Combined transcriptomic and proteomic analysis of flubendiamide resistance in Plutella xylostella

Li Jing‐Jing,Jin Ming‐Hui,Wang Nian‐Meng,Yu Qi‐Tong,Shang Ze‐Yu,Xue Chao‐Bin 한국곤충학회 2020 Entomological Research Vol.50 No.10

Diamondback moth (DBM), Plutella xylostella, is an important pest of crucifers worldwide. The extensive use of diamide insecticides has led to DBM resistance in the world, and this presents a serious threat to vegetable production. In the present study, transcriptomic and proteomic analyses were combined to investigate the potential flubendiamide‐resistance mechanism in DBM. The lab‐selected (Rh) and field‐collected (Rb) flubendiamide‐resistant lines of P. xylostella with resistance ratios of 1889.92‐fold and 1250.97‐fold, respectively, were used, as well as a lab‐reared flubendiamide‐susceptible line (S). Compared with the S group, the transcriptomic analysis revealed 151 upregulated and 287 downregulated gene messengers in the Rh group and 432 upregulated and 565 downregulated gene messengers in the Rb group. The most frequently enriched pathways of differentially expressed genes (DEGs) were mainly involved in metabolic pathways. Metabolism related genes, including two P450, two ABC transporters, and three trypsins, were upregulated in the Rh line. Additionally, some P450 genes, trypsin, juvenile hormone (JH), and mucin genes were also upregulated in the Rb line. In proteomic analysis comparisons with the S group, there were 78 upregulated and 90 downregulated proteins in the Rh group and 221 upregulated and 155 downregulated proteins in the Rb group. Further analyses found that three CYP and 11 CYP proteins were over‐expressed in Rh and Rb lines, respectively. Four glutathione S‐transferase (GST) and four UGTs were over‐expressed in Rb line. So, we deduced that the detoxification metabolism may be the main mechanism of flubendiamide resistance in P. xylostella.

Comparison of Several Polysaccharide-derived Chiral Stationary Phases for the Enantiomer Separation of N-Fluorenylmethoxy-carbonyl ${\alpha}$-Amino Acids by HPLC

Jin, Jing-Yu,Lee, Kyung-Ah,Kang, Jong-Seong,Kang, Young-Koo,Baek, Chae-Sun,Lee, Won-Jae 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.5

The liquid chromatographic enantiomer separation of H-fluorenylmethoxycarbonyl (FMOC) protected ${\alpha}$-amino acids was performed on nine polysaccharide-derived chiral stationary phases (CSPs). The cellulose derived coated CSPs, Chiralcel OD-H (separation factor = 1.09-2.70) and Chiralcel OD (separation factor = 1.08-2.55), had the best performance of all the CSPs for resolution of H-FMOC ${\alpha}$-amino acids and therefore, all analyte enantiomers were base-line separated on Chiralcel OD-H and/or Chiralcel OD. Enantioseparation on cellulosetris(3,5-dimethylpherylcarbamate) derived CSPs (Chiralcel OD-H, Chiralcel OD and Chiralpak IB) is generally greater than that on amylose tris(3,5-dimethylphenylcarbamate) derived CSPs (Chiralpak AD-RH, Chiralpak AD and Chiralpak IA). Additionally, coated type CSPs (Chiralcel OD-H or Chiralcel OD, and Chiralpak AD) generally provided better enantioseparation for these analytes than the covalently bonded type CSPs (Chiralpak IB and Chiralpak IA) with thesame chiral selector of cellulose tris(3,5-dimethylphenylcarbamate) and amylose tris(3,5-dime-thylphenylcarbamate), respectively. However, Chiralpak IB and Chiralpak IA had an advantage over the coated type CSPs in that a broader range of solvents could be used due to itscovalently bonded nature.

Cytoprotective Effect of Taurine against Hydrogen Peroxide- Induced Oxidative Stress in UMR-106 Cells through the Wnt/β- Catenin Signaling Pathway

( Jing Lou ),( Donghe Han ),( Huihui Yu ),( Guang Yu ),( Meihua Jin ),( Sung-jin Kim ) 한국응용약물학회 2018 Biomolecules & Therapeutics(구 응용약물학회지) Vol.26 No.6

Osteoporosis development is closely associated with oxidative stress and reactive oxygen species (ROS). Taurine has potential antioxidant effects, but its role in osteoblasts is not clearly understood. The aim of this study was to determine the protective effects and mechanisms of actions of taurine on hydrogen peroxide (H₂O₂)-induced oxidative stress in osteoblast cells. UMR-106 cells were treated with taurine prior to H₂O₂ exposure. After treatment, cell viability, apoptosis, intracellular ROS production, malondialdehyde content, and alkaline phosphate (ALP) activity were measured. We also investigated the protein levels of β-catenin, ERK, CHOP and NF-E2-related factor 2 (Nrf2) along with the mRNA levels of Nrf2 downstream antioxidants. The results showed that pretreatment of taurine could reverse the inhibition of cell viability and suppress the induced apoptosis in a dose-dependent manner: taurine significantly reduced H₂O₂-induced oxidative damage and expression of CHOP, while it induced protein expression of Nrf2 and β-catenin and activated ERK phosphorylation. DKK1, a Wnt/β-catenin signaling inhibitor, significantly suppressed the taurine-induced Nrf2 signaling pathway and increased CHOP. Activation of ERK signaling mediated by taurine in the presence of H₂O₂ was significantly inhibited by DKK1. These data demonstrated that taurine protects osteoblast cells against oxidative damage via Wnt/β-catenin-mediated activation of the ERK signaling pathway.

Development of a Validated HPLC Method for the Simultaneous Determination of D- and L-Thyroxine in Human Plasma

Jin, Jing-Yu,Baek, Chae-Sun,Lee, Won-Jae Korean Chemical Society 2007 Bulletin of the Korean Chemical Society Vol.28 No.6

Comparative studies between covalently immobilized and coated chiral stationary phases based on polysaccharide derivatives for enantiomer separation of N-protected α-amino acids and their ester derivatives

Jin, Jing Yu,Bae, Su Kyoung,Lee, Wonjae Wiley Subscription Services, Inc., A Wiley Company 2009 Chirality Vol.21 No.10

<P>Liquid chromatographic enantiomer separation of several N-benzyloxycarbonyl (CBZ) and N-tert-butoxycarbonyl (BOC) α-amino acids and their corresponding ethyl esters was performed on covalently immobilized chiral stationary phases (CSPs) (Chiralpak IA and Chiralpak IB) and coated-type CSPs (Chiralpak AD and Chiralcel OD) based on polysaccharide derivatives. The solvent versatility of the covalently immobilized CSPs in enantiomer separation of N-CBZ and BOC-α-amino acids and their ester derivatives was shown and the chromatographic parameters of their enantioselectivities and resolution factors were greatly influenced by the nature of the mobile phase. In general, standard mobile phases using 2-propanol and hexane on Chiralpak IA showed fairly good enantioselectivities for resolution of N-CBZ and BOC-α-amino acids and their esters. However, 50% MTBE/hexane (v/v) for resolution of N-CBZ-α-amino acids ethyl esters and 20% THF/hexane (v/v) for resolution of N-BOC-α-amino acids ethyl esters afforded the greatest enantioselectivities on Chiralpak IA. Also, liquid chromatographic comparisons of the enantiomer resolution of these analytes were made on amylose tris(3,5-dimethylphenylcarbamate)-derived CSPs (Chiralpak IA and Chiralpak AD) and cellulose tris(3,5-dimethylphenylcarbamate)-derived CSPs (Chiralpak IB and Chiralcel OD). Chiralpak AD and/or Chiralcel OD showed the highest enantioselectivities for resolution of N-CBZ-α-amino acids and esters, while Chiralpak AD or Chiralpak IA showed the highest resolution of N-BOC-α-amino acids and esters. Chirality 2009. © 2008 Wiley-Liss, Inc.</P>

Enantiomer Separation of N-Protected α-Amino Acids on Covalently Immobilized Cellulose Tris(3,5-chlorophenylcarbamate) Chiral Stationary Phase in HPLC

Jing Yu Jin,이원재 대한화학회 2008 Bulletin of the Korean Chemical Society Vol.29 No.2

Liquid Chromatographic Enantiomer Separation of N-Phthaloyl Protected α-Amino Acids on Coated and Immobilized Chiral Stationary Phases Derived from Polysaccharide Derivatives

Jin, Jing Yu,Lee, Wonjae,Park, Jung Hag,Ryoo, Jae Jeong Marcel Dekker 2007 JOURNAL OF LIQUID CHROMATOGRAPHY AND RELATED TECHN Vol.30 No.1

<P> Liquid chromatographic enantiomer separation of N-phthaloyl (PHT) protected α-amino acids on several coated and immobilized chiral stationary phases (CSPs) derived from polysaccharide derivatives was performed. The coated CSP of Chiralpak AD showed more or less enantioseparation than the covalently bonded CSP of Chiralpak IA with the same chiral selector of amylose tris(3,5-dimethylphenylcarbamate). However, the coated Chiralcel OD showed greater enantioseparation than the covalently bonded Chiralpak IB with the same chiral selector of cellulose tris(3,5-dimethylphenylcarbamate). Among all examined CSPs, Chiralcel OD afforded the greatest performance for enantiomer resolution of N-PHT α-amino acids and, therefore, all analytes enantiomers were baseline separated on Chiralcel OD. The chromatographic method developed in this study was usefully applied for determination of the enantiomeric purity of commercially available N-PHT α-amino acids analytes.</P>

Comparison of Several Polysaccharide-derived Chiral Stationary Phases for the Enantiomer Separation of N-Fluorenylmethoxycarbonyl α-Amino Acids by HPLC

Jing Yu Jin,Kyung-Ah Lee,Jong Seong Kang,Young Koo Kang,백채선,이원재 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.5

The liquid chromatographic enantiomer separation of N-fluorenylmethoxycarbonyl (FMOC) protected α-amino acids was performed on nine polysaccharide-derived chiral stationary phases (CSPs). The cellulose derived coated CSPs, Chiralcel OD-H (separation factor = 1.09- 2.70) and Chiralcel OD (separation factor = 1.08-2.55), had the best performance of all the CSPs for resolution of N-FMOC α-amino acids and therefore, all analyte enantiomers were base-line separated on Chiralcel OD-H and/or Chiralcel OD. Enantioseparation on cellulose tris(3,5-dimethylphenylcarbamate) derived CSPs (Chiralcel OD-H, Chiralcel OD and Chiralpak IB) is generally greater than that on amylose tris(3,5-dimethylphenylcarbamate) derived CSPs (Chiralpak AD-RH, Chiralpak AD and Chiralpak IA). Additionally, coated type CSPs (Chiralcel OD-H or Chiralcel OD, and Chiralpak AD) generally provided better enantioseparation for these analytes than the covalently bonded type CSPs (Chiralpak IB and Chiralpak IA) with the same chiral selector of cellulose tris(3,5-dimethylphenylcarbamate) and amylose tris(3,5-dimethylphenylcarbamate), respectively. However, Chiralpak IB and Chiralpak IA had an advantage over the coated type CSPs in that a broader range of solvents could be used due to its covalently bonded nature.

Cytoprotective Effect of Taurine against Hydrogen Peroxide-Induced Oxidative Stress in UMR-106 Cells through the Wnt/β-Catenin Signaling Pathway

Lou, Jing,Han, Donghe,Yu, Huihui,Yu, Guang,Jin, Meihua,Kim, Sung-Jin The Korean Society of Applied Pharmacology 2018 Biomolecules & Therapeutics(구 응용약물학회지) Vol.26 No.6

Osteoporosis development is closely associated with oxidative stress and reactive oxygen species (ROS). Taurine has potential antioxidant effects, but its role in osteoblasts is not clearly understood. The aim of this study was to determine the protective effects and mechanisms of actions of taurine on hydrogen peroxide ($H_2O_2$)-induced oxidative stress in osteoblast cells. UMR-106 cells were treated with taurine prior to $H_2O_2$ exposure. After treatment, cell viability, apoptosis, intracellular ROS production, malondialdehyde content, and alkaline phosphate (ALP) activity were measured. We also investigated the protein levels of ${\beta}-catenin$, ERK, CHOP and NF-E2-related factor 2 (Nrf2) along with the mRNA levels of Nrf2 downstream antioxidants. The results showed that pretreatment of taurine could reverse the inhibition of cell viability and suppress the induced apoptosis in a dose-dependent manner: taurine significantly reduced $H_2O_2$-induced oxidative damage and expression of CHOP, while it induced protein expression of Nrf2 and ${\beta}-catenin$ and activated ERK phosphorylation. DKK1, a Wnt/${\beta}-catenin$ signaling inhibitor, significantly suppressed the taurine-induced Nrf2 signaling pathway and increased CHOP. Activation of ERK signaling mediated by taurine in the presence of $H_2O_2$ was significantly inhibited by DKK1. These data demonstrated that taurine protects osteoblast cells against oxidative damage via Wnt/${\beta}-catenin$-mediated activation of the ERK signaling pathway.

Construction of a High-Quality Yeast Two-Hybrid Library and Its Application in Identification of Interacting Proteins with Brn1 in Curvularia lunata

Gao, Jin-Xin,Jing, Jing,Yu, Chuan-Jin,Chen, Jie The Korean Society of Plant Pathology 2015 Plant Pathology Journal Vol.31 No.2

Curvularia lunata is an important maize foliar fungal pathogen that distributes widely in maize growing area in China, and several key pathogenic factors have been isolated. An yeast two-hybrid (Y2H) library is a very useful platform to further unravel novel pathogenic factors in C. lunata. To construct a high-quality full length-expression cDNA library from the C. lunata for application to pathogenesis-related protein-protein interaction screening, total RNA was extracted. The SMART (Switching Mechanism At 5' end of the RNA Transcript) technique was used for cDNA synthesis. Double-stranded cDNA was ligated into the pGADT7-Rec vector with Herring Testes Carrier DNA using homologous recombination method. The ligation mixture was transformed into competent yeast AH109 cells to construct the primary cDNA library. Eventually, a high qualitative library was successfully established according to an evaluation on quality. The transformation efficiency was about $6.39{\times}10^5$ transformants/$3{\mu}g$ pGADT7-Rec. The titer of the primary cDNA library was $2.5{\times}10^8cfu/mL$. The numbers for the cDNA library was $2.46{\times}10^5$. Randomly picked clones show that the recombination rate was 88.24%. Gel electrophoresis results indicated that the fragments ranged from 0.4 kb to 3.0 kb. Melanin synthesis protein Brn1 (1,3,8-hydroxynaphthalene reductase) was used as a "bait" to test the sufficiency of the Y2H library. As a result, a cDNA clone encoding VelB protein that was known to be involved in the regulation of diverse cellular processes, including control of secondary metabolism containing melanin and toxin production in many filamentous fungi was identified. Further study on the exact role of the VelB gene is underway.