Protoplast Preparation and Polyethylene Glycol (PEG)-mediated Transformation of Candida glycerinogenes

Cheng Zhang,Hong Zong,Bin Zhuge,Xinyao Lu,Huiying Fang,Jiali Zhu,Jian Zhuge 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.1

The regeneration of Candida glycerinogenes protoplasts is a major step following genetic manipulations such as fusion and DNA-mediated transformation. An investigation of protoplast formation and cytological examination was used to gain further insight into the loss of protoplast viability in osmotically stabilized support media. Protoplasts with the highest regeneration frequency (98.6% protoplasts/mL) were isolated, using lysozyme dissolved in 1M sorbitol osmoticum. The commercial enzyme preparations, osmotic stabilisers, and growth phase were effective in raising the protoplast yield. Sodium chloride was effective for protoplast preparation; however, sugars and sugar alcohols were better for protoplast regeneration. Sorbitol at a concentration of 1 M was used in regeneration agar for further studies. Regeneration of colonies from protoplasts was maximal (11 ~ 15%) when protoplasts were incorporated in cooled agar containing 0.5% glucose, supplemented with 1M sorbitol as osmotic stabilizer. C. glycerinogenes strain was highly sensitive to zeocin, so transformation of protoplasts and PEG-mediated was achieved with an improved transformation system, using plasmid pURGAP-gfp containing zeocin gene driven by a PCgGAP promoter from C. glycerinogenes to express gfp gene and be transformed into the 5.8S rDNA site of C. glycerinogenes in order to test the system for studying the yeast osmoregulation. We developed an efficient method for transformation of C. glycerinogenes, and parameters involved in transformation efficiency were optimized. Expressions of gfp at different levels were conducted under osmotic stress containing NaCl, KCl, sorbitol or glycerol for the recombinant strains. These improved procedures for protoplast isolation, regeneration and transformation proved to be useful applications in genetic studies for other Candida species and industrial yeast.

Study on economic performances of multi-span suspension bridges part 1: simple estimation formulas

Zhang, Li-Wen,Xiao, Ru-Cheng,Sun, Bin,Jiang, Yang,Zhang, Xue-Yi,Zhuang, Dong-Li,Zhou, Yun-Gang,Tu, Xue Techno-Press 2013 Structural Engineering and Mechanics, An Int'l Jou Vol.47 No.2

A study on economic performances of consecutive multi-span suspension bridges is carried out. In this part of the study, material amount and structural cost estimation formulas of the bridges is derived based on the structural ultimate carrying capacity. The bridge cost includes the part of superstructure and the part of substructure. Three types of bridge foundations, bored piles, concrete caissons and floating foundations, are considered in substructure. These formulas are to be used for the parametric study of the bridge cost in order to define its more economical layout under different conditions in the part two of the study.

A synthetic decursin analog with increased in vivo stability suppresses androgen receptor signaling in vitro and in vivo.

Zhang, Yong,Shaik, Ahmad Ali,Xing, Chengguo,Chai, Yubo,Li, Li,Zhang, Jinhui,Zhang, Wei,Kim, Sung-Hoon,L?, Junxuan,Jiang, Cheng M. Nijhoff ; Kluwer Academic Publishers 2012 Investigational new drugs Vol.30 No.5

<P>Targeting androgen receptor (AR) signaling with agents distinct from current antagonist drugs remains a rational approach to the prevention and treatment of prostate cancer (PCa). Our previous studies have shown that decursin and isomer decursinol angelate (DA), isolated from the Korean medicinal herb Angelica gigas Nakai, interrupt AR signaling and possess anti-PCa activities in vitro. In the LNCaP PCa cell model, these pyranoccoumarin compounds exhibit properties distinct from currently used antagonists (e.g., Casodex). However, both are rapidly de-esterified to decursinol, a partial AR agonist. We report here that a synthetic decursin analog, decursinol phenylthiocarbamate (DPTC), has greater in vivo stability than the parent compounds. DPTC-decursinol conversion was undetectable in mice. Furthermore, in LNCaP cells, DPTC decreased prostate specific antigen (PSA) expression, down-regulated AR abundance and mRNA and inhibited AR nuclear translocation. The effect of DPTC on AR and PSA mRNA and protein abundance was also observed in VCaP cells expressing wild type AR. DPTC inhibited growth of both PCa cell lines through G(1) cell cycle arrest and apoptosis, as did decursin and DA. Furthermore, i.p. administration of DPTC for 3 weeks suppressed the expression of AR target genes probasin and Nkx3.1 in mouse prostate glands. Overall, our data suggest that DPTC represents a prototype lead compound for development of in vivo stable and active novel decursin analogs for the prevention or therapy of PCa.</P>

Influence of FeSe doping on superconducting properties of MgB2 by hybrid microwave method

Cheng Cheng,Zhenjie Feng,Qing Li,Xu Wang,Chuan Yu,Hao Chu,Ya Yang,Changqin Liu,Yiming Cao,Zhe Li,Jingzhe Chen,Chao Jing,Shixun Cao,Jincang Zhang 한국물리학회 2017 Current Applied Physics Vol.17 No.11

The effect of FeSe doping on the physical properties of MgB2 is studied. Bulk samples of the FeSe doped MgB2 with weight ratio x ðFeSe : MgB2Þ ¼ 0%; 3%; 7% and 10% were prepared by hybrid microwave method. It is proved that FeSe is not stable together with MgB2. Fe2þ enters into MgB2 lattice, some Mg2þ and Se2『 are combined into the new impurity compound MgSe. The superconducting transition temperature (Tc) slightly decreased with increasing doping content of FeSe from R-T and M-T curves, which results from the substitution of Mg2þ by Fe2þ in the MgB2 lattice. The Jc increase slightly with the FeSe doping content increasing from 3 wt % to 10 wt %, which results from the increasing MgSe impurity pinning centers.

K-doping effect of the superconductivity in K2xFeTe1-xSx (0.07 ≤ x ≤ 0.3)

Cheng Cheng,Zhenjie Feng,Qing Li,Tao Li,Qiang Hou,Fei Chen,Zhongmin Ou,Jun-Yi Ge,Shixun Cao,Jincang Zhang 한국물리학회 2019 Current Applied Physics Vol.19 No.4

Bulk samples of K doping K2xFeTe1-xSx with x=0.07, 0.1, 0.2, 0.3 are successfully prepared by using easy-to-use stable compound K2S as the reactant. The lattice constant calculated from X-ray diffraction patterns indicate that K ions enter the Fe-Te-S layers. K doping is beneficial enhance the superconductivity transition temperature from the R-T curves. The apparent diamagnetic signal is observed in M-T curves when the content of K is smaller than 0.1. However, differential curves (dM/dT) in K-rich samples appear sharp slope mutations, which means that the Meissner effect signal is covered by the increased excess ferromagnetic ions. The number of excess Fe magnetic ions is proportional to K content, which may play an important role in determining the superconductivity.

Peroxisome proliferator-activated receptor γ is essential for secretion of ANP induced by prostaglandin D₂ in the beating rat atrium

Zhang, Ying,Li, Xiang,Liu, Li-Ping,Hong, Lan,Liu, Xia,Zhang, Bo,Wu, Cheng-Zhe,Cui, Xun The Korean Society of Pharmacology 2017 The Korean Journal of Physiology & Pharmacology Vol.21 No.3

Prostaglandin $D_2$ ($PGD_2$) may act against myocardial ischemia-reperfusion (I/R) injury and play an anti-inflammatory role in the heart. Although the effect of $PGD_2$ in regulation of ANP secretion of the atrium was reported, the mechanisms involved are not clearly identified. The aim of the present study was to investigate whether $PGD_2$ can regulate ANP secretion in the isolated perfused beating rat atrium, and its underlying mechanisms. $PGD_2$ (0.1 to $10{\mu}M$) significantly increased atrial ANP secretion concomitantly with positive inotropy in a dose-dependent manner. Effects of $PGD_2$ on atrial ANP secretion and mechanical dynamics were abolished by AH-6809 ($1.0{\mu}M$) and AL-8810 ($1.0{\mu}M$), $PGD_2$ and prostaglandin $F2{\alpha}$ ($PGF2{\alpha}$) receptor antagonists, respectively. Moreover, $PGD_2$ clearly upregulated atrial peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and the $PGD_2$ metabolite 15-deoxy-${\Delta}12$, 14-$PGJ_2$ (15d-$PGJ_2$, $0.1{\mu}M$) dramatically increased atrial ANP secretion. Increased ANP secretions induced by $PGD_2$ and 15d-$PGJ_2$ were completely blocked by the $PPAR{\gamma}$ antagonist GW9662 ($0.1{\mu}M$). PD98059 ($10.0{\mu}M$) and LY294002 ($1.0{\mu}M$), antagonists of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) signaling, respectively, significantly attenuated the increase of atrial ANP secretion by $PGD_2$. These results indicated that $PGD_2$ stimulated atrial ANP secretion and promoted positive inotropy by activating $PPAR{\gamma}$ in beating rat atria. MAPK/ERK and PI3K/Akt signaling pathways were each partially involved in regulating $PGD_2$-induced atrial ANP secretion.

A Comparison of Natural (D-α-tocopherol) and Synthetic (DL-α-tocopherol Acetate) Vitamin E Supplementation on the Growth Performance, Meat Quality and Oxidative Status of Broilers

Cheng, K.,Niu, Y.,Zheng, X.C.,Zhang, H.,Chen, Y.P.,Zhang, M.,Huang, X.X.,Zhang, L.L.,Zhou, Y.M.,Wang, T. Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.5

The present study was conducted to compare the supplementation of natural (D-${\alpha}$-tocopherol) and synthetic (DL-${\alpha}$-tocopherol acetate) vitamin E on the growth performance, meat quality, muscular antioxidant capacity and genes expression related to oxidative status of broilers. A total of 144 1 day-old Arbor Acres broiler chicks were randomly allocated into 3 groups with 6 replicates of 8 birds each. Birds were given a basal diet (control group), and basal diet supplemented with either 20 IU D-${\alpha}$-tocopherol or DL-${\alpha}$-tocopherol acetate for 42 days, respectively. The results indicated that treatments did not alter growth performance of broilers (p>0.05). Compared with the control group, concentration of ${\alpha}$-tocopherol in the breast muscle was increased by the supplementation of vitamin E (p<0.05). In the thigh, ${\alpha}$-tocopherol content was also enhanced by vitamin E inclusion, and this effect was more pronounced in the natural vitamin E group (p<0.05). Vitamin E supplementation increased the redness of breast (p<0.05). In the contrast, the inclusion of synthetic vitamin E decreased lightness of thigh (p<0.05). Dietary vitamin E inclusion reduced drip loss at 24 h of thigh muscle (p<0.05), and this effect was maintained for drip loss at 48 h in the natural vitamin E group (p<0.05). Broilers given diet supplemented with vitamin E showed decreased malondialdehyde (MDA) content in the breast (p<0.05). Additionally, natural rather than synthetic vitamin E reduced MDA accumulation in the thigh (p<0.05). Neither natural nor synthetic vitamin E supplementation altered muscular mRNA abundance of genes related to oxidative stress (p>0.05). It was concluded that vitamin E supplementation, especially the natural vitamin E, can enhance the retention of muscular ${\alpha}$-tocopherol, improve meat quality and muscular antioxidant capacity of broilers.

A Cyclic Trinuclear 1,2,4-Triazole-containing Copper(II) Complex Involving a Pyramidal Cu₃(OH) Core: Synthesis, Structure and Fluorescence

Zhang, Xiu-Cheng,Liu, Bing Korean Chemical Society 2009 Bulletin of the Korean Chemical Society Vol.30 No.10

Distinctive Roles of Wnt Signaling in Chondrogenic Differentiation of BMSCs under Coupling of Pressure and Platelet-Rich Fibrin

Cheng Baixiang,Feng Fan,Shi Fan,Huang Jinmei,Zhang Songbai,Quan Yue,Tu Teng,Liu Yanli,Wang Junjun,Zhao Ying,Zhang Min 한국조직공학과 재생의학회 2022 조직공학과 재생의학 Vol.19 No.4

BACKGROUND: Although newly formed constructs of feasible pressure-preadjusted bone marrow mesenchymal stem cells (BMSCs) and platelet-rich fibrin (PRF) showed biomechanical flexibility and superior capacity for cartilage regeneration, it is still not very clear how BMSCs and seed cells feel mechanical stimuli and convert them into biological signals, and the difference in signal transduction underlying mechanical and chemical cues is also unclear. METHODS: To determine whether mechanical stimulation (hydrostatic pressure) and chemical cues (platelet-rich fibrin, PRF) activate canonical or noncanonical Wnt signaling in BMSCs, BMSCs cocultured with PRF were subjected to hydrostatic pressure loading, and the activation of the Wnt signaling molecules and expression of cartilage-associated proteins and genes were determined by western blotting and polymerase chain reaction (PCR). Inhibitors of canonical or noncanonical Wnt signaling, XVX-939 or L690,330, were adopted to investigate the role of Wnt signaling molecules in mechanically promoted chondrogenic differentiation of BMSCs. RESULTS: Hydrostatic pressure of 120 kPa activated both Wnt/b-catenin signaling and Wnt/Ca2? signaling, with the the maximum promotion effect at 60 min. PRF exerted no synergistic effect on Wnt/b-catenin signaling activation. However, the growth factors released by PRF might reverse the promotion effects of pressure on Wnt/Ca2? signaling. Real-time PCR and Western blotting results showed that pressure could activate the expression of Col-II, Sox9, and aggrecan in BMSCs cocultured with PRF. Blocking experiment found a positive role of Wnt/b-catenin signaling, and a negative role of Wnt/ Ca2? signaling in chondrogenic differentiation of the BMSCs. Mutual inhibition exists between canonical and noncanonical Wnt signaling in BMSCs under pressure. CONCLUSION: Wnt signaling participates in the pressure-promoted chondrogenesis of the BMSCs co-cultured with PRF, with canonical and noncanonical pathways playing distinct roles during the process.

Two Novel 3-D Zinc Compounds with Mixture Ligands of H3BTB and 4,4'-Bipy: Crystal Structures, Blue Fluorescence and Single Point Energy Calculation

Zhang, Xiu-Cheng,Xu, Ling,Liu, Wen-Guang,Liu, Bing Korean Chemical Society 2010 Bulletin of the Korean Chemical Society Vol.31 No.9

Two 3-D zinc framework compounds, $[Zn_6(BTB)_4(4,4'-bipy)_4(H_2O)_4]{\cdot}9H_2O$ (1) and $[Zn_3(BTB)_2(4,4'-bipy)_2(H_2O)_2]{\cdot}5H_2O$ (2) ($H_3BTB$ = 1,2,3-benzenetricarboxylic acid, 4,4'-bipy = 4,4'-bipyridine), are obtained from the diffusion method and hydrothermal reaction respectively. Though 1 and 2 has the same coordination geometries of zinc atoms and coordination mode of $BTB^{3-}$, their 2-D layers are different: mirror symmetric layers in 1; parallel ones in 2, further connecting by 4,4'-bipy into 3-D frameworks. The hydrothermal reaction of 2 results in a more stable 3-D framework than the one in 1, which is supported by the single point energy calculations. 1 and 2 show similar blue fluorescence at 417 nm, which can be assigned to LMCT.