Reports : Aurora-A kinase-inactive mutants disrupt the interaction with Ajuba and cause defects in mitotic spindle formation and G2/M phase arrest in HeLa cells

( Mei Rong Bai ),( Jun Ni ),( Su Qin Shen ),( Qiang Huang ),( Jia Xue Wu ),( Yi Chen Le ),( Long Yu ) 생화학분자생물학회(구 한국생화학분자생물학회) 2014 BMB Reports Vol.47 No.11

Aurora-A is a centrosome-localized serine/threonine kinase that is overexpressed in multiple human cancers. We previously reported an intramolecular inhibitory regulation of Aurora-A between its N-terminal regulatory domain (Nt, amino acids [aa] 1-128) and the C-terminal catalytic domain (Cd, aa 129-403). Here, we demonstrate that although both Aurora-A mutants (AurA-K250G and AurA-D294G/Y295G) lacked interactions between the Nt and Cd, they also failed to interact with Ajuba, an essential activator of Aurora-A, leading to loss of kinase activity. Additionally, overexpression of either of the mutants resulted in centrosome amplification and mitotic spindle formation defects. Both mutants were also able to cause G2/M arrest and apoptosis. These results indicate that both K250 and D294/Y295 are critical for direct interaction between Aurora-A and Ajuba and the function of the Aurora-A complex in cell cycle progression. [BMB Reports 2014; 47(11): 631-636]

Optomizing Transfection Efficiency of Cervical Cancer Cells Transfected by Cationic Liposomes Lipofectamine^TM2000

Huang, Fei,Zhao, Feng,Liang, Li-Ping,Zhou, Mei,Qu, Zhi-Ling,Cao, Yan-Zhen,Lin, Chen Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.17

Background: Currently, cationic liposome has become the commonly used vehicles for gene transfection. Furthermore, one of the most significant steps in microRNAs expression studies is transferring microRNAs into cell cultures successfully. In this study we aim to approach the feasibility of transfection of cervical cancer cell lines mediated by liposome and to obtain the optimized transfection condition for cervical cancer cell lines. Materials and Methods: $Lipofectamine^{TM}2000$ as the carrier, miR-101 mimic was transfected into Hela cells and Siha cells. Using green fluorescent protein as reporter gene, to set different groups according to cell seeding density, the amount of miRNA, miRNA and the proportion of Liposomes, Whether to add serum into medium to study their impact on the liposomal transfection efficiency. Finally, MTT assay was used to analyze the relative minimal cell toxicity of liposome reagents. Results: The seeding density of Hela cell line and Siha are $1.5{\times}10^4$ (per well of 24 well plates), miRNA amount is 1ul of both, the ratio of miRNA and liposome is 1:0.5 of Hela cell line; 1:0.7 of Siha cell line respectively, after 24 hours we can get the highest transfection efficiency. Compared with serum medium, only Siha cells cultured with serum-free medium obtained higher transfection efficiency before transfection (P<0.01). MTT assay showed that according to the above conditions which has the lowest cytotoxicity. Conclusions: The method of Liposome to transfected is a suitable way and it can be an efficient reagent for miRNA delivery for Hela cells and Siha cells in vitro. It may serve as a reference for the further research or application.

Expression and Underlying Roles of IGFBP-3 in Paclitaxel-Treated Gastric Cancer Sgc-7901 Cells

Huang, Gang,Dang, Zhong-Feng,Dang, Ya-Mei,Cai, Wei,Li, Yuan,Chen, Yi-Rong,Xie, Xiao-Dong Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.14

Purpose: To study the expression of insulin-like growth factor binding proteins (IGFBPs) in paclitaxel-treated gastric cancer SGC-7901 cells, and to further investigate underlying mechanisms. Materials and Methods: Real time PCR and Western blot assays were applied to detect the mRNA and protein expression of IGFBP-2, -3 and -5 after paclitaxel (10 nM) treatment of SGC-7901 cells. In addition IGFBP-3 expression was silenced by RNA interference to determine effects. Cell viability was determined by MTT assay. Cell cycling and apoptosis were assessed by flow cytometry. Results: Compared to the control group, only IGFBP-3 expression was elevated significantly after paclitaxel (10 nM) treatment (p<0.05). Paclitaxel treatment caused cell cycle arrest and apoptosis via downregulating Bcl-2 expression. However, the effect could be abrogated by IGFBP-3 silencing. Conclusions: IGFBP-3 exhibits anti-apoptotic effects on paclitaxel-treated SGC-7901 cells via elevating Bcl-2 expression.

Dissecting the meteorological and genetic factors affecting rice grain quality in Northeast China

Chen Mojun,Li Zhao,Huang Jie,Yan Yongfeng,Wu Tao,Bian Mingdi,Zhou Jinsong,Wang Yongjun,Lyv Yanjie,Hu Guanghui,Jin Yong-Mei,Huang Kai,Guo Liping,Jiang Wenzhu,Du Xinglin 한국유전학회 2021 Genes & Genomics Vol.43 No.8

Background The Northeast Plain of China, which is an important region for the production of high grain quality rice in China. However, the grain quality of the rice produced varies across this region, even for the same cultivar. Objective In order to explore the meteorological factors that have the greatest infuence on quality and the transcriptional level diferences between diferent cultivars and diferent locations at grain flling stage. Methods We grew eight rice cultivars in three locations in Northeast China during two growing seasons (2017 and 2018). We recorded meteorological conditions, including air temperature, air temperature range, and photosynthetically active radiation (PAR) during the grain-flling stage of each cultivar, and analyzed the grain quality of those eight cultivars. Results Across all eight cultivars, meteorological factors had a stronger efect on eating quality than genotype, while genotype had a stronger efect on milling quality. Of the three environmental factors assessed, PAR was signifcantly correlated with the most grain quality traits. Using RNA-sequencing analysis, we identifed 573 environment-specifc DEGs (Diferentially Expressed Genes), and 119 genotype-specifc DEGs; 11 DEGs were responsive to genotype×environment interactions. These DEGs were involved in many key metabolic processes. Conclusion Our results indicated that interactions among environmental factors, especially PAR, afected rice quality in Northeast China. Further analyses of the DEGs identifed herein may provide useful information for future breeding programs aiming to develop high grain quality rice varieties suitable for cultivation across Northeast China.

The Identification and Classification of Unknown Words in Chinese An N-Grams-Based Approach

( Mei Chu Wang ),( Chu Ren Huang ),( Keh Jiann Chen ) 한국언어정보학회 1994 국제 워크샵 Vol.1994 No.-

Multi-walled Carbon Nanotubes/Polypropylene-based Coating Layer on the Composite Metal Filaments: Characteristic Evaluations and Radiation-shielded Fabric

Mei-Feng Lai,Chen-Hung Huang,Ching-Wen Lou,Yu-Chun Chuang,Cyun-Yu Wei,Jia-Horng Lin 한국섬유공학회 2022 Fibers and polymers Vol.23 No.3

In this study, polypropylene (PP) and multi-walled carbon nanotubes (MWCNTs) are used to coat stainless steel(SS) wrapped yarns, the product of which is then fabricated into conductive textiles. Afterwards, the tensile properties,surface resistivity, and electromagnetic shielding effectiveness (EMSE) of conductive textiles are evaluated, therebydetermining the influences of the MWCNTs content. The test results show that using MWCNT can effectively improve themechanical properties of the coated yarns and conductive woven fabrics. In addition, 5 wt % of MWCNT provides the wovenfabrics with a lower surface resistivity and higher EMSE. The influences of the lamination angle and number of laminationlayers on EMSE are investigated, and the maximum EMSE of -49.89 dB occurs when the lamination angle is 0 °/90 °/0 °.

Anti-inflammatory and Anticancer Activities of Ethanol Extract of Pendulous Monkshood Root in vitro

Huang, Xian-Ju,Ren, Wei,Li, Jun,Chen, Lv-Yi,Mei, Zhi-Nan Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.6

Aim: Pendulous monkshood root is traditionally used for the treatment of several inflammatory pathologies such as rheumatisms, wounds, pain and tumors in China. In this study, the anti-inflammatory and anticancer activities and the mechanism of crude ethanol extract of pendulous monkshood root (EPMR) were evaluated and investigated in vitro. Materials and Methods: The cytotoxic effects of EPMR on different tumor cell lines were determined by the MTT method. Cell apoptosis and cell nucleus morphology were assessed by Hoechst 33258 staining. Moreover, nitric oxide (NO) levels and intracellular oxidative stress in peritoneal macrophages were determined to further elucidate mechanisms of action. Results: The data showed that EPMR could produce significant dose-dependent toxicity on three kinds of tumor cells. Furthermore, EPMR displayed obvious anti-inflammatory effects on LPS-induced mouse peritoneal macrophages at the dosage of 4 - 200 ${\mu}g/mL$. The results demonstrated the therapeutic potential of Pendulous Monkshood Root on cancer and inflammatory diseases. Conclusion: Our results indicate that EPMR has anti-inflammatory and anticancer properties, suggesting that pendulous monkshood root may be a useful anti-tumor and anti-inflammatory reagent in the clinic.

Human Papillomavirus Infection and Prognostic Predictors in Patients with Oropharyngeal Squamous Cell Carcinoma

Huang, Hui,Zhang, Bin,Chen, Wen,Zhou, Shuang-Mei,Zhang, Yong-Xia,Gao, Li,Xu, Zhen-Gang,Qiao, You-Lin,Tang, Ping-Zhang Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.3

This study focused on infection rates and subtypes of human papillomavirus (HPV) in patients with oropharyngeal squamous cell carcinoma (OSCC), and the relationship between HPV status and prognosis of the disease. We evaluated sixty-six OSCC patients who met the enrollment criteria during the period from January 1999 to December 2009. The presence or absence of oncogenic HPV types in tumors was determined using the SPF10 LiPA25 assay. Overall survival (OS) and disease specific survival (DSS) for HPV positive and HPV negative patients were estimated using Kaplan-Meier analysis. The Cox regression model was applied for multivariate analysis. HPV-DNA was detected in 11(16.7%) of all specimens. Among them, 7 were type HPV-16, while other types were HPV-16/11, HPV-35, HPV-58/52, and HPV-33/52/54. Patients with HPV positive tumors were more likely to be female, non-smokers and non-drinkers (p=0.002, 0.001 and 0.001, respectively). After a median follow-up of 24.5 months, patients with HPV positive tumors had significantly better overall survival (HR=0.106[95%CI=0.014-0.787], p=0.016,) and disease specific survival (HR=0.121[95%CI=0.016-0.906], p=0.030). Patients with HPV positive OSCC have significantly better prognosis than patients with HPV negative tumors. HPV infection is an independent prognostic factor.

Significant Association of Alpha-Methylacyl-CoA Racemase Gene Polymorphisms with Susceptibility to Prostate Cancer: a Meta-Analysis

Chen, Nan,Wang, Jia-Rong,Huang, Lin,Yang, Yang,Jiang, Ya-Mei,Guo, Xiao-Jiang,He, Ya-Zhou,Zhou, Yan-Hong Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.5

Background: Alpha-methylacyl-CoA racemase(AMACR) is thought to play key roles in diagnosis and prognosis of prostate cancer. However, studies of associations between AMACR gene polymorphisms and prostate cancer risk reported inconsistent results. Therefore, we conducted the present meta-analysis to clarify the link between AMACR gene polymorphisms and prostate cancer risk. Materials and Methods: A literature search was performed in PubMed, Embase, China National Knowledge Infrastructure (CNKI), Wanfang and Weipu databases. Odds ratios (ORs) and 95% confidence intervals (95%CIs) were calculated to assess the strength of any association between AMACR polymorphisms and prostate cancer risk. Subgroup analyses by ethnicity, source of controls, quality control and sample size were also conducted. Results: Five studies covering 3,313 cases and 3,676 controls on five polymorphisms (D175G, M9V, S201L, K277E and Q239H) were included in this meta-analysis. Significant associations were detected between prostate cancer and D175G (dominant model: OR=0.89, 95%CI=0.80-0.99, P=0.04) and M9V (dominant model: OR=0.87, 95%CI=0.78-0.97, P=0.01) polymorphisms as well as that in subgroup analyses. We also observed significant decreased prostate cancer risk in the dominant model (OR=0.90, 95%CI=0.81-0.99, P=0.04) for the S201L polymorphism. However, K277E and Q239H polymorphisms did not appear to be related to prostate cancer risk. Conclusions: The current meta-analysis indicated that D175G and M9V polymorphisms of the AMACR gene are related to prostate cancer. The S201L polymorphism might also be linked with prostate cancer risk to some extent. However, no association was observed between K277E or Q239H polymorphisms and susceptibility to prostate cancer.

Cloning, Expression, and Characterization of Endoglucanase Gene egIV from Trichoderma viride AS 3.3711

( Huang Xiao Mei ),( Jin Xia Fan ),( Qian Yang ),( Xiu Ling Chen ),( Zhi Hua Liu ),( Yun Wang ),( Da Qing Wang ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.3

Endoglucanase gene egIV was cloned from Trichoderma viride AS 3.3711, an important cellulose-producing fungus, by using an RT-PCR protocol. The egIV cDNA is 1,297 bp in length and contains a 1,035 bp open reading frame encoding a 344 amino acid protein with an estimated molecular mass of 35.5 kDa and isoelectronic point (pI) of 5.29. The expression of gene egIV in T. viride AS 3.3711 could be induced by sucrose, corn straw, carboxymethylcellulose (CMC), or microcrystalline cellulose, but especially by CMC. The transcripts of egIV were regulated under these substrates, but the expression level of the egIV gene could be inhibited by glucose and fructose. Three recombinant vectors, pYES2-xegIV, pYES2Mα-egIV, and pYES2Mα-xegIV, were constructed to express the egIV gene in Saccharomyces cerevisiae H158. The CMCase activity of yeast transformants IpYES2Mα-xegIV was higher than that of transformant IpYES2-xegIV or IpYES2Mα-egIV, with the highest activity of 0.13 U/ml at induction for 48 h, illustrating that the modified egIV gene could enhance CMCase activity and that MFα signal peptide from S. cerevisiae could regulate exogenous gene expression more effectively in S. cerevisiae. The recombinant EGIV enzyme was stable at pH 3.5 to 7.5 and temperature of 35oC to 65oC. The optimal reaction condition for EGIV enzyme activity was at the temperature of 55oC, pH of 5.0, 0.75 mM Ba2+, and using CMC as substrate. Under these conditions, the highest activity of EGIV enzyme in transformant IpYES2Mα-xegIV was 0.18 U/ml. These properties would provide technical parameters for utilizing cellulose in industrial bioethanol production.