Ebracteolatain A and Ebracteolatain B Induce Apoptosis of Human Hepatoma Cell Line (HepG2)

Purpose: To evaluate the effects of ebracteolatain A (EA) and ebracteolatain B (EB) from Euphorbia ebracteolata Hyata (Euphorbiaceae) on the proliferation of HepG2 cells and the possible mechanisms.Methods: EA and EB from E. ebracteolata were obtained by column chromatography. 3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assays were used to study the cytotoxic and pro-apoptotic activities of EA and EB against HepG2 cells. Western blot assay was used to investigate the possible mechanisms of action.Results: EA and EB were successfully isolated from E. ebracteolata by column chromatography. The results of MTT assay indicate that EA and EB have significant anti-proliferative activities against HepG2 cells in dose- and time-dependent manners with half maximal inhibitory concentration (IC50) of 28.48 and 31.72 μg/mL. respectively. The results of flow cytometry assay suggest that EA and EB significantly (p < 0.01) induced the apoptosis of HepG2 cells at the levels of 47.45 and 42.26 %, respectively. Western blot data indicate that EA and EB significantly (p < 0.05 or 0.01) down-regulated the expression levels of anti-apoptotic proteins (survivin and Bcl-2) and up-regulated the expression levels of proapoptotic proteins (Smac, Bax, c-caspase-3 and c-caspase-9) in mitochondria-mediated apoptotic pathway.Conclusion: EA and EB inhibit the proliferation of HepG2 cells, the probable mechanisms being associated with mitochondria-mediated apoptosis.Keywords: Euphorbia ebracteolata, Phloroglucinol derivatives, Mitochondria-mediated apoptosis, Flow cytometry, Western blo

Distribution, characterization, and induction of CD8+ regulatory T cells and IL-17-producing CD8+ T cells in nasopharyngeal carcinoma

<p>Abstract</p> <p>Background</p> <p>CD8⁺effector cells often have an antitumor function in patients with cancer. However, CD8⁺Foxp3⁺regulatory T cells (Tcregs) and interleukin (IL)-17-producing CD8⁺T cells (Tc17 cells) also derive from the CD8⁺T cell lineage. Their role in the antitumor response remains largely unknown. In the present study, we aimed to investigate the distribution, characterization, and generation of CD8⁺Tcregs and Tc17 cells in NPC patients.</p> <p>Methods</p> <p>Peripheral blood and tumor biopsy tissues from 21 newly diagnosed patients with nasopharyngeal carcinoma (NPC) were collected, along with peripheral blood from 21 healthy donors. The biological characteristics of Tcregs and Tc17 cells from blood and tumor tissues were examined by intracellular staining, tetramer staining and fluorescence-activated cell sorting (FACS) analysis. The suppressive function of Tcregs was investigated using a proliferation assay that involved co-culture of sorted CD8⁺CD25⁺T cells with naïve CD4⁺T cells <it>in vitro</it>.</p> <p>Results</p> <p>We observed an increased prevalence of Tcregs and Tc17 cells among tumor-infiltrating lymphocytes (TILs) and different distribution among peripheral blood mononuclear cells (PBMCs) in NPC patients. Cytokine profiles showed that the Tcregs expressed a high level of IL-10 and low level of transforming growth factor β, whereas Tc17 cells expressed a high level of tumor necrosis factor α. Interestingly, both subsets expressed a high level of interferon γ in TILs, and the Tcregs suppressed naïve CD4⁺T cell proliferation by a cell contact-dependent mechanism <it>in vitro</it>. Moreover, we demonstrated the existence of Epstein-Barr virus latent membrane protein (LMP) 1 and LMP2 antigen-specific Tcregs in NPC.</p> <p>Conclusions</p> <p>Our data provide new insights into the composition and function of CD8⁺T-cell subsets in NPC, which may have an important influence on NPC immunotherapy.</p

BigDataBench: a Big Data Benchmark Suite from Internet Services

As architecture, systems, and data management communities pay greater attention to innovative big data systems and architectures, the pressure of benchmarking and evaluating these systems rises. Considering the broad use of big data systems, big data benchmarks must include diversity of data and workloads. Most of the state-of-the-art big data benchmarking efforts target evaluating specific types of applications or system software stacks, and hence they are not qualified for serving the purposes mentioned above. This paper presents our joint research efforts on this issue with several industrial partners. Our big data benchmark suite BigDataBench not only covers broad application scenarios, but also includes diverse and representative data sets. BigDataBench is publicly available from http://prof.ict.ac.cn/BigDataBench . Also, we comprehensively characterize 19 big data workloads included in BigDataBench with varying data inputs. On a typical state-of-practice processor, Intel Xeon E5645, we have the following observations: First, in comparison with the traditional benchmarks: including PARSEC, HPCC, and SPECCPU, big data applications have very low operation intensity; Second, the volume of data input has non-negligible impact on micro-architecture characteristics, which may impose challenges for simulation-based big data architecture research; Last but not least, corroborating the observations in CloudSuite and DCBench (which use smaller data inputs), we find that the numbers of L1 instruction cache misses per 1000 instructions of the big data applications are higher than in the traditional benchmarks; also, we find that L3 caches are effective for the big data applications, corroborating the observation in DCBench.Comment: 12 pages, 6 figures, The 20th IEEE International Symposium On High Performance Computer Architecture (HPCA-2014), February 15-19, 2014, Orlando, Florida, US

The effects of (+)-Gossypol on 11β-HSD and the concentration of corticosterone and dehydrocorticosterone in mice serum and tissues

11β-hydroxysteroid dehydrogenase (11β-HSD) plays an important part in mediating glucocorticoid action, catalyzing the interconversion of corticosterone (B) and dehydrocorticosterone (A) in rodents. The aim of our study is to investigate the effects of (+)-gossypol (G+) on 11β-HSD. Adult ICR mice were given B and B + (G+) by intraperitoneal injection. The activity of 11β-HSD was evaluated by measuring the ratio of A and B, meanwhile the effects of (+)-gossypol on the conversion rate of B to A was determined with HPLC. Serum A/B levels of the B+(G+) group decreased by 2.42, 7.32, 17.85, 31.39, and 40.02 % compared to the B group at each measured time interval. A/B levels at 1 h for the B + (G+) group decreased by 43.78, 21.29 and 34.47% in liver, kidney and adrenal glands, respectively, in comparison to the B group. However, (+)-gossypol had no effect on brain and testis. (+)-Gossypol was an inhibitor of 11β-HSD.Colegio de Farmacéuticos de la Provincia de Buenos Aire

The effects of (+)-Gossypol on 11β-HSD and the concentration of corticosterone and dehydrocorticosterone in mice serum and tissues

11β-hydroxysteroid dehydrogenase (11β-HSD) plays an important part in mediating glucocorticoid action, catalyzing the interconversion of corticosterone (B) and dehydrocorticosterone (A) in rodents. The aim of our study is to investigate the effects of (+)-gossypol (G+) on 11β-HSD. Adult ICR mice were given B and B + (G+) by intraperitoneal injection. The activity of 11β-HSD was evaluated by measuring the ratio of A and B, meanwhile the effects of (+)-gossypol on the conversion rate of B to A was determined with HPLC. Serum A/B levels of the B+(G+) group decreased by 2.42, 7.32, 17.85, 31.39, and 40.02 % compared to the B group at each measured time interval. A/B levels at 1 h for the B + (G+) group decreased by 43.78, 21.29 and 34.47% in liver, kidney and adrenal glands, respectively, in comparison to the B group. However, (+)-gossypol had no effect on brain and testis. (+)-Gossypol was an inhibitor of 11β-HSD.Colegio de Farmacéuticos de la Provincia de Buenos Aire

High expression of transcriptional coactivator p300 correlates with aggressive features and poor prognosis of hepatocellular carcinoma

<p>Abstract</p> <p>Background</p> <p>It has been suggested that p300 participates in the regulation of a wide range of cell biological processes and mutation of p300 has been identified in certain types of human cancers. However, the expression dynamics of p300 in hepatocellular carcinoma (HCC) and its clinical/prognostic significance are unclear.</p> <p>Methods</p> <p>In this study, the methods of reverse transcription-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry (IHC) were utilized to investigate protein/mRNA expression of p300 in HCCs. Receiver operating characteristic (ROC) curve analysis, spearman's rank correlation, Kaplan-Meier plots and Cox proportional hazards regression model were used to analyze the data.</p> <p>Results</p> <p>Up-regulated expression of p300 mRNA and protein was observed in the majority of HCCs by RT-PCR and Western blotting, when compared with their adjacent non-malignant liver tissues. According to the ROC curves, the cutoff score for p300 high expression was defined when more than 60% of the tumor cells were positively stained. High expression of p300 was examined in 60/123 (48.8%) of HCCs and in 8/123 (6.5%) of adjacent non-malignant liver tissues. High expression of p300 was correlated with higher AFP level, larger tumor size, multiplicity, poorer differentiation and later stage (<it>P </it>< 0.05). In univariate survival analysis, a significant association between overexpression of p300 and shortened patients' survival was found (<it>P </it>= 0.001). In different subsets of HCC patients, p300 expression was also a prognostic indicator in patients with stage II (<it>P </it>= 0.007) and stage III (<it>P </it>= 0.011). Importantly, p300 expression was evaluated as an independent prognostic factor in multivariate analysis (<it>P </it>= 0.021). Consequently, a new clinicopathologic prognostic model with three poor prognostic factors (p300 expression, AFP level and vascular invasion) was constructed. The model could significantly stratify risk (low, intermediate and high) for overall survival (<it>P </it>< 0.0001).</p> <p>Conclusions</p> <p>Our findings provide a basis for the concept that high expression of p300 in HCC may be important in the acquisition of an aggressive phenotype, suggesting that p300 overexpression, as examined by IHC, is an independent biomarker for poor prognosis of patients with HCC. The combined clinicopathologic prognostic model may become a useful tool for identifying HCC patients with different clinical outcomes.</p

Angiosperm phylogeny inferred from sequences of four mitochondrial genes

An angiosperm phylogeny was reconstructed in a maximum likelihood analysis of sequences of four mitochondrial genes, atp1, matR, nad5 , and rps3 , from 380 species that represent 376 genera and 296 families of seed plants. It is largely congruent with the phylogeny of angiosperms reconstructed from chloroplast genes atpB, matK , and rbcL , and nuclear 18S rDNA. The basalmost lineage consists of Amborella and Nymphaeales (including Hydatellaceae). Austrobaileyales follow this clade and are sister to the mesangiosperms, which include Chloranthaceae, Ceratophyllum , magnoliids, monocots, and eudicots. With the exception of Chloranthaceae being sister to Ceratophyllum , relationships among these five lineages are not well supported. In eudicots, Ranunculales, Sabiales, Proteales, Trochodendrales, Buxales, Gunnerales, Saxifragales, Vitales, Berberidopsidales, and Dilleniales form a basal grade of lines that diverged before the diversification of rosids and asterids. Within rosids, the COM (Celastrales–Oxalidales–Malpighiales) clade is sister to malvids (or rosid II), instead of to the nitrogen-fixing clade as found in all previous large-scale molecular analyses of angiosperms. Santalales and Caryophyllales are members of an expanded asterid clade. This study shows that the mitochondrial genes are informative markers for resolving relationships among genera, families, or higher rank taxa across angiosperms. The low substitution rates and low homoplasy levels of the mitochondrial genes relative to the chloroplast genes, as found in this study, make them particularly useful for reconstructing ancient phylogenetic relationships. A mitochondrial gene-based angiosperm phylogeny provides an independent and essential reference for comparison with hypotheses of angiosperm phylogeny based on chloroplast genes, nuclear genes, and non-molecular data to reconstruct the underlying organismal phylogeny.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/79100/1/JSE_97_sm_FigS2-1.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/79100/2/JSE_97_sm_FigS2-2.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/79100/3/JSE_97_sm_FigS3.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/79100/4/j.1759-6831.2010.00097.x.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/79100/5/JSE_97_sm_FigS1.pd