siRNAs Derived from Cymbidium Mosaic Virus and Odontoglossum Ringspot Virus Down-modulated the Expression Levels of Endogenous Genes in Phalaenopsis equestris

Han-hong Lan,Cui-mei Wang,Shuang-shuang Chen,Jian-ying Zheng 한국식물병리학회 2019 Plant Pathology Journal Vol.35 No.5

Interplay between Cymbidium mosaic virus (CymMV)/ Odontoglossum ringspot virus (ORSV) and its host plant Phalaenopsis equestris remain largely unknown, which led to deficiency of effective measures to control disease of P. equestris caused by infecting viruses. In this study, for the first time, we characterized viral small interfering RNAs (vsiRNAs) profiles in P. equestris co-infected with CymMV and ORSV through small RNA sequencing technology. CymMV and ORSV small interfering RNAs (siRNAs) demonstrated several general and specific/new characteristics. vsiRNAs, with A/ U bias at the first nucleotide, were predominantly 21-nt long and they were derived predominantly (90%) from viral positive-strand RNA. 21-nt siRNA duplexes with 0-nt overhangs were the most abundant 21-nt duplexes, followed by 2-nt overhangs and then 1-nt overhangs 21-nt duplexes in infected P. equestris. Continuous but heterogeneous distribution and secondary structures prediction implied that vsiRNAs originate predominantly by direct Dicer-like enzymes cleavage of imperfect duplexes in the most folded regions of the positive strand of both viruses RNA molecular. Furthermore, we totally predicted 54 target genes by vsiRNAs with psRNATarget server, including disease/stress response– related genes, RNA interference core components, cytoskeleton- related genes, photosynthesis or energy supply related genes. Gene Ontology classification showed that a majority of the predicted targets were related to cellular components and cellular processes and performed a certain function. All target genes were down-regulated with different degree by vsiRNAs as shown by realtime reverse transcription polymerase chain reaction. Taken together, CymMV and ORSV siRNAs played important roles in interplay with P. equestris by down modulating the expression levels of endogenous genes in host plant.

siRNAs Derived from Cymbidium Mosaic Virus and Odontoglossum Ringspot Virus Down-modulated the Expression Levels of Endogenous Genes in Phalaenopsis equestris

Lan, Han-hong,Wang, Cui-mei,Chen, Shuang-shuang,Zheng, Jian-ying The Korean Society of Plant Pathology 2019 Plant Pathology Journal Vol.35 No.5

Interplay between Cymbidium mosaic virus (CymMV)/Odontoglossum ringspot virus (ORSV) and its host plant Phalaenopsis equestris remain largely unknown, which led to deficiency of effective measures to control disease of P. equestris caused by infecting viruses. In this study, for the first time, we characterized viral small interfering RNAs (vsiRNAs) profiles in P. equestris co-infected with CymMV and ORSV through small RNA sequencing technology. CymMV and ORSV small interfering RNAs (siRNAs) demonstrated several general and specific/new characteristics. vsiRNAs, with A/U bias at the first nucleotide, were predominantly 21-nt long and they were derived predominantly (90%) from viral positive-strand RNA. 21-nt siRNA duplexes with 0-nt overhangs were the most abundant 21-nt duplexes, followed by 2-nt overhangs and then 1-nt overhangs 21-nt duplexes in infected P. equestris. Continuous but heterogeneous distribution and secondary structures prediction implied that vsiRNAs originate predominantly by direct Dicer-like enzymes cleavage of imperfect duplexes in the most folded regions of the positive strand of both viruses RNA molecular. Furthermore, we totally predicted 54 target genes by vsiRNAs with psRNATarget server, including disease/stress response-related genes, RNA interference core components, cytoskeleton-related genes, photosynthesis or energy supply related genes. Gene Ontology classification showed that a majority of the predicted targets were related to cellular components and cellular processes and performed a certain function. All target genes were down-regulated with different degree by vsiRNAs as shown by real-time reverse transcription polymerase chain reaction. Taken together, CymMV and ORSV siRNAs played important roles in interplay with P. equestris by down modulating the expression levels of endogenous genes in host plant.

Studies of the effects and mechanisms of ginsenoside Re and Rk₃ on myelosuppression induced by cyclophosphamide

Han, Jiahong,Xia, Jing,Zhang, Lianxue,Cai, Enbo,Zhao, Yan,Fei, Xuan,Jia, Xiaohuan,Yang, He,Liu, Shuangli The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.4

Background: Ginsenoside Re (Re) is one of the major components of Panax ginseng Meyer. Ginsenoside $Rk_3$ ($Rk_3$) is a secondary metabolite of Re. The aim of this study was to investigate and compare the effects and underlying mechanisms of Re and $Rk_3$ on cyclophosphamide-induced myelosuppression. Methods: The mice myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide. Peripheral blood cells, bone marrow nucleated cells, and colony yield of hematopoietic progenitor cells in vitro were counted. The levels of erythropoietin, thrombopoietin, and granulocyte macrophage colony-stimulating factor in plasma were measured by enzyme-linked immunosorbent assay. Bone marrow cell cycle was performed by flow cytometry. The expression of apoptotic protein bcl-2, bax, and caspase-3 was detected by Western blotting. Results: Both Re and $Rk_3$ could improve peripheral blood cells, bone marrow nucleated cell counts, thymus index, and spleen index. Furthermore, they could enhance the yield of colonies cultured in vitro and make the levels of granulocyte macrophage colony-stimulating factor, erythropoietin, and thrombopoietin normal, reduce the ratio of $G_0/G_1$ phase cells, and increase the proliferation index. Finally, Re and $Rk_3$ could upregulate the expression of bcl-2, whereas they could downregulate the expression of bax and caspase-3. Conclusion: Re and $Rk_3$ could improve the hematopoietic function of myelosuppressed mice. The effect of $Rk_3$ was superior to that of Re at any dose. Regulating the levels of cytokines, promoting cells enter the normal cell cycle, regulating the balance of bcl-2/bax, and inhibiting the expression of caspase-3 may be the effects of Re and $Rk_3$ on myelosuppression.

Methylation Status of Transcriptional Modulatory Genes Associated with Colorectal Cancer in Northeast China

Han-Lu Gao,Xuan Wang,Hong-Ru Sun,Jun-De Zhou,Shang-Qun Lin,Yu-Hang Xing,Lin Zhu,Hai-Bo Zhou,Ya-Shuang Zhao,Qiang Chi,Yu-Peng Liu 거트앤리버 소화기연관학회협의회 2018 Gut and Liver Vol.12 No.2

Background/Aims: Methylation status plays a causal role in carcinogenesis in targeted tissues. However, the relationship between the DNA methylation status of multiple genes in blood leukocytes and colorectal cancer (CRC) susceptibility as well as interactions between dietary factors and CRC risks are unclear. Methods: We performed a case-control study with 466 CRC patients and 507 cancer-free controls to investigate the association among the methylation status of individual genes, multiple CpG site methylation (MCSM), multiple CpG site heterogeneous methylation and CRC susceptibility. Peripheral blood DNA methylation levels were detected by performing methylation-sensitive high-resolution melting. Results: Total heterogeneous methylation of CA10 and WT1 conferred a significantly higher risk of CRC (adjusted odds ratio [ORadjusted], 5.445; 95% confidence interval [CI], 3.075 to 9.643; ORadjusted, 1.831; 95% CI, 1.100 to 3.047; respectively). Subjects with high-level MCSM (MCSM-H) status demonstrated a higher risk of CRC (ORadjusted, 4.318; 95% CI, 1.529 to 12.197). Additionally, interactions between the high-level intake of fruit and CRH, WT1, and MCSM on CRC were statistically significant. Conclusions: The gene methylation status of blood leukocytes may be associated with CRC risk. MCSM-H of blood leukocytes was associated with CRC, especially in younger people. Some dietary factors may affect hypermethylation status and influence susceptibility to CRC.

The antibacterial activity of E. coli bacteriophage lysin lysep3 is enhanced by fusing the Bacillus amyloliquefaciens bacteriophage endolysin binding domain D8 to the C-terminal region

Shuang Wang,Jingmin Gu,Meng Lv,Zhimin Guo,Guangmou Yan,Ling Yu,Chongtao Du,Xin Feng,Wenyu Han,Changjiang Sun,Liancheng Lei 한국미생물학회 2017 The journal of microbiology Vol.55 No.5

Bacteriophage endolysin is one of the most promising antibioticsubstitutes, but in Gram-negative bacteria, the outermembrane prevents the lysin from hydrolyzing peptidoglycansand blocks the development of lysin applications. Theprime strategy for new antibiotic substitutes is allowing lysinto access the peptidoglycan from outside of the bacteria byreformation of the lysin. In this study, the novel Escherichiacoli (E. coli) phage lyase lysep3, which lacks outside-in catalyticability, was fused with the N-terminal region of theBacillus amyloliquefaciens lysin including its cell wall bindingdomain D8 through the best manner of protein fusionbased on the predicted tertiary structure of lysep3-D8 to obtainan engineered lysin that can lyse bacteria from the outside. Our results showed that lysep3-D8 could lyse both Gramnegativeand Gram-positive bacteria, whereas lysep3 and D8have no impact on bacterial growth. The MIC of lysep3-D8on E. coli CVCC1418 is 60 μg/ml; lysep3-D8 can inhibit thegrowth of bacteria up to 12 h at this concentration. The bactericidalspectrum of lysep3-D8 is broad, as it can lyse of allof 14 E. coli strains, 3 P. aeruginosa strains, 1 Acinetobacterbaumannii strain, and 1 Streptococcus strain. Lysep3-D8 hassufficient bactericidal effects on the 14 E. coli strains testedat the concentration of 100 μg/ml. The cell wall binding domainof the engineered lysin can destroy the integrity of theouter membrane of bacteria, thus allowing the catalytic domainto reach its target, peptidoglycan, to lyse the bacteria. Lysep3-D8 can be used as a preservative in fodder to benefitthe health of animals. The method we used here proved to bea successful exploration of the reformation of phage lysin.

A Vitis vinifera xanthine dehydrogenase gene, VvXDH, enhances salinity tolerance in transgenic Arabidopsis

Shuang-Hong You,Bo Zhu,Feibing Wang,Hong-Juan Han,Miao Sun,Hengweng Zhu,Ri-he Peng,Quan-Hong Yao 한국식물생명공학회 2017 Plant biotechnology reports Vol.11 No.3

Xanthine dehydrogenase (EC1.1.1.204; XDH) plays an important role in purine catabolism that catalyzes the oxidative hydroxylation of hypoxanthine to xanthine and of xanthine to uric acid. Long attributed to its role in recycling and remobilization of nitrogen, recently, XDH is implicated in plant stress responses and acclimation, such research efforts, however, have thus far been restricted to Arabidopsis XDH-knockdown/knockout studies. This study, using an ectopic overexpression approach, is expected to provide novel findings. In this study, a XDH gene from Vitis vinifera, named VvXDH, was synthesized and overexpressed in Arabidopsis, the transgenic Arabidopsis showed enhanced salt tolerance. The VvXDH gene was investigated and the results demonstrated the explicit role of VvXDH in conferring salt stress by increasing allantoin accumulation and activating ABA signaling pathway, enhancing ROS scavenging in transgenic Arabidopsis. In addition, the water loss and chlorophyll content loss were reduced in transgenic plants; the transgenic plants showed higher proline level and lower MDA content than that of wild-type Arabidopsis, respectively. In conclusion, the VvXDH gene has the potential to be applied in increasing allantoin accumulation and enhancing the tolerance to abiotic stresses in Arabidopsis and other plants.

Two-Stream Convolutional Neural Network for Video Action Recognition

( Han Qiao ),( Shuang Liu ),( Qingzhen Xu ),( Shouqiang Liu ),( Wanggan Yang ) 한국인터넷정보학회 2021 KSII Transactions on Internet and Information Syst Vol.15 No.10

Video action recognition is widely used in video surveillance, behavior detection, human-computer interaction, medically assisted diagnosis and motion analysis. However, video action recognition can be disturbed by many factors, such as background, illumination and so on. Two-stream convolutional neural network uses the video spatial and temporal models to train separately, and performs fusion at the output end. The multi segment Two-Stream convolutional neural network model trains temporal and spatial information from the video to extract their feature and fuse them, then determine the category of video action. Google Xception model and the transfer learning is adopted in this paper, and the Xception model which trained on ImageNet is used as the initial weight. It greatly overcomes the problem of model underfitting caused by insufficient video behavior dataset, and it can effectively reduce the influence of various factors in the video. This way also greatly improves the accuracy and reduces the training time. What’s more, to make up for the shortage of dataset, the kinetics400 dataset was used for pre-training, which greatly improved the accuracy of the model. In this applied research, through continuous efforts, the expected goal is basically achieved, and according to the study and research, the design of the original dual-flow model is improved.

Phytoremediation of 2,4,6-trinitrotoluene by Arabidopsis plants expressing a NAD(P)H-flavin nitroreductase from Enterobacter cloacae

Shuang-Hong You,Bo Zhu,Hong-Juan Han,Bo Wang,Ri-He Peng,Quan-Hong Yao 한국식물생명공학회 2015 Plant biotechnology reports Vol.9 No.6

2,4,6-Trinitrotoluene (TNT) is released into natural environment from demilitarization facilities, manufacturing, and explosive remnants of war; this compound is one of the most recalcitrant explosives. TNT contamination is associated with human health risks because TNT strongly causes mutagenicity and carcinogenicity. Unfortunately, effective and affordable technologies to remediate TNT-contaminated environments are insufficient. As such, studies have been conducted to develop strategies using plants to extract and detoxify TNT from environment. In this study, a system was designed to overcome high phytotoxicity of TNT by expressing a NAD(P)H-flavin nitroreductase from Enterobacter cloacae to investigate the possibility of TNT phytoremediation. The resulting transgenic Arabidopsis showed a remarkable improvement in the ability to tolerate, absorb, and detoxify TNT as evidenced by their growth condition. This study can be used as reference to facilitate the effective cleanup of TNTcontaminated sites.

Investigation of muscle-specific beef color stability at different ultimate pHs

Wu Shuang,Han Jina,Liang Rongrong,Dong Pengcheng,Zhu Lixian,Hopkins David L.,Zhang Yimin,Luo Xin 아세아·태평양축산학회 2020 Animal Bioscience Vol.33 No.12

Objective: This study was aimed to investigate the muscle-specific beef color stability at normal and high ultimate pHs. Methods: The impact of muscle (Longissimus lumborum [LL] vs psoas major [PM]) and pH (normal ultimate pH [Np] vs high pH dark cutting beef [Hp]) on color stability, indicated by basic color traits, metmyoglobin reducing activity (MRA) and oxygen consumption (OC), as well as the lipid oxidation, were determined over 7 days of display at 4°C. Results: Hp-LL had the highest pH (6.92), followed by Hp-PM (6.01), Np-PM (5.76), and Np-LL (5.52). Hp-LL had increased (p<0.05) a*, chroma and % oxymyoglobin during display. Hp-LL also had the highest metmyoglobin (MMb) reducing activity and OC among all the samples, thus, the greatest color stability, although very dark throughout storage, with lowest values for lightness (L*) and yellowness (b*). Np-LL also exhibited relatively high color stability, as a result of its lower % MMb and OC and higher MRA than psoas muscle samples. The 0.2 unit difference of the pH between Hp and Np psoas muscle, resulted in the difference of the color intensity, not the color stability. Interestingly, high pH psoas muscle (Hp-PM) did not have better color stability than Np-PM, and in fact had lower color stability than even Np-LL. The similar level of OC and lipid oxidation cannot explain the difference in color stability between Hp-PM and Np-LL. Conclusion: The Hp does not always show better color stability compared with Np beef, which depends on the muscle type. The balance of MRA and OC is important to keep the color in great intensity and stability in the meantime.

Studies of the effects and mechanisms of ginsenoside Re and Rk₃ on myelosuppression induced by cyclophosphamide

Jiahong Han,Jing Xia,Lianxue Zhang,Enbo Cai,Yan Zhao,Xuan Fei,Xiaohuan Jia,He Yang,Shuangli Liu 고려인삼학회 2019 Journal of Ginseng Research Vol.43 No.4

Background: Ginsenoside Re (Re) is one of the major components of Panax ginseng Meyer. Ginsenoside Rk₃ (Rk₃) is a secondary metabolite of Re. The aim of this study was to investigate and compare the effects and underlying mechanisms of Re and Rk₃ on cyclophosphamide-induced myelosuppression. Methods: The mice myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide. Peripheral blood cells, bone marrow nucleated cells, and colony yield of hematopoietic progenitor cells in vitro were counted. The levels of erythropoietin, thrombopoietin, and granulocyte macrophage colony-stimulating factor in plasma were measured by enzyme-linked immunosorbent assay. Bone marrow cell cycle was performed by flow cytometry. The expression of apoptotic protein bcl-2, bax, and caspase-3 was detected by Western blotting. Results: Both Re and Rk₃ could improve peripheral blood cells, bone marrow nucleated cell counts, thymus index, and spleen index. Furthermore, they could enhance the yield of colonies cultured in vitro and make the levels of granulocyte macrophage colony-stimulating factor, erythropoietin, and thrombopoietin normal, reduce the ratio of G₀/G₁ phase cells, and increase the proliferation index. Finally, Re and Rk₃ could upregulate the expression of bcl-2, whereas they could downregulate the expression of bax and caspase-3. Conclusion: Re and Rk₃ could improve the hematopoietic function of myelosuppressed mice. The effect of Rk₃ was superior to that of Re at any dose. Regulating the levels of cytokines, promoting cells enter the normal cell cycle, regulating the balance of bcl-2/bax, and inhibiting the expression of caspase-3 may be the effects of Re and Rk₃ on myelosuppression.