BackgroundThis study investigated the dual control mechanism of the Shuanghuang Shengbai granule in modulating the cell cycle in Lewis-bearing mice with cyclophosphamide induced myelosuppression.MethodsThirty Lewis-bearing mice were randomly grouped into an untreated group, control group, and treated group. Both treated and untreated groups were intraperitoneally injected with cyclophosphamide to produce a myelosuppression model. Mice in the treated group were fed with the Shuanghuang Shengbai granule (40 g/day) for 6 consecutive days. Standard blood tests and the count of bone marrow nuclear cells were performed, and the cell reproductive cycles of bone marrow and tumors were measured in these mice. In addition, the western blot approach was used to measure the upstream activating signals of CyclinD-CDK4/6 such as c-Myc and CDC25A, the upstream suppression signals such as p16INK4a, and the expression of downstream activated signals such as Rb, pRB, and E2F. All of the tested results were validated by reverse transcription quantitative real-time polymerase chain reaction.ResultsThe results showed that the Shuanghuang Shengbai granule could elevate the count of leukocyte and bone marrow nuclear cells of Lewis-bearing mice with cyclophosphamide induced myelosuppression. It could also stimulate bone marrow cells to move from G0/G1 phases to S phase, accelerating the progress of the cell reproductive cycle and increasing the cell proliferation index. Simultaneously, the Shuanghuang Shengbai granule could also suppress cancer cells moving from G0/G1 phase to S phase, reducing the proliferation index. The tumor weight of Lewis-bearing mice in the treated group was much less than those of the control group. Expression levels of c-Myc, CDC25A, Rb, pRb, and E2F of bone marrow in Shuanghuang Shengbai granule-treated mice was higher compared to the control group, whereas they were lower in the cancer cells.ConclusionThe experimental results demonstrate that the Shuanghuang Shengbai granule has dual control on the cell reproductive cycles in cancer cells and bone marrow nuclear cells in Lewis-bearing mice.

BackgroundRecent studies have suggested that variables related to host adaptive immunity and the tumor microenvironment may predict the outcome in patients with non-Hodgkin's lymphoma. This study was undertaken to determine the prognostic value of peripheral blood leucocyte subpopulations in diffuse large-B-cell lymphoma patients.MethodsWe prospectively analyzed the 16 leukocyte subpopulations using Cytodiff flow cytometric technique in a cohort of 45 diffuse large-B-cell lymphoma patients at a single institution between February and December 2014. The Cox proportional hazards model was used to evaluate prognostic factors for overall survival and progression free survival.ResultsDiffuse large-B-cell lymphoma patients had decreased cytotoxic and non-cytotoxic NK&T cells as well as increased CD16+ monocytes, CD16- monocytes and mature neutrophils. The decreased CD16- monocyte/CD16+ monocyte ratio and increased mature neutrophil/cytotoxic NK&T cell ratio were related to poor progression-free and overall survival outcome in single and multivariate analysis. The co-constructed model using International Prognostic Index and mature neutrophil/cytotoxic NK&T cell ratio can also help discriminate the clinical outcome.ConclusionsThe decreased CD16-monocyte/CD16+monocyte ratio and increased mature neutrophil/cytotoxic NK&T cell ratio predict poor prognosis in diffuse large-B-cell lymphoma patients. This finding provides a strong rationale for the study of cellular immunotherapy in B-cell lymphoma.

Single nucleotide polymorphisms (SNPs) of vascular endothelial growth factor receptor (VEGFR) may have effects on the MAPK/ERK/STAT3 signaling pathway, and the resulting phenotypes may influence the response to sunitinib-targeted therapy for renal cell carcinoma. In order to test this hypothesis patients with advanced renal cell carcinoma treated with sunitinib, were enrolled in our study. Peripheral blood samples were used to run a polymerase chain reaction-restriction fragment length polymorphism protocol to type candidate nucleotide polymorphism loci (VEGFR1, VEGFR2 and VEGFR3). The samples were also used in western blots to determine p-MAPK/ERK/STAT3 protein expression levels. The clinical responses to treatment were recorded and then a logistic regression method was applied to analyze the correlation between polymorphism of loci and effectiveness of sunitinib therapy. According to a follow-up visit (on average after 15 months of treatment) there were 16 complete responses (CR), 29 partial responses (PR) and 23 stable disease (SD) and progression of disease (PD) cases. Tests were carried out for 5 SNPs: VEGFR1 (rs664393), VEGFR2 (rsl870377 and rs7667298) and VEGFR3 (rs448012 and rs72816988). Mutation rates of rsl870377 and rs448012 loci in the CR+PR group were lower than those in the SD+PD group. No such differences were found for the other 3 loci. Relative expression levels of p-MAPk, p-ERK and p-STAT3 in the CR+PR group were significantly lower than those in the SD+PD group (P<0.05). The median progression-free survival and overall survival (OS) in the CR+PR group were higher than those in the SD+PD group (P<0.001). The median OS of the TT rsl870377 genotype was higher than that of the AA genotype, and the median OS of the GG rs448012 genotype was higher than that of the CC genotype (P<0.001). It was concluded through a logistic regression model that rsl870377 (AA) and rs448012 (GG) are independent risk factors closely associated with the effectiveness of sunitinib-targeted therapy on renal cell carcinoma. VEGFR SNPs are able to mediate the MAPK/ERK/STAT3 signaling pathway and therefore influence the effectiveness of sunitinib-targeted therapy, which makes them possible new therapeutic targets.

Leukemia is a malignant tissue-forming disease, which induces the overproduction of large numbers of immature blood cells entering the peripheral blood. It is well documented that inflammation plays a crucial role in the expansion of leukemia. Daphnetin has confirmed anti-inflammatory effects against various diseases. In this experimental study, we evaluated the anti-leukemia and anti-inflammatory effect of daphnetin against benzene-induced leukemia in rats and explored the underlying mechanism. Benzene was used for inducing leukemia in experimental rats. The rats were divided into different groups and the body weight, hematological parameters, bone marrow cells, cytokines, and inflammatory mediators were estimated. Reverse transcription polymerase chain reaction (RT-PCR) was used for estimating the messenger RNA (mRNA) expression of sphingosine-1-phosphate receptor-1. Daphnetin-treated rats showed upregulation of body weight compared to other groups. Moreover, Daphnetin reduced blasts in leukemic rats. It also altered hematological parameters such as red blood cells, white blood cells, lymphocytes, neutrophils, monocytes, eosinophils, monocytes, and basophils, respectively. Daphnetin-treated rats showed a reduction of pro-inflammatory cytokines such as tumor necrosis factor-α, interleukin-1β (IL-1β), IL-2, IL-6, and inflammatory mediators including nuclear factor-κB. RT-PCR showed upregulated mRNA expression of sphingosine-1phosphate receptor-1 of daphnetin-treated group rats compared to other groups.The current study showed that the anti-inflammatory effect of daphnetin against the benzene-induced leukemia via alteration of cytokines.