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Feifei Chen Jianren Ye Wanhui Liu Chonlong Chio Wendy Wang Wensheng Qin 《Molecular Plant Pathology》2021,22(7):843-857
Genetic transformation is a valuable and essential method that provides powerful insights into the gene function of microorganisms and contributes to the construction of engineered bacteria. Here, we developed a novel genetic transformation system to easily knock out a highly GC-rich gene (74.71% GC) from Burkholderia pyrrocinia JK-SH007, a biocontrol strain of poplar canker disease. This system revealed a reliable selectable marker (trimethoprim resistance gene, Tmp) and a simplified, efficient transformation method (6,363.64 CFU/μg, pHKT2) that was developed via freeze-thawing. The knockout recombineering of B. pyrrocinia JK-SH007 was achieved through a suicide plasmid with a three-fragment mutagenesis construct. The three-fragment cassette for mutagenesis was generated by overlap extension and touchdown PCRs and composed of Tmp flanked by GC-rich upstream and downstream fragments from B. pyrrocinia JK-SH007. The mutant strain (ΔBpEG), which was verified by PCR, lost 93.3% of its ability to degrade carboxymethyl cellulose over 40 days. Overall, this system may contribute to future research on B. pyrrocinia traits. 相似文献
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ShuChun Kuo ChungChing Chio ChaoHung Yeh JuiTi Ma WenPin Liu MaoTsun Lin KaoChang Lin ChingPing Chang 《Aging cell》2021,20(5)
Amyloid‐beta (Aβ) oligomer is known to contribute to the pathophysiology of age‐related macular degeneration. Herein, we aimed to elucidate the in vivo and in vitro effects of Aβ1‐42 application on retinal morphology in rats. Our in vivo studies revealed that intracerebroventricular administration of Aβ1‐42 oligomer caused dysmorphological changes in both retinal ganglion cells and retinal pigment epithelium. In addition, in vitro studies revealed that ARPE‐19 cells following Aβ1‐42 oligomer application had decreased viability along with apoptosis and decreased expression of the tight junction proteins, increased expression of both phosphor‐AKT and phosphor‐GSK3β and decreased expression of both SIRT1 and β‐catenin. Application of conditioned medium (CM) obtained from mesenchymal stem cells (MSC) protected against Aβ1‐42 oligomer‐induced retinal pathology in both rats and ARPE‐19 cells. In order to explore the potential role of peptides secreted from the MSCs, we applied mass spectrometry to compare the peptidomics profiles of the MSC‐CM. Gene ontology enrichment analysis and String analysis were performed to explore the differentially expressed peptides by predicting the functions of their precursor proteins. Bioinformatics analysis showed that 3‐8 out of 155–163 proteins in the MSC‐CM maybe associated with SIRT1/pAKT/pGSK3β/β‐catenin, tight junction proteins, and apoptosis pathway. In particular, the secretomes information on the MSC‐CM may be helpful for the prevention and treatment of retinal pathology in age‐related macular degeneration. 相似文献
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Edmond Pui-Hang Choi Janet Yuen-Ha Wong Herman Hay-Ming Lo Wendy Wong Jasmine Hin-Man Chio Daniel Yee-Tak Fong 《PloS one》2016,11(11)
Dating applications (apps) on smartphones have become increasingly popular. The aim of this study was to explore the association between the use of dating apps and risky sexual behaviours. Data were collected in four university campuses in Hong Kong. Subjects completed a structured questionnaire asking about the use of dating apps, sexual behaviours, and sociodemographics. Multiple linear and logistics regressions were used to explore factors associated with sexual risk behaviours. Six hundred sixty-six subjects were included in the data analysis. Factors associated with having unprotected sexual intercourse with more lifetime sexual partners included use of dating apps (β = 0.93, p<0.01), having one’s first sexual intercourse before 16 years of age (β = 1.74, p<0.01), being older (β = 0.4, p<0.01), currently being in a relationship (= 0.69, p<0.05), having a monthly income at least HKD$5,000 (β = 1.34, p<0.01), being a current smoker (β = 1.52, p<0.01), and being a current drinker (β = 0.7, p<0.01). The results of a multiple logistic regression analysis found that users of dating apps (adjust odds ratio: 0.52, p<0.05) and current drinkers (adjust odds ratio: 0.40, p<0.01) were less likely to have consistent condom use. Users of dating apps (adjust odds ratio: 1.93, p<0.05), bisexual/homosexual subjects (adjust odds ratio: 2.57, p<0.01) and female subjects (adjust odds ratio: 2.00, p<0.05) were more likely not to have used condoms the last time they had sexual intercourse. The present study found a robust association between using dating apps and sexual risk behaviours, suggesting that app users had greater sexual risks. Interventions that can target app users so that they can stay safe when seeking sexual partners through dating apps should be developed. 相似文献
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Di Meglio P Di Cesare A Laggner U Chu CC Napolitano L Villanova F Tosi I Capon F Trembath RC Peris K Nestle FO 《PloS one》2011,6(2):e17160
IL-23 and Th17 cells are key players in tissue immunosurveillance and are implicated in human immune-mediated diseases. Genome-wide association studies have shown that the IL23R R381Q gene variant protects against psoriasis, Crohn's disease and ankylosing spondylitis. We investigated the immunological consequences of the protective IL23R R381Q gene variant in healthy donors. The IL23R R381Q gene variant had no major effect on Th17 cell differentiation as the frequency of circulating Th17 cells was similar in carriers of the IL23R protective (A) and common (G) allele. Accordingly, Th17 cells generated from A and G donors produced similar amounts of Th17 cytokines. However, IL-23-mediated Th17 cell effector function was impaired, as Th17 cells from A allele carriers had significantly reduced IL-23-induced IL-17A production and STAT3 phosphorylation compared to G allele carriers. Our functional analysis of a human disease-associated gene variant demonstrates that IL23R R381Q exerts its protective effects through selective attenuation of IL-23-induced Th17 cell effector function without interfering with Th17 differentiation, and highlights its importance in the protection against IL-23-induced tissue pathologies. 相似文献
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Boys ML Bian F Kramer JB Chio CL Ren XD Chen H Barrett SD Sexton KE Iula DM Filzen GF Nguyen MN Angell P Downs VL Wang Z Raheja N Ellsworth EL Fakhoury S Bratton LD Keller PR Gowan R Drummond EM Maiti SN Hena MA Lu L McConnell P Knafels JD Thanabal V Sun F Alessi D McCarthy A Zhang E Finzel BC Patel S Ciotti SM Eisma R Payne NA Gilbertsen RB Kostlan CR Pocalyko DJ Lala DS 《Bioorganic & medicinal chemistry letters》2012,22(10):3392-3397
A series of 2-(1H-pyrazol-1-yl)pyridines are described as inhibitors of ALK5 (TGFβ receptor I kinase). Modeling compounds in the ALK5 kinase domain enabled some optimization of potency via substitutions on the pyrazole core. One of these compounds PF-03671148 gave a dose dependent reduction in TGFβ induced fibrotic gene expression in human fibroblasts. A similar reduction in fibrotic gene expression was observed when PF-03671148 was applied topically in a rat wound repair model. Thus these compounds have potential utility for the prevention of dermal scarring. 相似文献
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McKimmie CS Fraser AR Hansell C Gutiérrez L Philipsen S Connell L Rot A Kurowska-Stolarska M Carreno P Pruenster M Chu CC Lombardi G Halsey C McInnes IB Liew FY Nibbs RJ Graham GJ 《Journal of immunology (Baltimore, Md. : 1950)》2008,181(5):3353-3363
D6 scavenges inflammatory chemokines and is essential for the regulation of inflammatory and immune responses. Mechanisms explaining the cellular basis for D6 function have been based on D6 expression by lymphatic endothelial cells. In this study, we demonstrate that functional D6 is also expressed by murine and human hemopoietic cells and that this expression can be regulated by pro- and anti-inflammatory agents. D6 expression was highest in B cells and dendritic cells (DCs). In myeloid cells, LPS down-regulated expression, while TGF-beta up-regulated expression. Activation of T cells with anti-CD3 and soluble CD28 up-regulated mRNA expression 20-fold, while maturation of human macrophage and megakaryocyte precursors also up-regulated D6 expression. Competition assays demonstrated that chemokine uptake was D6 dependent in human leukocytes, whereas mouse D6-null cells failed to uptake and clear inflammatory chemokines. Furthermore, we present evidence indicating that D6 expression is GATA1 dependent, thus explaining D6 expression in myeloid progenitor cells, mast cells, megakaryocytes, and DCs. We propose a model for D6 function in which leukocytes, within inflamed sites, activate D6 expression and thus trigger resolution of inflammatory responses. Our data on D6 expression by circulating DCs and B cells also suggest alternative roles for D6, perhaps in the coordination of innate and adaptive immune responses. These data therefore alter our models of in vivo D6 function and suggest possible discrete, and novel, roles for D6 on lymphatic endothelial cells and leukocytes. 相似文献
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Jen-Chieh Liao Chung-Han Ho Fu-Wen Liang Jhi-Joung Wang Kao-Chang Lin Chung-Ching Chio Jinn-Rung Kuo 《PloS one》2014,9(4)