1,8-Cineole Ameliorates LPS-Induced Vascular Endothelium Dysfunction in Mice via PPAR-γ Dependent Regulation of NF-κB.

1,8-Cineole (eucalyptol), a monoterpene, has been widely reported for the anti-inflammatory effects. Our previous data confirmed that 1,8-cineole ameliorated the inflammatory phenotype of human umbilical vein endothelial cells (HUVECs) by mediating NF-κB expression in vitro. At present, we investigated the protection effects of 1,8-cineole on vascular endothelium in lipopolysaccharide (LPS)-induced acute inflammatory injury mice and the potential mechanisms involved in the protection in HUVECs. Results from enzyme linked immunosorbent assays revealed that 1,8-cineole suppressed the secretion of interleukin (IL)-6 and IL-8 and increased the expression of IL-10 in the serum of LPS-induced mice. 1,8-Cineole reduced the inflammatory infiltration and the expression of vascular cell adhesion molecular 1 (VCAM-1) in the sections of thoracic aorta in LPS-induced acute inflammatory mice. Western blotting indicated that 1,8-cineole significantly decreased the phosphorylation of NF-κB p65 and increased the expression of PPAR-γ in the thoracic aorta tissue. 1,8-Cineole increased the expression of PPAR-γ in LPS-induced HUVECs. 1,8-Cineole and rosiglitazone reduced the protein and mRNA levels of VCAM-1, E-selectin, IL-6, and IL-8 in LPS-induced HUVECs, which could be reversed by the action of GW9662 (inhibitor of PPAR-γ). 1,8-Cineole and rosiglitazone blocked the LPS-induced IκBα degradation and NF-κB p65 nucleus translocation, which could be reversed by the pretreatment of GW9662 or silence of PPAR-γ gene. In conclusion, 1,8-cineole attenuated LPS-induced vascular endothelial cells injury via PPAR-γ dependent modulation of NF-κB.

Suspension culture combined with chemotherapeutic agents for sorting of breast cancer stem cells.

BACKGROUND: Cancer stem cell (CSC) hypothesis has not been well demonstrated by the lack of the most convincing evidence concerning a single cell capable of giving rise to a tumor. The scarcity in quantity and improper approaches for isolation and purification of CSCs have become the major obstacles for great development in CSCs. Here we adopted suspension culture combined with anticancer regimens as a strategy for screening breast cancer stem cells (BrCSCs). BrCSCs could survive and be highly enriched in non-adherent suspension culture while chemotherapeutic agents could destroy most rapidly dividing cancer cells and spare relatively quiescent BrCSCs. METHODS: TM40D murine breast cancer cells were cultured in serum-free medium. The expression of CD44+CD24- was measured by flow cytometry. Cells of passage 10 were treated in combination with anticancer agents pacilitaxel and epirubicin at different peak plasma concentrations for 24 hours, and then maintained under suspension culture. The rate of apoptosis was examined by flow cytometry with Annexin-V fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining method. Selected cells in different amounts were injected subcutaneously into BALB/C mice to observe tumor formation. RESULTS: Cells of passage 10 in suspension culture had the highest percentage of CD44+CD24- (about 77 percent). A single tumor cell in 0.35 PPC could generate tumors in 3 of 20 BALB/C mice. CONCLUSION: Suspension culture combined with anticancer regimens provides an effective means of isolating, culturing and purifying BrCSCs.

Suspension culture combined with anticancer regimens for screening breast cancer stem cells.

Breast cancer stem cells (BrCSCs)have been first identified in solid tumors. To be more exact, they are defined as tumorigenic cancer cells which exhibit the capacity to form tumors distinct from the majority non-tumorigenic cancer cells. However it is only a minor subset that cancer stem cells are highly enriched in. It is still unknown how to find the definitive research subject - "a single breast cancer stem cell" - that can cause a new tumor by itself. Since recent studies suggest that BrCSCs have a higher proportion in suspension culture and have a greater resistance to chemotherapy regimens, we propose a novel strategy to isolate and identify BrCSCs:suspension culture combined with anticancer regimens. This double sorting method by means of the unique properties of BrCSCs may be helpful to screen genuine cancer stem cells(CSCs).

[Evaluation of the minimal invasiveness of laparoscopic operation for colorectal carcinoma].

OBJECTIVE: To investigate the minimal invasiveness of laparoscopic operation for colorectal carcinoma. METHODS: Forty cases with pathologically proven colorectal carcinoma were divided into laparoscopic group (n=20) and open surgical group (n=20). Perioperative alterations of peripheral blood IL-6, IL-8, TNF-alpha, CRP, sICAM-1 and WBC CD11b were compared between the two groups. TNF-alpha, IL- 6, IL- 8 and sICAM-1 were determined by ELISA, CRP by scattered radiation turbidity comparison and WBC CD11b by flow cytometry with monoclonal antibody PS- CD11b, M2Ab. RESULTS: The postoperative cytokine levels of TNF-alpha, IL-6 and IL-8 in open surgery group were significantly higher than those in laparoscopic group (P< 0.05). Dynamic level of sICAM-1 at 6 and 24 hours after operation in open surgery group were significantly higher than those in laparoscopic group. Peripheral WBC CD11b decreased to the lowest level at 6 hours after operation in open surgery group,significantly lower than that in laparoscopic group (P< 0.05). CONCLUSION: Laparoscopic surgery for colorectal carcinoma exerts less effects on patients than traditional open surgery, and can maintain patients defense function,therefore it is less invasive.