Liu,Chong-Yan Zhai,Yi Liu,Ke-Qin Zhang 한국미생물학회 2009 The journal of microbiology Vol.47 No.3

Many species of Paecilomyces are entomogenous fungi and several are efficacious toward nematodes. To study the potential of Paecilomyces species in controlling nematodes, fungal extracts of 40 Paecilomyces spp. were evaluated for their nematicidal activity against Bursaphelenchus xylophilus and Panagrellus redivivus. The extracts of six Paecilomyces spp. exhibited the nematicidal activity against P. redivivus, and 11 species exhibited the nematicidal activity against B. xylophilus. The methanol extract of strain 1.01761 incubating on Czapek solid medium killed more than 95% P. redivivus in 24 h at 5 mg/ml, and the filtrate of strain 1.01788 cultured in Sabouraud's broth medium resulted in 90% mortality of B. xylophilus in 24 h at 5 mg/ml. A novel nematicidal compound, 4-(4’-carboxy-2’-ethyl-hydroxypentyl)-5,6-dihydro-6-methylcyclobuta[ b]pyridine-3,6-dicarboxylic acid, was isolated from Paecilomyces sp. YMF1.01761. The LD50 value of the compound within 24 h against P. redivivus was 50.86 mg/L, against Meloidogyne incognita was 47.1 mg/L, and against B. xylophilus was 167.7 mg/L.

Performance improvement of amorphous indium-gallium-zinc oxide ReRAM with SiO2 inserting layer

Yanli Pei,Biaoren Mai,Xiaoke Zhang,Ruiqin Hu,Ya Li,Zimin Chen,Bingfeng Fan,Jun Liang,Gang Wang 한국물리학회 2015 Current Applied Physics Vol.15 No.4

In this study, the resistive switching performance of amorphous indiumegalliumezinc oxide (a-IGZO) resistive switching random-access memory (ReRAM) was improved by inserting a thin silicon oxide layer between silver (Ag) top electrode and a-IGZO resistive switching layer. Compared with the single a-IGZO layer structure, the SiO2/a-IGZO bi-layer structure exhibits the higher On/Off resistance ratio larger than 103, and the lower operation power using a smaller SET compliance current. In addition, good endurance and excellent retention characteristics were achieved. Furthermore, multilevel resistance states are obtained through adjusting SET compliance current and RESET stop voltage, which shows a promise for high-performance nonvolatile multilevel memory application.

Senescence Effects of Angelica sinensis Polysaccharides on Human Acute Myelogenous Leukemia Stem and Progenitor Cells

Liu, Jun,Xu, Chun-Yan,Cai, Shi-Zhong,Zhou, Yue,Li, Jing,Jiang, Rong,Wang, Ya-Ping Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.11

Leukemia stem cells (LSCs) play important roles in leukemia initiation, progression and relapse, and thus represent a critical target for therapeutic intervention. Hence, it is extremely urgent to explore new therapeutic strategies directly targeting LSCs for acute myelogenous leukemia (AML) therapy. We show here that Angelica sinensis polysaccharide (ASP), a major active component in Dong quai (Chinese Angelica sinensis), effectively inhibited human AML $CD34^+CD38^-$ cell proliferation in vitro culture in a dose-dependent manner while sparing normal hematopoietic stem and progenitor cells at physiologically achievable concentrations. Furthermore, ASP exerted cytotoxic effects on AML K562 cells, especially LSC-enriched $CD34^+CD38^-$ cells. Colony formation assays further showed that ASP significantly suppressed the formation of colonies derived from AML $CD34^+CD38^-$ cells but not those from normal $CD34^+CD38^-$ cells. Examination of the underlying mechanisms revealed that ASP induced $CD34^+CD38^-$ cell senescence, which was strongly associated with a series of characteristic events, including up-regulation of p53, p16, p21, and Rb genes and changes of related cell cycle regulation proteins P16, P21, cyclin E and CDK4, telomere end attrition as well as repression of telomerase activity. On the basis of these findings, we propose that ASP represents a potentially important agent for leukemia stem cell-targeted therapy.

Synthesis, Crystal Structure and Characterization of Cu(II) and Cd(II) Coordination Compounds Based on Ligand 2-(3-(Pyridin-2-yl)-1H-pyrazol-1-yl)acetic Acid

Zhang, Ya-Jun,Wang, Cui-Juan,Mao, Kai-Li,Liu, Xiao-Lei,Huang, Shuai,Tong, Yan,Zhou, Xian-Li Korean Chemical Society 2014 Bulletin of the Korean Chemical Society Vol.35 No.7

Two novel coordination compounds $[Cu_2(pypya)_3(H_2O)_2]{\cdot}Cl{\cdot}(H_2O)_5$ (1) and $\{[Cd(pypya)(ta)_{1/2}]{\cdot}H_2O\}_n$ (2) (Hpypya=2-(3-(pyridin-2-yl)-1H-pyrazol-1-yl)acetic acid, $H_2ta$=terephthalic acid) were synthesized and characterized by single X-ray diffraction. Structure determination reveals that complex 1 and complex 2 crystallize in the triclinic system, with the P-1 space group. The asymmetric unit of 1 contains two Cu(II) ions, and their coordination modes are different. These units of complex 1 are linked together via hydrogen bonds and ${\pi}-{\pi}$ interactions, and the 3D structure of complex 1 was formed. Complex 2, a mononuclear Cd(II) coordination compound, has a 2D structure which was constructed via coordination bonds. TGA and fluorescence spectra analysis of complex 1 and complex 2 have also been studied. In addition, the geometry parameters of complex 1 have been optimized with the B3LYP method of density functional theory (DFT) to explain its coordination behavior. The electronic properties of the complex 1 and ligand Hpypya have been investigated based on the nature bond orbital (NBO) analysis at the B3LYP level of theory. The result verifies that the synergistic effect have occurred in the compound.

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>