MDSC-targeted liposomal all-trans retinoic acid suppresses mMdscs and improves immunotherapy in HBV infection.

BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are evolving as a prominent determinant in cancer occurrence and development and are functionally found to suppress T cells in cancer. Not much research is done regarding its involvement in viral infections. This research was designed to investigate the role of MDSCs in hepatitis B virus (HBV) infection and how targeting these cells with our novel all-trans retinoic acid encapsulated liposomal formulation could improve immunotherapy in C57BL/6 mice. METHODS: Ten micrograms (10 µg) of plasmid adeno-associated virus (pAAV/HBV 1.2, genotype A) was injected hydrodynamically via the tail vein of C57BL/6 mice. An all-trans retinoic acid encapsulated liposomal formulation (L-ATRA) with sustained release properties was used in combination with tenofovir disoproxil fumarate (TDF), a nucleotide analog reverse transcriptase inhibitor (nRTI) to treat the HBV infection. The L-ATRA formulation was given at a dose of 5 mg/kg intravenously (IV) twice a week. The TDF was given orally at 30 mg/kg daily. RESULTS: Our results revealed that L-ATRA suppresses MDSCs in HBV infected mice and enhanced T-cell proliferation in vitro. In vivo studies showed higher and improved immunotherapeutic effect in mice that received L-ATRA and TDF concurrently in comparison with the groups that received monotherapy. Lower HBV DNA copies, lower concentrations of HBsAg and HBeAg, lower levels of ALT and AST and less liver damage were seen in the mice that received the combination therapy of L-ATRA + TDF. CONCLUSIONS: In effect, targeting MDSCs with the combination of L-ATRA and TDF effectively reduced mMDSC and improved immunotherapy in the HBV infected mice. Targeting MDSCs could provide a breakthrough in the fight against hepatitis B virus infection.

An Unusual Bismuth-Antimony-Europium Cluster-Imbedded Polyoxotungstate and Its Bidirectional Luminescence Detection.

An unprecedented aggregate formed by two bismuth-antimony-europium cluster-imbedded tungsten-oxo clusters and one Krebs-type polyoxotungstate linker [H2N(CH3)2]14Na30H6[W4O10][B-ß-BiW9O33]2{[Bi5.35Sb0.65Eu3O9(H2O)9][B-α-SbW9O33]3}2·124H2O (1) was prepared. The polyoxoanion skeleton of 1 contains a Krebs-type polyoxotungstate [W4O10][B-ß-BiW9O33]2}14- ({Bi2W22}) (1a) as a linker that offers six active coordinate O atoms (two µ3-O and four µ2-O atoms) to grasp two Bi-Sb-Eu cluster-imbedded tungsten-oxo clusters {[Bi5.35Sb0.65Eu3O9(H2O)9][B-α-SbW9O33]3}18- (1b) through Bi-O-W and Sb-O-W bonds. 1b comprises an unprecedented nona-nuclearity Bi-Sb-Eu [Bi5.35Sb0.65Eu3O9(H2O)9]9+ cluster encircled by three trivacant [B-α-SbW9O33]9-segments in a triangular motif through Eu-O-W, Sb-O-W, and Bi-O-W linkages into a trilobal trimer. Moreover, a bidirectional detection method by using 1 as an effective luminescence probe was proposed to recognize both Mn2+ and CO32- through an "on-off-on" mode. 1 can be used as an "on-off" luminescent sensor to detect Mn2+ ions in aqueous solution. The limit of detection was 0.05 µM (9 × 10-6 mg L-1), which is much lower than the World Health Organization (WHO) guideline for Mn2+ concentration in drinking water (0.05 mg L-1). Then the Mn2+-quenching system can be used as an "off-on" sensor to detect CO32- in water system. This work provides a new research idea for the application of rare-earth-imbedded polyoxotungstate-based materials in the field of optical smart detection.

Intracellular distribution and mechanisms of actions of photosensitizer Zinc(II)-phthalocyanine solubilized in Cremophor EL against human hepatocellular carcinoma HepG2 cells.

Zinc(II)-phthalocyanine (ZnPc) is a metal photosensitizer. In the present study, we formulated the poorly-soluble ZnPc in Cremophor EL solution to enhance its solubility and determined its intracellular distribution and mechanisms of action on human hepatocellular carcinoma HepG2 cells. ZnPc uptake by the cells reached a plateau by 8h. ZnPc primarily located in mitochondria, lysosome and endoplasmic reticulum. The concentration-growth inhibition curves of ZnPc on the cell lines were pharmacodynamically enhanced by 10-50 folds by irradiation. Once irradiated, ZnPc produced significant amount of reactive oxygen species (ROS), activated caspase-3 and caspase-9, arrested cell cycle mainly at G2/M stage, and decreased membrane potential (ΔΨm) of HepG2 cells. In conclusion, the present study first elucidated cellular and molecular mechanisms of ZnPc on HepG2 cells.

Inhibition of human hepatocellular carcinoma HepG2 by phthalocyanine photosensitiser PHOTOCYANINE: ROS production, apoptosis, cell cycle arrest.

Photodynamic therapy (PDT) has been accepted as an alternative treatment for cancer. The rationale for the development of PDT for cancer is that target specificity can be achieved by controlling the location at which light activates the drug, i.e. photosensitiser. Metal phthalocyanines represent a new class of photosensitisers developed for cancer treatment. In the present study, we focused on exploring molecular mechanisms of the lead photosensitiser PHOTOCYANINE on hepatocellular carcinoma (HCC) HepG2 cells to guide our future development of PHOTOCYANINE. Growth inhibition potency of PHOTOCYANINE and its analogues was tested in vitro with and without irradiation at wavelength 670 nm. Irradiation shifted the concentration-growth inhibition curves of PHOTOCYANINE to the left and decreased the IC50s of PHOTOCYANINE required to produce equivalent inhibition by 200-fold on various cell lines. The amphipathic PHOTOCYANINE permeated through HepG2 cell membrane and predominately distributed to lysosome and mitochondria, where it significantly reduced mitochondrial membrane potential (ΔΨm) and increased caspase-3 activity in a concentration-dependent manner after irradiation. Early apoptosis of HepG2 occurred followed by necrosis when concentrations of PHOTOCYANINE were increased in the presence of irradiation. Reactive oxygen species (ROS) production was significant following PHOTOCYANINE plus irradiation treatment and cell cycle was mainly arrested at G2/M stage. In conclusion, PHOTOCYANINE, once irradiated, induces HepG2 cells into apoptosis via reducing ΔΨm, producing ROS, activating caspase-3, and causing cell arrest at G2/M stage. This study provides important insights into molecular mechanisms of the anti-cancer PHOTOCYANINE, which now is being applied for in the clinical trials II in China.

In vitro and in vivo anticancer activity evaluation of ursolic acid derivatives.

Twenty-three ursolic acid (1) derivatives 2-24 (ten novel compounds 8-10, 14-17 and 22-24) modified at the C-3 and the C-28 positions were synthesized, and their structures were confirmed by IR, (1)H NMR, MS, and elemental analysis. The single crystals of compounds 15 and 17 were obtained. The cytotoxic activity of the derivatives was evaluated against HepG2, BGC-823, SH-SY5Y, HeLa and HELF cells by the MTT assay. The induction of apoptosis and affects on the cell cycle distribution with compound 14 were assessed by fluorescence microscopy, flow cytometry and the activity of caspase-3 in HepG2 cells. Compounds 14-17 had more significant antiproliferative ability against the four cancer cell lines and low cytotoxicity to human embryonic lung fibroblast cells (HELF). Compounds 11, 14-16, 21 and 23 were particularly active against HepG2 cell growth. Compound 14 was selected to investigate cell apoptosis and cell cycle distribution. Flow cytometric analysis and morphologic changes of the cell exhibited that treatment of HepG2 cells with compound 14 led to cell apoptosis accompanied by cell cycle arrest at the S phase in a dose-dependent manner. Furthermore, the activity of the caspase-3 enzyme was increased in the treated cells. In vivo studies using H22 xenografts in Kunming mice were conducted with compound 14 at doses of 50, 100 and 150 mg/kg body weight. The results revealed that the medium dosage group (100 mg/kg) showed significant anticancer activity (45.6 ± 4.3%) compared to the control group.

[The effect of two leukocyte depletion in-line filters on the efficiency of whole blood filtration].

The aim of this study was to observe the difference in respect to the leukocyte reduction efficiency and quality of fresh-frozen plasma (FFP) from filtered whole blood between two types of in-line filters wherein only filter materials were surface modified by the two methods respectively. Whole blood was kept in refrigerator and filtered within 6 h of collection at ambient temperature. Samples were taken pre- and post filtration for analysis of WBC numbers, coagulation factors and complement activation (n = 8 for each type of filter). All filtered units contained < 2. 5 x 10(6) residual leucocytes. RBCs recovery was over 93%. No significant difference between group A and B was seen. But group B appeared to take longer time for filtration than did group A (9'29" vs. 8'01"). Neither group A nor group B showed statistically significant losses of total protein, album, IgG, IgM, fibrin, factors VIII, IX, vWF and C3 (P > 0.05). Factor V, XI and AT-III decreased significantly in two group filters. Group B showed more significant losses of IgA content and factor V activity than did group A, which appeared to be related to the difference in surface character between group A and group B filters. These two types of filters could remove leukocytes effectively, and no significant changes were observed in the quality of FFP from the filtered whole blood. It is presumed that the filter material with better bio-compatibility will give a high recovery of plasma protein and coagulation factors after filtration.