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Dun, Yaoyan,Liu, Min,Chen, Jing,Peng, Danli,Zhao, Haixia,Zhou, Zhiyong,Wang, Ting,Liu, Chaoqi,Guo, Yuhui,Zhang, Changcheng,Yuan, Ding The Korean Society of Ginseng 2018 Journal of Ginseng Research Vol.42 No.1
Background: Saponins from Panax japonicus (SPJ) are the most abundant and main active components of P. japonicus, which replaces ginseng roots in treatment for many kinds of diseases in the minority ethnic group in China. Our previous studies have demonstrated that SPJ has the effects of anti-inflammation through the mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-${\kappa}B$) signaling pathways. The present study was designed to investigate whether SPJ can modulate intestinal tight junction barrier in aging rats and further to explore the potential mechanism. Methods: Aging rats had been treated with different doses (10 mg/kg, 30 mg/kg, and 60 mg/kg) of SPJ for 6 mo since they were 18 mo old. After the rats were euthanized, the colonic samples were harvested. Levels of tight junctions (claudin-1 and occludin) were determined by immunohistochemical staining. Levels of proinflammatory cytokines (interleukin-$1{\beta}$ and tumor necrosis factor-${\alpha}$) were examined by Western blot. NF-${\kappa}B$ and phosphorylation of MAPK signaling pathways were also determined by Western blot. Results: We found that SPJ increased the expression of the tight junction proteins claudin-1 and occludin in the colon of aging rats. Treatment with SPJ decreased the levels of interleukin-$1{\beta}$ and tumor necrosis factor-${\alpha}$, reduced the phosphorylation of three MAPK isoforms, and inhibited the expression of NF-${\kappa}B$ in the colon of aging rats. Conclusion: The studies demonstrated that SPJ modulates the damage of intestinal epithelial tight junction in aging rats, inhibits inflammation, and downregulates the phosphorylation of the MAPK and $NF-{\kappa}B$ signaling pathways.
Cloning and expression of a Chk1 gene in Daphnia pulex during different modes of reproduction
Xiaoge Guo,Shanliang Xu,Xuemei Yan,Wei Zhou,Xinyu Dai,Xiu Zou,Chunlin Wang,Danli Wang,Yunlong Zhao 한국유전학회 2015 Genes & Genomics Vol.37 No.9
The full-length cDNA of a Chk1 gene (DpChk1) was cloned from Daphnia pulex using RACE method. This cDNA is 1767 bp in length, consisting of a 1497 bp open reading frame encoding 498 amino acid protein. The deduced DpChk1 protein sequence contains phosphorylation sites, three conserved Ser-Gln and Thr- Gln motifs and shares 51–55 % similarity with A. pisum, M. rotundata, D. melanogaster, A. echinatior, B. terrestris, R. pulchellus and M. occidentalis, respectively. Gene expression analysis showed that Chk1 is differentially expressed in D. pulex during different reproductive modes. DpChk1 expression in sexually-reproducing females and males is significantly higher than in parthenogenetic Daphnia. Whole-mount in situ hybridization revealed that DpChk1 was expressed during two kinds of reproductive modes. In parthenogenetic females, hybridization signals were found in the antennae and thoracic limb, whereas expression levels in the corresponding sites of sexuallyreproducing females were relatively strong, DpChk1 could not be detected in the gonads of ephippial and parthenogenetic females. Taken together, these different reproductive stages and sex specific expression patterns are regulated temporally and spatially. We speculate that DpChk1 may be involved in conversion between different stages of reproduction and in sexual differentiation in D. pulex.
Wang Limei,Yan Haijing,Chen Xiaomeng,Han Lin,Liu Guibo,Yang Hua,Lu Danli,Liu Wenting,Che Chengye 한국미생물·생명공학회 2023 Journal of microbiology and biotechnology Vol.33 No.1
Fungal keratitis is a refractory kind of keratopathy. We attempted to investigate the antiinflammatory role of thymol on Aspergillus fumigatus (A. fumigatus) keratitis. Wound healing and fluorescein staining of the cornea were applied to verify thymol’s safety. Mice models of A. fumigatus keratitis underwent subconjunctival injection of thymol. The anti-inflammatory roles of thymol were verified by hematoxylin-eosin (HE) staining, slit lamp observation, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blotting. In contrast with the DMSO group, more transparent corneas and less inflammatory cells infiltration were detected in mice treated with 50 μg/ml thymol. Thymol downregulated the synthesis of TLR4, MyD88, NF-kB, IL-1β, NLRP3, caspase 1, caspase 8, GSDMD, RIPK3 and MLKL. In summary, we proved that thymol played a protective part in A. fumigatus keratitis by cutting down inflammatory cells aggregation, downregulating the TLR4/ MyD88/ NF-kB/ IL-1β signal expression and reducing necroptosis and pyroptosis.
Metabolic Regulation of Gene Expression by Histone Lysine β-Hydroxybutyrylation
Xie, Zhongyu,Zhang, Di,Chung, Dongjun,Tang, Zhanyun,Huang, He,Dai, Lunzhi,Qi, Shankang,Li, Jingya,Colak, Gozde,Chen, Yue,Xia, Chunmei,Peng, Chao,Ruan, Haibin,Kirkey, Matt,Wang, Danli,Jensen, Lindy M. Elsevier 2016 Molecular cell Vol.62 No.2
<P><B>Summary</B></P> <P>Here we report the identification and verification of a β-hydroxybutyrate-derived protein modification, lysine β-hydroxybutyrylation (Kbhb), as a new type of histone mark. Histone Kbhb marks are dramatically induced in response to elevated β-hydroxybutyrate levels in cultured cells and in livers from mice subjected to prolonged fasting or streptozotocin-induced diabetic ketoacidosis. In total, we identified 44 histone Kbhb sites, a figure comparable to the known number of histone acetylation sites. By ChIP-seq and RNA-seq analysis, we demonstrate that histone Kbhb is a mark enriched in active gene promoters and that the increased H3K9bhb levels that occur during starvation are associated with genes upregulated in starvation-responsive metabolic pathways. Histone β-hydroxybutyrylation thus represents a new epigenetic regulatory mark that couples metabolism to gene expression, offering a new avenue to study chromatin regulation and diverse functions of β-hydroxybutyrate in the context of important human pathophysiological states, including diabetes, epilepsy, and neoplasia.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Lysine β-hydroxybutyrylation (Kbhb) is a new type of histone mark </LI> <LI> 44 non-redundant histone Kbhb sites are identified in human and mouse cells </LI> <LI> Histone Kbhb increases under starvation and STZ-induced ketoacidosis </LI> <LI> Starvation-induced H3K9bhb is associated with active gene expression </LI> </UL> </P> <P><B>Graphical Abstract</B></P> <P>[DISPLAY OMISSION]</P>
Gentic overexpression increases production of hypocrellin A in Shiraia bambusicola S4201
Dan Li,Ning Zhao,Bing-Jing Guo,Xi Lin,Shuang-Lin Chen,Shu-Zhen Yan 한국미생물학회 2019 The journal of microbiology Vol.57 No.2
Hypocrellin A (HA) is a perylenequinone (PQ) isolated from Shiraia bambusicola that shows antiviral and antitumor activities, but its application is limited by the low production from wild fruiting body. A gene overexpressing method was expected to augment the production rate of HA in S. bambusicola. However, the application of this molecular biology technology in S. bambusicola was impeded by a low genetic transformation efficiency and little genomic information. To enhance the plasmid transformant ratio, the Polyethylene Glycol-mediated transformation system was established and optimized. The following green fluorescent protein (GFP) analysis showed that the gene fusion expression system we constructed with a GAPDH promoter Pgpd1 and a rapid 2A peptide was successfully expressed in the S. bambusicola S4201 strain. We successfully obtained the HA high-producing strains by overexpressing O-methyltransferase/FAD-dependent monooxygenase gene (mono) and the hydroxylase gene (hyd), which were the essential genes involved in our putative HA biosynthetic pathway. The overexpression of these two genes increased the production of HA by about 200% and 100%, respectively. In general, this study will provide a basis to identify the genes involved in the hypocrellin A biosynthesis. This improved transformation method can also be used in genetic transformation studies of other fungi.
Dan Li,Xiaolin Ding,Xiaolei Li 한국식품과학회 2010 Food Science and Biotechnology Vol.19 No.3
Tartary buckwheat, a healthy food, is associated with a reduced risk of certain human chronic diseases;however, its bioactive components are unknown. This study used high performance liquid chromatographyphotodiode array-mass spectroscopy (HPLC-PDA-MS) to identify the flavonoids in flavonoid-rich fractions (FRFs)from tartary buckwheat grain and tested antioxidative capacity of FRFs to confirm their bioactivity. Ultraviolet (UV) absorption and electrospray ionization (ESI−) MS/MS spectra identified several flavonols in the embryo,endosperm, testa, and hull, including the predominant flavonoid rutin and minor flavonoids quercetin 3-Orutinoside-3'-O-β-glucopyranoside, kaempferol 3-O-rutinoside,and quercetin. Tartary buckwheat FRFs extended the Rancimat induction period of lard less than common buckwheat FRF did, while their scavenging rate of DPPH free radicals exceeded that of common buckwheat FRF and rutin. These results indicate that tartary buckwheat FRFs from the grains have potential health benefits.
Dan Li,He Xiu Quan,Jin Fu Wen,Jing Yu Jin,,박성훈,김선영,김성주,조경우 대한약리학회 2005 The Korean Journal of Physiology & Pharmacology Vol.9 No.2
It is not clear whether Ca2+-induced Ca2+ release from the sarcoplasmic reticulum (SR) is involved in the regulation of atrial natriuretic peptide (ANP) release. Previously, we have shown that nifedipine increased ANP release, indicating that Ca2+ entry via voltage-gated L-type Ca2+ channel activation decreases ANP release. The purpose of the present study was two-fold: to define the role of SR Ca2+ release in the regulation of ANP release and whether Ca2+ entry via L-type Ca2+ channel is prerequisite for the SR-related effect on ANP release. Experiments were performed in perfused beating rabbit atria. Ryanodine, an inhibitor of SR Ca2+ release, increased atrial myocytic ANP release (8.69±3.05, 19.55±1.09, 27.31±3.51, and 18.91±4.76% for 1, 2, 3, and 6μM ryanodine, respectively; all P<0.01) with concomitant decrease in atrial stroke volume and pulse pressure in a dose-dependent manner. In the presence of thapsigargin, an inhibitor of SR Ca2+ pump, ryanodine-induced increase in ANP release was not observed. Thapsigargin attenuated ryanodine-induced decrease in atrial dynamic changes. Blockade of L-type Ca2+ channel with nifedipine abolished ryanodine-induced increase in ANP release (0.69±5.58% vs. 27.31±3.51%; P<0.001). In the presence of thapsigargin and ryanodine, nifedipine increased ANP release and decreased atrial dynamics. These data suggest that Ca2+-induced Ca2+ release from the SR is inversely involved in the regulation of atrial myocytic ANP release.
( Dan Li ),( Dongwei Fu ),( Yue Zhang ),( Xueling Ma ),( Liguo Gao ),( Xiaohua Wang ),( Dongpo Zhou ),( Kai Zhao ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.8
The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10- deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by <sup>1</sup>H NMR, <sup>13</sup>C NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (β-sitosterol and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger, which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger.