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Optimization of biotransformation from phytosterol to androstenedione by a mutant Mycobacterium neoaurum ZJUVN-08

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Journal of Zhejiang University-Science B(Biomedicine & biotechnology) 2013年第2期14卷 132-143页

作者： Xiao-yan ZHANG Yong PENG Zhong-rui SU Qi-he CHEN Hui RUAN Guo-qing HE department of Food Science and Nutrition Zhejiang University department of Food Science and Technology School of Agriculture and BiologyShanghai Jiao Tong University

Biotransformation of phytosterol(PS) by a newly isolated mutant Mycobacterium neoaurum ZJUVN-08 to produce androstenedione has been investigated in this paper.The parameters of the biotransformation process were optimized using fractional factorial design and response surface methodology.Androstenedione was the sole product in the fermentation broth catalyzed by the mutant M.neoaurum ZJUVN-08 strain.Results showed that molar ratio of hydroxypropyl-β-cyclodextrin(HP-β-CD) to PS and substrate concentrations were the two most significant factors affecting androstenedione production.By analyzing the statistical model of three-dimensional surface plot,the optimal process conditions were observed at 0.1 g/L inducer,pH 7.0,molar ratio of HP-β-CD to PS 1.92:1,8.98 g/L PS,and at 120 h of incubation time.Under these conditions,the maximum androstenedione yield was 5.96 g/L and nearly the same with the non-optimized(5.99 g/L),while the maximum PS conversion rate was 94.69% which increased by 10.66% compared with the non-optimized(84.03%).The predicted optimum conditions from the mathematical model were in agreement with the verification experimental results.It is considered that response surface methodology was a powerful and efficient method to optimize the parameters of PS biotransformation process.

关键词：晶状体上皮细胞蛋白质组分细胞增殖姜黄素

Delayed treatment of secondary degeneration following acute optic nerve transection using a combination of ion channel inhibitors

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Neural Regeneration Research 2017年第2期12卷 307-316页

作者： Nathanael J.Yates Marcus K.Giacci Ryan L. O'Hare Doig Wissam Chiha Bethany E. Ashworth Jade Kenna Carole A. Bartlett Melinda Fitzgerald department of Experimental and Regenerative Neurosciences School of Animal Biology The University of Western Australia Crawley Western Australia Australia department of Experimental and Regenerative Neurosciences School of Anatomy Physiology and Human Biology The University of Western Australia Crawley Western Australia Australia

Studies have shown that a combined application of several ion channel inhibitors immediately after central nervous system injury can inhibit secondary degeneration. However, for clinical use, it is necessary to determine how long after injury the combined treatment of several ion channel inhibitors can be delayed and efficacy maintained. In this study, we delivered Ca^2＋ entry-inhibiting P2X7 receptor antagonist oxidized-ATP and AMPA receptor antagonist YM872 to the optic nerve injury site via an iPRECIO-@ pump immediately, 6 hours, 24 hours and 7 days after partial optic nerve transection surgery. In addition, all of the ion channel inhibitor treated rats were administered with calcium channel antagonist lomerizine hydrochloride. It is important to note that as a result of implantation of the particular pumps required for programmable delivery of therapeutics directly to the injury site, seromas occurred in a significant proportion of animals, indicating infection around the pumps in these animals. Improvements in visual function were observed only when treatment was delayed by 6 hours; phosphorylated Tau was reduced when treatment was delayed by 24 hours or 7 days. Improvements in structure of node/paranode of Ranvier and reductions in oxidative stress indicators were also only observed when treatment was delayed for 6 hours, 24 hours, or 7 days. Benefits of ion channel inhibitors were only observed with time-delayed treatment, suggesting that delayed therapy of Ca^2＋ ion channel inhibitors produces better neuroprotective effects on secondary degeneration, at least in the presence of seromas.

关键词： nerve regeneration optic nerve injury neurotrauma secondary degeneration seromas calcium channel inhibitor node of Ranvier Tau phosphorylation lipid peroxidation oxidative stress neural regeneration

Developing an efficient and visible prime editing system to restore tobacco 8-hydroxy-copalyl diphosphate gene for labdane diterpene Z-abienol biosynthesis

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Science China(Life Sciences) 2023年第12期66卷 2910-2921页

作者： Jianduo Zhang Lu Zhang Chengwei Zhang Yongxing Yang Huayin Liu Lu Li Shengxue Zhang Xianggan Li Xinxiang Liu Ya Liu Jin Wang Guangyu Yang Qingyou Xia Weiguang Wang Jinxiao Yang Yunnan Key Laboratory of Tobacco Yunnan Academy of Tobacco ScienceKunming 650231China Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding Beijing Academy of Agriculture&Forestry SciencesBeijing 100089China Key Laboratory of Natural Products Synthetic biology of Ethnic Medicinal Endophytes State Ethnic Affairs CommissionYunnan Minzu UniversityKunming 650031China Cropedit biotechnology Co.Ltd. Beijing 102206China Biological Science Research Center Southwest UniversityChongqing 400716China

Prime editing(PE)is a versatile CRISPR-Cas based precise genome-editing platform widely used to introduce a range of possible base conversions in various organisms.However,no PE systems have been shown to induce heritable mutations in tobacco,nor in any other dicot.In this study,we generated an efficient PE system in tobacco that not only introduced heritable mutations,but also enabled anthocyanin-based reporter selection of transgene-free T_(1) plants.This system was used to confer Zabienol biosynthesis in the allotetraploid tobacco cultivar HHDJY by restoring a G>T conversion in the NtCPS2 gene.High levels of Z-abienol were detected in the leaves of homozygous T_(1) plants at two weeks after topping.This study describes an advance in PE systems and expands genome-editing toolbox in tobacco,even in dicots,for use in basic research and molecular breeding.And restoring biosynthesis of Z-abienol in tobacco might provide an efficient way to obtain Z-abienol in plants.

关键词： prime editing Z-abienol biosynthesis visible marker CRISPR Cas9 tobacco

OVOL1 inhibits breast cancer cell invasion by enhancing the degradation of TGF-β type I receptor

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Signal Transduction and Targeted Therapy 2022年第5期7卷 1842-1859页

作者： Chuannan Fan Qian Wang Gerard van der Zon Jiang Ren Cedrick Agaser Roderick C.Slieker Prasanna Vasudevan Iyengar Hailiang Mei Peter ten Dijke department of Cell and Chemical biology Leiden University Medical CenterLeiden 2300 RCThe Netherlands Oncode Institute Leiden University Medical CenterLeiden 2300 RCThe Netherlands department of Biomedical Data Sciences Sequencing Analysis Support CoreLeiden University Medical CenterLeiden 2300 RCThe Netherlands department of Epidemiology and Data Science Amsterdam Public Health InstituteAmsterdam Cardiovascular Sciences InstituteAmsterdam UMClocation VUmcAmsterdam 1081 HVThe Netherlands

Ovo-like transcriptional repressor 1(OVOL1)is a key mediator of epithelial lineage determination and mesenchymal–epithelial transition(MET).The cytokines transforming growth factor-β(TGF-β)and bone morphogenetic proteins(BMP)control the epithelial–mesenchymal plasticity(EMP)of cancer cells,but whether this occurs through interplay with OVOL1 is not known.Here,we show that OVOL1 is inversely correlated with the epithelial–mesenchymal transition(EMT)signature,and is an indicator of a favorable prognosis for breast cancer patients.OVOL1 suppresses EMT.

关键词： invasion breast cancer

Ccdc57 is required for straightening the body axis by regulating ciliary motility in the brain ventricle of zebrafish

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Journal of Genetics and Genomics 2023年第4期50卷 253-263页

作者： Lu Li Juan Li Yuan Ou Jiaxin Wu Huilin Li Xin Wang Liying Tang Xiangyan Dai Conghui Yang Zehong Wei Zhan Yin Yuqin Shu State Key Laboratory of Developmental biology of Freshwater Fish College of Life SciencesHunan Normal UniversityChangshaHunan 410081China College of Life Sciences Hunan Normal UniversityChangshaHunan 410081China School of Life Sciences East China Normal UniversityShanghai 200241China Key Laboratory of Freshwater Fish Reproduction and Development Ministry of EducationSouthwest UniversityChongqing 40715China State Key Laboratory of Freshwater Ecology and biotechnology Institute of HydrobiologyChinese Academy of SciencesWuhanHubei 430072China

Recently,cilia defects have been proposed to contribute to scoliosis.Here,we demonstrate that coiled-coil domain-containing 57(Ccdc57)plays an essential role in straightening the body axis of zebrafish by regulating ciliary beating in the brain ventricle(BV).Zygotic ccdc57(Zccdc57)mutant zebrafish developes scoliosis without significant changes in their bone density and calcification,and the maternal-zygotic ccdc57(MZccdc57)mutant embryos display curved bodies since the long-pec stage.The expression of ccdc57 is enriched in ciliated tissues and immunofluorescence analysis reveals colocalization of Ccdc57-HA with acetylated a-tubulin,implicating it in having a role in ciliary function.Further examination reveals that it is the coordinated cilia beating of multiple cilia bundles(MCB)in the MZccdc57 mutant embryos that is affected at 48 hours post fertilization,when the compromised cerebrospinal fluid flow and curved body axis have already occurred.Either ccdc57 m RNA injection or epinephrine treatment reverses the spinal curvature in MZccdc57 mutant larvae from ventrally curly to straight or even dorsally curly and significantly upregulates urotensin signaling.This study reveals the role of ccdc57 in maintaining coordinated cilia beating of MCB in the BV.

关键词： Scoliosis CSF ccdc57 Cilia Epinephrine Urotensin

Plastins:versatile modulators of actin organization in (patho)physiological cellular processes

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Acta Pharmacologica Sinica 2005年第7期26卷 769-779页

作者： Veerle DELANOTE Jo■l VANDEKERCKHOVE Jan GETTEMANS Faculty of Medicine and Health Sciences Albert Baertsoenkaai 3 B-9000 Ghent Belgium department of Medical Protein Research Flanders Interuniversity Institute for biotechnology(VIB) Ghent University Faculty of Medicine and Health Sciences Albert Baertsoenkaai 3 B-9000 Ghent department of Medical Protein Research Flanders Interuniversity Institute for biotechnology(VIB) Belgium Ghent University Faculty of Medicine and Health Sciences Albert Baertsoenkaai 3 B-9000 Ghent Belgium department of Medical Protein Research Flanders Interuniversity Institute for biotechnology(VIB) Ghent University

Many actin-binding proteins are expressed in eukaryotic cells.These polypep-tides assist in stabilizing and rearranging the organization of the actin cytoskel-eton in response to external stimuli,or during cell migration and adhesion.Herewe review a particular set of actin-binding proteins called plastins.Plastins(alsocalled fimbrins)belong to a subclass of actin-binding proteins known as actinbundling proteins.Three isoforms have been characterized in mammals:T-plastinis expressed in cells from solid tissue,whereas L-plastin occurs predominantly inhematopoietic cells.The third isoform,I-plastin,is specifically expressed in thesmall intestine,colon and kidney.These proteins share the unique property ofcross-linking actin filaments into fight bundles.Although plastins are primarilyinvolved in regulation of the actin cytoskeleton,they possess some uniquefeatures.For instance,they are implicated in invasion by pathogenic bacteriasuch as Shigellaflexneri and Salmonella typhimurium.Also,L-plastin plays animportant role in leukocyte function.T-plastin,on the other hand,is possiblyinvolved in DNA repair.Finally,both T-and L-plastin are implicated in severaldiseases,and L-plastin is considered to be a valuable marker for cancer.

关键词： actin actin-binding protein plastin cytoskeleton

Effects of diabetes type 2 and metformin treatment in Swedish patients with colorectal cancer

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World Journal of Gastroenterology 2022年第19期28卷 2148-2151页

作者： Jan Dimberg Levar Shamoun Kalle Landerholm Dick Wågsäter department of Natural Science and Biomedicine School of Health and WelfareJönköping UniversityJönköping 55111Sweden department of Laboratory Medicine and Pathology Region Jönköping CountyJönköping 55305Sweden department of Medical Cell biology Uppsala UniversityUppsala 75123Sweden department of Surgery Region Jönköping CountyJönköping 55305Sweden department of Biomedical and Clinical Sciences Linköping UniversityLinköping 58185Sweden

The association between type 2 diabetes mellitus(DM)and colorectal cancer(CRC)has been thoroughly investigated and reports have demonstrated that the risk of CRC is increased in DM patients.The association between DM and the survival of patients with CRC is controversial.Evidence suggests that metformin with its anti-inflammatory effects is a protective factor against the development of CRC among DM patients and that metformin therapy is associated with a better prognosis in patients with DM.In our cohort,we did not find any associations between the presence of DM or metformin and cancer specific survival or any relation to plasma levels of a panel of 40 inflammatory factors and irisin.On the other hand,we identified that the insulin-like growth factor binding protein 7 single nucleotide polymorphism rs2041437 was associated with DM in CRC patients.The dominance of the T bearing genotypes in patients with DM was statistically significant(P=0.038),with an odds ratio of 1.66(95%confidence interval:1.03-2.69).

关键词： Diabetes Metformin Cytokines Survival Colorectal cancer Polymorphism

Disruption of MIR396e and MIR396f improves rice yield under nitrogen-deficient conditions

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National Science Review 2020年第1期7卷 102-112页

作者： Jinshan Zhang Zhenyu Zhou Jinjuan Bai Xiaoping Tao Ling Wang Hui Zhang Jian-Kang Zhu Shanghai Center for Plant Stress biology and Center for Excellence in Molecular Plant Sciences Chinese Academy of Sciences University of Chinese Academy of Sciences School of biotechnology Jiangsu University of Science and Technology Shanghai Engineering Research Center of Plant Germplasm Resources College of Life Science Shanghai Normal University department of Horticulture and Landscape Architecture Purdue UniversityWest Lafayette

The microRNA miR396 directly represses GROWTH-REGULATING FACTORs(OsGRFs) and has been implicated in regulating rice yield and in nitrogen assimilation. Overexpressing the miR396 targets OsGRF4 and OsGRF6 improves rice yield via increased grain size and panicle branching, respectively. Here, we used CRISPR/Cas9 to assess the function of miR396 genes in rice. Knockout of MIR396 ef(MIR396e and MIR396f), but not other isoforms, enhanced both grain size and panicle branching, resulting in increased grain yield. Importantly, under nitrogen-deficient conditions, mir396ef mutants showed an even higher relative increase in grain yield as well as elevated above-ground biomass. Furthermore, we identified OsGRF8 as a new target of miR396, in addition to the known targets OsGRF4 and OsGRF6. Disruption of the miR396-targeting site in OsGRF8 was sufficient to both enlarge grain size and elongate panicles. Our results suggest that rice-seed and panicle development are regulated by miR396ef-GRF4/6/8-GIF1/2/3 modules and that mi R396ef are promising targets of genome editing for breeding environmentally friendly rice varieties that require less nitrogen fertilization.

关键词： microRNA miR396 OsGRF4/6/8 OsGIF1/2/3 nitrogen deficiency grain yield

Dust mite allergen-specific immunotherapy increases IL4 DNA methylation and induces Der p-specific T cell tolerance in children with allergic asthma

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Cellular & Molecular Immunology 2018年第11期15卷 963-972页

作者： Chuang-Ming Wang Chia-Bin Chang Michael WY Chan Zhi-Hong Wen Shu-Fen Wu department of Pediatrics Ditmanson Medical Foundation Chia-Yi Christian Hospital60002Chia-YiTaiwanChina department of Nursing Chang Gung University of Science and Technology61363Chia-YiTaiwanChina department of Life Science and Institute of Molecular biology National Chung-Cheng University62102Chia-YiTaiwanChina department of Marine biotechnology and Resources Asia-Pacific Ocean Research CenterNational Sun Yat-Sen University80424KaohsiungTaiwanChina

Allergen-specific immunotherapy(allergen-SIT)is a highly effective treatment for children with allergic asthma(AA),an immune-mediated chronic disease leading to bronchial muscle hypertrophy and airway obstruction in response to specific allergens.T helper cells and secreted cytokines play important roles in the pathogenesis of asthma,and epigenetic modulation controls genes important for T cell development and cytokine expression.This study evaluated T helper cell-secreted cytokines and DNA methylation patterns in children treated with Dermatophagoides pteronyssinus(Der p)allergen-SIT.Our results showed that after Der p challenge,peripheral blood mononuclear cells(PBMCs)from the SIT group,compared with the non-SIT AA group,produced lower levels of IL-4,IL-5 and IL-2.The SIT group,compared with the AA group,exhibited decreased sensitivity to the Der p allergen,concurrent with IL-4 down-modulation due to increased promoter DNA methylation,as estimated in PBMCs.Our results showed that SIT decreased IL-4 and IL-5,and inhibited T cell proliferation,by inhibiting IL-2 production after the specific allergen challenge.These results suggest that decreased IL-2 production and increased IL-4 cytokine promoter methylation is a potential mechanism of Der p-specific allergen desensitization immunotherapy.

关键词： allergic asthma allergen-specific immunotherapy dust mite Dermatophagoides pteronyssinus DNA methylation interleukin-4