• Title/Summary/Keyword: Key Technology

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Insertion Mutation in HMG-CoA Lyase Increases the Production Yield of MPA through Agrobacterium tumefaciens-Mediated Transformation

  • Dong, Yuguo;Zhang, Jian;Xu, Rui;Lv, Xinxin;Wang, Lihua;Sun, Aiyou;Wei, Dongzhi
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1924-1932
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    • 2016
  • Mycophenolic acid (MPA) is an antibiotic produced by Penicillium brevicompactum. MPA has antifungal, antineoplastic, and immunosuppressive functions, among others. ${\beta}-Hydroxy-{\beta}-methylglutaryl-CoA$ (HMG-CoA) lyase is a key enzyme in the bypass metabolic pathway. The inhibitory activity of HMG-CoA lyase increases the MPA biosynthetic flux by reducing the generation of by-products. In this study, we cloned the P. brevicompactum HMG-CoA lyase gene using the thermal asymmetric interlaced polymerase chain reaction and gene walking technology. Agrobacterium tumefaciens-mediated transformation (ATMT) was used to insert a mutated HMG-CoA lyase gene into P. brevicompactum. Successful insertion of the HMG-CoA lyase gene was confirmed by hygromycin screening, PCR, Southern blot analysis, and enzyme content assay. The maximum MPA production by transformants was 2.94 g/l. This was 71% higher than wild-type ATCC 16024. Our results demonstrate that ATMT may be an alternative practical genetic tool for directional transformation of P. brevicompactum.

Scanning Rayleigh Doppler Lidar for Wind Profiling Based on Non-polarized Beam Splitter Cube Optically Contacted FPI

  • Zheng, Jun;Sun, Dongsong;Chen, Tingdi;Zhao, Ruocan;Han, Yuli;Li, Zimu;Zhou, Anran;Zhang, Nannan
    • Current Optics and Photonics
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    • v.2 no.2
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    • pp.195-202
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    • 2018
  • A Scanning Rayleigh Doppler lidar for wind profiling based on a non-polarized beam splitter cube optically contacted FPI is developed for wind measurement from high troposphere to low stratosphere in 5-35 km. Non-polarized beam splitter cube optically contacted to the FPI are used for a stable optical receiver. Zero Doppler shift correction is used to correct for laser or FPI frequency jitter and drift and the timing sequence is designed. Stability of the receiver for Doppler shift discrimination is validated by measuring the transmissions of FPI in different days and analyzed the response functions. The maximal relative wind deviation due to the stability of the optical receiver is about 4.1% and the standard deviation of wind velocity is 1.6% due to the stability. Wind measurement comparison experiments were carried out in Jiuquan ($39.741^{\circ}N$, $98.495^{\circ}E$), Gansu province of China in 2015, showing good agreement with radiosonde result data. Continuous wind field observation was performed from October 16th to November 12th and semi-continuous wind field of 19 nights are presented.

Fumigaclavine C attenuates adipogenesis in 3T3-L1 adipocytes and ameliorates lipid accumulation in high-fat diet-induced obese mice

  • Yu, Wan-Guo;He, Yun;Chen, Yun-Fang;Gao, Xiao-Yao;Ning, Wan-E;Liu, Chun-You;Tang, Ting-Fan;Liu, Quan;Huang, Xiao-Cheng
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.3
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    • pp.161-169
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    • 2019
  • Fumigaclavine C (FC), an active indole alkaloid, is obtained from endophytic Aspergillus terreus (strain No. FC118) by the root of Rhizophora stylosa (Rhizophoraceae). This study is designed to evaluate whether FC has anti-adipogenic effects in 3T3-L1 adipocytes and whether it ameliorates lipid accumulation in high-fat diet (HFD)-induced obese mice. FC notably increased the levels of glycerol in the culture supernatants and markedly reduced lipid accumulation in 3T3-L1 adipocytes. FC differentially inhibited the expressions of adipogenesis-related genes, including the peroxisome proliferator-activated receptor proteins, CCAAT/enhancer-binding proteins, and sterol regulatory element-binding proteins. FC markedly reduced the expressions of lipid synthesis-related genes, such as the fatty acid binding protein, lipoprotein lipase, and fatty acid synthase. Furthermore, FC significantly increased the expressions of lipolysis-related genes, such as the hormone-sensitive lipase, Aquaporin-7, and adipose triglyceride lipase. In HFD-induced obese mice, intraperitoneal injections of FC decreased both the body weight and visceral adipose tissue weight. FC administration significantly reduced lipid accumulation. Moreover, FC could dose-dependently and differentially regulate the expressions of lipid metabolism-related transcription factors. All these data indicated that FC exhibited anti-obesity effects through modulating adipogenesis and lipolysis.

Comparison of immunoadjuvant activities of four bursal peptides combined with H9N2 avian influenza virus vaccine

  • Zhang, Cong;Zhou, Jiangfei;Liu, Zhixin;Liu, Yongqing;Cai, Kairui;Shen, Tengfei;Liao, Chengshui;Wang, Chen
    • Journal of Veterinary Science
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    • v.19 no.6
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    • pp.817-826
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    • 2018
  • The bursa of Fabricius (BF) is a central humoral immune organ unique to birds. Four bursal peptides (BP-I, BP-II, BP-III, and BP-IV) have been isolated and identified from the BF. In this study, the immunoadjuvant activities of BPs I to IV were examined in mice immunized with H9N2 avian influenza virus (AIV) vaccine. The results suggested that BP-I effectively enhanced cell-mediated immune responses, increased the secretion of Th1 (interferon gamma)- and Th2 (interleukin-4)-type cytokines, and induced an improved cytotoxic T-lymphocyte (CTL) response to the H9N2 virus. BP-II mainly elevated specific antibody production, especially neutralizing antibodies, and increased Th1- and Th2-type cytokine secretion. BP-III had no significant effect on antibody production or cell-mediated immune responses compared to those in the control group. A strong immune response at both the humoral and cellular levels was induced by BP-IV. Furthermore, a virus challenge experiment followed by H&E staining revealed that BP-I and BP-II promoted removal of the virus and conferred protection in mouse lungs. BP-IV significantly reduced viral titers and histopathological changes and contributed to protection against H9N2 AIV challenge in mouse lungs. This study further elucidated the immunoadjuvant activities of BPs I to IV, providing a novel insight into immunoadjuvants for use in vaccine design.

Physiological Characteristics of Lactobacillus casei Strains and Their Alleviation Effects against Inflammatory Bowel Disease

  • Liu, Yang;Li, Yifeng;Yu, Xinjie;Yu, Leilei;Tian, Fengwei;Zhao, Jianxin;Zhang, Hao;Zhai, Qixiao;Chen, Wei
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.92-103
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    • 2021
  • Lactobacillus casei, one of the most widely used probiotics, has been reported to alleviate multiple diseases. However, the effects of this species on intestinal diseases are strain-specific. Here, we aimed to screen L. casei strains with inflammatory bowel disease (IBD)-alleviating effects based on in vitro physiological characteristics. Therefore, the physiological characteristics of 29 L. casei strains were determined, including gastrointestinal transit tolerance, oligosaccharide fermentation, HT-29 cell adhesion, generation time, exopolysaccharide production, acetic acid production, and conjugated linoleic acid synthesis. The effects of five candidate strains on mice with induced colitis were also evaluated. The results showed that among all tested L. casei strains, only Lactobacillus casei M2S01 effectively relieved colitis. This strain recovered body weight, restored disease activity index score, and promoted anti-inflammatory cytokine expression. Gut microbiota sequencing showed that L. casei M2S01 restored a healthy gut microbiome composition. The western blotting showed that the alleviating effects of L. casei M2S01 on IBD were related to the inhibition of the NF-κB pathway. A good gastrointestinal tolerance ability may be one of the prerequisites for the IBD-alleviating effects of L. casei. Our results verified the efficacy of L. casei in alleviating IBD and lay the foundation for the rapid screening of L. casei strain with IBD-alleviating effects.

A Fiber Optic Sensor for Determination of 2,4-Dichlorophenol Based on Oxygen Oxidation Catalyzed by Iron(III) Tetrasulfophthalocyanine

  • Tong, Yilin;Li, Dapeng;Huang, Jun;Zhang, Cong;Li, Kun;Ding, Liyun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.11
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    • pp.3307-3311
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    • 2013
  • A new fiber optical sensor was developed for the determination of 2,4-dichlorophenol (DCP). The sensor was based on DCP oxidation by oxygen with the catalysis of iron(III) tetrasulfophthalocyanine (Fe(III)PcTs). The optical oxygen sensing film with $Ru(bpy)_3Cl_2$ as the fluorescence indicator was used to determine the consumption of oxygen in solution. A lock-in amplifier was used for detecting the lifetime of the oxygen sensing film by measuring the phase delay change of the sensor head. The different variables affecting the sensor performance were evaluated and optimized. Under the optimal conditions (i.e. pH 6.0, $25^{\circ}C$, Fe(III)PcTs concentration of 0.62 mg/mL), the linear detection range and response time of the sensor are $1.0{\times}10^{-6}-9.0{\times}10^{-6}$ mol/L and 250 s, respectively. The sensor displays high selectivity, good repeatability and stability, and can be used as an effective tool in analyzing DCP concentration in practical samples.

MoJMJD6, a Nuclear Protein, Regulates Conidial Germination and Appressorium Formation at the Early Stage of Pathogenesis in Magnaporthe oryzae

  • Li Zhang;Dong Li;Min Lu;Zechi Wu;Chaotian Liu;Yingying Shi;Mengyu Zhang;Zhangjie Nan;Weixiang Wang
    • The Plant Pathology Journal
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    • v.39 no.4
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    • pp.361-373
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    • 2023
  • In plant-pathogen interactions, Magnaporthe oryzae causes blast disease on more than 50 species of 14 monocot plants, including important crops such as rice, millet, and most 15 recently wheat. M. oryzae is a model fungus for studying plant-microbe interaction, and the main source for fungal pathogenesis in the field. Here we report that MoJMJD6 is required for conidium germination and appressorium formation in M. oryzae. We obtained MoJMJD6 mutants (ΔMojmjd6) using a target gene replacement strategy. The MoJMD6 deletion mutants were delayed for conidium germination, glycogen, and lipid droplets utilization and consequently had decreased virulence. In the ΔMojmjd6 null mutants, global histone methyltransferase modifications (H3K4me3, H3K9me3, H3K27me3, and H3K36me2/3) of the genome were unaffected. Taken together, our results indicated that MoJMJD6 function as a nuclear protein which plays an important role in conidium germination and appressorium formation in the M. oryzae. Our work provides insights into MoJMJD6-mediated regulation in the early stage of pathogenesis in plant fungi.

Effect of fermented blueberry on the oxidative stability and volatile molecule profiles of emulsion-type sausage during refrigerated storage

  • Zhou, Hengyue;Zhuang, Xinbo;Zhou, Changyu;Ding, Daming;Li, Chunbao;Bai, Yun;Zhou, Guanghong
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.5
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    • pp.812-824
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    • 2020
  • Objective: The aim of this work was to assess the effect of fermented blueberry (FB; 2%, 4%, and 6%) on the oxidative stability and volatile molecule profiles of emulsion-type sausage stored at 4℃ for 28 days. Methods: The antioxidant activity of FB was determined through radical-scavenging activity against 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals. Four formulations of sausage treatments with different FB levels (0%, 2%, 4%, 6%) were prepared, then peroxide value (POVs), thiobarbituric acid-reactive substances (TBARS) values, protein carbonyls and thiol groups were measured. The aroma profiles of sausages for each treatment was also determined. Results: The half maximal inhibitory concentration indicated that FB had greater scavenging ability than ascorbic acid against DPPH and hydroxyl radicals. Sausages with FB significantly retarded increases in POVs and TBARS, as well as in the content of protein carbonyls during all storage days (p<0.05). Particularly, 4% and 6% FB-treated sausages had better oxidation inhibition effects. However, FB accelerated the reduction in thiol groups (p<0.05). Additionally, FB inhibits the excessive formation of aldehyde compounds; for example, hexanal, which may cause rancid flavors, decreased from 58.25% to 19.41%. FB also created 6 alcohols (i.e., 2-methyl-1-propanol, 3-methyl-1-butanol, and phenylethyl alcohol), 5 ester compounds (i.e., ethyl acetate, ethyl lactate, and ethyl hexanoate) and 3-hydroxy-2-butanone in the sausages that contribute to sausage flavors. The principal component analysis showed that the aroma profiles of sausages with and without FB are easily identified. Conclusion: The addition of FB could significantly reduce the lipid and protein oxidation and improve oxidative stability for storage. Also, adding FB could inhibit rancid flavors and contribute to sausage flavors.

The Integrins Involved in Soybean Agglutinin-Induced Cell Cycle Alterations in IPEC-J2

  • Pan, Li;Zhao, Yuan;Yuan, Zhijie;Farouk, Mohammed Hamdy;Zhang, Shiyao;Bao, Nan;Qin, Guixin
    • Molecules and Cells
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    • v.40 no.2
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    • pp.109-116
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    • 2017
  • Soybean agglutinin (SBA) is an anti-nutritional factor of soybean, affecting cell proliferation and inducing cytotoxicity. Integrins are transmembrane receptors, mediating a variety of cell biological processes. This research aims to study the effects of SBA on cell proliferation and cell cycle progression of the intestinal epithelial cell line from piglets (IPEC-J2), to identify the integrin subunits especially expressed in IPEC-J2s, and to analyze the functions of these integrins on IPEC-J2 cell cycle progression and SBA-induced IPEC-J2 cell cycle alteration. The results showed that SBA lowered cell proliferation rate as the cell cycle progression from G0/G1 to S phase (P < 0.05) was inhibited. Moreover, SBA lowered mRNA expression of cell cycle-related gene CDK4, Cyclin E and Cyclin D1 (P < 0.05). We successfully identified integrins ${\alpha}2$, ${\alpha}3$, ${\alpha}6$, ${\beta}1$, and ${\beta}4$ in IPEC-J2s. These five subunits were crucial to maintain normal cell proliferation and cell cycle progression in IPEC-J2s. Restrain of either these five subunits by their inhibitors, lowered cell proliferation rate, and arrested the cells at G0/G1 phase of cell cycle (P < 0.05). Further analysis indicated that integrin ${\alpha}2$, ${\alpha}6$, and ${\beta}1$ were involved in the blocking of G0/G1 phase induced by SBA. In conclusion, these results suggested that SBA lowered the IPEC-J2 cell proliferation rate through the perturbation of cell cycle progression. Furthermore, integrins were important for IPEC-J2 cell cycle progression, and they were involved in the process of SBA-induced cell cycle progression alteration, which provide a basis for further revealing SBA anti-proliferation and anti-nutritional mechanism.

Real Time Related Key Attack on Hummingbird-2

  • Zhang, Kai;Ding, Lin;Li, Junzhi;Guan, Jie
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.6 no.8
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    • pp.1946-1963
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    • 2012
  • Hummingbird is a lightweight encryption and message authentication primitive published in RISC'09 and WLC'10. In FSE'11, Markku-Juhani O.Saarinen presented a differential divide-and-conquer method which has complexity upper bounded by $2^{64}$ operations and requires processing of few megabytes of chosen messages under two related nonces (IVs). The improved version, Hummingbird-2, was presented in RFIDSec 2011. Based on the idea of differential collision, this paper discovers some weaknesses of the round function WD16. Combining with the simple key loading algorithm, a related-key chosen-IV attack which can recover the full secret key is proposed. Under 15 pairs of related keys, the 128 bit initial key can be recovered, requiring $2^{27}$ chosen IV and the computational complexity is $O(2^{27})$. In average, the attack needs several minutes to recover the full 128-bit secret key on a PC. The experimental result corroborates our attack. The result shows that the Hummingbird-2 cipher can't resist related key attack.