Pickering emulsion co-delivery system: Stimuli-responsive biomineralized particles act as particulate emulsifiers and bioactive carriers.

Pickering emulsions provide a promising platform for the efficient delivery of bioactive. However, co-delivery of fragile bioactives with different physicochemical properties for comprehensive effects still faces practical challenges due to the limited protection for bioactives and the lack of stimuli-responsive property for on-demand release. Herein, a stimuli-responsive co-delivery system is developed based on biomineralized particles stabilized Pickering emulsions. In this tailor co-delivery system, hydrophilic bioactive (pepsin) with the fragile structure is encapsulated and immobilized by biomineralization, the obtained biomineralized particles (PPS@CaCO3) are further utilized as emulsifiers to form O/W Pickering emulsions, in which the hydrophobic oxidizable bioactive (curcumin) is stably trapped into the dispersed phase. The results show that two bioactives are successfully co-encapsulated in Pickering emulsions, and benefiting from the protection capacities of biomineralization and Pickering emulsions, the activity of pepsin and curcumin shows a 7.33-fold and 144.83-fold enhancement compared to the free state, respectively. Moreover, In vitro study demonstrates that Pickering emulsions enable to co-release of two bioactives with high activity retention by the acid-induced hydrolyzation of biomineralized particles. This work provides a powerful stimuli-responsive platform for the co-delivery of multiple bioactive compounds, enabling high activity of bioactives for the comprehensive health effects.

Precision diagnosis and antidiastole on supernumerary cusp of tooth by CBCT.

PURPOSE: Based on a case of supernumerary cusp on the bucca of left maxillary second molar diagnosed by cone beam computed tomography (CBCT), its genesis, diagnosis and antidiastole are to be analysed. The clinic implication of CBCT is correspondingly discussed. METHODS: The supernumerary cusp was diagnosed by oral general examination, intra-oral radiograph and CBCT. The features of supernumerary cusp, fused tooth, geminated tooth and concrescence tooth, especially differentiate points among them were discussed. RESULTS: The case of supernumerary cusp on the bucca of left maxillary second molar was diagnosed definitely by the combined application of oral general examination, periapical radiograph and CBCT. CONCLUSION: Supernumerary cusp on the bucca of left maxillary second molar is a rare phenomenon, which is difficult to be differentiated from other tooth deformities. CBCT can improve accuracy of diagnosis.

[Study on preparation of ampelopsin liposomes].

OBJECTIVE: To study the formulation and preparation of ampelopsin liposomes and evaluate their quality. METHOD: The liposomes were prepared by a film-ultrasonic dispersion technique. Served as quota with the entrapment ratio and appearance and diameter of the liposomes, the optimal formulation and preparation were selected by means of an uniform design test. The appearance of liposomes was observed by micrography. The diameter and electric charge of surface were determined by granularity mensuration instrument. The entrapment ratio and the leakage rate of ampelopsin liposome were determined by means of dialyze. The content of ampelopsin was determined by UV. RESULT: The result of electron micrography and the size distribution showed that the liposomes were similar to spherical small unilamellar vesicles. The mean diameter was (258.2 +/- 51.2) nm and the electric charge of surface is 19.0 mV. The entrapment ratio of ampelopsin liposomes was 62. 3% and the lecithoid oxidative rate was 0.83% (n = 3). CONCLUSION: The selected formulation and preparation of ampelopsin liposomes is efficient and practicable.

Ampelopsin prevents apoptosis induced by H2O2 in MT-4 lymphocytes.

Human immunodeficiency virus (HIV) infection can result in oxidative stress through production of reactive oxygen species (ROS). Simultaneously, oxidative stress is able to activate the replication of virus and lead to the apoptosis of T lymphocytes which is the defense of the immune function. Ampelopsin, belonging to the flavonoids, is a purified component from the root of a Chinese medicinal herb. Our previous studies revealed that ampelopsin could protect sensitive cells against HIV-1 infection and reduce HIV-1 antigen P24 expression. In this study, we determined whether ampelopsin, as an antioxidant, has protective effects on oxidant stress-induced apoptosis in MT-4 cells, a CD4 T lymphocyte cell line. The results indicate that ampelopsin scavenged hydroxyl radicals (.OH) and superoxide radicals (O(2).-) in a concentration-dependent manner. It significantly increased MT-4 cells viability after treatment with H(2)O(2) and inhibited H(2)O(2)-induced DNA laddering. The data from flow cytometry analysis showed that ampelopsin remarkably decreased the percentage of apoptotic cells induced by H(2)O(2). In addition, activation of caspase-3 was detected during the course of apoptosis induction. Western blot analysis showed that ampelopsin inhibited the cleavage of caspase-3 induced by H(2)O(2). All these findings might shed new light on the understanding of the anti-AIDS functions of ampelopsin by protecting T cells of persons infected with HIV.

[Optimizing the extracting technique of ampelopsin from Ampelopsis cantoniensis Planch by a uniform design method].

OBJECTIVE: To optimize the preparation of ampelopsin from Ampelopsis Cantoniensis Planch. METHODS: The extraction and purification process was studied by the uniform design with the extract of ampelopsin content and purity as markers. The facters which influence the extraction and the purification of ampelopsin content were studied by uniform design. RESULTS: The optimum extraction and purification process: the concentration for alcohol was 90%, and refluxing quartic, 1.5 h each time; extraction by petroleum ether quintic, the mount of active carbon was 1 g/100 g of the medicine material, and recrystaling thrice. CONCLUSION: This extraction process has higher yield of ampelopsin and is available for production.

[The chemotaxis effect of ampelopsin on the immunocytes].

OBJECTIVE: To study the chemotaxis effect of ampelopsin with different concentration on monocytes and neutrophilic granulocytes. METHODS: Chemokinesis and chemotaxis tests were proceed in agarose gel comparing with chemokine IL-8 or MCP-1. RESULTS: At 25.6 microg/ml or 51.2 microg/ml, ampelopsin could strongly enhance the migration of neutrophilic granulocytes and monocytes. The chemotaxis effect induced by 25.6 microg/ml of ampelopsin had no significant differences with that induced by 150 ng/ml of IL-8 or 50 ng/ml of MCP-1 (P > 0.05). At a concentration of 12.8 microg/ml, the chemokime effect of ampelopsin was more potent than that of 150 ng/ml of IL-8 or 100 ng/ml of MCP-1 (P < 0.05). Ampelopsin exerted a synergistic action with IL-8 or MCP-1 on its chemotaxis effect to neutrophilic granulocytes and monocytes. CONCLUSION: Ampelopsin can strongly enhance the chemokinesis and chemotaxis effects of neutrophilic granulocytes and moncytes and exert a synergistic action with IL-8 or MCP-1 on its chemotaxis effect to neutrophilic granulocytes and monocytes.

[Inhibitory effects of ampelopsin on angiogenesis].

OBJECTIVE: To study the effect of ampelopsin on angiogenesis. METHODS: The anti-angiogenic effect was evaluated by MTT assay for proliferation of endothelial cells. The concentration of vascular endothelial growth factor (VEGF) and basic-fibroblast growth factor (bFGF) from human hepatocellular carcinoma Bel-7402 cells were detected by enzyme linked immunosorbant assay (ELISA). Immunohistochemical staining was conducted to detect the expression of VEGF and bFGF. The VEGF and bFGF in the cancer cells were examined by flow cytometry. The inhibitory effect of ampelopsin on the growth of human hepatocellular carcinoma Bel-7402 in nude mice was studied. RESULTS: Ampelopsin was shown to inhibit the proliferation of primary cultured bovine aortic endothelial cells in a concentration dependent manner in range of 6.4 - 51.2 microg/ml. The IC50 (50% inhibition concentration) value was 22.0 +/- 4.0 microg/ml. ELISA assay was shown that treatment with 12.8 microl/m1, 25.6 microl/ml and 38.4 microg/ml of ampelopsin resulted in an inhibition of VEGF production released by Bel-7402, and the inbibtitory rate was 14.2%, 40.0% and 49.6%, respectively. After exposure to 12.8 microg/ml of ampelopsin, a decrease in the expression and activity of VEGF and bFGF was observed by immunohistochemical staining. The concentration of VEGF and bFGF secretion by Bel-7402 cells were lower following ampelopsin treatment as shown by flow cytometry. Treatment with 25.6 microg/mL and 38.4 microg/ml of ampelopsin, the inbibitory rates were 32.2% and 57.4% for VEGF, and 54.9% and 62.6% for bFGF, respectively. The inhibitory rate of ampelopsin to the growth of the transplant tumor in nude mice were 24.3%, 41.4% and 45.75 respectively at the dose of 100 mg/kg, 150 mg/kg and 200 mg/kg. CONCLUSION: Ampelopsin is a potent inhibitor of VEGF and bFGF expression and production in human hepatocellular carcinoma Bel-7402 cell, and may be a promising angiogenesis inhibitor.

Ampelopsin, a small molecule inhibitor of HIV-1 infection targeting HIV entry.

OBJECTIVE: To investigate the anti-HIV effects of ampelopsin and its interaction with HIV-1 coreceptor CXCR4. METHODS: Through anti-virus experiments in vitro, the inhibitory effect of ampelopsin on HIV-1 infection was verified. Chemotaxis assay was performed to show the ability to induce PBMCs migration by ampelopsin, RANTES and SDF-1alpha. Fluorescence labelling monoclonal antibody was utilized to observe the interaction of ampelopsin and CXCR4. Mice immunosuppressant model was also established to detail the role ampelopsin played in regulating cellular immunological functions. RESULTS: Ampelopsin could protect sensitive cells against HIV-1 infection and dramatically reduce HIV-1 antigen P24 expression. HIV-1SF33 attaching to MT-4 cells was interfered by ampelopsin, and the EC50 was 0.175 mg/mL for cellular protection and 0.024 mg/mL for P24 inhibition. At co-cultivating phase, EC50 was 0.229 mg/mL and 0.197 mg/mL respectively. Furthermore, the EC50 was 0.179 mg/mL and 0.348 mg/mL in acute infection. Human PBMCs migration was induced after being challenged with ampelopsin or chemokines, and synergistic action was observed during co-treatment. Ampelopsin alone resulted in maximal chemotaxis at 1 mg/mL. HIV-1 co-receptor CXCR4 on the surface of PBMCs was decreased by internalization, which indicated the effect of ampelopsin on CXCR4. About 70% CXCR4 was reduced by ampelopsin at 1 mg/mL. Ampelopsin also augmented cellular immunological functions in immunosuppressive mice. CONCLUSION: Ampelopsin displays a strong inhibitive role during HIV-1 absorption, incubation and acute infection. These results are coincident with its immune enhancement.

[Anti-tumor effects of ampelopsin on human lung cancer GLC-82 implanted in nude mice].

OBJECTIVE: To investigate the effects of ampelopsin on the growth of human lung cancer cell line GLC-82 in nude mice. METHODS: GLC-82 cells were inoculated into the armpit of nude mice to establish lung cancer model. Then the BALB/C nude mice 6 with xenograft tumor were randomized into 6 groups. Ampelopsin was administered at 3 dosages by intraperitoneal injection daily. The anti-tumor effect of ampelopsin was evaluated on tumor volume, relative tumor volume, tumor weight, relative tumor proliferative rale, and tumor growth curve. RESULTS: At the 250 mg/kg dose of ampelopsin, the growth inhibition of the transplant tumor was 35.5% (P < 0.01) and 37.1% (P < 0.01), and the relative tumor proliferative rate was 55.24% (P < 0.01) and 57.71% (P < 0.05), respectively in two independent experiments by compared with the normal saline control. CONCLUSION: Ampelopsin appears to be a potent anti-tumor agent that was firstly discovered in the transplant human lung cancer cell line GLC-82 in nude mice in vivo.