Anti-Inflammatory Protein of Schistosoma japonicum Directs the Differentiation of the WEHI-3B JCS Cells and Mouse Bone Marrow Cells to Macrophages

Sj16 is an anti-inflammatory protein identified from Schistosoma japonicum. Our previous studies showed that recombinant Sj16 (rSj16) could suppress host's inflammatory responses and inhibit macrophage maturation. In the present study, the effects of rSj16 on the differentiation of the murine myeloid leukemia WEHI-3B JCS cell line and on mouse hematopoiesis were investigated. Our data demonstrated that rSj16 expressed and purified from Escherichia coli could suppress the proliferation of the WEHI-3B JCS cells in a time- and concentration-dependent manner, while not affect the viability of the cells. Further studies indicated that rSj16 induced macrophage differentiation of the WEHI-3B JCS cells, and arrested the cell cycle in the G1/G0 and G2/M phases. The macrophage differentiation of the rSj16-treated WEHI-3B JCS cells was confirmed by their expression of macrophage specific antigen F4/80 and phagocytic activity. Furthermore, our results revealed that rSj16 biased the colony formation of mouse bone marrow cells towards macrophage linage

Inflammatory and Tumor Stimulating Responses after Laparoscopic Sigmoidectomy

PURPOSE: Laparoscopic colectomy has clinical benefits such as short hospital stay, less postoperative pain, and early return of bowel function. However, objective evidence of its immunologic and oncologic benefits is scarce. We compared functional recovery after open versus laparoscopic sigmoidectomy and investigated the effect of open versus laparoscopic surgery on acute inflammation as well as tumor stimulation. MATERIALS AND METHODS: A total of 57 patients who were diagnosed with sigmoid colon cancer were randomized for elective conventional or laparoscopically assisted sigmoidectomy. Serum samples were obtained preoperatively and on postoperative day 1. C-reactive protein (CRP) and interleukin-6 (IL-6) were measured as inflammation markers, and vascular endothelial growth factor (VEGF) and insulin-like growth factor binding protein-3 (IGFBP-3) were used as tumor stimulation factors. Clinical parameters and serum markers were compared. RESULTS: Postoperative hospital stay (p=0.031), the first day of gas out (p=0.016), and the first day of soft diet (p<0.001) were significantly shorter for the laparoscopic surgery group than the open surgery group. The levels of CRP, IL-6, and VEGF rose significantly, and the concentration of IGFBP-3 fell significantly after both open and laparoscopic surgery. However, there were no significant differences in the preoperative and postoperative levels of CRP, IL-6, VEGF, and IGFBP-3 between the two groups. CONCLUSION: Our data suggest that both open and laparoscopic surgeries are accompanied by significant changes in IL-6, CRP, IGFBP-3, and VEGF levels. Acute inflammation markers and tumor stimulating factors may not reflect clinical benefits of laparoscopic surgery.ope

Photolumniscence/Fluorescence Spectroscopic Technique for Nanomaterials Characterizations

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The Immunomodulatory Activity of Jacaric Acid, a Conjugated Linolenic Acid Isomer, on Murine Peritoneal Macrophages.

This study aims at demonstrating the immunomodulatory property of jacaric acid, a conjugated linolenic acid (CLNA) isomer that is present in jacaranda seed oil, on murine peritoneal macrophages. Our results showed that jacaric acid exhibited no significant cytotoxicity on the thioglycollate-elicited murine peritoneal macrophages as revealed by the neutral red uptake assay, but markedly increased their cytostatic activity on the T-cell lymphoma MBL-2 cells as measured by the fluorometric CyQuant® NF Cell Proliferation Assay Kit. Flow cytometric analysis indicated that jacaric acid could enhance the endocytic activity of macrophages and elevated their intracellular production of superoxide anion. Moreover, jacaric acid-treated macrophages showed an increase in the production of nitric oxide which was accompanied by an increase in the expression level of inducible nitric oxide synthase protein. In addition, the secretion of several pro-inflammatory cytokines, including interferon-γ, interleukin-1β and tumor necrosis factor-α, was up-regulated. Collectively, our results indicated that the naturally-occurring CLNA isomer, jacaric acid, could exhibit immunomodulating activity on the murine peritoneal macrophages in vitro, suggesting that this CLNA isomer may act as an immunopotentiator which can be exploited for the treatment of some immunological disorders with minimal toxicity and fewer side effects

Omega-3 Polyunsaturated Fatty Acids Trigger Cell Cycle Arrest and Induce Apoptosis in Human Neuroblastoma LA-N-1 Cells

Omega-3 (n-3) fatty acids are dietary long-chain fatty acids with an array of health benefits. Previous research has demonstrated the growth-inhibitory effect of n-3 fatty acids on different cancer cell lines in vitro, yet their anti-tumor effects and underlying action mechanisms on human neuroblastoma LA-N-1 cells have not yet been reported. In this study, we showed that docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) exhibited time- and concentration-dependent anti-proliferative effect on the human neuroblastoma LA-N-1 cells, but had minimal cytotoxicity on the normal or non-tumorigenic cells, as measured by MTT reduction assay. Mechanistic studies indicated that DHA and EPA triggered G0/G1 cell cycle arrest in LA-N-1 cells, as detected by flow cytometry, which was accompanied by a decrease in the expression of CDK2 and cyclin E proteins. Moreover, DHA and EPA could also induce apoptosis in LA-N-1 cells as revealed by an increase in DNA fragmentation, phosphatidylserine externalization and mitochondrial membrane depolarization. Up-regulation of Bax, activated caspase-3 and caspase-9 proteins, and down-regulation of Bcl-XL protein, might account for the occurrence of apoptotic events. Collectively, our results suggest that the growth-inhibitory effect of DHA and EPA on LA-N-1 cells might be mediated, at least in part, via triggering of cell cycle arrest and apoptosis. Therefore, DHA and EPA are potential anti-cancer agents which might be used for the adjuvant therapy or combination therapy with the conventional anti-cancer drugs for the treatment of some forms of human neuroblastoma with minimal toxicity

Jacaric acid enhanced NO production in murine peritoneal macrophages.

<p>Macrophages were incubated with different concentrations of jacaric acid (either 50 or 100 μM) in the presence of LPS (60 ng/ml) at 37°C for 72 h. Cells treated with 0.1% ethanol acted as the control. (A) Cell-free supernatant was transferred to another 96-well plate and the amount of NO production was detected by Griess reagent. The results were expressed as the mean ± SE. *** <i>p</i>< 0.001. (B and C) Protein expression level of iNOS was assayed by Western blotting with β-actin protein as an internal control. The relative protein expression level of iNOS compared to β-actin was quantified. Results represent mean ± SE. * <i>p</i>< 0.05.</p

Cytostatic activity of jacaric acid-treated murine peritoneal macrophages on T-cell lymphoma MBL-2 cells.

<p>Macrophages were pre-treated with two different concentrations of jacaric acid (either 50 or 100 μM) in the absence (A) or presence (B) of 60 ng/ml LPS at 37°C for 72 h. Cells treated with 0.1% ethanol acted as the control. The pre-treated macrophages were incubated with MBL-2 cells at 37°C for 48 h and the cytostatic activity of macrophages was determined by the CyQuant^® NF Cell Proliferation Assay Kit. The results were expressed as the mean percentage inhibition of cell proliferation ± SE. * <i>p</i>< 0.05; ** <i>p</i>< 0.01.</p

Jacaric acid enhanced the secretion of pro-inflammatory cytokines in murine peritoneal macrophages.

<p>Macrophages were incubated with different concentrations of jacaric acid (either 50 or 100 μM) in the presence of LPS (60 ng/ml) at 37°C for 72 h. Cells treated with 0.1% ethanol acted as the control. The concentrations of IFN-γ (A), IL-1β (B) and TNF-α (C) were quantified by ELISA kit according to manufacturers’ instructions, and the results were expressed as mean ± SE. * <i>p</i>< 0.05; *** <i>p</i>< 0.001.</p

Effect of N-acetyl-L-cysteine on the jacaric acid-induced ROS generation and cytostatic activity in murine peritoneal macrophages.

<p>Macrophages were incubated with different concentrations of jacaric acid (either 50 or 100 μM) and N-acetyl-L-cysteine (either 10 or 100 μM), in the presence of LPS (60 ng/ml) at 37°C for 72 h. Cells treated with 0.1% ethanol acted as the control. (A) The intracellular levels of superoxide anion in the treated cells were measured by staining cells with DHE at 37°C for 30 min and analyzed for red fluorescence (FL-3) by flow cytometry. Results were expressed as mean ± SE. * <i>p</i>< 0.05; ** <i>p</i>< 0.01. (B) The pre-treated macrophages were incubated with MBL-2 cells at 37°C for 48 h and the cytostatic activity of macrophages was determined by the CyQuant^® NF Cell Proliferation Assay Kit. The results were expressed as the mean percentage inhibition of cell proliferation ± SE. * <i>p</i>< 0.05; ** <i>p</i>< 0.01.</p