Purification and Characterization of Extracellular Inulinase from a Marine Yeast Pichia guilliermondii and Inulin Hydrolysis by the Purified Inulinase

Fang Gong,Tong Zhang,Jun Sheng,Jing Li,Xianghong Wang,Zhenming Chi 한국생물공학회 2008 Biotechnology and Bioprocess Engineering Vol.13 No.5

The extracellular inulinase of the marine yeast Pichia guilliermondii strain 1 was purified to homogeneity resulting in a 7.2-fold increase in specific inulinase activity. The molecular mass of the purified enzyme was estimated to be 50.0 kDa. The op-timal pH and temperature for the purified enzyme were 6.0 and 60C, respectively. The enzyme was activated by Mn²+, Ca²+, K+, Li+, Na+, Fe³+, Fe²+, Cu²+, and Co²+, but Mg²+, Hg²+, and Ag+ inhibited activity. The enzyme was strongly inhibited by phenylmethanesulphonyl fluoride (PMSF), iodoacetic acid, EDTA, and 1, 10-phenanthroline. The Km and Vmax values of the purified inulinase for inulin were 21.1 mg/mL and 0.08 mg/min, respectively. A large number of monosaccharides were de-tected after the hydrolysis of inulin. The deduced protein sequence from the cloned P. guilliermondii strain 1 inulinase gene contained the consensus motifs R-D-P-K-V-F-W-H and W-M-N-D-P-N-G, which are conserved among the inulinases from other microorganisms.

Preparation of lithium-doped NaV6O15 thin film cathodes with high cycling performance in SIBs

Xu Hai-Yan,Ruan Jun Hai,Liu Fang Lin,Li Dong-Cai,Zhang Feng-Jun,Wang Ai-Guo,Sun Dao-Sheng,오원춘 한국세라믹학회 2022 한국세라믹학회지 Vol.59 No.3

Lithium ions-doped NaV6O15 thin films have been prepared using a simple low temperature liquid phase deposition method and subsequent annealing process. X-ray diffraction (XRD), Fourier transform infrared spectrometer (FTIR), scanning elec- tron microscopy (SEM), and photoelectron spectroscopy (XPS) have been used to study the structural and physicochemical characteristics of the NaV6O15 film. The films were grown on the FTO conductive glass and used directly as an electrode of sodium ion batteries. The prepared lithium ions-doped NaV6O15 thin film electrodes showed an excellent cycling stability and discharge capacity, which may be attributed to the stability of the Li+ embedded into the gap between the V–O layers to maintain the structure and its stable β-phase structure transformed after the first cycle. The cycling stability greatly improved with increasing annealing temperature, while the discharge capacity decreased. The capacities of the film electrodes annealed at 400 °C and 450 °C maintained above 97% after 100 cycles. The lithium-doped NaV6O15 underwent a phase transition dur- ing the first charge/discharge cycle. The new transformed phase has perfect crystal structure stability undergoing insertion and deinsertion of Na+. Therefore, the lithium-doped NaV6O15 thin film possesses good cycling stability and is expected to be a promising thin film cathode for sodium-ion batteries.

A large-area free-standing graphene oxide multilayer membrane with high stability for nanofiltration applications

Chen, Long,Li, Yanhui,Chen, Lina,Li, Na,Dong, Chenglong,Chen, Qiong,Liu, Beibei,Ai, Qing,Si, Pengchao,Feng, Jinkui,Zhang, Lin,Suhr, Jonghwan,Lou, Jun,Ci, Lijie Elsevier 2018 CHEMICAL ENGINEERING JOURNAL -LAUSANNE- Vol.345 No.-

<P><B>Abstract</B></P> <P>A flexible and free-standing graphene oxide and nylon 6 (GO@nylon 6) multilayer nanofiltration membrane was prepared by a layer-by-layer assembly process. The combination of electrospinning and electrospraying technique was employed, which can facilely prepare large-area membrane with size of 20 × 30 cm. The mechanical stability of multilayer membrane has enhanced significantly due to the tightly locked structure achieved by nylon 6 nanofibers network. The novel GO@nylon 6–13 multilayer nanofiltration membrane demonstrated a pure water flux up to 11.15 L m<SUP>−2</SUP> h<SUP>−1</SUP> bar<SUP>−1</SUP>, while keeping high organic dye rejection rate (>95% for methylene blue, and >99% for methyl orange). The rejections rate of the Na<SUB>2</SUB>SO<SUB>4</SUB>, NaCl, CuSO<SUB>4</SUB>, and Pb(NO<SUB>3</SUB>)<SUB>2</SUB> were 56.5%, 27.6%, 36.7%, and 18.9%, respectively. Furthermore, GO@nylon 6–13 multilayer nanofiltration membrane also demonstrated a high flux of some common organic solvents (8.4, 5.3, and 0.8 L m<SUP>−2</SUP> h<SUP>−1</SUP> bar<SUP>−1</SUP> for methanol, ethanol, and NMP, respectively), showing excellent chemical stability for separation process in those solvents.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Large-area GO@nylon 6 multilayer nanofiltration membrane was prepared. </LI> <LI> The multilayer structure enhances the mechanical stability. </LI> <LI> The multilayer membrane demonstrates a high water flux. </LI> <LI> The multilayer membrane shows high rejection rate for organic dyes. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>Photograph and cross-section SEM image of GO@nylon 6 multilayer nanofiltration membrane, the inset shows the water contact angle.</P> <P>[DISPLAY OMISSION]</P>

Pharmacological blockade of cholesterol trafficking by cepharanthine in endothelial cells suppresses angiogenesis and tumor growth

Lyu, Junfang,Yang, Eun Ju,Head, Sarah A.,Ai, Nana,Zhang, Baoyuan,Wu, Changjie,Li, Ruo-Jing,Liu, Yifan,Yang, Chen,Dang, Yongjun,Kwon, Ho Jeong,Ge, Wei,Liu, Jun O.,Shim, Joong Sup Elsevier 2017 Cancer letters Vol.409 No.-

<P><B>Abstract</B></P> <P>Cholesterol is an important modulator of membrane protein function and signaling in endothelial cells, thus making it an emerging target for anti-angiogenic agents. In this study, we employed a phenotypic screen that detects intracellular cholesterol distribution in endothelial cells (HUVEC) and identified 13 existing drugs as cholesterol trafficking inhibitors. Cepharanthine, an approved drug for anti-inflammatory and cancer management use, was amongst the candidates, which was selected for in-depth mechanistic studies to link cholesterol trafficking and angiogenesis. Cepharanthine inhibited the endolysosomal trafficking of free-cholesterol and low-density lipoprotein in HUVEC by binding to Niemann-Pick disease, type C1 (NPC1) protein and increasing the lysosomal pH. The blockade of cholesterol trafficking led to a cholesterol-dependent dissociation of mTOR from the lysosomes and inhibition of its downstream signaling. Cepharanthine inhibited angiogenesis in HUVEC and in zebrafish in a cholesterol-dependent manner. Furthermore, cepharanthine suppressed tumor growth in vivo by inhibiting angiogenesis and it enhanced the antitumor activity of the standard chemotherapy cisplatin in lung and breast cancer xenografts in mice. Altogether, these results strongly support the idea that cholesterol trafficking is a viable drug target for anti-angiogenesis and that the inhibitors identified among existing drugs, such as cepharanthine, could be potential anti-angiogenic and antitumor agents.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A phenotypic screen identified 13 existing drugs, including cepharanthine, as cholesterol trafficking inhibitors. </LI> <LI> Cepharanthine inhibited lysosomal cholesterol trafficking by binding to NPC1 protein and increasing the lysosomal pH. </LI> <LI> The blockade of cholesterol trafficking led to a cholesterol-dependent dissociation of mTOR from the lysosomes. </LI> <LI> Cepharanthine inhibited angiogenesis in HUVEC and in zebrafish in a cholesterol-dependent manner. </LI> <LI> Cepharanthine treatment enhanced the antitumor activity of cisplatin in lung and breast cancer xenografts in mice. </LI> </UL> </P>

Silencing of long noncoding RNA HOXA11-AS inhibits the Wnt signaling pathway via the upregulation of HOXA11 and thereby inhibits the proliferation, invasion, and self-renewal of hepatocellular carcinoma stem cells

Jun-Cheng Guo,Yi-Jun Yang,Jin-Fang Zheng,Jian-Quan Zhang,Min Guo,Xiang Yang,Xiang-Ling Jiang,Li Xiang,You Li,Huang Ping,Liu Zhuo 생화학분자생물학회 2019 Experimental and molecular medicine Vol.51 No.-

Hepatocellular carcinoma (HCC) is a major cause of cancer-related deaths, but its molecular mechanisms are not yet well characterized. Long noncoding RNAs (lncRNAs) play crucial roles in tumorigenesis, including that of HCC. However, the role of homeobox A11 antisense (HOXA11-AS) in determining HCC stem cell characteristics remains to be explained; hence, this study aimed to investigate the effects of HOXA11-AS on HCC stem cell characteristics. Initially, the expression patterns of HOXA11-AS and HOXA11 in HCC tissues, cells, and stem cells were determined. HCC stem cells, successfully sorted from Hep3B and Huh7 cells, were transfected with short hairpin or overexpression plasmids for HOXA11-AS or HOXA11 overexpression and depletion, with an aim to study the influences of these mediators on the self-renewal, proliferation, migration, and tumorigenicity of HCC stem cells in vivo. Additionally, the potential relationship and the regulatory mechanisms that link HOXA11-AS, HOXA11, and the Wnt signaling pathway were explored through treatment with Dickkopf-1 (a Wnt signaling pathway inhibitor). HCC stem cells showed high expression of HOXA11-AS and low expression of HOXA11. Both HOXA11-AS silencing and HOXA11 overexpression suppressed the self-renewal, proliferation, migration, and tumorigenicity of HCC stem cells in vivo, as evidenced by the decreased expression of cancer stem cell surface markers (CD133 and CD44) and stemness-related transcription factors (Nanog, Sox2, and Oct4). Moreover, silencing HOXA11-AS inactivated the Wnt signaling pathway by decreasing the methylation level of the HOXA11 promoter, thereby inhibiting HCC stem cell characteristics. Collectively, this study suggested that HOXA11-AS silencing exerts an antitumor effect, suppressing HCC development via Wnt signaling pathway inactivation by decreasing the methylation level of the HOXA11 promoter.

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>

Physical properties and chemical composition of the cores in the California molecular cloud

Zhang, Guo-Yin,Xu, Jin-Long,Vasyunin, A. I.,Semenov, D. A.,Wang, Jun-Jie,Dib, Sami,Liu, Tie,Liu, Sheng-Yuan,Zhang, Chuan-Peng,Liu, Xiao-Lan,Wang, Ke,Li, Di,Wu, Zhong-Zu,Yuan, Jing-Hua,Li, Da-Lei,Gao, Springer-Verlag 2018 Astronomy and astrophysics Vol.620 No.-

<P><I>Aims.</I> We aim to reveal the physical properties and chemical composition of the cores in the California molecular cloud (CMC), so as to better understand the initial conditions of star formation.</P><P><I>Methods.</I> We made a high-resolution column density map (18.2′′) with <I>Herschel</I> data, and extracted a complete sample of the cores in the CMC with the fellwalker algorithm. We performed new single-pointing observations of molecular lines near 90 GHz with the IRAM 30m telescope along the main filament of the CMC. In addition, we also performed a numerical modeling of chemical evolution for the cores under the physical conditions.</P><P><I>Results.</I> We extracted 300 cores, of which 33 are protostellar and 267 are starless cores. About 51% (137 of 267) of the starless cores are prestellar cores. Three cores have the potential to evolve into high-mass stars. The prestellar core mass function (CMF) can be well fit by a log-normal form. The high-mass end of the prestellar CMF shows a power-law form with an index <I>α</I> = −0.9 ± 0.1 that is shallower than that of the Galactic field stellar mass function. Combining the mass transformation efficiency (<I>ε</I>) from the prestellar core to the star of 15 ± 1% and the core formation efficiency (CFE) of 5.5%, we suggest an overall star formation efficiency of about 1% in the CMC. In the single-pointing observations with the IRAM 30m telescope, we find that 6 cores show blue-skewed profile, while 4 cores show red-skewed profile. [HCO<SUP>+</SUP>]/[HNC] and [HCO<SUP>+</SUP>]/[N2H<SUP>+</SUP>] in protostellar cores are higher than those in prestellar cores; this can be used as chemical clocks. The best-fit chemical age of the cores with line observations is ~5 × 10<SUP>4</SUP> yr.</P>

Proanthocyanidins accelerate the germination of cucumber (Cucumis sativus L.) seeds

Li-jun Zhu,Xing-guang Deng,Li-juan Zou,Jun-qiang Wu,Da-wei Zhang,Honghui Lin 한국식물학회 2016 Journal of Plant Biology Vol.59 No.2

Proanthocyanidins (PAs) are the end products of the flavonoid biosynthetic pathway in many seeds, but their biological function is rarely unknown during seed germination. In the present study, we observed that PAs pretreatment accelerated cucumber seeds germination with maximum efficiency at 0.15% by measuring germination percentage and radical length. Using inhibitors of abscisic acid (ABA), gibberellins (GA) and alternative oxidase (AOX) and H2O2 scavenger pretreatment and gene expression analysis, we found that the accelerated effect of 0.15% PAs on seed germination was due to the decreased ABA biogenesis and enhanced GA production. ROS are induced by PAs pretreatment. Then, the enhanced ROS contributed to GA and ethylene accumulation and ABA decrease in seeds. Moreover, the improvement of GA was involved in the further induction of antioxidant enzymes activities. Therefore, our findings uncover a novel role of PAs in seed germination and clarify the relationships between ROS, ABA, GA and ethylene during seed germination.

Celastrol, produced by Tripterygium wilfordii Hook F. enhances defense response in cucumber seedlings against diverse environmental stresses

Li-jun Zhu,Xing-guang Deng,Li-juan Zou,Peng-xu Li,Jun-qiang Wu,Da-wei Zhang,Honghui Lin 한국식물학회 2017 Journal of Plant Biology Vol.60 No.1

Celastrol is an active triterpenoid compound isolated from Tripterygium wilfordii Hook F.. Many reports have highlighted that celastrol is an effective, safe and desirable approach to the treatment of cancers. However, their biological function during environmental stresses in plants is rarely reported. In the present study, the effects of celastrol on the tolerance against high light (HL), polyethylene glycol (PEG) and cold stress in cucumber (Cucumis sativus L.) were investigated. Celastrol pretreatment could enhance cucumber seedlings stress tolerance at a concentration of 1 μg ml–1. The results showed that pretreatment with 1 μg ml–1 celastrol clearly induced the activities of antioxidative enzymes, which subsequently alleviated stress-induced oxidative damage in plant cells. We also provided evidence that celastrol upregulated ABA biosynthetic gene NCED2 expression and ABA accumulation in cucumber seedlings, which resulted to the enhanced tolerance in response to environmental stresses. Furthermore, the celastrol-pretreated seedlings showed less photosystem damaged caused by the stress conditions, when compared with the control. Therefore, our findings provide a novel role of celastrol in plant against environmental stresses and indicate that the celastrol-induced activities of antioxidative enzymes and ABA content might contribute to the stress tolerance.

A New Unsymmetrical Zinc Phthalocyanine as Photosensitizers for Dye-sensitized Solar Cells

Zhang, Dan,Zhang, Xue-Jun,Zhang, Lei,Mao, Li-Jun Korean Chemical Society 2012 Bulletin of the Korean Chemical Society Vol.33 No.4

A new unsymmetrical zinc phthalocyanine has been designed and synthesized based on the 'push-pull' and extended ${\pi}$-conjugation concept for the dye-sensitized solar cells. Three tert-butoxy groups, which act as electron releasing ('push'), enhance the solubility of phthalocyanine in common organic solvents and reduce the aggregation. Hydroxy substituted 9,10-anthraquinones act as electron acceptors ('pull') for the study of photoinduced electron transfer processes as well as grafting onto nanocrystalline $TiO_2$. The new unsymmetrical zinc phthalocyanine was fully characterized by FTIR, UV-vis, $^1H$ NMR, cyclic voltammetry and differential pulse voltammetry. The new sensitizer was tested in dye-sensitized solar cells, and gave a better performance.