A Novel EYA1 Mutation Causing Alternative RNA Splicing in a Chinese Family With Branchio-Oto Syndrome: Implications for Molecular Diagnosis and Clinical Application

Anhai Chen,Chufeng He,Yong Feng,Jie Ling,Xin Peng,Xianlin Liu,Shuang Mao,Yongjia Chen,Mengyao Qin,Shuai Zhang,Yijiang Bai,Jian Song,Zhili Feng,Lu Ma,Dinghua He,Lingyun Mei1 대한이비인후과학회 2023 Clinical and Experimental Otorhinolaryngology Vol.16 No.4

Objectives. Branchio-oto syndrome (BOS) primarily manifests as hearing loss, preauricular pits, and branchial defects. EYA1is the most common pathogenic gene, and splicing mutations account for a substantial proportion of cases. However,few studies have addressed the structural changes in the protein caused by splicing mutations and potential pathogenicfactors, and several studies have shown that middle-ear surgery has limited effectiveness in improving hearing in thesepatients. BOS has also been relatively infrequently reported in the Chinese population. This study explored the ge-netic etiology in the family of a proband with BOS and provided clinical treatment to improve the patient’s hearing. Methods. We collected detailed clinical features and peripheral blood samples from the patients and unaffected individualswithin the family. Pathogenic mutations were identified by whole-exome sequencing and cosegregation analysis andclassified according to the American College of Medical Genetics and Genomics guidelines. Alternative splicing wasverified through a minigene assay. The predicted three-dimensional protein structure and biochemical experimentswere used to investigate the pathogenicity of the mutation. The proband underwent middle-ear surgery and was fol-lowed up at 1 month and 6 months postoperatively to monitor auditory improvement. Results. A novel heterozygous EYA1 splicing variant (c.1050+4 A >C) was identified and classified as pathogenic (PVS1(RNA),PM2, PP1). Skipping of exon 11 of the EYA1 pre-mRNA was confirmed using a minigene assay. This mutation mayimpair EYA1-SIX1 interactions, as shown by an immunoprecipitation assay. The EYA1-Mut protein exhibited cellularmislocalization and decreased protein expression in cytological experiments. Middle-ear surgery significantly improvedhearing loss caused by bone-conduction abnormalities in the proband. Conclusion. We reported a novel splicing variant of EYA1 in a Chinese family with BOS and revealed the potential molec-ular pathogenic mechanism. The significant hearing improvement observed in the proband after middle-ear surgeryprovides a reference for auditory rehabilitation in similar patients.

Bioconversion of ginsenoside Rc into Rd by a novel α-L-arabinofuranosidase, Abf22-3 from Leuconostoc sp. 22-3: cloning, expression, and enzyme characterization.

Liu, Qing-Mei,Jung, Hae-Min,Cui, Chang-Hao,Sung, Bong-Hyun,Kim, Jin-Kwang,Kim, Song-Gun,Lee, Sung-Taik,Kim, Sun-Chang,Im, Wan-Taek N.V. Swets en Zeitlinger 2013 Antonie van Leeuwenhoek Vol.103 No.4

<P>A novel α-L-arabinofuranosidase (Abf22-3) that could biotransform ginsenoside Rc into Rd was obtained from the ginsenoside converting Leuconostoc sp. strain 22-3, isolated from the Korean fermented food kimchi. The gene, termed abf22-3, consisting of 1,527 bp and encoding a protein with a predicted molecular mass of 58,486 Da was cloned into the pMAL-c2x (TEV) vector. A BLAST search using the Abf22-3's amino acid sequence revealed significant homology to that of family 51 glycoside hydrolases. The over-expressed recombinant Abf22-3 in Escherichia coli BL21 (DE3) catalyzed the hydrolysis of the arabinofuranoside moiety attached to the C-20 position of ginsenoside Rc under optimal conditions of pH 6.0 and 30 C. This result indicated that Abf22-3 selectively converts ginsenoside Rc into Rd, but did not catalyze the hydrolysis of glucopyranosyl groups from Rc or other ginsenosides such as Rb1 and Rb2. Over-expressed recombinant enzymes were purified by two steps with amylose-affinity and DEAE-cellulose chromatography and then characterized. The kinetic parameters for α-L-arabinofuranosidase showed apparent Km and Vmax values of 0.95 0.02 μM and 1.2 0.1 μmol min(-1) mg of protein(-1) against p-nitrophenyl-α-L-arabinofuranoside, respectively. Using a purified MBP-Abf22-3 (10 μg/ml), 0.1 % of ginsenoside Rc was completely converted to ginsenoside Rd within 20 min.</P>

Introduction of Long-Term Care need Assessment Tool

Mei-Yin Liu,Shyh-Dye Lee 한국간호과학회 2011 한국간호과학회 학술대회 Vol.2011 No.10

Multi-Sensor Multi-Target Passive Locating and Tracking

Mei Liu,Nuo Xu,Haihao Li 대한전기학회 2007 International Journal of Control, Automation, and Vol.5 No.2

The passive direction finding cross localization method is widely adopted in passive tracking, therefore there will exist masses of false intersection points. Eliminating these false intersection points correctly and quickly is a key technique in passive localization. A new method is proposed for passive locating and tracking multi-jammer target in this paper. It not only solves the difficulty of determining the number of targets when masses of false intersection points existing, but also solves the initialization problem of elastic network. Thus this method solves the problem of multi-jammer target correlation and the elimination of static false intersection points. The method which dynamically establishes multiple hypothesis trajectory trees solves the problem of eliminating the remaining false intersection points. Simulation results show that computational burden of the method is lower, the elastic network can more quickly find all or most of the targets and have a more probability of locking the real targets. This method can eliminate more false intersection points.

Ultra-sensitive graphene based mid-infrared plasmonic bio-chemical sensing using dielectric beads as a medium

Liu, Xiao,Zhang, Duan,Wu, Ye-Cun,Yang, Mei,Wang, Qian,Coileá,in, Cormac Ó,.,Xu, Hongjun,Yang, Chen,Abid, Mohamed,Abid, Mourad,Liu, Huajun,Chun, Byong Sun,Shi, Qingfan,Wu, Han-Chun Elsevier 2017 Carbon Vol.122 No.-

<P>Graphene is moving beyond the realm of simple electronic devices toward areas such as advanced biochemical sensing. The infrared (IR) response of graphene, characterized by collective long-lived charge-carrier oscillations, could be applied in IR-absorption spectroscopy, typically used for bio-chemical analysis. However, direct light absorption by propagating plasmons in graphene is forbidden due to the large momentum mismatch. Proposed methods to overcome this bottleneck come at a cost, the use of noble metal particles on graphene reduces the spectral bandwidth and nano-structuring graphene is expensive. Here, we propose a simple and cheap method to fabricate large scale ultra-sensitive graphene based mid-IR biosensors, by introducing dielectric beads to excite mid-IR range plasmons. Interference from waves scattered by the beads excite surface plasmon polaritons, which propagate several micrometers in graphene and enhance the interaction between the molecules and mid-IR light. This method opens an interesting window for the application of graphene in bio-chemical sensing. (C) 2017 Elsevier Ltd. All rights reserved.</P>

How Is Organization Capital Priced in the Korean Market?

Mei Liu,박세열 조선대학교 지식경영연구원 2022 기업과 혁신연구 Vol.45 No.3

In this paper, we test whether the positive relationship between organization capital and stock return documented in the US stock market holds in the Korean stock market, and whether the difference is caused by human capital risk. The results show that organization capital does not appear to affect stock returns in Korea. Although a recursive stock proxy based on total SG&A generates a significant difference in excess return between high and low portfolios, the difference does not seem to be attributable to organization capital and human capital risk. The hypothesis that the difference may be caused by higher downside risk from sticky SG&A costs is also ruled out. The results provide evidence against a human capital risk explanation for the effect of organization capital on stock returns.

Efficacy and safety of herbal medicine (Binafuxi granules) for the common cold with fever: A multicenter, randomized, double-blind, placebo-controlled, phase II clinical trial

Liu Xuemei,Min Jie,She Bin,Chen Yang,Li Jun,Huang Lei,Chen Ju,Luo Ai,Mei Yang,Li Ting,Wu Yanqing,Chen Daohong,Hongli Zhong,Liu Wei,Mao Bing,Jiang Hongli 한국한의학연구원 2023 Integrative Medicine Research Vol.12 No.3

Background: Binafuxi granules are a traditional Uighur medicine (TUM) for treating the common cold with fever. However, high-quality clinical studies supporting its efficacy and safety are lacking. Methods: In this multicenter, randomized, double-blind, placebo-controlled, phase II clinical trial, patients with common cold and fever were randomly assigned to a high-dose group, low-dose group, and placebo group in a 1:1:1 ratio. Outcomes were time to fever relief, time to fever clearance, proportion of afebrile patients, time to symptom disappearance, rate of symptom disappearance, effective rate, emergency drug usage and safety assessment. Results: A total of 235 patients were recruited. Of these, 234 were included in the full analysis set (FAS), and 217 were included in the per-protocol set (PPS). In the FAS analysis, the median time to fever relief was 6.00 h, 5.54 h and 10.65 h (P = 0.31) in the high-dose group, low-dose group and placebo group, respectively. The median time to fever clearance was 18.29 h, 20.08 h and 25.00 h (P = 0.0018), respectively, and the proportion of afebrile patients was 92.4%, 89.7% and 71.4% (P = 0.0002), respectively. There was a significant difference in the disappearance time and disappearance rate of all symptoms and of individual symptoms. No serious adverse events were found. Conclusions: Binafuxi granules can dose-dependently shorten the fever course and improve clinical symptoms in patients suffering from the common cold with fever.

Trichloroethylene induces dopaminergic neurodegeneration in Fisher 344 rats

Liu, Mei,Choi, Dong-Young,Hunter, Randy L.,Pandya, Jignesh D.,Cass, Wayne A.,Sullivan, Patrick G.,Kim, Hyoung-Chun,Gash, Don M.,Bing, Guoying Blackwell Publishing Ltd 2010 Journal of Neurochemistry Vol.112 No.3

<P><I>J. Neurochem.</I> (2010) <B>112</B>, 773–783.</P><P>Abstract</P><P>Trichloroethylene, a chlorinated solvent widely used as a degreasing agent, is a common environmental contaminant. Emerging evidence suggests that chronic exposure to trichloroethylene may contribute to the development of Parkinson’s disease. The purpose of this study was to determine if selective loss of nigrostriatal dopaminergic neurons could be reproduced by systemic exposure of adult Fisher 344 rats to trichloroethylene. In our experiments, oral administration of trichloroethylene induced a significant loss of dopaminergic neurons in the substantia nigra pars compacta in a dose-dependent manner, whereas the number of both cholinergic and GABAergic neurons were not decreased in the striatum. There was a robust decline in striatal levels of 3, 4-dihydroxyphenylacetic acid without a significant depletion of striatal dopamine. Rats treated with trichloroethylene showed defects in rotarod behavior test. We also found a significantly reduced mitochondrial complex I activity with elevated oxidative stress markers and activated microglia in the nigral area. In addition, we observed intracellular &agr;-synuclein accumulation in the dorsal motor nucleus of the vagus nerve, with some in nigral neurons, but little in neurons of cerebral cortex. Overall, our animal model exhibits some important features of Parkinsonism, and further supports that trichloroethylene may be an environmental risk factors for Parkinson’s disease.</P>

Mining Proteins Associated with Oral Squamous Cell Carcinoma in Complex Networks

Liu, Ying,Liu, Chuan-Xia,Wu, Zhong-Ting,Ge, Lin,Zhou, Hong-Mei Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.8

The purpose of this study was to construct a protein-protein interaction (PPI) network related to oral squamous cell carcinoma (OSCC). Each protein was ranked and those most associated with OSCC were mined within the network. First, OSCC-related genes were retrieved from the Online Mendelian Inheritance in Man (OMIM) database. Then they were mapped to their protein identifiers and a seed set of proteins was built. The seed proteins were expanded using the nearest neighbor expansion method to construct a PPI network through the Online Predicated Human Interaction Database (OPHID). The network was verified to be statistically significant, the score of each protein was evaluated by algorithm, then the OSCC-related proteins were ranked. 38 OSCC related seed proteins were expanded to 750 protein pairs. A protein-protein interaction nerwork was then constructed and the 30 top-ranked proteins listed. The four highest-scoring seed proteins were SMAD4, CTNNB1, HRAS, NOTCH1, and four non-seed proteins P53, EP300, SMAD3, SRC were mined using the nearest neighbor expansion method. The methods shown here may facilitate the discovery of important OSCC proteins and guide medical researchers in further pertinent studies.

PBK/TOPK Expression During TPA-Induced HL-60 Leukemic Cell Differentiation

Liu, Yu-Hong,Gao, Xue-Mei,Ge, Fan-Mei,Wang, Zhe,Wang, Wen-Qing,Li, Xiao-Yong Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.5

Objective: This study concerns expression of PBK/TOPK during differentiation of HL-60 leukemic cells induced by tetradecanoyl phorbol acetate (TPA). Methods: Wright-Giemsa staining was performed to observe morphological changes in the HL-60 cells, and flow cytometry was used to assess the cell cycle and CD11b, CD14, CD13, and CD33 expression. PBK/TOPK levels were determined by Western blot analysis. Results: After treating HL60 cells with $5.1{\times}10^{-9}$ mmol/L of TPA for three days, the number of nitroblue-tetrazolium-positive cells and CD11b, CD13, and CD14 expression increased, whereas the PBK/TOPK levels decreased. Conclusions: TPA can inhibit proliferation and induce differentiation of HL60 cells of the granulocytic or monocytic lineage. PBK/TOPK expression was downregulated during this process, whereas the Pho-PBK/TOPK expression was increased.