Analysis of differentially expressed proteins between the spinetoram-susceptible and -resistant strains of Plutella xylostella (L.)

Fei Yin,Qing Sheng Lin,Xia Feng,Huan-yu Chen,Zhen-yu Li,Zhen-di Hu 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.1

Plutella xylostella (L.), aworldwide vegetable pest, has developed resistance to spinetoram,whichwas previously effective for the control of P. xylostella (L.). The insecticidal resistance mechanismis essential to develop effective resistance management strategies. To explore the spinetoram resistance mechanism, a comparative proteomics approach was used to investigate the proteomic differences between the spinetoram-susceptible strain (SS) and spinetoram-resistant strain (RS) of P. xylostella (L.). Approximately 280 protein spotswere detected on each SDSPAGE gel. Of these, 19 proteinswere successfully identified byMALDI-TOF-MS.·Therewere 6 significantly downregulated spots and 13 up-regulated spots in RS, which showed significantly difference compared to that in SS. Based on the gene ontology(GO) system and KEGG database, the 19 identified proteins were classified into 6 groups includingmetabolisms, signal transduction, chaperones, transcriptional, protein synthesis, structural protein. Meanwhile, the expression profiles of 5 resistant related protein were further analysed by qRT-PCR. The results showed that 60S acidic ribosomal protein P2, glutathione S-transferase isozyme 3 and glutathione Stransferase deltawere significantly up-regulated,while phosphoglyceratemutase and receptor for activated protein kinase C homolog were significantly down-regulated. The expression tendency of mRNA was in accordance with which of protein. This study provided evidences that spinetoram induces proteomic changes in P. xylostella (L.), and it is contributed to help us understand the resistance mechanism of P. xylostella (L.) to spinetoram.

Isolation of Probiotic Piliated Lactobacillus rhamnosus Strains from Human Fecal Microbiota Using SpaA Antiserum-Based Colony Immunoblotting

( Zhen-quan Yang ),( Yu Xue ),( Sheng-qi Rao ),( Mi Zhang ),( Lu Gao ),( Yong-qi Yin ),( Da-wei Chen ),( Xiao-hui Zhou ),( Xin-an Jiao ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.11

Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti- SpaA-based CIB procedure could quantitatively detect 2.5 × 10³ CFU/ml of pLR colonies spiked in 10⁶ CFU/ml of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.

Generation of Cloned Transgenic Cats Expressing Red Fluorescence Protein1

Yin, Xi Jun,Lee, Hyo Sang,Yu, Xian Feng,Choi, Eugene,Koo, Bon Chul,Kwon, Mo Sun,Lee, Young S.,Cho, Su Jin,Jin, Guang Zhen,Kim, Lyoung Hyo,Shin, Hyoung Doo,Kim, Teoan,Kim, Nam Hyung,Kong, Il Keun Oxford University Press 2008 BIOLOGY OF REPRODUCTION Vol.78 No.3

<P>A method for engineering and producing genetically modified cats is important for generating biomedical models of human diseases. Here we describe the use of somatic cell nuclear transfer to produce cloned transgenic cats that systemically express red fluorescent protein. Immature oocytes were collected from superovulating cat ovaries. Donor fibroblasts were obtained from an ear skin biopsy of a white male Turkish Angora cat, cultured for one to two passages, and subjected to transduction with a retrovirus vector designed to transfer and express the red fluorescent protein (RFP) gene. A total of 176 RFP cloned embryos were transferred into 11 surrogate mothers (mean = 16 +/- 7.5 per recipient). Three surrogate mothers were successfully impregnated (27.3%) and delivered two liveborn and one stillborn kitten at 65 to 66 days of gestation. Analysis of nine feline-specific microsatellite loci confirmed that the cloned cats were genetically identical to the donor cat. Presence of the RFP gene in the transgenic cat genome was confirmed by PCR and Southern blot analyses. Whole-body red fluorescence was detected 60 days after birth in the liveborn transgenic (TG) cat but not in the surrogate mother cat. Red fluorescence was detected in tissue samples, including hair, muscle, brain, heart, liver, kidney, spleen, bronchus, lung, stomach, intestine, tongue, and even excrement of the stillborn TG cat. These results suggest that this nuclear transfer procedure using genetically modified somatic cells could be useful for the efficient production of transgenic cats.</P>

Identifying Creep and Destructuration Related Soil Parameters by Optimization Methods

Yin-Fu Jin,Zhen-Yu Yin,Yvon Riou,Pierre-Yves Hicher 대한토목학회 2017 KSCE JOURNAL OF CIVIL ENGINEERING Vol.21 No.4

The paper aims to construct an efficient optimization method for identifying creep and destructuration related governing parameters of soft structured clay. An elastic viscoplastic model has been developed and adopted. Different optimization processes, by genetic algorithm or particle swarm optimization with uniform or random samplings initialization methods, are carried out to obtain the material parameters from conventional undrained triaxial tests performed on a K0-concolidated natural soft clay. All comparisons demonstrate that the uniqueness of the solution is better guaranteed with the genetic algorithm rather than with the particle swarm optimization method. Furthermore, the efficiency of genetic algorithm has been verified by simulating other tests.

Numerical Modeling of Creep Degradation of Natural Soft Clays under One-dimensional Condition

Qi-Yin Zhu,Zhen-Yu Yin,Dong-Mei Zhang,Hongwei Huang 대한토목학회 2017 KSCE JOURNAL OF CIVIL ENGINEERING Vol.21 No.5

Creep degradation is a common phenomenon of natural soft clays. This paper focuses on developing a one-dimensional constitutive model considering the influence of bond degradation on the creep behavior for natural soft clays. First, conventional oedometer creep tests are studied and a creep based structure indicator ϖ denoting the difference between the creep coefficient of the reconstituted samples and that of intact samples is proposed. Then, the creep coefficient of intact clay is formulated by this indicator and the intrinsic creep coefficient corresponding to reconstituted clay. This formula is then incorporated into a one-dimensional creep model to describe the creep degradation behavior induced by bond degradation. The model parameters can be determined in a straightforward way from oedometer tests which leads to an easy application of the model for practice. Furthermore, coupled with the consolidation theory, the model is used to simulate oedometer tests at constant strain rate as well as long-term creep tests on sensitive Batiscan clay. The destructuration effects on the evolution of creep coefficient are analyzed. The comparisons between experimental and numerical results show that the proposed model can precisely describe the creep degradation behavior induced by destructuration of natural soft clay under one-dimensional loading.

Preliminary Evaluation of the in vitro Efficacy of 1, 2-di (Quinazolin-4-yl) Diselane against SiHa Cervical Cancer Cells

Huang, Yin-Jiu,Zhang, Yu-Yuan,Liu, Gang,Tang, Jie,Hu, Jian-Guo,Feng, Zhen-Zhong,Liu, Fang,Wang, Qi-Yi,Li, Dan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.15

Cervical cancer is one the most common malignancies among females. In recent years, its incidence rate has shown a rising trend in some countries so that development of anticancer drugs for cervical cancer is an urgent priority. In our recent anticancer drug discovery screen, 1, 2-di (quinazolin-4-yl)diselane (LG003) was found to possess wide spectrum anticancer efficacy. In the present work, the in vitro anticancer activity of LG003 was evaluated in the SiHa cervical cancer cell line. Compared with commercial anticancer drugs 10-hydroxycamptothecin, epirubicin hydrochloride, taxol and oxaliplatin, LG003 showed better anticancer activity. Furthermore, inhibition effects were time- and dose-dependent. Morphological observation exhibited LG003 treatment results in apoptosis like shrinking and blebbing, and cell membrane damage. Lactate dehydrogenase release assay revealed that LG003 exerts such effects in SiHa cells through a physiology pathway rather than cytotoxicity, which suggests that title compound LG003 can be a potential candidate agent for cervical cancer.

Isoindolin-1-ones from the stems of Nicotiana tabacum and their antiviral activities

Guang-Yu Yang,Jia-Meng Dai,Zhen-Jie Li,Jin Wang,Feng-Xian Yang,Xin Liu,Jing Li,Qian Gao,Xue-Mei Li,Yin-Ke Li,Wei-Guang Wang,Min Zhou,Qiu-Fen Hu 대한약학회 2022 Archives of Pharmacal Research Vol.45 No.8

In previous studies, several isoindolin-1-oneanalogs that exhibited signifi cant anti-tobacco mosaic virus(anti-TMV) activities were isolated from Nicotiana tabacum . Since gene-editing mutants provide a new sample for thediscovery of active metabolites, we focused on the stems ofYN-18–23 (a mutant N. tabacum for gene editing with thealkaloid metabolic pathway cultivated by Yunnan TobaccoCompany), which led to the isolation of four new ( 1–4 )and four known ( 5–8 ) isoindolin-1-ones. To the best of ourknowledge, nicindole C ( 3 ) is the fi rst subclass of isoindolin-1-one bearing a pentacyclic ketone, while nicindole D ( 4 )is the fi rst example of isoindolin-1-one bearing a methylpyridin-2-(1 H )-one moiety. Compounds 1–4 were testedfor their anti-TMV activities, and the results revealed thatcompounds 1 , 3 , and 4 exhibited high anti-TMV activities atconcentrations of 20 μM with inhibition rates of 48.6, 42.8,and 71.5%, respectively. These rates are higher than the inhibitionrate of the positive control (33.2%). The mechanisticstudy of compound 4 , which had the highest anti-TMV activityrevealed that increased potentiation of defense-related enzyme activities and downregulation of expression of theNtHsp70 protein may induce resistance in tobacco againstthe viral pathogen TMV. Molecular docking studies alsorevealed that the isoindolin-1-one substructure is fundamentalfor anti-TMV activity. The methyl-pyridin-2-(1 H )-onemoiety in compound 4 and the 2-oxopropyl groups in compounds1 and 3 at the N -2 position may increase inhibitoryactivities. This study of the structure–activity relationshipis helpful for fi nding new anti-TMV activity inhibitors. Tostudy whether the isoindolin-1-ones have broader antiviralactivities, compounds 1–4 were also tested for their antirotavirusactivities. Compound 4 exhibited high anti-rotavirusactivity with a therapeutic index (TI) value of 20.7. This TI value is close to that of the positive control (20.2).

Differences in the methanogen community between the nearshore and offshore sediments of the South Yellow Sea

Chen Ye,Zhen Yu,Wan Jili,Yin Xia,Li Siqi,Liu Jiayin,Zhang Guodong,Mi Tiezhu 한국미생물학회 2022 The journal of microbiology Vol.60 No.8

The differences in methanogen abundance and community composition were investigated between nearshore and offshore sediments in the South Yellow Sea (SYS). Shannon, Simpson, and Chao1 indices revealed a higher diversity of methanogens in the nearshore sediments than in the offshore sediments. The Mann–Whitney U test demonstrated that the relative abundance of Methanococcoides was significantly higher in the offshore sediments, while the relative abundances of Methanogenium, Methanosarcina, Methanosaeta, Methanolinea, and Methanomassiliicoccus were significantly higher in the nearshore sediments (P < 0.05). The abundance of the mcrA gene in the nearshore sediments was significantly higher than that in the offshore sediments. Furthermore, a similar vertical distribution of the methanogen and sulfatereducing bacteria (SRB) abundances was observed in the SYS sediments, implying there is potential cooperation between these two functional microbes in this environment. Finally, total organic carbon (TOC) was significantly correlated with methanogen community composition.

Enhanced tyrosine hydroxylase expression in PC12 cells co-cultured with feline mesenchymal stem cells

공일근,Guang-Zhen Jin,Xi-Jun Yin,Xian-Feng Yu,Su-Jin Cho,Hyo-Sang Lee,이효종 대한수의학회 2007 Journal of Veterinary Science Vol.8 No.4

Mesenchymal stem cells (MSCs) secrete a variety of neuroregulatory molecules, such as nerve growth factor, brain-derived neurotrophic factor, and glial cell-derived neurotrophic factor, which upregulate tyrosine hydroxylase (TH) gene expression in PC12 cells. Enhancing TH gene expression is a critical step for treatment of Parkinson's disease (PD). The objective of this study was to assess the effects of co-culturing PC12 cells with MSCs from feline bone marrow on TH protein expression. We divided the study into three groups: an MSC group, a PC12 cell group, and the combined MSC + PC12 cell group (the co-culture group). All cells were cultured in DMEM-HG medium supplemented with 10% fetal bovine serum for three days. Thereafter, the cells were examined using western blot analysis and immunocytochemistry. In western blots, the co-culture group demonstrated a stronger signal at 60 kDa than the PC12 cell group (p < 0.001). TH was not expressed in the MSC group, either in western blot or immunocytochemistry. Thus, the MSCs of feline bone marrow can up-regulate TH expression in PC12 cells. This implies a new role for MSCs in the neurodegenerative disease process.

Generation of Neuronal-Like Cells from Umbilical Cord Blood-Derived Mesenchymal Stem Cells of a RFP-Transgenic Cloned Cat

JIN, Guang-Zhen,YIN, Xi-Jun,YU, Xian-Feng,CHO, Su-Jin,CHOI, Eu-Gene,LEE, Young-S,JEON, Jin-Tae,YEE, Sung-Tae,KONG, Il-Keun Japanese Society of Veterinary Science 2008 The Journal of veterinary medical science Vol.70 No.7

<P>Umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) are multipotent adult stem cells, which can differentiation into cells of connective tissue and neural lineages. This study investigated the potential for neuronal differentiation of red fluorescent protein (RFP)-transgenic cat UCB-derived MSCs. The cells were cultured in pre-induction medium for 24 hr and in neuronal-induction medium for 72 hr. Immunofluorescent staining showed that 6.85% of the total cells were β III-tubulin-positive, 3.37% were neurofilament light (NF-L)-positive and 7.04% were neurofilament medium (NF-M)-positive. A β III-tubulin band was detected by western blot analysis. Our results demonstrate that RFP-transgenic UCB-derived MSCs can be differentiated into neuronal cells <I>in vitro</I>. Thus, RFP-transgenic MSCs could provide alternative tracing material for stem cell transplantation.</P>