Isolation, Culture and Identification of Porcine Skeletal Muscle Satellite Cells

Li, Bo-jiang,Li, Ping-hua,Huang, Rui-hua,Sun, Wen-xing,Wang, Han,Li, Qi-fa,Chen, Jie,Wu, Wang-jun,Liu, Hong-lin Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.8

The objective of this study was to establish the optimum protocol for the isolation and culture of porcine muscle satellite cells. Mononuclear muscle satellite cells are a kind of adult stem cell, which is located between the basal lamina and sarcolemma of muscle fibers and is the primary source of myogenic precursor cells in postnatal muscle. Muscle satellite cells are a useful model to investigate the mechanisms of muscle growth and development. Although the isolation and culture protocols of muscle satellite cells in some species (e.g. mouse) have been established successfully, the culture system for porcine muscle satellite cells is very limited. In this study, we optimized the isolation procedure of porcine muscle satellite cells and elaborated the isolation and culture process in detail. Furthermore, we characterized the porcine muscle satellite cells using the immunofluorecence. Our study provides a reference for the isolation of porcine muscle satellite cells and will be useful for studying the molecular mechanisms in these cells.

Two New Bibenzyl Glucosides from Dendrobium chrysotoxum

Fa-Wu Dong,Huai-Rong Luo,Qin-Li Wan,Feng-Qing Xu,Weiwei Fan,Kai-Jin Wang,Ning Li,Jiang-Miao Hu 대한화학회 2012 Bulletin of the Korean Chemical Society Vol.33 No.7

Two new bibenzyl glucosides, 3,3',4',5-tetramethoxybibenzyl-4-O-β-D-glucopyranoside (1) and 3,4,4',5- tetramethoxybibenzyl-3'-O-β-D-glucopyranoside (2), together with five known ones, chrysotobibenzyl (3), erianin (4), chrysotoxine (5), gigantol (6) and tristin (7) were isolated from the stems of Dendrobium chrysotoxum. The structures of those compounds were elucidated by extensive spectroscopic analysis. Moreover, compounds 1-7 were assessed for inhibitory activity of two enzymes-AChE (acetylcholine esterase) and BChE (butyrylcholine esterase).

Two New Bibenzyl Glucosides from Dendrobium chrysotoxum

Dong, Fa-Wu,Luo, Huai-Rong,Wan, Qin-Li,Xu, Feng-Qing,Fan, Wei-Wei,Wang, Kai-Jin,Li, Ning,Hu, Jiang-Miao Korean Chemical Society 2012 Bulletin of the Korean Chemical Society Vol.33 No.7

Two new bibenzyl glucosides, 3,3',4',5-tetramethoxybibenzyl-4-O-${\beta}$-D-glucopyranoside (1) and 3,4,4',5-tetramethoxybibenzyl-3'-O-${\beta}$-D-glucopyranoside (2), together with five known ones, chrysotobibenzyl (3), erianin (4), chrysotoxine (5), gigantol (6) and tristin (7) were isolated from the stems of Dendrobium chrysotoxum. The structures of those compounds were elucidated by extensive spectroscopic analysis. Moreover, compounds 1-7 were assessed for inhibitory activity of two enzymes-AChE (acetylcholine esterase) and BChE (butyrylcholine esterase).

Isolation, Culture and Identification of Porcine Skeletal Muscle Satellite Cells

Bo-jiang Li,Ping-hua Li,Rui=hua Huang,Wen-xing Sun,Han Wang,Qi-fa Li,Jie Chen,Wang Jun Wu,Honglin Liu 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.8

The objective of this study was to establish the optimum protocol for the isolation and culture of porcine muscle satellite cells. Mononuclear muscle satellite cells are a kind of adult stem cell, which is located between the basal lamina and sarcolemma of muscle fibers and is the primary source of myogenic precursor cells in postnatal muscle. Muscle satellite cells are a useful model to investigate the mechanisms of muscle growth and development. Although the isolation and culture protocols of muscle satellite cells in some species (e.g. mouse) have been established successfully, the culture system for porcine muscle satellite cells is very limited. In this study, we optimized the isolation procedure of porcine muscle satellite cells and elaborated the isolation and culture process in detail. Furthermore, we characterized the porcine muscle satellite cells using the immunofluorecence. Our study provides a reference for the isolation of porcine muscle satellite cells and will be useful for studying the molecular mechanisms in these cells.

Synthesis and Structural Studies of an Organic Complex and its Association with BSA

Fa-Yan Meng,Jin-Mei Zhu,Li Wang,Xue-Ping Zhong,Li-Xi Liang,Sheng-Rong Yu,Cui-Wu Lin 대한화학회 2011 Bulletin of the Korean Chemical Society Vol.32 No.7

The self-assembly of one novel organic complex based on chlorogenic acid (HCA) and 2,2'-bipyridine (2,2'-bipy) has been synthesized and characterized. The complex achieved by hydrogen-bonding interactions,adopted a 1:1 stoichiometry in a solid state. The proton transfer occurred from the carboxyl oxygen to the aromatic nitrogen atom to form salts CA·(2,2'-Hbipy), the 2,2'-Hbipy molecule individually occupies the pseudo-tetragonum that is formed with CA. In this paper, the interactions of CA·(2,2'-Hbipy) with bovine serum albumin (BSA) were studied by fluorescence spectrometry. For CA·(2,2'-Hbipy), HCA and 2,2'-bipy,the average quenching constants for BSA were 2.4384 × 10^4, 4.653 × 10^3, and 3.059 × 10^3 L·mol^−1, respectively. The mechanism for protein fluorescence quenching is apparently governed by a static quenching process. The Stern-Volmer quenching constants and corresponding thermodynamic parameters ΔH, ΔG and ΔS were calculated. The binding constants and the number of binding sites were also investigated. The conformational changes of BSA were observed from synchronous fluorescence spectra.

Overexpression of a Nitrile Hydratase from Klebsiella oxytoca KCTC 1686 in Escherichia coli and Its Biochemical Characterization

Fa-Mou Guo,Jian-ping Wu,Li-Rong Yang,Gang Xu 한국생물공학회 2015 Biotechnology and Bioprocess Engineering Vol.20 No.6

The putative Co-type nitrile hydratase (NHaseK, consisting of α- and β-subunits) genes and the putative activator (17K) gene adjacent to the β subunit region were cloned from Klebsiella oxytoca KCTC 1686. 17K is essential for the functional expression of recombinant NHaseK in Escherichia coli; however, the expression level of 17K was very low when the 17K gene and NHaseK structural genes were expressed as a gene cluster in E. coli BL21(DE3). To improve the 17K expression level and NHaseK activity, the expression cassette was redesigned by placing the 17K gene and NHaseK structural genes under the control of different promoters in the pETDuet-1 expression vector, co-expressing the 17K gene with the gene cluster in a double plasmid or a single plasmid with a double promoter, and introducing an efficient Shine- Dalgarno sequence 5' to the17K gene. The specific activity of NHaseK was improved when 17K was co-expressed with the gene cluster, whereas the production of NHaseK protein decreased. The maximum activity was achieved when an efficient Shine-Dalgarno sequence was introduced 5' to the 17K gene: the expression level of 17K was significantly improved and the expression level of NHaseK did not decrease significantly. The maximum activity was about 63,480 ± 1915.6 U/L broth towards 3-Cyanopyridine. Recombinant NHaseK could hydrolyze a wide range of aliphatic, aromatic, and heterocyclic nitriles, and convert racemic nitriles to the corresponding S-amides, with E values ranging from 9 to 17. The enzyme had a temperature optimum of 35°C and exhibited remarkably stability below 35°C.

A Combination of a Microemulsion and a Phospholipid Complex for Topical Delivery of Oxymatrine

Fa-Hao Cao,Wu-Qing OuYang,Yan-Ping Wang,Peng-Fei Yue,Shu-Peng Li 대한약학회 2011 Archives of Pharmacal Research Vol.34 No.4

Oxymatrine (OMT), a water-soluble drug, has a very low oral bioavailability because of its low membrane permeability and its biotransformation in the gastrointestinal tract. Formulated as an oxymatrine-phospholipid complex (OMT-PLC) can improve the lipid solubility and effectiveness of OMT. The purpose of this study was to explore the utility of the combination of a microemulsion and an OMT-PLC as a topical delivery vehicle for enhancing the absorption and efficacy of OMT. The solubility of OMT-PLC was determined and phase diagrams of microemulsions were constructed. Various microemulsion formulations were developed and characterized by their physicochemical properties, and their in vitro and in vivo permeability through skin. An optimal microemulsion (ME4), which presented as spherical droplets and consisted of 10.0% OMT-PLC, 8.0% isopropyl myristate, 30.0% Cremophor RH40/polyethylene glycol 400 (1:1) and 52.0% water, was selected. It possessed an average droplet size of 32.4 nm, a low viscosity of 113.7 mPa · s, and a high cloud point of 88°C. Compared to the control solution, ME4 provided better skin permeability in vitro and a higher retention ratio of OMT in skin in vivo. Moreover, ME4 significantly enhanced the antiproliferative activity of OMT on scar fibroblasts. These results indicate that the combination of a microemulsion and a phospholipid complex represents an effective vehicle for topical delivery of OMT.

Syntheses, Crystal Structures, and Spectral Properties of Two Coordination Compounds Based on 1,2-Bis(benzimidazolyl)benzene

Fa-YanMeng,Bing-Li Jiang,Cui-Wu Lin,Li Wang,Xiao-He Tan 대한화학회 2011 Bulletin of the Korean Chemical Society Vol.32 No.3

Two new benzimidazolyl-containing complexes have been synthesized by reactions of Cu^II salts and 1,2-bis(benzimidazolyl)benzene (H_2bbbz) with two different dicarboxylate ligands.When phthalate acid (H2pt) was employed as secondary ligand, a 0Dmolecular complex Cu(H_2bbbz)(pt)·(H_2pt) (1)was furnished and when the secondary ligand was instead by a linear bridging ligand of terephthalic acid (H_2tp) a 1D zipper-like coordination polymer [Cu(H_2bbbz)(tp)·2(C_2H_5OH)·H_2O]n (2) was obtained, suggesting the structure-direction effect of the secondary dicarbxylate ligand. The preliminary investigation on the spectral properties of the complexes was also presented.

Syntheses, Crystal Structures, and Spectral Properties of Two Coordination Compounds Based on 1,2-Bis(benzimidazolyl)benzene

Meng, Fa-Yan,Jiang, Bing-Li,Lin, Cui-Wu,Wang, Li,Tan, Xiao-He Korean Chemical Society 2011 Bulletin of the Korean Chemical Society Vol.32 No.3

Two new benzimidazolyl-containing complexes have been synthesized by reactions of $Cu^{II}$ salts and 1,2-bis(benzimidazolyl) benzene ($H_2bbbz$) with two different dicarboxylate ligands. When phthalate acid ($H_2pt$) was employed as secondary ligand, a 0D molecular complex Cu$(H_2bbbz)(pt){\cdot}(H_2pt)$ (1)was furnished and when the secondary ligand was instead by a linear bridging ligand of terephthalic acid ($H_2tp$) a 1D zipper-like coordination polymer $[Cu(H_2bbbz)(tp){\cdot}2(C_2H_5OH){\cdot}H_2O]_n$ (2) was obtained, suggesting the structure-direction effect of the secondary dicarbxylate ligand. The preliminary investigation on the spectral properties of the complexes was also presented.

Synthesis and Structural Studies of an Organic Complex and its Association with BSA

Meng, Fa-Yan,Yu, Sheng-Rong,Liang, Li-Xi,Zhong, Xue-Ping,Wang, Li,Zhu, Jin-Mei,Lin, Cui-Wu Korean Chemical Society 2011 Bulletin of the Korean Chemical Society Vol.32 No.7

The self-assembly of one novel organic complex based on chlorogenic acid (HCA) and 2,2'-bipyridine (2,2'-bipy) has been synthesized and characterized. The complex achieved by hydrogen-bonding interactions, adopted a 1:1 stoichiometry in a solid state. The proton transfer occurred from the carboxyl oxygen to the aromatic nitrogen atom to form salts CA${\cdot}$(2,2'-Hbipy), the 2,2'-Hbipy molecule individually occupies the pseudo-tetragonum that is formed with CA. In this paper, the interactions of CA${\cdot}$(2,2'-Hbipy) with bovine serum albumin (BSA) were studied by fluorescence spectrometry. For CA${\cdot}$(2,2'-Hbipy), HCA and 2,2'-bipy, the average quenching constants for BSA were $2.4384{\times}10^4$, $4.653{\times}10^3$, and $3.059{\times}10^3\;L{\cdot}mol^{-1}$, respectively. The mechanism for protein fluorescence quenching is apparently governed by a static quenching process. The Stern-Volmer quenching constants and corresponding thermodynamic parameters ${\Delta}$H, ${\Delta}$G and ${\Delta}$S were calculated. The binding constants and the number of binding sites were also investigated. The conformational changes of BSA were observed from synchronous fluorescence spectra.