Cloning of Phospholipase D from Grape Berry and Its Expression under Heat Acclimation

Wan, Si-Bao,Wang, Wei,Wen, Peng-Fei,Chen, Jian-Ye,Kong, Wei-Fu,Pan, Qiu-Hong,Zhan, Ji-Cheng,Tian, Li,Liu, Hong-Tao,Huang, Wei-Dong Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.4

To investigate whether phospholipase D (PLD, EC 3.1.4.4) plays a role in adaptive response of post-harvest fruit to environment, a PLD gene was firstly cloned from grape berry (Vitis Vinifera L. cv. Chardonnay) using RT-PCR and 3'- and 5'-RACE. The deduced amino acid sequence (809 residues) showed 84.7% identity with that of PLD from Ricinus communis. The secondary structures of this protein showed the characteristic C2 domain and two active sites of a phospholipid-metabolizing enzyme. The PLD activity and its expression in response to heat acclimation were then assayed. The results indicated PLD was significantly activated at enzyme activity, as well as accumulation of PLD mRNA and synthesis of new PLD protein during the early of heat acclimation, primary suggesting that the grape berry PLD may be involved in the heat response in post-harvest grape berry. This work offers an important basis for further investigating the mechanism of post-harvest fruit adaptation to environmental stresses.

POLARIZATION PROPERTIES OF LARGE-AREA GaN-BASED LIGHT-EMITTING DIODES WITH AN OMNIDIRECTIONAL NANOSTRUCTURE

WAN-WEI WANG,LUNG-CHIEN CHEN,YA-HSIN WANG 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2012 NANO Vol.7 No.5

Large-area GaN-based blue light-emitting diodes (LEDs) on sapphire substrates with an omnidirectional nanostructure consisted of Ag nanoparticles and TiO2/SiO2 Bragg re°ector were fabricated. For the LEDs without the omnidirectional nanostructure, the spectra of the P- and S-polarization light are lightly di®erent and the peak wavelength of P- and S-polarization light was estimated to be 464 nm and 463 nm, respectively at forward current of 20 mA. In contrast, for the LEDs with the omnidirectional nanostructure, the spectra of the P- and S-polarization light are nearly the same at forward current of 20mA and 100 mA.

Cloning and characterization of the cardiac-specific Lrrc10 promoter

( Xiong Wei Fan ),( Qing Yang ),( You Liang Wang ),( Yan Zhang ),( Jian Wang ),( Jia Jia Yuan ),( Yong Qing Li ),( Yue Qun Wang ),( Yun Deng ),( Wu Zhou Yuan ),( Xiao Yang Mo ),( Yong Qi Wan ),( Karen 생화학분자생물학회 2011 BMB Reports Vol.44 No.2

Leucine-rich repeat containing protein 10 (LRRC10) is characterized as a cardiac-specific gene, suggesting a role in heart development and disease. A severe cardiac morphogenic defect in zebrafish morphants was recently reported but a contradictory result was found in mice, suggesting a more complicated molecular mechanism exists during mouse embryonic development. To elucidate how LRRC10 is regulated, we analyzed the 5`enhancer region approximately 3 kilo bases (kb) upstream of the Lrrc10 start site using luciferase reporter gene assays. Our characterization of the Lrrc10 promoter indicates it possesses complicated cis-and trans-acting elements. We show that GATA4 and MEF2C could both increase transcriptional activity of Lrrc10 promoter individually but that they do not act synergistically, suggesting that there exists a more complex regulation pattern. Surprisingly, knockout of Gata4 and Mef2c binding sites in the 5`enhancer region (-2,894/-2,889) didn`t change the transcriptional activity of the Lrrc10 promoter and the likely GATA4 binding site identified was located in a region only 100 base pair (bp) upstream of the promoter. Our data provides insight into the molecular regulation of Lrrc10 expression, which probably also contributes to its tissue-specific expression. [BMB reports 2011; 44(2): 123-128]

Cloning of Phospholipase D from Grape Berry and Its Expression under Heat Acclimation

( Si Bao Wan ),( Wei Wang ),( Peng Fei Wen ),( Jian Ye Chen ),( Wei Fu Kong ),( Qiu Hong Pan ),( Ji Cheng Zhan ),( Li Tian ),( Hong Tao Liu ),( Wei Dong Huang ) 생화학분자생물학회 2007 BMB Reports Vol.40 No.4

IL-33/ST2 axis mediates hyperplasia of intrarenal urothelium in obstructive renal injury

Wei-Yu Chen,Jenq-Lin Yan,Yi-Hsiu Wu,Lung-Chih Li,Ru-Fang Li,Ya-Ting Chang,Lo-Hsin Dai,Wan-Chen Wang,Ya-Jen Chang 생화학분자생물학회 2018 Experimental and molecular medicine Vol.50 No.-

The monolayered intrarenal urothelium covers the renal papilla and ureteropelvic junction (UPJ). In response to increased renal pressure during obstruction or ischemic injuries, intrarenal urothelial cells begin to proliferate and form a multilayered urothelium. Little is known regarding the mechanism and pathophysiological role of urothelium hyperplasia during renal obstruction. In this study, we investigated the expression of interleukin (IL)-33, an IL-1 family cytokine, in kidneys with unilateral ureteral obstruction (UUO)-induced obstructive injury. IL-33 levels in hydronephrotic urine and serum were upregulated 2 days after UUO. The number of ST2-expressing immune cells was increased in the UUO kidney. We found that IL-33 was upregulated in vimentin-positive cells in the cortical and medullar layers and the UPJ stroma. Moreover, IL-33 expression was predominantly induced in multilayered keratin 5- positive urothelial cells in the UPJ. IL-33 was not detected in terminally differentiated superficial umbrella cells expressing uroplakin 3a. In vivo, we confirmed that deficiency of IL33 or its receptor ST2 attenuated UUO-induced hyperplasia of the UPJ urothelium. Deficiency of IL33 attenuated the expression of UUO-induced type 2 inflammatory cytokines and upregulated uroplakins and urothelial differentiation signaling in UPJ tissues. Our results collectively suggest that the IL-33/ST2 axis mediates the activation of innate immune responses and contributes to urothelial hyperplasia by regulating urothelial differentiation in obstructive kidney injury.

Gene Silencing of β-catenin by RNAi Inhibits Proliferation of Human Esophageal Cancer Cells by Inducing G0/G1 Cell Cycle Arrest

Wang, Jin-Sheng,Ji, Ai-Fang,Wan, Hong-Jun,Lu, Ya-Li,Yang, Jian-Zhou,Ma, Li-Li,Wang, Yong-Jin,Wei, Wu Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.6

Objectives: The aim of the present study was to explore mechanisms underlying the effects of down-regulating ${\beta}$-catenin expression on esophageal carcinoma (EC) cells. Methods: Cell cycle distribution and apoptosis were determined using flow cytometry and annexin V apoptosis assay, respectively. Transmission electron microscopy (TEM) was used to examine changes in ultrastructure, while expression of cyclin D1 protein and mRNA was detected by western blot and real-time PCR. Proliferating cell nuclear antigen (PCNA) and extracellular signal-regulated kinase (ERK) 1-2 were evaluated by Western blot analysis. PCNA labeling index (LI) was determined by immunocytochemistry. Results: Compared with pGen-3-con transfected and Eca-109 cells, the percentage of G0/G1-phase pGen-3-CTNNB1 transfected cells was obviously increased (P<0.05), with no significant difference among the three groups with regard to apoptosis (P>0.05). pGen-3-CTNNB1 transfected cells exhibited obvious decrease in cyclin D1 mRNA and protein expression (P<0.05) and the ultrastructure of Eca-109 cells underwent a significant change after being transfected with pGen-3-CTNNB1, suggesting that down-regulating ${\beta}$-catenin expression can promote the differentiation and maturation. The expression of PCNA and the ERKI/2 phosphorylation state were also down-regulated in pGen-3-CTNNB1 transfected cells (P<0.05). At the same time, the PCNA labeling index was decreased accordingly (P<0.05). Conclusion: Inhibition of EC Eca-109 cellproliferation by down-regulating ${\beta}$-catenin expression could improve cell ultrastructure by mediating blockade in G0/G1 through inhibiting cyclin D1, PCNA and the MAPK pathway (p-ERK1/2).

Study on aerodynamic coefficients and responses of the integrated catwalk of Halogaland Bridge

Wan, Jia-wei,Wang, Qi,Liao, Hai-li,Li, Ming-shui Techno-Press 2017 Wind and Structures, An International Journal (WAS Vol.25 No.3

Wind tunnel tests and numerical aerodynamic analyses were conducted for an integrated catwalk structure under strong winds. From the wind tunnel tests, it is found that the aerodynamic coefficients were different from those of the typical type. The drag coefficient was larger than typical and was sensitive to number of vertical meshes installed rather than the solidity ratio. Comparing with typical catwalk, the integrated one showed larger deformation under strong wind, and the large torsional deformation are mainly caused by drag force. It did not show aerodynamic divergence even the torsional deformation reaching $20^{\circ}$. The reason could be that the stiffness is smaller and thus the catwalk is able to deform to the shape compactable with higher loading. Considering safety for construction, storm rope system is introduced to the catwalk to reduce the deformation to acceptable level.

ATAD2 is Highly Expressed in Ovarian Carcinomas and Indicates Poor Prognosis

Wan, Wei-Na,Zhang, Yi-Xia,Wang, Xue-Mei,Liu, Yan-Jun,Zhang, Yu-Qin,Que, Yan-Hong,Zhao, Wen-Jing Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

The purpose of this study was to explore the expression of ATAD2 in ovarian tumor tissue as well as its relationship with degree of malignancy. Tumor tissue from 110 cases of ovarian cancer was collected in accordance with the Declaration of Helsinki for evaluation of ATAD2 expression iimmunohistochemistry, quantitative PCR (qPCR) and Western blotting. The correlation between the ATAD2 expression and and the prognosis of ovarian cancer was evaluated by Cox regression model. In addition, HO-8910 and OVCAR-3 cells were transfected with two siRNAs targeting ATAD2. Cell viability was evaluated with MTT assay, and cell migration by transwell migration assay. ATAD2 was shown to be highly expressed in 65.5% (72/110) of ovarian cancer cases, both at transcriptional and protein levels. Moreover, highly expression was positively correlated with degree of malignancy. Knock-down of ATAD2 in HO-8910 and OVCAR-3 cells was found to reduce cell migration. In addition, follow-up visits of the patients demonstrated that the 5-year survival rate was lower in patients with high expression of ATAD2. Our study suggested that ovarian tumor tissue may have highly expressed ATAD2, which is associated with tumor stage, omentum-metastasis, ascites and CA-125. Increased ATAD2 may play important roles in tumor proliferation and migration. ATAD2 could serve in particular as a prognostic marker and a therapeutic target for ovarian cancer.

Mechanistic Analysis of Taxol-induced Multidrug Resistance in an Ovarian Cancer Cell Line

Wang, Ning-Ning,Zhao, Li-Jun,Wu, Li-Nan,He, Ming-Feng,Qu, Jun-Wei,Zhao, Yi-Bing,Zhao, Wan-Zhou,Li, Jie-Shou,Wang, Jin-Hua Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.9

Objectives: To establish a taxol-resistant cell line of human ovarian carcinoma (A2780/Taxol) and investigate its biological features. Methods: The drug-resistant cell line (A2780/Taxol) was established by continuous stepwise selection with increasing concentrations of Taxol. Cell morphology was assessed by microscopy and growth curves were generated with in vitro and in vivo tumor xenograft models. With rhodamine123 (Rh123) assays, cell cycle distribution and the apoptotic rate were analyzed by flow cytometry (FCM). Drug resistance-related and signal associated proteins, including P-gp, MRPs, caveolin-1, PKC-${\alpha}$, Akt, ERK1/2, were detected by Western blotting. Results: A2780/Taxol cells were established with stable resistance to taxol. The drug resistance index (RI) was 430.7. Cross-resistance to other drugs was also shown, but there was no significant change to radioresistance. Compared with parental cells, A2780/Taxol cells were significantly heteromorphous, with a significant delay in population doubling time and reduced uptake of Rh123 (p<0.01). In vivo, tumor take by A2780 cells was 80%, and tumor volume increased gradually. In contrast, with A2780/Taxol cells in xenograft models there was no tumor development. FCM analysis revealed that A2780/Taxol cells had a higher percentage of G0/G1 and lower S phase, but no changes of G2 phase and the apoptosis rate. Expression of P-gp, MRP1, MRP2, BCRP, LRP, caveolin-1, PKC-${\alpha}$, Phospho-ERK1/2 and Phospho-JNK protein was significantly up-regulated, while Akt and p38 MARK protein expression was not changed in A2780/Taxol cells. Conclusion: The A2780/Taxol cell line is an ideal model to investigate the mechanism of muti-drug resistance related to overexpression of drug-resistance associated proteins and activation of the PKC-${\alpha}/ERK$ (JNK) signaling pathway.

Analysis of the Genome Sequence of Strain GiC-126 of Gloeostereum incarnatum with Genetic Linkage Map

( Wan-zhu Jiang ),( Fang-jie Yao ),( Ming Fang ),( Li-xin Lu ),( You-min Zhang ),( Peng Wang ),( Jing-jing Meng ),( Jia Lu ),( Xiao-xu Ma ),( Qi He ),( Kai-sheng Shao ),( Asif Ali Khan ),( Yun-hui Wei 한국균학회 2021 Mycobiology Vol.49 No.4

Gloeostereum incarnatum has edible and medicinal value and was first cultivated and domesticated in China. We sequenced the G. incarnatum monokaryotic strain GiC-126 on an Illumina HiSeq X Ten system and obtained a 34.52-Mb genome assembly sequence that encoded 16,895 predicted genes. We combined the GiC-126 genome with the published genome of G. incarnatum strain CCMJ2665 to construct a genetic linkage map (GiC-126 genome) that had 10 linkage groups (LGs), and the 15 assembly sequences of CCMJ2665 were integrated into 8 LGs. We identified 1912 simple sequence repeat (SSR) loci and detected 700 genes containing 768 SSRs in the genome; 65 and 100 of them were annotated with gene ontology (GO) terms and KEGG pathways, respectively. Carbohydrate-active enzymes (CAZymes) were identified in 20 fungal genomes and annotated; among them, 144 CAZymes were annotated in the GiC-126 genome. The A mating-type locus (MAT-A) of G. incarnatum was located on scaffold885 at 38.9 cM of LG1 and was flanked by two homeodomain (HD1) genes, mip and beta-fg. Fourteen segregation distortion markers were detected in the genetic linkage map, all of which were skewed toward the parent GiC-126. They formed three segregation distortion regions (SDR1-SDR3), and 22 predictive genes were found in scaffold1920 where three segregation distortion markers were located in SDR1. In this study, we corrected and updated the genomic information of G. incarnatum. Our results will provide a theoretical basis for fine gene mapping, functional gene cloning, and genetic breeding the follow-up of G. incarnatum.