Publications by authors named "Sanyang Gao"

5 Publications

  • Page 1 of 1

Applications of loop-mediated isothermal DNA amplification.

Appl Biochem Biotechnol 2011 Apr 16;163(7):845-50. Epub 2010 Sep 16.

Key Laboratory of Preventive Veterinary Medicine and Animal Biotechnology, Binzhou Animal Husbandry and Veterinary Research Academy, Binzhou 256600, People's Republic of China.

During the last 10 years, with the development of loop-mediated isothermal amplification (LAMP) method, it has been widely applied in nucleic acid analysis because of its simplicity, rapidity, high efficiency, and outstanding specificity. This method employs a DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA. Expensive equipment are not necessary to acquire a high level of precision, and there are fewer preparation steps compared to conventional PCR and real-time PCR assays. This paper briefly summarized the applications of LAMP method in pathogenic microorganisms, genetically modified ingredients, tumor detection, and embryo sex identification.
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http://dx.doi.org/10.1007/s12010-010-9088-8DOI Listing
April 2011

Involvement of N-terminal-extended form of sphingosine kinase 2 in serum-dependent regulation of cell proliferation and apoptosis.

J Biol Chem 2005 Oct 15;280(43):36318-25. Epub 2005 Aug 15.

Division of Biochemistry, Department of Molecular and Cellular Biology, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan.

Sphingosine kinase (SPHK) 1 is implicated in the regulation of cell proliferation and anti-apoptotic processes by catalyzing the formation of an important bioactive messenger, sphingosine 1-phosphate. Unlike the proliferative action of SPHK1, another isozyme, SPHK2, has been shown to possess anti-proliferative or pro-apoptotic action. Molecular mechanisms of SPHK2 action, however, are largely unknown. The present studies were undertaken to characterize the N-terminal-extended form of SPHK2 (SPHK2-L) by comparing it with the originally reported form, SPHK2-S. Real-time quantitative PCR analysis revealed that SPHK2-L mRNA is the major form in several human cell lines and tissues. From sequence analyses it was concluded that SPHK2-L is a species-specific isoform that is expressed in human but not in mouse. At the protein level it has been demonstrated by immunoprecipitation studies that SPHK2-L is the major isoform in human hepatoma HepG2 cells. SPHK2-L, when expressed in human embryonic kidney (HEK) 293 cells, did not show any inhibition of DNA synthesis in the presence of serum, whereas it showed marked inhibition in the absence of serum. Moreover, serum deprivation resulted in the translocation of SPHK2-L into the nuclei. In addition, serum deprivation induced SPHK2-L expression in HEK293 cells. Furthermore, suppression of SPHK2 by small interfering RNA treatment prevented serum deprivation- or drug-induced apoptosis in HEK293 cells. Taken together, these results indicate that a major form of SPHK2 splice variant, SPHK2-L, in human cells does not inhibit DNA synthesis under normal conditions and that SPHK2-L accumulation in the nucleus induced by serum deprivation may be involved in the cessation of cell proliferation or apoptosis depending on the cell type.
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http://dx.doi.org/10.1074/jbc.M504507200DOI Listing
October 2005

Phosphorylation of extracellular signal-regulated protein kinase in cultured keloid fibroblasts when stimulated by platelet-derived growth factor BB.

Scand J Plast Reconstr Surg Hand Surg 2003 ;37(6):321-4

Department of Plastic Surgery, Kobe University School of Medicine, Kobe, Hyogo, Japan.

Abnormal scars result in distressing symptoms and disfiguring blemishes; an understanding of the molecular events that cause such scars, particularly keloids, would make possible the optimisation of both wound healing and treatment. Extracellular signal-regulated protein kinase (ERK) has a crucial role in distinct signalling pathways in different cells, but to date we know of no study on its signalling events in keloid fibroblasts. The purpose of this study was to characterise the expression of tyrosine phosphorylation kinases, particularly that of ERK, in keloids at the protein level by immunoblotting analysis. Studies on phosphorylation were made on cell lysates of three cultures of five different keloid fibroblasts (n = 5), their relatively 'normal' fibroblasts in adjacent skin (rNHDF, n = 5), and normal human dermal fibroblasts (n = 1, standard control). The result showed that ERK signalling molecular protein was more highly phosphorylated in keloid fibroblast culture than in the other two cultures.
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http://dx.doi.org/10.1080/02844310310004677DOI Listing
January 2005

A novel phenoxazine derivative suppresses surface IgM expression in DT40 B cell line.

Br J Pharmacol 2002 Nov;137(6):749-55

Department of Genome Sciences, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan.

1. 2-amino-4, 4alpha-dihydro-4alpha, 7-dimethyl-3H-phenoxazine-3-one (Phx) has been demonstrated to be an actinomycin D-like phenoxazine, and to display anti-tumour activity. 2. In this study, we report on the effect of Phx on B cell antigen receptor (BCR) and receptor-mediated signalling in DT40 B cells. 3. Treatment of B cells with Phx for 12 h inhibited BCR-stimulated tyrosine phosphorylation of cellular proteins. 4. B cells exposed to Phx exhibited down-regulation of surface IgM which is part of BCR. In contracts with actinomycin D, Phx rapidly reduced the expression of IgM without decreasing the expression of other signalling molecules. 5. Analysis with confocal microscopy demonstrated that Phx treatment reduced IgM expression both at the cell surface and inside the cell. 6. Treatment of B cells with Phx resulted in the reduction of IgM secretion. Since MG-132, a proteasomal inhibitor, restored IgM contents to the control levels, Phx has the specific effect of accelerating IgM degradation. 7. These results suggest that Phx down-regulates the expression of IgM and inhibits BCR-mediated signalling and IgM secretion. Phx may be useful as an immunosuppressive agent for therapeutic purposes.
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http://dx.doi.org/10.1038/sj.bjp.0704939DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1573560PMC
November 2002

Syk is required for p38 activation and G2/M arrest in B cells exposed to oxidative stress.

Antioxid Redox Signal 2002 Jun;4(3):509-15

Department of Genome Sciences, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan.

Syk has been demonstrated to play a crucial role in oxidative stress signaling in B cells. In this study, we have investigated the role of Syk in p38 activation and the regulation of cell-cycle progression upon oxidative stress. In B cells, p38 is activated by hydrogen peroxide (H(2)O(2)) stimulation. Syk is required for p38 activation following stimulation with 10-100 microM H(2)O(2), but not with 1 mM H(2)O(2). H(2)O(2)-induced p38 activation is abrogated in phospholipase C-gamma2 (PLC-gamma2)-deficient as well as Syk-deficient cells, suggesting that Syk activates p38 through PLC-gamma2 upon H(2)O(2) stimulation. Although stimulation with 20-100 microM H(2)O(2) induces cellular apoptosis in B cells, pretreatment with SB203580, a p38-specific inhibitor, has no effect on H(2)O(2)-induced apoptosis. Flow cytometric analysis reveals that B cells exposed to 10-20 microM H(2)O(2) exhibit cell-cycle profile of G2/M arrest, and pretreatment with SB203580 inhibits only a little H(2)O(2)-induced G2/M arrest. On the other hand, Syk-deficient cells show no induction of G2/M arrest following H(2)O(2) stimulation. These findings indicate that Syk plays a role in the regulation of cell-cycle progression in G2/M phase via p38-dependent and -independent pathways after oxidative stress.
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http://dx.doi.org/10.1089/15230860260196317DOI Listing
June 2002