Whole-genome identification and salt- and ABA-induced expression trends of the Nicotiana tabacum CKX gene family

Chen Wenwen,Wang Genhong,Yi Meiqin,Gao Tianke,Song Chunman,Gao Qian,Xia Qingyou,Xiang Haiying 한국식물생명공학회 2023 Plant biotechnology reports Vol.17 No.3

Cytokinin hormones are indispensable for plant growth and development. Cytokinin oxidase/dehydrogenase (CKX) helps regulate the dynamic balance of endogenous cytokinin levels. However, little is known about the CKX genes of Nicotiana tabacum (NtCKXs). In silico analyses were used to isolate, characterize, and elucidate the phylogenetic relationships of 15 NtCKX genes. Multi-species phylogenetic tree construction placed NtCKX1–15 on five of the eight branches of the CKX phylogenetic tree. Protein structure and network analyses revealed that NtCKX genes located on the same phylogenetic branch generally contained several conserved motifs and have highly similar structures, with structural domains related to flavin adenine dinucleotide (FAD) and cytokinin-binding found on all of the predicted NtCKX-encoded proteins. The upstream promotor region of NtCKX genes contained many abiotic stress-responsive cis-acting elements, including DRE, ERE, MBS, MYB, and MYC. Gene expression analysis revealed that each NtCKX gene responded differently to salt stress and exogenous abscisic acid (ABA) treatment. Four NtCKX genes exhibited ABA-induced expression trends with varying peak times. Under salt stress, NtCKX expression was significantly suppressed in two genes and upregulated in five others. In summary, we provided basic information regarding the CKX gene family of N. tabacum and elucidated their gene expression patterns under abiotic stresses, including ABA treatment and salt stress. The findings of this work can serve as a foundation for future study of the functions of NtCKX genes.

Involvement of Lysosome Membrane Permeabilization and Reactive Oxygen Species Production in the Necrosis Induced by Chlamydia muridarum Infection in L929 Cells

( Lixiang Chen ),( Cong Wang ),( Shun Li ),( Xin Yu ),( Xue Liu ),( Rongrong Ren ),( Wenwen Liu ),( Xiaojing Zhou ),( Xiaonan Zhang ),( Xiaohui Zhou ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.4

Chlamydiae, obligate intracellular bacteria, are associated with a variety of human diseases. The chlamydial life cycle undergoes a biphasic development: replicative reticulate bodies (RBs) phase and infectious elementary bodies (EBs) phase. At the end of the chlamydial intracellular life cycle, EBs have to be released to the surrounded cells. Therefore, the interactions between Chlamydiae and cell death pathways could greatly influence the outcomes of Chlamydia infection. However, the underlying molecular mechanisms remain elusive. Here, we investigated host cell death after Chlamydia infection in vitro, in L929 cells, and showed that Chlamydia infection induces cell necrosis, as detected by the propidium iodide (PI)-Annexin V double-staining flow-cytometric assay and Lactate dehydrogenase (LDH) release assay. The production of reactive oxygen species (ROS), an important factor in induction of necrosis, was increased after Chlamydia infection, and inhibition of ROS with specific pharmacological inhibitors, diphenylene iodonium (DPI) or butylated hydroxyanisole (BHA), led to significant suppression of necrosis. Interestingly, live-cell imaging revealed that Chlamydia infection induced lysosome membrane permeabilization (LMP). When an inhibitor upstream of LMP, CA-074-Me, was added to cells, the production of ROS was reduced with concomitant inhibition of necrosis. Taken together, our results indicate that Chlamydia infection elicits the production of ROS, which is dependent on LMP at least partially, followed by induction of host-cell necrosis. To our best knowledge, this is the first live-cell-imaging observation of LMP post Chlamydia infection and report on the link of LMP to ROS to necrosis during Chlamydia infection.

Metabolite profiles of ginsenosides Rk1 and Rg5 in zebrafish using ultraperformance liquid chromatography/quadrupole-time-of-flight MS

Shen, Wenwen,Wei, Yingjie,Tang, Daoquan,Jia, Xiaobin,Chen, Bin The Korean Society of Ginseng 2017 Journal of Ginseng Research Vol.41 No.1

Background: In the present study, metabolite profiles of ginsenosides Rk1 and Rg5 from red ginseng or red notoginseng in zebrafish were qualitatively analyzed with ultraperformance liquid chromatography/quadrupole-time-of-flight MS, and the possible metabolic were pathways proposed. Methods: After exposing to zebrafish for 24 h, we determined the metabolites of ginsenosides Rk1 and Rg5. The chromatography was accomplished on UPLC BEH C18 column using a binary gradient elution of 0.1% formic acetonitrile-0.1% formic acid water. The quasimolecular ions of compounds were analyzed in the negative mode. With reference to quasimolecular ions and MS2 spectra, by comparing with reference standards and matching the empirical molecular formula with that of known published compounds, and then the potential structures of metabolites of ginsenosides Rk1 and Rg5 were acquired. Results: Four and seven metabolites of ginsenoside Rk1 and ginsenoside Rg5, respectively, were identified in zebrafish. The mechanisms involved were further deduced to be desugarization, glucuronidation, sulfation, and dehydroxymethylation pathways. Dehydroxylation and loss of C-17 residue were also metabolic pathways of ginsenoside Rg5 in zebrafish. Conclusion: Loss of glucose at position C-3 and glucuronidation at position C-12 in zebrafish were regarded as the primary physiological processes of ginsenosides Rk1 and Rg5.

Identification of a Novel Human Lysophosphatidic Acid Acyltransferase, LPAAT-theta, Which Activates mTOR Pathway

Tang, Wenwen,Yuan, Jian,Chen, Xinya,Gu, Xiuting,Luo, Kuntian,Li, Jie,Wan, Bo,Wang, Yingli,Yu, Long Korean Society for Biochemistry and Molecular Biol 2006 Journal of biochemistry and molecular biology Vol.39 No.5

Lysophosphatidic acid acyltransferase (LPAAT) is an intrinsic membrane protein that catalyzes the synthesis of phosphatidic acid (PA) from lysophosphatidic acid (LPA). It is well known that LPAAT is involved in lipid biosynthesis, while its role in tumour progression has been of emerging interest in the last few years. To date, seven members of the LPAAT gene family have been found in human. Here we report a novel LPAAT member, designated as LPAAT-theta, which was 2728 base pairs in length and contained an open reading frame (ORF) encoding 434 amino acids. The LPAAT-theta gene consisted of 12 exons and 11 introns, and mapped to chromosome 4q21.23. LPAAT-theta was ubiquitously expressed in 18 human tissues by RT-PCR analysis. Subcellular localization of LPAAT-theta-EGFP fusion protein revealed that LPAAT-theta was distributed primarily in the endoplasmic reticulum (ER) of COS-7 cells. Furthermore, we found that the overexpression of LPAAT-theta can induce mTOR-dependent p70S6K phosphorylation on Thr389 and 4EBP1 phosphorylation on Ser65 in HEK293T cells.

Fabrication of silica/PVA‑co‑PE nanofiber membrane for oil/water separation

Yuanli Chen,Hui Fan,Xinlin Zha,Wenwen Wang,Yi Wu,Yi Xiong,Kun Yan,Yuedan Wang,Dong Wang 한국의류학회 2021 Fashion and Textiles Vol.8 No.1

High efficiency and anti-pollution oil/water separation membrane has been widely explored and researched. There are a large number of hydroxyl groups on the surface of silica, which has good wettability and can be used for oil-water separation membranes. Hydrophilic silica nanostructures with different morphologies were synthesized by changing templates and contents of trimethylbenzene (TMB). Here, silica nanospheres with radical pores, hollow silica nanospheres and worm-like silica nanotubes were separately sprayed on the PVA-co-PE nanofiber membrane (PM). The abundance of hydroxyl groups and porous structures on PM surfaces enabled the absorption of silica nanospheres through hydrogen bonds. Compared with different silica nanostructures, it was found that the silica/PM exhibited excellent super-hydrophilicity in air and underwater “oil-hating” properties. The PM was mass-produced in our lab through meltextrusion- phase-separation technique. Therefore, the obtained membranes not only have excellent underwater superoleophobicity but also have a low-cost production. The prepared silica/PM composites were used to separate n-hexane/water, silicone oil/ water and peanut oil water mixtures via filtration. As a result, they all exhibited efficient separation of oil/water mixture through gravity-driven filtration.

Self-assembled nanoparticles based on linoleic-acid modified carboxymethyl-chitosan as carrier of adriamycin (ADR)

Chenguang Liu,Wenwen Fan,Xiguang Chen,Chengsheng Liu,Xianghong Meng,Hyun Jin Park 한국물리학회 2007 Current Applied Physics Vol.7 No.s1

Linoleic-acid (LA) is covalently conjugated to carboxymethyl-chitosan via a 1-ethyl-3-(3-dimethylaminopropyyl)-carbodiimide-med-iated (EDC-mediated) reaction to generate self-aggregated chitosan nanoparticles by sonication. The average diameter of the particles isloading experiments indicate that the loading capacity and eciency increase with increasing concentration of ADR. ADR is slowlyreleased from chitosan self-aggregates for about 3 days. Furthermore, the eects of drug controlled release become more obvious onincreasing the pH value.

Cryptotanshinone inhibits TNF-α-induced LOX-1 expression by suppressing reactive oxygen species (ROS) formation in endothelial cells

Xiaoli Ran,Wenwen Zhao,Wenping Li,Jingshan Shi,Xiuping Chen 대한생리학회-대한약리학회 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.4

Cryptotanshinone (CPT) is a natural compound isolated from traditional Chinese medicine Salvia miltiorrhiza Bunge. In the present study, the regulatory effect and potential mechanisms of CPT on tumor necrosis factor alpha (TNF-α) induced lectin-like receptor for oxidized low density lipoprotein (LOX-1) were investigated. Human umbilical vein endothelial cells (HUVECs) were cultured and the effect of TNF-α on LOX-1 expression at mRNA and protein levels was determined by Real-time PCR and Western blotting respectively. The formation of intracellular ROS was determined with fluorescence probe CM-DCFH2-DA. The endothelial ox-LDL uptake was evaluated with DiI-ox-LDL. The effect of CPT on LOX-1 expression was also evaluated with SD rats. TNF-α induced LOX-1 expression in a dose- and time- dependent manner in endothelial cells. TNF-α induced ROS formation, phosphorylation of NF-κB p65 and ERK, and LOX-1 expression, which were suppressed by rotenone, DPI, NAC, and CPT. NF-κB inhibitor BAY11-7082 and ERK inhibitor PD98059 inhibited TNF-α-induced LOX-1 expression. CPT and NAC suppressed TNF-α-induced LOX-1 expression and phosphorylation of NF-κB p65 and ERK in rat aorta. These data suggested that TNF-α induced LOX-1 expression via ROS activated NF-κB/ERK pathway, which could be inhibited by CPT. This study provides new insights for the anti-atherosclerotic effect of CPT.

Evaluation of the sp3/sp2 ratio of DLC films by RF-PECVD and its quantitative relationship with optical band gap

Sheng Hongxun,Xiong Wenwen,Zheng Songsheng,Chen Chao,He Song,Cheng Qijin 한국탄소학회 2021 Carbon Letters Vol.31 No.5

DLC has been attractive as semiconductor materials for solar cell due to its biological friendliness, fexible microstructures, and especially its tunable band gap. In order to fabricate high-efciency multiband gap solar cell, it is important to control the sp3 / sp2 bonds ratio of DLC flm corresponding to optical band gap (Eg). There are many references reporting the relations among the fabrication conditions, Eg, sp3 /sp2 , and ID/IG. However, a more comprehensive database is needed for controllable fabrication. Especially, the quantitative relationship of sp3 /sp2 ratio to Eg of DLC flm by PECVD is unclear. In this paper, 36 sets of DLC flms were fabricated by RF-PECVD. Characterization methods of XPS, Raman spectroscopy, and IR absorption have been used to determine the sp3 /sp2 ratio of DLC flms. UV/visible light absorption method has applied to evaluate Eg. The Eg obtained is in the range 1.45–3.0 eV. Our results agree well with the references. The XPS spectra gives a linear relationship as Eg=− 0.161 (±0.136)+26.095 (±1.704) · {sp3 (XPS)/sp2 }, the Raman spectra shows a linear function that Eg=1.327 (±0.046)+0.428 (±0.036) · (ID/IG), as well as the FTIR analysis demonstrates that Eg=− 0.492 (±0.093)+0.464 (±0.044) · {sp3 (FTIR)/sp2 }.

Isocryptotanshinone Induced Apoptosis and Activated MAPK Signaling in Human Breast Cancer MCF-7 Cells

Xuenong Zhang,Weiwei Luo,Wenwen Zhao,Jinjian Lu,Xiuping Chen 한국유방암학회 2015 Journal of breast cancer Vol.18 No.2

Purpose: Isocryptotanshinone (ICTS) is a natural bioactive product that is isolated from the roots of the widely used medical herb Salvia miltiorrhiza. However, few reports exist on the mechanisms underlying the therapeutic effects of ICTS. Here, we report that ICTS has anticancer activity and describe the mechanism underlying this effect. Methods: The antiproliferative effect of ICTS was determined using 3-(4,5-dimethyl-2-thiazolyl)-2,5- diphenyl-2-H-tetrazolium bromide (MTT) and clonogenic assays. The effect of ICTS on the cell cycle was measured using flow cytometry. Apoptosis was determined by Hoechst 33342 staining, DNA fragmentation assays, and Western blotting for apoptotic proteins. Finally, the effect of ICTS on mitogen-activated protein kinases (MAPKs) was determined by Western blotting. Results: ICTS significantly inhibited proliferation of MCF-7 and MDA-MB-231 human breast cancer cells, HepG2 human liver cancer cells, and A549 human lung cancer cells in vitro. Among the tested cell lines, MCF-7 cells showed the highest sensitivity to ICTS. ICTS significantly inhibited colony formation by MCF-7 cells. Furthermore, exposure of MCF-7 cells to ICTS induced cell cycle arrest at the G1 phase and decreased mitochondrial membrane potential. Hoechst 33342 staining and Western blot analysis for apoptotic proteins suggested that ICTS induced apoptosis in MCF-7 cells. In addition, ICTS activated MAPK signaling in MCF-7 cells by inducing time- and concentration-dependent phosphorylation of JNK, ERK, and p38 MAPK. Conclusion: Our results suggest that ICTS inhibited MCF-7 cell proliferation by inducing apoptosis and activating MAPK signaling pathways.

A post-genome-wide association study validating the association of the glycophorin C gene with serum hemoglobin level in pig

Yang Liu,Zhengzheng Hu,Chen Yang,Shiwei Wang,Wenwen Wang,Qin Zhang 아세아·태평양축산학회 2017 Animal Bioscience Vol.30 No.5

Objective: This study aimed to validate the statistical evidence from the genome-wide association study (GWAS) as true-positive and to better understand the effects of the glycophorin C (GYPC) gene on serum hemoglobin traits. Methods: Our initial GWAS revealed the presence of two single nucleotide polymorphisms (SNPs) (ASGA0069038 and ALGA0084612) for the hemoglobin concentration trait (HGB) in the 2.48 Mb region of SSC15. From this target region, GYPC was selected as a promising gene that associated with serum HGB traits in pigs. SNPs within the GYPC gene were detected by sequencing. Thereafter, we performed association analysis of the variant with the serum hemoglobin level in three pig populations. Results: We identified one SNP (g.29625094 T>C) in exon 3 of the GYPC gene. Statistical analysis showed a significant association of the SNP with the serum hemoglobin level on day 20 (p<0.05). By quantitative real-time polymerase chain reaction, the GYPC gene was expressed in eight different tissues. Conclusion: These results might improve our understanding of GYPC function and provide evidence for its association with serum hemoglobin traits in the pig. These results also indicate that the GYPC gene might serve as a useful marker in pig breeding programs.