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Wang J.-Q.,Guangdong Pharmaceutical University | Wang J.-Q.,Guangdong Provincial Key Laboratory of Biotechnology Candidate Drug Research | Zhao Z.-Z.,Sun Yat Sen University | Bo H.-B.,Guangdong Pharmaceutical University | Chen Q.-Z.,Guangdong Pharmaceutical University
Journal of Coordination Chemistry | Year: 2016

Three new Ru(II) complexes, [Ru(dmb)2(ipad)](ClO4)2 (dmb = 4,4′-dimethyl-2,2′-bipyridine, ipad = 2-(anthracene-9,10-dione-2-yl) imidazo[4,5-f][1,10]phenanthroline, 1), [Ru(dmp)2(ipad)](ClO4)2 (dmp = 2,9-dimethyl-1,10-phenanthroline, 2), and [Ru(dip)2(ipad)](ClO4)2 (dip = 4,7-diphenyl-1,10-phenanthroline, 3), have been synthesized and characterized. The three Ru(II) complexes intercalate with the base pairs of DNA. The in vitro antiproliferative activities and apoptosis-inducing characteristics of these complexes were investigated. The complexes exhibited cytotoxicity against various human cancer cell lines. BEL-7402 cells displayed the highest sensitivity to 1, accounted for by the greatest cellular uptake. Complex 1 was shown to accumulate preferentially in the nuclei of BEL-7402 cells and cause DNA damage and induce apoptosis, which involved cell cycle arrest and reactive oxygen species generation. © 2015 Taylor & Francis. Source

Wang Y.,South China Agricultural University | Jia W.-Z.,Guangdong Pharmaceutical University | Jia W.-Z.,Guangdong Provincial Key Laboratory of Biotechnology Candidate Drug Research | Tan M.-J.,Guangdong Pharmaceutical University | And 3 more authors.
Chinese Traditional and Herbal Drugs | Year: 2013

Objective: To obtain the indispensable key enzyme involved in the MEP pathway, the 2C-methyl-D-erythritol-2, 4-cyclodi-phosphate synthase gene (MCS) was cloned from Artemisia annua, and bioinformatic analysis, prokaryotic expression, and tissue-specific expression were conducted. Methods: According to AaMCS EST sequence, the full length of cDNA and genomic sequences were obtained by the design of specific primers, rapid-amplification of cDNA ends (RACE) and genome amplification. The coding region of MCS gene was cloned into the expression vector pET-21a (+) by gene recombination technology, the recombinant plasmid pET-21a (+)-MCS was transformed into E. coli BL21 (DE3), and the expression of recombinant protein was induced by IPTG. Semi-quantitative RT-PCR was used to detect the expression of AaMCS transcripts. Results: The AaMCS cDNA was found to be 994 bp containing an open reading frame (ORF) of 681 bp that translated into a putative peptide of 226 amino acid, The full length of MCS was 2540 bp consisting of three exons and two introns. A recombinant pET-21a (+)-MCS was constructed by genetically fusing the MCS to pET-21a (+) vector system, and was successfully expressed in E. coli BL21. The tissue expression patterns indicated that the expression level of AaMCS transcripts in flowers was higher than that in the roots and stems. Conclusion: The MCS gene is cloned from A. annua, and the stable prokaryotic expression system of pET-21a (+)-MCS is constructed. This work is helpful for investigating the activities and other physiological functions of MCS protein. Source

Jia W.,Guangdong Pharmaceutical University | Jia W.,Guangdong Provincial Key Laboratory of Biotechnology Candidate Drug Research | Wang Y.,South China Agricultural University | Yu X.,Guangdong Pharmaceutical University | And 7 more authors.
Plant OMICS | Year: 2014

Terpene synthases (TPSs) are responsible for the synthesis of the various terpene molecules which play important roles in the accumulation of secondary metabolites. However, systematic analysis of Artemisia annua TPS family has not been carried out, and little is known about the differences of transcript levels for TPS genes in A. annua (AaTPSs) and the regulation thereof. Though plant TPSs are generally divided into seven clades, a family of TPS in A. annua was characterized with lineages having a majority of TPS in TPS-a and TPS-b clades. Semi-quantitative RT-PCR demonstrates that 17 AaTPS genes are functional or potentially functional and express in at least some tissues or organs of the plant. In trichomes, the transcripts of majority of AaTPSs and artemisinin synthesis-related genes are express at a higher level than in leaves containing trichomes, except that AaBFS, AaCPS and AaSTS1 appear to be lower in trichomes but higher in leaves. AaECS is mainly express in trichomes, but although the highest expression level was detected in old leaf trichomes. The expression of AaSQS in leaves was slightly higher than that of trichomes, indicating that this gene is also express mainly in other parts of leaves instead of trichomes. Characteristics of sequence motifs, phylogenetic relationships and transcript levels of AaTPSs provide a foundation for future functional analysis. The present study contributes more data on the tissue-specific expression of AaTPSs in A. annua, and provides clues to dissect the concerted regulation for AaTPSs expression and the specific competitor for farnesyl diphosphate (FDP) in A. annua trichomes. Source

Wu F.,Guangdong Pharmaceutical University | Wu F.,Southern Medical University | Wu F.,Guangdong Provincial Key Laboratory of Biotechnology Candidate Drug Research | Zhang W.,Southern Medical University | And 15 more authors.
Cancer Letters | Year: 2013

Adoptive cell therapy provides an attractive treatment of cancer, and our expanding capacity to target tumor antigens is driven by genetically engineered human T lymphocytes that express genes encoding tumor-specific T cell receptors (TCRs). The intrinsic properties of cultured T cells used for therapy were reported to have tremendous influences on their persistence and antitumor efficacy in vivo. In this study, we isolated CD8+ central memory T cells from peripheral blood lymphocytes of healthy donors, and then transferred with the gene encoding TCR specific for tumor antigen using recombinant adenovirus vector Ad5F35-TRAV-TRBV. We found effector T cells derived from central memory T cells improved cell viability, maintained certain level of CD62L expression, and reacquired the CD62L+CD44high phenotype of central memory T cells after effector T cells differentiation. We then compared the antitumor reactivity of central memory T cells and CD8+T cells after TCR gene transferred. The results indicated that tumor-specific TCR gene being transferred to central memory T cells effectively increased the specific killing of antigen positive tumor cells and the expression of cytolytic granule protein. Furthermore, TCR gene transferred central memory T cells were more effective than TCR gene transferred CD8+T cells in CTL activity and effector cytokine secretion. These results implicated that isolating central memory T cells rather than CD8+T cells for insertion of gene encoding tumor-specific TCR may provide a superior tumor-reactive T cell population for adoptive transfer. © 2013 Elsevier Ireland Ltd. Source

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