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Xuemei Zhang,Hongya Hei,Jianjun Gao,Jibin Dong,Jie Tao,Lulu Tian,Wanma Pan,Hongyu Wang 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.7
Large conductance calcium-activated potassium (BK) channels participate in many important physiological functions in excitable tissues such as neurons, cardiac and smooth muscles, whereas the knowledge of BK channels in bone tissues and osteoblasts remains elusive. To investigate the role of BK channels in osteoblasts, we used transcription activator-like effector nuclease (TALEN) to establish a BK knockout cell line on rat ROS17/2.8 osteoblast, and detected the proliferation and mineralization of the BK-knockout cells. Our study found that the BK-knockout cells significantly decreased the ability of proliferation and mineralization as osteoblasts, compared to the wild type cells. The overall expression of osteoblast differentiation marker genes in the BK-knockout cells was significantly lower than that in wild type osteoblast cells. The BK-knockout osteoblast cell line in our study displays a phenotype decrease in osteoblast function which can mimic the pathological state of osteoblast and thus provide a working cell line as a tool for study of osteoblast function and bone related diseases.
Hei, Hongya,Gao, Jianjun,Dong, Jibin,Tao, Jie,Tian, Lulu,Pan, Wanma,Wang, Hongyu,Zhang, Xuemei Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.7
Large conductance calcium-activated potassium (BK) channels participate in many important physiological functions in excitable tissues such as neurons, cardiac and smooth muscles, whereas the knowledge of BK channels in bone tissues and osteoblasts remains elusive. To investigate the role of BK channels in osteoblasts, we used transcription activator-like effector nuclease (TALEN) to establish a BK knockout cell line on rat ROS17/2.8 osteoblast, and detected the proliferation and mineralization of the BK-knockout cells. Our study found that the BKknockout cells significantly decreased the ability of proliferation and mineralization as osteoblasts, compared to the wild type cells. The overall expression of osteoblast differentiation marker genes in the BK-knockout cells was significantly lower than that in wild type osteoblast cells. The BK-knockout osteoblast cell line in our study displays a phenotype decrease in osteoblast function which can mimic the pathological state of osteoblast and thus provide a working cell line as a tool for study of osteoblast function and bone related diseases.
Hu Boran,Gao Jin,Xu Shaochen,Zhu Jiangyu,Fan Xuemei,Zhou Xiaoyan 한국응용생명화학회 2020 Applied Biological Chemistry (Appl Biol Chem) Vol.63 No.3
The metabolites that provide the aroma and flavor to wine are the products of several influences, such as grape cultivar, geographic location and associated environmental features, viticultural practices, and vinification techniques, which are central to production protocols, quality evaluation and development of wine regions. Accordingly, we initiated the requisite studies to investigate the differences in the dry red wine metabolites of different grape varieties. The proton-nuclear magnetic resonance technique (1H-NMR) combined with multivariate statistical analysis was used to investigate the changes of metabolite levels in Cabernet Sauvignon, Merlot and Cabernet Gernischt dry red wines vinified in Changli, Hebei province, China, in 2017. The results showed that the types of metabolites in different varieties of dry red wines were similar, but the content was significantly different. The main contributors to the differences in Cabernet Sauvignon, Merlot and Cabernet Gernischt dry red wines were ethyl acetate, lactic acid, alanine, succinic acid, proline, malic acid, and gallic acid, indicating 1H-NMR method combined with multivariate statistical analysis can distinguish these three types of dry red wines from each other. It provides a benchmark for further comparative study on wine quality and the verification of wine authenticity.
BK Channel Deficiency in Osteoblasts Reduces Bone Formation via the Wnt/β-Catenin Pathway
Jiang, Lan,Yang, Qianhong,Gao, Jianjun,Yang, Jiahong,He, Jiaqi,Xin, Hong,Zhang, Xuemei Korean Society for Molecular and Cellular Biology 2021 Molecules and cells Vol.44 No.8
Global knockout of the BK channel has been proven to affect bone formation; however, whether it directly affects osteoblast differentiation and the mechanism are elusive. In the current study, we further investigated the role of BK channels in bone development and explored whether BK channels impacted the differentiation and proliferation of osteoblasts via the canonical Wnt signaling pathway. Our findings demonstrated that knockout of Kcnma1 disrupted the osteogenesis of osteoblasts and inhibited the stabilization of β-catenin. Western blot analysis showed that the protein levels of Axin1 and USP7 increased when Kcnma1 was deficient. Together, this study confirmed that BK ablation decreased bone mass via the Wnt/β-catenin signaling pathway. Our findings also showed that USP7 might have the ability to stabilize the activity of Axin1, which would increase the degradation of β-catenin in osteoblasts.
Tan Yahong,Song Wenxia,Gao Lijuan,Zhang Weican,Lu Xuemei 한국미생물학회 2022 The journal of microbiology Vol.60 No.4
Cytophaga hutchinsonii can efficiently degrade crystalline cellulose, in which the cell surface cellulases secreted by the type IX secretion system (T9SS) play important roles, but the degradation mechanism remains unclear, and the anchor mechanism of cellulases on the outer membrane in C. hutchinsonii has not been studied. Here, chu_2177 was identified by transposon mutagenesis and was proved to be indispensable for cellulose utilization in C. hutchinsonii. Disruption of chu_2177 resulted in O-antigen deficiency and chu_ 177 could confer O-antigen ligase activity upon an Escherichia coli waal mutant, indicating that chu_2177 encoded the Ontigen ligase. Moreover, deletion of chu_2177 caused defects in cellulose utilization, cell motility, biofilm formation, and stress resistance. Further study showed that the endoglucanase activity was markedly decreased in the outer membrane but was increased in the culture fluid without chu_2177. Western blot proved that endoglucanase CHU_1336 was not located on the outer membrane but was released in the culture fluid of the Δ2177 mutant. Further proteomics analysis showed that many cargo proteins of T9SS were missing in the outer membrane of the Δ2177 mutant. Our study revealed that the deletion of chu_2177 affected the localization of many T9SS cargo proteins including cellulases on the outer membrane of C. hutchinsonii.
Xiaorong Yu,Rui Zhang,Cunsheng Wei,Yuanyuan Gao,Yanhua Yu,Lin Wang,Junying Jiang,Xuemei Zhang,Junrong Li,Xuemei Chen 한국통합생물학회 2021 Animal cells and systems Vol.25 No.2
Monocarboxylate transporter 2 (MCT2) is the predominant monocarboxylate transporter expressed by neurons. MCT2 plays an important role in brain energy metabolism. Stroke survivors are at high risk of cognitive impairment. We reported previously that stroke-induced cognitive impairment was related to impaired energy metabolism. In the present study, we report that cognitive function was impaired after stroke in rats. We found that MCT2 expression, but not that of MCT1 or MCT4, was markedly decreased in the rat hippocampus at 7 and 28 days after transient middle cerebral artery occlusion (tMCAO). Moreover, MCT2 overexpression promoted recovery of cognitive function after stroke. The molecular mechanism underlying these effects may be related to an increase in adenosine monophosphate-activated protein kinase-mediated mitochondrial biogenesis induced by overexpression of MCT2. Our findings suggest that MCT2 activation ameliorates cognitive impairment after stroke.
Fengping Ye,Qili Mi,Ning Zhang,Xuemei Li,Jing Yu,Zhongping Gao,Ying Zheng,Quanshui Fan,Jie Wang,Jinglin Wang 대한화학회 2016 Bulletin of the Korean Chemical Society Vol.37 No.5
Chemical modifications of the nucleotides can improve the stability of aptamers against enzyme degradation in serum, but it is not clear whether these methods are effective in snake venom. In this study, a DNA aptamer, βB-1, which specifically recognize β-bungarotoxin and Bungarus multicinctus venom was chosen, and the key binding sequence of the aptamer was determined. Based on the secondary structure of the truncated aptamer, locked nucleic acids and 2′-O-methyl nucleotides were applied to modify the stem and loop sequences, respectively. In addition, a 3′-3′-thymidine cap was also adopted to block the 3′ end. It was shown that these chemical modifications can all enhance the stability of the aptamer in snake venom. Simultaneously, modified aptamer with the above modifications in one sequence exhibited a significantly elevated biostability, with the half-life improved from several minutes to 210 min while maintaining its binding affinity to the target.
Yan-Qing Ye,Cong-Fang Xia,Juan-Xia Yang,Yu-Chun Yang,Ying Qin,Xue-Mei Gao,Gang Du,Xuemei Li,Qiu-Fen Hu 대한화학회 2014 Bulletin of the Korean Chemical Society Vol.35 No.10
Two new butyrolactones, asperphenol A (1) and B (2), together with four known butyrolactones (3-6) were isolated from the fermentation products of an endophytic fungus Aspergillus versicolor. Their structures were elucidated by spectroscopic methods including extensive 1D- and 2D-NMR techniques. Compounds 1-6 were also tested for their anti-tobacco mosaic virus (anti-TMV) activities. The results showed that compound 2 exhibited high anti-TMV activity with inhibition rate of 46.7%. The other compounds also exhibited potential anti-TMV activities with inhibition rates in the range of 21.8-28.4%.