OBJECTIVES: To investigate the synergistic inhibitory effects of AKBA and doxorubicin on malignant phenotype of triple-negative breast cancer (TNBC) MDA-MB-231 cells. METHODS: CCK-8 assay was used to determine the 48-h IC₅₀ of AKBA and doxorubicin in MDA-MB-231 cells, and SynergyFinder was employed to calculate the synergistic index and the optimal concentrations of the two agents. MDA-MB-231 cells treated with AKBA (22.5 μmol/L), doxorubicin (0.84 μmol/L) or their combination were examined for changes in cell proliferation, migration, invasion and apoptosis using Transwell migration, scratch assay, clone generation, RT-qPCR and Western blotting. Network pharmacology analysis was conducted to identify the downstream targets of AKBA in TNBC. In nude mouse models bearing subcutaneous MDA-MB-231 cell xenografts, the effects of normal saline, AKBA (50 mg/kg), doxorubicin (2.5 mg/kg), and AKBA combined with doxorubicin on xenograft growth and histopathology were observed. RESULTS: The IC₅₀ of AKBA and doxorubicin in MDA-MB-231 cells at 48 h was 45.15±0.97 μmol/L and 0.42±0.99 μmol/L, respectively. SynergyFinder confirmed the synergistic effect of AKBA and ADR with a ZIP>10. The combined treatment with AKBA and doxorubicin significantly inhibited the proliferation, migration and invasion, promoted apoptosis of MDA-MB-231 cells, and effectively suppressed xenograft growth in nude mice. Network pharmacology analysis predicted that AKBA affects the progression of TNBC through its downstream target AKBA. CONCLUSIONS AKBA combined with doxorubicin inhibits proliferation, migration and invasion, promotes apoptosis of MDA-MB-231 cells and suppresses MDA-MB-231 cell xenograft growth in nude mice. The combined use of AKBA can attenuate the toxic effects of doxorubicin in nude mice.
Animals ; Doxorubicin/pharmacology* ; Triple Negative Breast Neoplasms/pathology* ; Mice, Nude ; Mice ; Cell Proliferation/drug effects* ; Cell Line, Tumor ; Humans ; Female ; Apoptosis/drug effects* ; Cell Movement/drug effects* ; Xenograft Model Antitumor Assays ; Drug Synergism ; MDA-MB-231 Cells

OBJECTIVETo investigate the mechanisms underlying the ability ofheparin-treated dendritic cells (DCs) to promote Th0 to Th1 differentiation in chronic hepatitis B (CHB).

METHODSPeripheral blood mononuclear cells (PBMCs) were isolated from CHB patients and cultured in RPMI-1640 with recombinant GM-CSF and IL-4 with or without heparin to obtain DCs for study. The levels of Toll-like receptors (TLRs) on the DCs were measured using FACS and qPCR techniques.DC subsets with high expression of TLRs were selected for analysis of functional changes by treatment with the corresponding TLR-siRNA. The CD4+ T cell subpopulation was purified from peripheral blood by Dynal immunomagnetic beads, and then the production of IL-12 by DCs in the presence of poly(I:C) or R848 and ofIFN and IL-4 by Th cells co-cultured with DCs was evaluated by ELISA. The t-test was used for statistical analysis.

RESULTSTLR3 expression, and not expression of TLR 7 or TLR8,was significantly increased in heparin-treated DCs as compared to levels detected in the DCs without heparin treatment (t =2.849,P less than 0.05;t =3.027,P less than 0.05). The level of IL-12 produced by heparin-treated DCs stimulated with poly(I:C) was obviously higher than that produced by DCs without heparin treatment and stimulated with poly(I: C) (t =8.68,P less than 0.01) or with R848 (t =19.01,P less than 0.01). However, the IL-12 production by TLR3-siRNA transfected-DCs was significantly reduced (t =31.49, P less than 0.01).When Th cells from allogenic patients with CHB were co-cultured with the TLR3-siRNA transfectedDCs, the frequency ofCD4+ IFN+ cells was significantly reduced (1.64+/-0.57% vs.6.31+/-0.88%,P less than 0.01),as was the capability of Thl to generate IFNg (t =20.83,Pless than 0.01).

CONCLUSIONHeparin may have up-regulated the TLR3 expression level of DCs, and sequentially promoted Th0 to Th1 differentiation.

CD4-Positive T-Lymphocytes ; cytology ; Cell Differentiation ; Coculture Techniques ; Dendritic Cells ; cytology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Heparin ; pharmacology ; Hepatitis B, Chronic ; immunology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-4 ; pharmacology ; Monocytes ; cytology ; Recombinant Proteins ; pharmacology ; Toll-Like Receptor 3 ; metabolism

BACKGROUND: Membrane materials of bioreactor have exchange of substance and good physiochemical characteristics as well as good biocompatibility.OBJECTIVE: To evaluate the biocompatibility of interface of human hepatocyte/microporous polypropylene, i.e. graft modified microporous polypropylene semipermeable ultrafiltration membrane (MPP).DESIGN, TIME AND SETTING: Animal observation was performed at the Organ Transplantation Center, Ruijin Hospital of Shanghai Jiao Tong University Medical School and Polymers Institute of Zhejiang University between September 2005 and October 2007.MATERIALS: The microporous polypropylene ultrafiltration plane thin membranes, 0.2 μm diameter, M<,r> 50 000-100 000 molecular blockage, were used. Photochemical graft polymerization modification technique was adopted to graft hydrophilic acrylamide group through chemical bonds on MPP surface and succeeded in constructing an interface of human hepatocyte/microporous polypropylene, i.e. bioreactor membrane of bioartificial liver, graft modified MPP.METHODS: The biocompatibility of modified MPP was evaluated by hemolysis test, cytotoxicity test, acute systemic toxicity test, pyrogen test, skin sensitization and percutaneous stimulation test according to the requirements and biological evaluation criteria of medical device of ISO10993-1:1992.MAIN OUTCOME MEAURES: The experimental results of hemolysis, cytotoxicity, general acute toxicity, pyrogen, skin sensitization and percutaneous stimulation of modified MPP.RESULTS: The hemolytic rate of modified MPP was 1.90% (<5%), which showed that modified MPP did not lead to hemolysis. The extract solution of modified MPP exhibited no significant inhibition on the proliferative activity of L929 cells. At 24, 48 and 72 hours after MPP injection, no mice death, significant changes in body mass, or acute systemic toxicity were observed, such as ptosis, dyspnea, eyanosis, abdominal stimulation, diarrhea, decreased movement or tremor. In rabbit pyrogen test, the body temperature changed in a range from -0.2 to 0.4, which was consistent with the evaluation criteria of biomedical materials without pyrogen. Only one case was found with very slight erythema in skin sensitization test; its integral was 1 and primary stimulation index was 0.25 (<0.4), and the primary stimulation index of percutaneous stimulation test was 0.2; the average primary stimulation index was 0.068, indicating that modified MPP had no skin irritation.CONCLUSION: Modified MPP has no haemolytieus, cytotoxicity, pyrogenicity or skin sensitization, suggesting good biocompatibility by photochemical graft acrylamide on the surface of MPP.

OBJECTIVE:To prepare ambroxol hydrochloride orally disintegrating tablets and to evaluate its quality.METHODS:Mannitol was used as main adjuvant to prepare ambroxol hydrochloride orally disintegrating tablet,the UV spec-trometry was adopted to determine ambroxol content,the disintegration time,taste and dissolution rate were evaluated.RESULTS:The optimal formulation was composed of MCC 50g,mannitol 20g,L-HPC 10g,and NaHCO3 2g;The detected concentration of ambroxol was linear in the range of 0.005～0.030mg/ml(r=0.9 999),the average recovery was 100.50%and RSD=0.71;The prepared tablets disintegrated completely within 30s.CONCLUSION:The orally disintegrating tablets is quick to disintegrate and is as effective as market tablets.

Science and technology is known as a two-blade sword. For on one hand, the scientific development has brought human incredible advantages, while on the other hand, it has also lead to many dilemmas between science and ethics. For instance, technologies such as test-tube baby, artificial fertilization and in vitro fertilization, and cloning have drawn a lot of attention. A serous reflection should be given to ethics-related issues caused by these high-tech products.