Construction of a Transposon-mediated Baculovirus Vector Hanpvid and a New Cell Line for Expressing Barnase

Qin, Qin,Liu, Ying-Le,Zhu, Ying,Li, Shun-Yi,Qi, Yi-Peng Korean Society for Biochemistry and Molecular Biol 2005 Journal of biochemistry and molecular biology Vol.38 No.1

In this study we developed the transposon-mediated shuttle vector 'Hanpvid', which composed of HaNPV (Heliothis armigera nuclear polyhedrosis virus) genomic DNA and a transposon cassette from Bacmid of Bac-to-Bac system. Hanpvid replicates in E. coli in the same way as Bacmid and retains infective function in cotton bollworm cells (Hz-AM1). Using Hanpvid we constructed a recombinant virus, which could infect Hz-AM1 cells and generate recombinant HaNPV (rHa-Bar) containing the barnase gene, a ribonuclease gene from Bacillus amyloliquefaciens. Since the expression vector carrying barnase gene cannot replicate in the absence of barstar, a specific inhibitor of barnase, we constructed a new cotton bollworm cell line (AM1-NB) using the marker rescue method. In AM1-NB barstar was integrated into the cellular chromosome to sustain the replication of rHa-Bar. To screen out recombinant HaNPV for potential use as biopesticide, Hz-AM1 and AM1-NB cell lines were infected with rHa-Bar, respectively. The results obtained indicate that Viral progenies in AM1-NB were 23 and 160 times greater than those in Hz-AM1 48 h and 72 h after infection, respectively. With additional insertion of the polyhedron gene from AcNPV (Autographa californica nuclear polyhedrosis virus) into the Hanpvid genome, rHa-Bar regained the polyhedron phenotype and its pest-killing rate greatly improved. Toxic analysis showed that the lethal dosages ($LD_{50}$) and the lethal time(s) ($LT_{50}$) of rHa-Bar were reduced by 20% and 30%, respectively, compared to wt-HaNPV in the third instar larvae of cotton bollworm. This study shows that in AM1-NB barnase can be effectively produced and used as pest-killing agent for the biological control of cotton pests.

Flexural Behavior of Anchor Horizontal Boundary Element in Steel Plate Shear Wall

Ying Qin,JinYu Lu,Li-Cheng-Xi Huang,Shi Cao 한국강구조학회 2017 International Journal of Steel Structures Vol.17 No.3

Steel plate shear walls (SPSWs) have been increasingly used in lateral load resisting system since the post-buckling strength of web plates was realized and considered in practical design. Diagonal tension field is formed in the web plate to resist the lateral force induced by winds and earthquakes. The force is anchored by the surrounded horizontal boundary elements (HBEs) and vertical boundary elements (VBEs) and eventually transferred to the ground. Therefore, the response of HBEs, especially anchor HBEs, is essential to ensure that SPSWs could exhibit the required strength, ductility, and energy dissipation capacity. This paper presents the results of theoretical studies into the flexural behavior of the anchor HBEs of SPSWs and can be regarded as an extension to the previous work by Qu and Bruneau (2011) and Qin et al. (2017). The boundary effect was considered to reflect the actual stress state at the HBE-to-VBE connection, which assembled the method by Qin et al. (2017). Furthermore, more proper distributions of the vertical component of tension field for the positive and negative flexure cases, respectively, were proposed comparing to the work by Qu and Bruneau (2011). The hand calculation approach for the plastic flexural capacity of the anchor HBEs was given by the summation of the contributions from the flanges and the web. The developed equations are compared with previous data and good agreement was found between them. Meanwhile, comprehensively discussions are conducted to evaluate the influence of key parameters on the flexural behavior of anchor HBE. It was found that the plastic flexural capacity of anchor HBE decreases from unity to the minimum as a result of the increase in shear force, axial force and vertical stresses. Moreover, the flexural response of anchor HBE is most vulnerable to the change in shear force. This indicates that boundary effect is significantly important and cannot be ignored in the analysis. The research in this paper provides basis for the capacity design of anchor HBE.

Construction of a Transposon-mediated Baculovirus Vector Hanpvid and a New Cell Line for Expressing Barnase

( Qin Qin ),( Ying Le Liu ),( Ying Zhu ),( Shun Yi Li ),( Yi Peng Qi ) 생화학분자생물학회 2005 BMB Reports Vol.38 No.1

In this study we developed the transposon-mediated shuttle vector `Hanpvid`, which composed of HaNPV (Heliothis armigera nuclear polyhedrosis virus) genomic DNA and a transposon cassette from Bacmid of Bac-to-Bac system. Hanpvid replicates in E. coli in the same way as Bacmid and retains infective function in cotton bollworm cells (Hz-AMU). Using Hanpvid we constructed a recombinant virus, which could infect Hz-AM1 cells and generate recombinant HaNPV (rHa-Bar) containing the barnase gene, a ribonuclease gene from Bacillus amyloliquefaciens. Since the expression vector carrying barnase gene cannot replicate in the absence of barstar, a specific inhibitor of barnase, we constructed a new cotton bollworm cell line (AMl-NB) using the marker rescue method. In AM1-NB barstar was integrated into the cellular chromosome to sustain the replication of rHa-Bar. To screen out recombinant HaNPV for potential use as biopesticide, Hz-AM! and AM1-NB cell lines were infected with rHa-Bar, respectively. The results obtained indicate that Viral progenies in AM1-NB were 23 and 160 times greater than those in Hz-AM1 48 h and 72 h after infection, respectively. With additional insertion of the polyhedron gene from AcNPV (Autographa californica nuclear polyhedrosis virus) into the Hanpvid genome, rHa-Bar regained the polyhedron phenotype and its pest-killing rate greatly improved. Toxic analysis showed that the lethal dosages (LD_(50)) and the lethal time(s) (LT_(50)) of rHa-Bar were reduced by 20% and 30%, respectively, compared to wt-HaNPV in the third instar larvae of cotton bollworm. This study shows that in AM1-NB barnase can be effectively produced and used as pest-killing agent for the biological control of cotton pests.

Buckling analysis of elastically-restrained steel plates under eccentric compression

Ying Qin,Gan-Ping Shu,Er-Feng Du,Rui-Hua Lu 국제구조공학회 2018 Steel and Composite Structures, An International J Vol.29 No.3

In this research, the explicit closed-form local buckling solution of steel plates in contact with concrete, with both loaded and unloaded edges elastically restrained against rotation and subjected to eccentric compression is presented. The Rayleigh-Rize approach is applied to establish the eigenvalue problem for the local buckling performance. Buckling shape which combines trigonometric and biquadratic functions is introduced according to that used by Qin <i>et al</i>. (2017) on steel plate buckling under uniform compression. Explicit solutions for predicting the local buckling stress of steel plate are obtained in terms of the rotational stiffness. Based on different boundary conditions, simply yet explicit local buckling solutions are discussed in details. The proposed formulas are validated against previous research and finite element results. The influences of the loading stress gradient parameter, the aspect ratio, and the rotational stiffness on the local buckling stress resultants of steel plates with different boundary conditions were evaluated. This work can be considered as an alternative to apply a different buckling shape function to study the buckling problem of steel plate under eccentric compression comparing to the work by Qin <i>et al</i>. (2018), and the results are found to be in consistent with those in Qin <i>et al</i>. (2018).

Anti-inflammatory effect of hispidin on LPS induced macrophage inflammation through MAPK and JAK1/STAT3 signaling pathways

Han Ying-Hao,Chen Dong-Qin,Jin Mei-Hua,Jin Ying-Hua,Li Jing,Shen Gui-Nan,Li Wei-Long,Gong Yi-Xi,Mao Ying-Ying,Xie Dan-Ping,Lee Dong-Seok,Yu Li-Yun,Kim Sun-Uk,김지수,권태호,Cui Yu-Dong,Sun Hu-Nan 한국응용생명화학회 2020 Applied Biological Chemistry (Appl Biol Chem) Vol.63 No.3

Severe inflammatory reactions caused by macrophage activation can trigger a systemic immune response. In the present study, we observed the anti-inflammatory properties of hispidin on LPS induced RAW264.7 macrophage cells. Our results showed that hispidin treatment significantly reduced the production of cellular NO, IL-6 and reactive oxygen species (ROS) while has not inhibitory effect on TNF-α productions. Excitingly, hispidin treatment retains the phagocytosis ability of macrophages which enabling them to perform the function of removing foreign invaders. Signaling studies showed, hispidin treatment dramatic suppressed the LPS induced mitogen activated protein kinases (MAPK) and JAK/STAT activations. In conclusion, our findings suggest that hispidin may be a new therapeutic target for clinical treatment of macrophages-mediated inflammatory responses.

Structural behavior of the stiffened double-skin profiled composite walls under compression

Ying Qin,Yong-Wei Li,Xu-Zhao Lan,Yu-Sen Su,Xiang-Yu Wang,Yuan-De Wu 국제구조공학회 2019 Steel and Composite Structures, An International J Vol.31 No.1

Steel-concrete composite walls have been proposed and developed for applications in various types of structures. The double-skin profiled composite walls, as a natural development of composite flooring, provide structural and architectural merits. However, adequate intermediate fasteners between profiled steel plates and concrete core are required to fully mobilize the composite action and to improve the structural behavior of the wall. In this research, two new types of fasteners (i.e., threaded rods and vertical plates) were proposed and three specimens with different fastener types or fastener arrangements were tested under axial compression. The experimental results were evaluated in terms of failure modes, axial load versus axial displacement response, strength index, ductility index, and load-strain relationship. It was found that specimen with symmetrically arranged thread rods sustained more stable axial strain than that with staggered arranged threaded rods. Meanwhile, vertical plates are more suitable for practical use since they provide stronger confinement to profiled steel plate and effectively prevent the steel plate from early local buckling, which eventually enhance the composite action and increase the axial compressive capacity of the wall. The calculation methods were then proposed and good agreement was observed between the test results and the predicted results.

Behavior of improved through-diaphragm connection to square tubular column under tensile loading

Qin, Ying,Zhang, Jing-Chen,Shi, Peng,Chen, Yi-Fu,Xu, Yao-Han,Shi, Zuo-Zheng 국제구조공학회 2018 Structural Engineering and Mechanics, An Int'l Jou Vol.68 No.4

Square tubular columns are commonly used in moment resisting frames, while through-diaphragm connection is the most typical configuration detail to connect the H-shaped beam to the column. However, brittle fracture normally occurs at the complete joint penetration weld between the beam flange and the through-diaphragm due to the stress concentration caused by the geometrical discontinuity. Accordingly, three improved types of through-diaphragm are presented in this paper to provide smooth force flow path comparing to that of conventional connections. Tensile tests were conducted on four specimens and the results were analyzed in terms of failure modes, load-displacement response, yield and ultimate capacity, and initial stiffness. Furthermore, strain distributions on the through-diaphragm, the beam flange plate, and the column face were comprehensively evaluated and discussed. It was found that all the proposed three types of improved through-diaphragm connections were able to reduce the stress concentration in the welds between the beam flange and the through-diaphragm. Furthermore, the stress distribution in connection with longer tapered through-diaphragm was more uniform.

Behavior of L-shaped double-skin composite walls under compression and biaxial bending

Ying Qin,Xin Chen,Wang Xi,Xingyu Zhu,Yuanze Chen 국제구조공학회 2020 Steel and Composite Structures, An International J Vol.37 No.4

The application of double-skin composite wall should meet different layout plans. However, most available research focused on the rectangular section with uniform axial compression. In this research, the structural behavior of double-skin composite wall with L section was studied. Due to the unsymmetric geometric characteristics, the considered loading condition combined the axial compression and biaxial bending. Five specimens were designed and tested under eccentric compression. The variables in the test included the width of the web wall, the truss spacing, the thickness of the steel faceplate, and the thickness of the web wall. The test results were discussed in terms of the load-displacement responses, buckling behavior, stiffness, ductility, strength utilization, strain distribution. Two modern codes were employed to predict the interaction between the axial compression and the biaxial bending. The method to calculate the available bending moment along the two directions was proposed. It was found that CECS 159:2004 offers more suitable results than AISC 360.

Deficiency of Follistatin-Like Protein 1 Accelerates the Growth of Breast Cancer Cells at Lung Metastatic Sites

Ying Zhang,Xiaozhou Xu,Ying Yang,Jie Ma,Lulu Wang,Xiangzhi Meng,Bing Chen,Ling Qin,Tao Lu,Yan Gao 한국유방암학회 2018 Journal of breast cancer Vol.21 No.3

Purpose: Follistatin-like protein 1 (FSTL1) is a secreted glycoprotein that has been shown to play a role in various types of cancer. However, the clinical significance and function of FSTL1 in breast cancer have not been reported. We investigated the role of FSTL1 in breast cancer in this study. Methods: Enzyme-linked immunosorbent assays, western blot analysis, and reverse transcription polymerase chain reaction were used to monitor the expression of FSTL1 in breast cancer tissue and in serum samples from breast cancer patients. We employed a 4T1 breast cancer model and Fstl1+/− mice for in vivo studies. Hematoxylin and eosin staining, western blot analysis, and RNA sequencing were used to analyze the effect of FSTL1 on primary tumor growth and lung metastasis. Results: We demonstrated that the expression of FSTL1 is reduced in both the breast cancer tissue and the serum of breast cancer patients. We showed that reduced levels of FSTL1 in serum correlate with elevated expression of Ki-67 and epidermal growth factor receptor (EGFR) in cancer tissues. Moreover, lowered expression of FSTL1 was associated with decreased survival in breast cancer patients. Experiments on the Fstl1+/− mouse model established that FSTL1 deficiency had no effect on primary tumor growth, but increased the lung metastases of breast cancer cells, resulting in reduced survival of tumor-bearing mice. RNA sequencing found significantly reduced expression of Egln3 and increased expression of EGFR in Fstl1+/− mice. Thus, our results suggest that FSTL1 may affect the expression of EGFR through Egln3, inhibiting the proliferation of breast cancer cells at lung metastatic sites. Conclusion: In conclusion, we suggest a suppressor role of FSTL1 in breast cancer lung metastasis. Furthermore, FSTL1 may represent a potential prognostic biomarker and a candidate therapeutic target in breast cancer patients.

Effect of height-to-width ratio on composite wall under compression

Ying Qin,Xin Yan,Guangen Zhou,Gan-Ping Shu 국제구조공학회 2020 Steel and Composite Structures, An International J Vol.36 No.5

Double skin composite walls are increasingly popular and have been applied to many safety-related facilities. They come from the concept of composite slabs. Conventional connectors such as shear studs and binding bars were used in previous studies to act as the internal mechanical connectors to lock the external steel faceplates to the concrete core. However, the restraint effects of these connectors were sometimes not strong enough. In this research, a recently proposed unique type of steel truss was employed along the wall height to enhance the composite action between the two materials. Concrete-filled tube columns were used as the boundary elements. Due to the existence of boundary columns, the restraints of steel faceplates to the concrete differ significantly for the walls with different widths. Therefore, there is a need to explore the effect of height-to-width ratio on the structural behavior of the wall. In the test program, three specimens were designed with the height of 3000 mm, the thickness of 150 mm, and different widths, to simulate the real walls in practice. Axial compression was applied by two actuators on the tested walls. The axial behavior of the walls was evaluated based on the analysis of test results. The influences of height-to-width ratio on structural performance were evaluated. Finally, discussion was made on code-based design.