Silencing downstream of receptor kinase gene (drk) impairs larval-pupal ecdysis in Leptinotarsa decemlineata (Say)

Pan Deng,Jun-Li Du,Li-Li Mu,Kai-Yun Fu,Wen-Chao Guo,Guo-Qing Li 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.1

In insects, an insulin-like peptide (ILP) triggers the formation of the insulin receptor (InR)/the insulin receptor substrate Chico complex. The complex then recruits downstream of receptor kinase (Drk) and phosphatidylinositol-3-kinase (PI3K) to initiate two signaling branches, i.e., Drk-mitogen-activated protein kinase (MAPK) and Pi3K-protein kinase B subdivisions. Previous findings reveal that RNA interference (RNAi) of LdILP2 or Ldchico, rather than Ldpi3k92E, impairs larval-pupal and pupal-adult molting in the Colorado potato beetle Leptinotarsa decemlineata. It is accordingly hypothesized that the Drk-MAPK branch regulates larval metamorphosis. In the present paper, we first found that silencing LdILP2, Ldchico or Ldpi3k92E did not decrease the expression level of Lddrk, indicating other receptor tyrosine kinases’ signaling except insulin pathway is not affected in the RNAi larvae. Moreover, two InRs and Torso were highly expressed in the final larval instars. Furthermore, RNAi of either Lddrk or Ldchico, or both of them equally affected larval-pupal and pupal-adult molts, and similarly repressed the expression of representative MAPK (Ldras and Ldraf), ecdysteroidogenesis (Ldphm and Ldsad), and 20E signaling (LdEcR, LdUSP, LdHR3 and LdE75) genes. 20E feeding by Lddrk RNAi larvae completely restored the reduced mRNA levels of LdEcR, LdHR3 and LdE75, but did not rescued the decreased Lddrk and LdUSP levels and the lowered pupation and emergence rates. Therefore, our findings suggest that the Drk-MAPK branch is involved in metamorphosis regulation in L. decemlineata.

Experimental Study of Cooling Speed for Ultra-Thick Steel Plate during the Jet Impinging and Quenching Process

Tian-Liang Fu,Xiang-Tao Deng,Guo-Huai Liu,Zhao-Dong Wang,Guo-Dong Wang 한국정밀공학회 2016 International Journal of Precision Engineering and Vol.17 No.11

The quenching temperature drop curve for Q345B steel plate with 84 mm and 170 mm thickness was tested to analyze the distributing regularities and influencing factors of cooling speed for ultra-thick steel plate during the jet impinging and quenching process. The influences for temperature drop, temperature gradient and cooling speed were analyzed under the conditions of 60~100 m3/h water amount, 0.4~1.0 MPa water pressure, transient switching of quenching mode and the distribution of heat exchanger. Threedimensional heat anti transfer model, surface heat transfer coefficient model and thermal physical parameter model were built up by finite element and optimization. The results showed that the deviation of calculated and measured values was less than 4% for temperature drop curve model. The cooling speed of vertical section for 84 mm-thick steel plate was approximately proportional to surface heat transfer coefficient. The influence of surface heat transfer to cooling speed became weak when the thickness was increased. The influences of temperature effect when switching different quenching modes and temperature gradient of vertical section to cooling speed were stronger. The minimum value of cooling speed was about 1.0~1.8oC/s, between H/6 and H/3 region. These data provide the key information for increasing the cooling speed and uniformity.

Biocatalysis and Fermentation Technology : Secretory Expression, Functional Characterization, and Molecular Genetic Analysis of Novel Halo-Solvent-Tolerant Protease from Bacillus gibsonii

( Ai Hua Deng ),( Guo Qiang Zhang ),( Na Na Shi ),( Jie Wu ),( Fu Ping Lu ),( Ting Yi Wen ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.2

A novel protease gene from Bacillus gibsonii, aprBG, was cloned, expressed in B. subtilis, and characterized. High-level expression of aprBG was achieved in the recombinant strain when a junction was present between the promoter and the target gene. The purified recombinant enzyme exhibited similar N-terminal sequences and catalytic properties to the native enzyme, including high affinity and hydrolytic efficiency toward various substrates and a superior performance when exposed to various metal ions, surfactants, oxidants, and commercial detergents. AprBG was remarkably stable in 50% organic solvents and retained 100% activity and stability in 0-4 M NaCl, which is better than the characteristics of previously reported proteases. AprBG was most closely related to the high-alkaline proteases of the subtilisin family with a 57-68% identity. The secretion and maturation mechanism of AprBG was dependent on the enzyme activity, as analyzed by site-directed mutagenesis. Thus, when taken together, the results revealed that the halo-solvent-tolerant protease AprBG displays significant activity and stability under various extreme conditions, indicating its potential for use in many biotechnology applications.

Deterministic Secure Quantum Communication Without Maximally Entangled States

Xi-Han Li,Fu-Guo Deng,Chun-Yan Li,Hong-Yu Zhou,Ping Zhou,Yu-Jie Liang 한국물리학회 2006 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.49 No.4

Two deterministic secure quantum communication schemes are proposed, one based on pure entangled states and the other on d-dimensional single-photon states. In these two schemes, only single-photon measurements are required for the two authorized users, which makes the schemes more convenient than others in practical applications. Although each qubit can be read out after a transmission of additional classical bit, it is unnecessary for the users to transmit qubits double the distance between the sender and the receiver, which will increase their bit rate and their security. The parties use decoy photons to check eavesdropping efficiently. The obvious advantage in the first scheme is that the pure entangled source is feasible with present techniques.

Transcriptional response of Methoprene-tolerant (Met) gene to three insect growth disruptors in Leptinotarsa decemlineata (Say)

Qing-Wei Meng,Qing-Yu Xu,Pan Deng,Kai-Yun Fu,Wen-Chao Guo,Guo-Qing Li 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.2

Some insect growth disruptors (IGDs), such as pyriproxyfen and halofenozide, may be used to control Leptinotarsa decemlineata. However, their mechanism of action remains elusive. Methoprene-tolerant (Met) mediates juvenile hormone (JH) signal to control numerous essential physiological processes. In the present paper, we identified a Met gene (LdMet). LdMet protein was a typical basic helix-loop-helix/Per-Arnt-Sim (bHLHPAS) transcription factor with a bHLH domain, two PAS domains (PAS-A and PAS-B) and a region called PAS associated C terminal (PAC). Eight conserved amino acids critical for JH binding were located in PAS-B and PAC domains. The temporal expression pattern of LdMet was in accordance with the variation of circulating JH titers. Feeding of juvenoid methoprene or pyriproxyfen, or provide for JH dose-dependently stimulated the expression of LdMet. RNA interference-mediated knockdown of two JH degradation genes increased the transcription of LdMet, while silencing of a JH biosynthesis gene repressed the transcription. Conversely, ingestion of an ecdysteroid agonist halofenozide or 20-hydroxyecdysone (20E) reduced the mRNA levels of LdMet, in a dosedependent manner, whereas knockdown of either ecdysteroidogenesis or 20E signaling genes increased the mRNA accumulation. Providing that the expression of LdMet can be disturbed by methoprene, pyriproxyfen and halofenozide, LdMet may be a potential target of these IGDs in L. decemlineata larvae.

Characteristics Comparison before and after Lyophilization of Transferrin Modified Procationic-Liposome-Protamine-DNA Complexes (Tf-PLPD)

Zhi-Rong Zhong,Zhi-rong Zhang,Ji Liu,Yong Deng,Hong-wei Zhang,Yao Fu,Qing-guo Song,Qin He 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.1

A novel non-viral gene delivery system, Procationic-Liposome-Protamine-DNA complexes (PLPD) which could further adsorb transferrin on the surface as a targeting ligand to form Tf- PLPD, was prepared and characterized before and after lyophilization. The size distribution of Tf-PLPD was in the range of 240 ± 12 nm and the zeta potential was -24.10 ± 2.5 mV. The transfection efficiencies of PLPD and Tf-PLPD were 12.18 ± 3.8 and 24.26 ± 2.6 mU β-galactosidase/ mg protein respectively. The lyophilization and the presence of serum didn’t affect the tansfectivities of PLPD or Tf-PLPD. Compared to LipofectamineTM 2000 (Invitrogen, U.S.A.), the procationic liposomes had less cytotoxicity to cells. In summary the procationic lipoplex described here, combining the condensing effect of protamine and the targeting capability of Tf, was a perspective non-viral vector for gene delivery system.

Characteristics Comparison before and after Lyophilization of Transferrin Modified Procationic-Liposome-Protamine-DNA Complexes (Tf-PLPD)

Zhong, Zhi-Rong,Zhang, Zhi-Rong,Liu, Ji,Deng, Yong,Zhang, Hong-Wei,Fu, Yao,Song, Qing-Guo,He, Qin 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.1

A novel non-viral gene delivery system, Procationic-Liposome-Protamine-DNA complexes (PLPD) which could further adsorb transferrin on the surface as a targeting ligand to form Tf-PLPD, was prepared and characterized before and after lyophilization. The size distribution of Tf-PLPD was in the range of $240{\pm}12nm$ and the zeta potential was $-24.10{\pm}2.5mV$. The transfection efficiencies of PLPD and Tf-PLPD were $12.18{\pm}3.8\;and\;24.26{\pm}2.6mU\;{\beta}-galactosidase/mg$ protein respectively. The lyophilization and the presence of serum didn't affect the tansfectivities of PLPD or Tf-PLPD. Compared to $Lipofectamine^{TM}$ 2000 (Invitrogen, U.S.A.), the procationic liposomes had less cytotoxicity to cells. In summary the procationic lipoplex described here, combining the condensing effect of protamine and the targeting capability of Tf, was a perspective non-viral vector for gene delivery system.

Expression of Transcription Factor FOXC2 in Cervical Cancer and Effects of Silencing on Cervical Cancer Cell Proliferation

Zheng, Chun-Hua,Quan, Yuan,Li, Yi-Yang,Deng, Wei-Guo,Shao, Wen-Jing,Fu, Yan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.4

Objective: Forkhead box C2 (FOXC2) is a member of the winged helix/forkhead box (Fox) family of transcription factors. It has been suggested to regulate tumor vasculature, growth, invasion and metastasis, although it has not been studied in cervical cancer. Here, we analyzed FOXC2 expression in cervical tissues corresponding to different stages of cervical cancer development and examined its correlation with clinicopathological characteristics. In addition, we examined the effects of targeting FOXC2 on the biological behavior of human cervical cancer cells. Methods: The expression of FOXC2 in normal human cervix, CIN I-III and cervical cancer was examined by immunohistochemistry and compared among the three groups and between cervical cancers with different pathological subtypes. Endogenous expression of FOXC2 was transiently knocked down in human Hela and SiHa cervical cells by siRNA, and cell viability and migration were examined by scratch and CCK8 assays, respectively. Results: In normal cervical tissue the frequency of positive staining was 25% (10/40 cases), with a staining intensity (PI) of $0.297{\pm}0.520$, in CIN was 65% (26/40cases), with a PI of $3.00{\pm}3.29$, and in cancer was 91.8% (68/74 cases), with a PI of $5.568 {\pm}3.449$. The frequency was 100% in adenocarcinoma (5/5 cases) and 91.3% in SCCs (63/69 cases). The FOXC2 positive expression rate was 88.5% in patients with cervical SCC stage I and 100% in stage II, showing significant differences compared with normal cervix and CIN. With age, pathologic differentiation degree and tumor size, FOXC2 expression showed no significant variation. On transient transfection of Hela and SiHa cells, FOXC2-siRNA inhibition rates were 76.2% and 75.7%; CCK8 results showed reduced proliferation and relative migration (in Hela cells from $64.5{\pm}3.16$ to $49.5{\pm}9.24$ and in SiHa cells from $60.1{\pm}3.05$ to $44.3{\pm}3.98$) (P < 0.05). Conclusion: FOXC2 gene expression increases with malignancy, especially with blood vessel hyperplasia and invasion degree. Targeted silencing was associated with reduced cell proliferation as well as invasion potential.

QTL mapping and identification of candidate genes for cold tolerance at the germination stage in wild rice

Pan Ying-Hua,Nong Bao-Xuan,Chen Lei,Yang Xing-Hai,Xia Xiu-Zhong,Zhang Zong-Qiong,Qing Dong-Jin,Gao Ju,Huang Cheng-Cui,Li Dan-Ting,Deng Guo-Fu 한국유전학회 2023 Genes & Genomics Vol.45 No.7

Background Cold damage stress significantly affects rice growth (germination and seedling) and causes serious losses in yield in temperate and high-altitude areas around the globe. Objective This study aimed to explore the cold tolerance (CT) locus of rice and create new cold-tolerant germplasm. We constructed a chromosome segment substitution line (CSSL) with strong CT and fine mapped quantitative trait loci (QTLs) associated with CT by performing the whole-genome resequencing of CSSL with phenotypes under cold treatment. Methods A chromosome CSSL, including 271 lines from a cross between the cold-tolerant wild rice Y11 (Oryza rufipogon Griff.) and the cold-sensitive rice variety GH998, was developed to map QTLs conferring CT at the germination stage. The whole-genome resequencing was performed on CSSL for mapping QTLs of associated with CT at the germination stage. Results A high-density linkage map of the CSSLs was developed using the whole-genome resequencing of 1484 bins. The QTL analysis using 615,466 single-nucleotide polymorphisms (SNPs) led to the identification of 2 QTLs related to germination rate at low-temperature on chromosome 8 (qCTG-8) and chromosome 11 (qCTG-11). The qCTG-8 and qCTG-11 explained 14.55% and 14.31% of the total phenotypic variation, respectively. We narrowed down qCTG-8 and qCTG-11 to 195.5 and 78.83-kb regions, respectively. The expression patterns of important candidate genes in different tissues, and of RNA-sequencing (RNA-seq) in CSSLs, were identified based on gene sequences in qCTG-8 and qCTG-11 cold-induced expression analysis. LOC_Os08g01120 and LOC_Os08g01390 were identified as candidate genes in qCTG-8, and LOC_Os11g32880 was identified as a candidate gene in qCTG-11. Conclusions This study demonstrated a general method that could be used to identify useful loci and genes in wild rice and aid in the future cloning of candidate genes of qCTG-8 and qCTG-11. The CSSLs with strong CT were supported for breeding cold-tolerant rice varieties.