In Vitro Evaluation and Monitoring of the Expression Level and Localization of Aldose Reductase Using Functionalized Quantum Dots and EGFP

Xiaomin Liu,Chengbin Yang,Jing Liu,Jianwei Liu,Rui Hu,Hongwei Lian,Guimiao Lin,Liwei Liu,Ken-Tye Yong,Ling Ye 한국생물공학회 2015 Biotechnology and Bioprocess Engineering Vol.20 No.4

The optimization of aldose reductase (AR) expression levels and tracking of the AR expression sites within the cell is an essential step in developing a platform for the effective production of aldose reductase inhibitors (ARIs). In this study, we have demonstrated the use of both immunocytochemistry and quantum dots-based immunofluorescence techniques for observing and detecting the expression level and localization of AR in the cytoplasm and cell membrane of a eukaryotic cell model with high levels of AR protein expression. Our results show that high expression levels of human AR can be achieved using the eukaryotic cell model that we have developed. The overexpressed AR can be used for translational studies of hAR and the screening of ARIs. More importantly, the use of the established quantum dots-based immunofluorescence technique in the intracellular labeling of AR allows the determination of the expression and distribution of the AR gene. Overall, the use of the interdisciplinary approach of both genetic engineering and quantum dot-based immunofluorescence allows not only the effective production of a desired protein, but also the determination of the cellular localization of such an expressed protein.

Clusterin negatively modulates mechanical stress-mediated ligamentum flavum hypertrophy through TGF-β1 signaling

Liu Chunlei,Li Peng,Ao Xiang,Lian Zhengnan,Liu Jie,Li Chenglong,Huang Minjun,Wang Liang,Zhang Zhongmin 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-

Ligamentum flavum hypertrophy (LFH) is a major cause of lumbar spinal canal stenosis (LSCS). The pathomechanisms for LFH have not been fully elucidated. Isobaric tags for relative and absolute quantitation (iTRAQ) technology, proteomics assessments of human ligamentum flavum (LF), and successive assays were performed to explore the effect of clusterin (CLU) upregulation on LFH pathogenesis. LFH samples exhibited higher cell positive rates of the CLU, TGF-β1, α-SMA, ALK5 and p-SMAD3 proteins than non-LFH samples. Mechanical stress and TGF-β1 initiated CLU expression in LF cells. Notably, CLU inhibited the expression of mechanical stress-stimulated and TGF-β1-stimulated COL1A2 and α-SMA. Mechanistic studies showed that CLU inhibited mechanical stress-stimulated and TGF-β1-induced SMAD3 activities through suppression of the phosphorylation of SMAD3 and by inhibiting its nuclear translocation by competitively binding to ALK5. PRKD3 stabilized CLU protein by inhibiting lysosomal distribution and degradation of CLU. CLU attenuated mechanical stress-induced LFH in vivo. In summary, the findings showed that CLU attenuates mechanical stress-induced LFH by modulating the TGF-β1 pathways in vitro and in vivo. These findings imply that CLU is induced by mechanical stress and TGF-β1 and inhibits LF fibrotic responses via negative feedback regulation of the TGF-β1 pathway. These findings indicate that CLU is a potential treatment target for LFH.

Improving the Self-adaptive Rate Control to Wireless Multicast for Virtual Environment

Liu Li,Chen Hong-qian,ChenYi,YaoChun-Lian 보안공학연구지원센터 2014 International Journal of Multimedia and Ubiquitous Vol.9 No.11

Distributed virtual environment requires high stability and low delay to support real-time data transmission for large number of users. In wireless network-based virtual environment, traditional transmission protocol fails to meet the requirements of virtual environment due to variety of nodes distribution, complexity, and difference of bandwidth. Based on our research of rate control for distributed virtual environment, we hereby propose a bandwidth self-adaptive rate control algorithm based on layers of interest. According to effective bandwidth of proxy gateway in distributed virtual environment, this method solving the problem of time inconsistency between publishers and subscribers, constraining occupied bandwidth within the range of effective bandwidth of gateways. Our experiments prove that this algorithm can solve the problem that users’needs of interactive with each other very frequently in wireless network virtual environment. The algorithm automatically match data transmission based on bandwidth and the received datagram is smooth and much more stable. The prime performances meet our expected research objectives.

Synthesis and Anti-HCV Activity of 3',5'-cyclic SATE Phosphonodiester Nucleoside as a Novel Prodrug

Lian Jin Liu,Rac Seok Seo,Seung Won Yoo,Jin Choi,홍준희 대한화학회 2010 Bulletin of the Korean Chemical Society Vol.31 No.4

A novel 2',4'-dimethyl carbocyclic adenosine 5'-phosphonic acid analogue (20) was prepared using acyclic stereoselective route from commercially available 4-hydroxybutan-2-one (4). To improve cellular permeability and enhance the anti-HCV activity of this phosphonic acid, a 3',5'-cyclic SATE phosphonodiester nucleoside prodrug (22) was prepared. The synthesized phosphonic nucleoside analogues, (20) and (22), were assayed for their ability to inhibit HCV RNA replication in a subgenomic replicon Huh7 cell line.

A Braking Force Distribution Strategy for Four-in-Wheel-Motor-Driven Electric Vehicles on Roads with Different Friction Coefficients

Lian Yufeng,Liu Shuaishi,Zhongbo Sun,Liu Keping,Nie Zhigen,Tian Chongwen 한국자동차공학회 2021 International journal of automotive technology Vol.22 No.4

This paper presents an active collision avoidance system based on a braking force distribution strategy for four-in-wheel-motor-driven electric vehicles (FIWMD-EVs) on roads with different friction coefficients. There are three major contributions in the proposed braking force distribution strategy. Firstly, the braking force distribution strategy based on constrained regenerative braking strength continuity (CRBSC) is further improved, and its general analytic expressions are derived. It provides the theoretical basis of braking force distribution between front and rear wheels. Secondly, the braking forces between front and rear wheels can be redistributed by considering power demand efficiency (PDE) to protect energy storage system from overcharge. Finally, the braking forces between left and right wheels can be distributed with different adhesion coefficients to adapt to complex roads. Simulations using rapid control prototyping (RCP) and hardware-in-the-loop (HIL) simulator are performed to demonstrate the effectiveness of control scheme and adaptability of the active collision avoidance system based on the proposed braking force distribution strategy on complex roads.

Construction of fat1 Gene Expression Vector and Its Catalysis Efficiency in Bovine Fetal Fibroblast Cells

Liu, Boyang,Yang, Runjun,Li, Junya,Zhang, Lupei,Liu, Jing,Lu, Chunyan,Lian, Chuanjiang,Li, Zezhong,Zhang, Yong-Hong,Zhang, Liying,Zhao, Zhihui Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.5

The FAT-1 protein is an n-3 fatty acid desaturase, which can recognize a range of 18- and 20-carbon n-6 substrates and transform n-6 polyunsaturated fatty acids (PUFAs) into n-3 PUFAs while n-3 PUFAs have beneficial effect on human health. Fat1 gene is the coding sequence from Caenorhabditis elegans which might play an important role on lipometabolism. To reveal the function of fat1 gene in bovine fetal fibroblast cells and gain the best cell nuclear donor for transgenic bovines, the codon of fat1 sequence was optimized based on the codon usage frequency preference of bovine muscle protein, and directionally cloned into the eukaryotic expression vector pEF-GFP. After identifying by restrictive enzyme digests with AatII/XbaI and sequencing, the fusion plasmid pEF-GFP-fat1 was identified successfully. The pEF-GFP-fat1 vector was transfected into bovine fetal fibroblast cells mediated by Lipofectamine2000$^{TM}$. The positive bovine fetal fibroblast cells were selected by G418 and detected by RT-PCR. The results showed that a 1,234 bp transcription was amplified by reverse transcription PCR and the positive transgenic fat1 cell line was successfully established. Then the expression level of fat1 gene in positive cells was detected using quantitative PCR, and the catalysis efficiency was detected by gas chromatography. The results demonstrated that the catalysis efficiency of fat1 was significantly high, which can improve the total PUFAs rich in EPA, DHA and DPA. Construction and expression of pEF-GFP-fat1 vector should be helpful for further understanding the mechanism of regulation of fat1 in vitro. It could also be the first step in the production of fat1 transgenic cattle.

Decision-Making of Determining the Start Time of Charging / Discharging of Electrical Vehicle Based on Prospect Theory

Lian Liu,Xiang Lyu,Chuanwen Jiang,Da Xie 대한전기학회 2014 Journal of Electrical Engineering & Technology Vol.9 No.3

The moment when Electrical Vehicle (EV) starts charging or discharging is one of the most important parameters in estimating the impact of EV load on the grid. In this paper, a decisionmaking problem of determining the start time of charging and discharging during allowed period is proposed and studied under the uncertainty of real-time price. Prospect theory is utilized in the decision-making problem of this paper for it describes a kind of decision making behaviors under uncertainty. The case study uses the parameters of Springo SGM7001EV and adopts the historical realtime locational marginal pricing (LMP) data of PJM market for scenario reduction. Prospect values are calculated for every possible start time in the allowed charging or discharging period. By comparing the calculated prospect values, the optimal decisions are obtained accordingly and the results are compared with those based on Expected Utility Theory. Results show that with different initial Stateof-Charge ( 0 SoC ) and different reference points, the optimal start time of charging can be the one between 12 a.m. to 3 a.m. and optimal discharging starts at 2 p.m. or 3p.m. Moreover, the decision results of Prospect Theory may differ from that of the Expected Utility Theory with the reference points changing.

Global Disease Burden and Attributable Risk Factor Analysis of Asthma in 204 Countries and Territories From 1990 to 2019

Liu Hailing,Zhang Jing,Liu Li,Lian Guoli,Shi Ruiming,Xu Man,Yang Juan,Liu Xiaohong 대한천식알레르기학회 2023 Allergy, Asthma & Immunology Research Vol.15 No.4

Purpose: Asthma is a common chronic inflammatory respiratory tract disease with high morbidity and mortality. The global trends in asthma burden remain poorly understood, and asthma incidence has increased during the worldwide coronavirus disease 2019 (COVID-19) pandemic. This study aimed to provide a comprehensive view of the global distribution of asthma burden and its attributable risk factors from 1990 to 2019. Methods: Based on the Global Burden of Disease Study 2019 Database, asthma incidence, deaths, disability-adjusted life years (DALYs), the corresponding age-standardized incidence rate (ASIR), age-standardized death rate (ASDR), age-standardized DALY rate, and estimated annual percentage change were analyzed according to age, sex, sociodemographic index (SDI) quintiles, and locations. Risk factors contributing to asthma deaths and DALYs were also investigated. Results: Globally, the asthma incidence increased by 15%, but deaths and DALYs decreased. The corresponding ASIR, ASDR, and age-standardized DALY rate also decreased. The high SDI region had the highest ASIR, and the low SDI region had the highest ASDR. The ASDR and age-standardized DALY rate were negatively correlated with the SDI. The low-middle SDI region, particularly South Asia, showed the highest asthma-related deaths and DALYs. The incidence peak was under 9 years old, and more than 70% of all deaths occurred in the population over 60 years old. Smoking, occupational asthmagens, and a high body mass index were the main risk factors for asthma-related mortality and DALYs, and their distributions varied between sexes. Conclusions: Globally, the asthma incidence has increased since 1990. The greatest asthma burden is borne by the low-middle SDI region. The 2 groups that need special attention are those under 9 years old and those over 60 years old. Targeted strategies are needed to reduce the asthma burden based on geographic and sex-age characteristics. Our findings also provide a platform for further investigation into the asthma burden in the era of COVID-19.

The Internal Force Relationship of Rectangular and I-section for Bi-linear Hardening Material with Limit Strain

Liu-Lian Li,Guo-Qiang Li 한국강구조학회 2016 International Journal of Steel Structures Vol.16 No.1

Based on bilinear stress-strain constitutive law σ = f (ε), the elastic to fully plastic analysis of bending of rectangular-section and bi-symmetrical I-section beams with the influence of axial load is presented for hardening material with limit strain. The variation of the applied bending moment with the axial force for the fully elastic, elastic-plastic, and fully plastic conditions is given in analytical form. The Internal force relationship of the elastic limit is the same for both hardening and non-hardening material and independent of the geometry of the beam section. However, for the elastic-plastic and plastic limits, the relationships are dependent of the hardening parameter βq, limit strain ε lim and the geometry of the beam section for neutral axis (N.A) inside the cross section. When N.A outside the cross section, the relationships are dependent of hardening parameter βq and limit strain ε lim but independent of the geometry of the beam section. The results given by the analytical expressions reduce to the ones for non-hardening material are in good agreement with the existing results.

Short Low Concentration Cisplatin Treatment Leads to an Epithelial Mesenchymal Transition-like Response in DU145 Prostate Cancer Cells

Liu, Yi-Qing,Zhang, Guo-An,Zhang, Bing-Chang,Wang, Yong,Liu, Zheng,Jiao, Yu-Lian,Liu, Ning,Zhao, Yue-Ran Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.3

Background: Prostate cancer is one of the main causes of cancer death, and drug resistance is the leading reason for therapy failure. However, how this occurs is largely unknown. We therrfore aimed to study the response of DU145 cells to cisplatin. Materials and Methods: Du145 prostate cancer cells were treated with a low dose of cisplatin for 24 h and cell viability and number were determined by MTT assay and trypan blue exclusion assay, respectively. The real time polymerase chain reaction (PCR) was used to assess responses to cisplatin treatment. Results: After 24h $2{\mu}g/ml$ treatment did not result in significant reduction in cell viability or number. However, it led to enhanced cancer cell invasiveness. E-cadherin mRNA was reduced, and vimentin, Snail, Slug, metalloproteinase 9 (MMP9) mRNA expression increased significantly, a feature of epithelial-mesenchymal transition (EMT). Conclusions: Short time low concentration cisplatin treatment leads to elevated invasiveness of DU145 cancer cells and this is possibly due to EMT.