Molecular Cloning and Function Analysis of an Anthocyanidin Synthase Gene from Ginkgo biloba, and Its Expression in Abiotic Stress Responses

Feng Xu,Hua Cheng,Rong Cai,Lin Ling Li,Jie Chang,Jun Zhu,Feng Xia Zhang,Liu Ji Chen,Yan Wang,Shu Han Cheng,Shui Yuan Cheng 한국분자세포생물학회 2008 Molecules and cells Vol.26 No.6

Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase), a 2-oxoglutarate iron-dependent oxygenase, catalyzed the penultimate step in the biosynthesis of the anthocyanin class of flavonoids, from the colorless leucoanthocyanidins to the colored anthocyanidins. The full-length cDNA and genomic DNA sequences of ANS gene (designated as GbANS) were isolated from Ginkgo biloba for the first time. The full-length cDNA of GbANS contained a 1062-bp open reading frame (ORF) encoding a 354-amino-acid protein. The genomic DNA analysis showed that GbANS gene had three exons and two introns. The deduced GbANS protein showed high identities to other plant ANSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2-oxoglutarate binding were found in GbANS at the similar positions like other ANSs. Southern blot analysis indicated that GbANS belonged to a multi-gene family. The expression analysis by real-time PCR showed that GbANS expressed in a tissue-specific manner in G. biloba. GbANS was also found to be up-regulated by all of the six tested abiotic stresses, UV-B, abscisic acid, sucrose, salicylic acid, cold and ethylene, consistent with the promoter region analysis of GbANS. The recombinant protein was successfully expressed in E. coli strain with pET-28a vector. The in vitro enzyme activity assay by HPLC indicated that recombinant GbANS protein could catalyze the formation the cyanidin from leucocyanidin and conversion of dihydroquercetin to quercetin, suggesting GbANS is a bifunctional enzyme within the anthocyanidin and flavonol biosynthetic pathway.

Long Noncoding RNA HOXA11-AS Modulates the Resistance of Nasopharyngeal Carcinoma Cells to Cisplatin via miR-454-3p/c-Met

Feng-Jie Lin,Xian-Dong Lin,Lu-Ying Xu,Shi-Quan Zhu 한국분자세포생물학회 2020 Molecules and cells Vol.43 No.10

To elucidate the mechanism of action of HOXA11-AS in modulating the cisplatin resistance of nasopharyngeal carcinoma (NPC) cells. HOXA11-AS and miR-454-3p expression in NPC tissue and cisplatin-resistant NPC cells were measured via quantitative reverse transcriptase polymerase chain reaction. NPC parental cells (C666-1 and HNE1) and cisplatin-resistant cells (C666-1/DDP and HNE1/DDP) were transfected and divided into different groups, after which the MTT method was used to determine the inhibitory concentration 50 (IC50) of cells treated with different concentrations of cisplatin. Additionally, a clone formation assay, flow cytometry and Western blotting were used to detect DDP-induced changes. Thereafter, xenograft mouse models were constructed to verify the in vitro results. Obviously elevated HOXA11-AS and reduced miR-454-3p were found in NPC tissue and cisplatin-resistant NPC cells. Compared to the control cells, cells in the si-HOXA11-AS group showed sharp decreases in cell viability and IC50, and these results were reversed in the miR-454-3p inhibitor group. Furthermore, HOXA11-AS targeted miR-454-3p, which further targeted c-Met. In comparison with cells in the control group, HNE1/DDP and C666-1/DDP cells in the si-HOXA11-AS group demonstrated fewer colonies, with an increase in the apoptotic rate, while the expression levels of c-Met, p-Akt/Akt and p-mTOR/mTOR decreased. Moreover, the si-HOXA11-AS-induced enhancement in sensitivity to cisplatin was abolished by miR-454-3p inhibitor transfection. The in vivo experiment showed that DDP in combination with si-HOXA11-AS treatment could inhibit the growth of xenograft tumors. Silencing HOXA11-AS can inhibit the c-Met/AKT/mTOR pathway by specifically upregulating miR-454-3p, thus promoting cell apoptosis and enhancing the sensitivity of cisplatin-resistant NPC cells to cisplatin.

A Data Compensation Model for Predicting SOH and RUL of Lithium–Ion Battery

Feng Hai-Lin,Xu An-Ke 대한전기학회 2024 Journal of Electrical Engineering & Technology Vol.19 No.1

Accurately estimating the state of health and remaining useful life of lithium–ion batteries is particularly critical to ensure safety and reliability. In this paper, ES-EDM-DCM, a data compensation model which fusing a new empirical model ES-EDM and a feature-driven model is established. The ES-EDM describes the degradation trajectory of li–ion batteries better. The feature driven model is established based on support vector regression which driven by a single health feature extracted and optimized from partial battery data. Finally, two datasets are used to verify that the prediction error of the proposed model is reduced by 0.1–3.68% compared with two single models in this paper. And compared with other literature methods, the prediction accuracy of the proposed model is improved by 0.9–52.98%. The proposed model has lower computational cost and higher robustness, and the modeling based on partial degradation data is more convenient for practical.

An improved horizontally reversible plow design based on virtual assembly semantics and constraint

Lin Zhu,Shuang-Shuang Peng,Yin-Yin Qi,Xi Cheng,Wen-Feng Zhang,Liang-Yuan Xu,De-Quan Zhu 대한기계학회 2016 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.30 No.1

There exit model errors in the constructed Horizontally reversible plough (HRP) structure through Interference check technique (ICT). This is basically due to missing parts and geometric interference and, hence, has significantly adverse effects in improving HRP. In thispaper an improved design to refine the three dimensional (3D) model of HRP is implemented by using Virtual assembly technology(VAT). To achieve this, the authors propose a combined virtual assembly semantics and constraint for assembly planning and simulationof HRP in the commercial software, e.g., SolidWorks. First, the assembly planning of HRP was captured with a semantics-based model,including spatial position, assembly orientation, type and parameters of the semantic entities; secondly, the assembly simulation of HRPwas performed by using constraint-based VAT, e.g. component grasping, moving and releasing; finally, the obtained HRP model wasverified through ICT again. The results demonstrate that not any model error exists in the refined 3D HRP model any longer and thatsemantics and constrain based VAT can support the interactive operation more effectively and accurately than those with geometric constraints. This technique was previously used for HRP model. Based on the refined 3D HRP model, the design improvement of Remotecylinder base (RCB), an important component of HRP, will be forthcoming in a future paper.

Influence of Preparation Conditions on the Formation of Copper (II) Architectures with Pyrazine-2,3,5-tricarboxylic Acid

Feng-Qin Wang,Shu Lin,Ming-Lin Guo,Jun-Jian Xu,Xiao-Qing Wang,Yong-Nan Zhao 대한화학회 2011 Bulletin of the Korean Chemical Society Vol.32 No.7

Three new metal-organic copper(II) complexes, [Cu(H_2PZTC)_2]_n·2nH_2O (1), [Cu(HPZTC)·2H_2O]_n·2nH_2O (2),and Cu_2[(PZHD)(OH)(H_2O)_2]_n (3) (H_3PZTC = pyrazine-2,3,5-tricarboxylic acid, PZHD^3− = 2-hydroxypyrazine-3,5-dicarboxylate), have been synthesized from Cu(II)/H3PZTC system under different synthetic conditions, and characterized by single-crystal X-ray diffraction, elemental analysis, IR spectroscopy and thermogravimetric analysis. In complexes 1 and 2, H_3PZTC ligands loose one and two protons, which were transformed into H_2PZTC− anion and HPZTC^2− dianion under different preparation condition, respectively. Furthermore, two ligands coordinate with Cu(II) cations in different modes, leading to the formation of the different chain structures. In complex 3, H_3PZTC ligand was converted into a new ligand-PZHD by in situ decarboxylation and hydroxylation under a higher pH value than that for complexes 1 and 2. PZHD ligands link the Cu(II) cations to form a 2D layer structure. These results demonstrate that the preparation conditions,including pH value and reaction temperature etc, play an important role in the construction of complexes based on H_3PZTC ligand.

Identification of mountain-cultivated ginseng and cultivated ginseng using UPLC/oa-TOF MSE with a multivariate statistical sample-profiling strategy

Xu, Xin-fang,Cheng, Xian-long,Lin, Qing-hua,Li, Sha-sha,Jia, Zhe,Han, Ting,Lin, Rui-chao,Wang, Dan,Wei, Feng,Li, Xiang-ri The Korean Society of Ginseng 2016 Journal of Ginseng Research Vol.40 No.4

Background: Mountain-cultivated ginseng (MCG) and cultivated ginseng (CG) both belong to Panax ginseng and have similar ingredients. However, their pharmacological activities are different due to their significantly different growth environments. Methods: An ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS)-based approach was developed to distinguish MCG and CG. Multivariate statistical methods, such as principal component analysis and supervised orthogonal partial-least-squares discrimination analysis were used to select the influential components. Results: Under optimized UPLC-QTOF-MS/MS conditions, 40 ginsenosides in both MCG and CG were unambiguously identified and tentatively assigned. The results showed that the characteristic components of CG and MCG included ginsenoside Ra3/isomer, gypenoside XVII, quinquenoside R1, ginsenoside Ra7, notoginsenoside Fe, ginsenoside Ra2, ginsenoside Rs6/Rs7, malonyl ginsenoside Rc, malonyl ginsenoside Rb1, malonyl ginsenoside Rb2, palmitoleic acid, and ethyl linoleate. The malony ginsenosides are abundant in CG, but higher levels of the minor ginsenosides were detected in MCG. Conclusion: This is the first time that the differences between CG and MCG have been observed systematically at the chemical level. Our results suggested that using the identified characteristic components as chemical markers to identify different ginseng products is effective and viable.

Distinctions Between Clinicopathological Factors and Prognosis of Alpha-fetoprotein Negative and Positive Hepatocelluar Carcinoma Patients

Xu, Jia,Liu, Chang,Zhou, Lei,Tian, Feng,Tai, Ming-Hui,Wei, Ji-Chao,Qu, Kai,Meng, Fan-Di,Zhang, Ling-Qiang,Wang, Zhi-Xin,Zhang, Jing-Yao,Chang, Hu-Lin,Liu, Si-Nan,Xu, Xin-Shen,Song, Yan-Zhou,Liu, Jun,Z Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.2

Serum alpha-fetoprotein (AFP) is a significant marker for clinical diagnosis and prognosis evaluation in hepatocellular carcinoma (HCC) patients. However, some proportion of liver cancer patients are AFP-negative (AFP ${\leq}$20ng/ml). In order to study the differences between clinicopathological factors and prognosis of alpha-fetoprotein negative and positive patients, a total of 114 cases (41 AFP-negative and 73 AFP-positive) were selected for our research. By systematically statistical analysis, the results demonstrated that compared with AFP-negative patients, AFP-positive examples were more likely to feature cirrhosis nodules, non-complete neoplasm capsules, and a poor Edmondson-steiner grade. Furthermore, AFP-negative patients demonstrated a favorable long-term prognosis. By univariate analysis and multivariate analysis with Cox's proportional hazards model, multiple tumors were found to be independent risk factors for worse survival of AFP negative patients; however, less tumor-free margins, multiple tumors and Edmondson-steiner grades III/IV, proved to be independent risk factors leading to a poor prognosis of AFP positive cases. Finally, we can infer that high levels of AFP signify a highly malignant tumor and unfavorable prognosis.

Effects of Montmorillonite Nanocomposite on Mercury Residues in Growing/Finishing Pigs

Lin, Xianglin,Xu, Zirong,Zou, Xiaoting,Wang, Feng,Yan, Xianghua,Jiang, Junfang Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.10

The study was conducted to evaluate the effects of montmorillonite anocomposite (MNC) on mercury residues in growing/finishing pigs. A total of 96 cross bred pigs ($Duroc{\times}Landrace{\times}large$ white, 48 barrows and gilts respectively), with similar initial weight (27.87${\pm}$1.15 kg), were used in this study. The animals were randomly assigned to two concentrations of mercury (0.1 and 0.3 ppm from $HgCl_2$) and two levels (0 and 0.3%) of MNC in a $2{\times}2$factorial arrangement of treatments. Each group has 3 pens (replications), and each pen has 8 pigs (4 barrows and 4 gilts). The experiment lasted for 90 days. The results showed that pig growth performances were not affected significantly by inclusion of Hg and addition of MNC (p$\geq$0.05). It indicated that the extent of intoxication in these pigs were not severe enough to impair growth performances. Both on the bases of 0.1 ppm and 0.3 ppm mercury supplementations, addition of 0.3% MNC markedly decreased mercury levels of blood, muscle, kidney and liver tissue (p<0.05). These results implied that the addition of non-nutritive sorptive material, MNC, could effectively reduce the gastrointestinal absorption of mercury via its specific adsorption, with a consequent reduction of mercury residues in body tissues. MNC had offered an encouraging solution to produce safe animal products with mercury contaminated feed.

Alum sludge conditioning with ferrous iron/peroxymonosulfate oxidation: Characterization and mechanism

Xu Zhou,Wenbiao Jin,Lan Wang,Lin Che,Chuan Chen,Shao-feng Li,Xue-Ting Wang,Renjie Tu,Song-fang Han,Xiaochi Feng,Nan-Qi Ren 한국화학공학회 2020 Korean Journal of Chemical Engineering Vol.37 No.4

Alum sludge produced by drinking water plants needs to be conditioned and dewatered before final disposal. In this study, a novel ferrous iron/peroxymonosulfate (PMS) oxidation process was employed to enhance alum sludge dewaterability. The effect of oxidative sulfate radicals generated by Fe2+ activated HSO5  on alum sludge was studied. The results showed that the optimal conditioning conditions for addition of Fe2+ and PMS were 0.5 g/g and 0.1 g/g TSS, respectively. Meanwhile, the capillary suction time (CST) and specific resistance to filtration (SRF) of alum sludge was reduced by 66% and 88%. Also found was that the absolute value of the zeta potential increased and the particle size decreased in alum sludge after Fe2+-PMS conditioning, which indicated that oxidative sulfate radicals destroyed the floc structure of alum sludge and smaller particles were generated. At the same time, the water contained in sludge flocs was released and enhanced sludge dewaterability, while leaching of aluminum ions also characterized decomposition of alum sludge.

Development of High-specificity Antibodies against Renal Urate Transporters Using Genetic Immunization

Xu, Guoshuang,Chen, Xiangmei,Wu, Di,Shi, Suozhu,Wang, Jianzhong,Ding, Rui,Hong, Quan,Feng, Zhe,Lin, Shupeng,Lu, Yang Korean Society for Biochemistry and Molecular Biol 2006 Journal of biochemistry and molecular biology Vol.39 No.6

Recently three proteins, playing central roles in the bidirectional transport of urate in renal proximal tubules, were identified: two members of the organic anion transporter (OAT) family, OAT1 and OAT3, and a protein that designated renal urate-anion exchanger (URAT1). Antibodies against these transporters are very important for investigating their expressions and functions. With the cytokine gene as a molecular adjuvant, genetic immunization-based antibody production offers several advantages including high specificity and high recognition to the native protein compared with current methods. We fused high antigenicity fragments of the three transporters to the plasmids pBQAP-TT containing T-cell epitopes and flanking regions from tetanus toxin, respectively. Gene gun immunization with these recombinant plasmids and two other adjuvant plasmids, which express granulocyte/macrophage colony-stimulating factor and FMS-like tyrosine kinase 3 ligand, induced high level immunoglobulin G antibodies, respectively. The native corresponding proteins of URAT1, OAT1 and OAT3, in human kidney can be recognized by their specific antibodies, respectively, with Western blot analysis and immunohistochemistry. Besides, URAT1 expression in Xenopus oocytes can also be recognized by its corresponding antibody with immuno-fluorescence. The successful production of the antibodies has provided an important tool for the study of UA transporters.