Objective: To isolate new endophytic fungus from Phaleria macrocarpa (P. macrocarpa) that is able to produce E2.2 compound.
Methods: Endophytic fungi were isolated from P. macrocarpa. Morphological and molecular identification was done to determine the species of the endophytic fungus. High performance liquid chromatography was used to determine the ability of this fungus to produce E2.2 compound and to quantify the total yield of E2.2 from fungal fermen-tation. Fermentation process was optimized by observing suitable medium, pH and length of fermentation process. Phloroglucinol and gallic acid addition were examined to determine the effect of each compound on E2.2 production.
Results: One endophytic fungus was successfully isolated from P. macrocarpa plant. Morphological and molecular identification showed that it was a Colletotrichum gloeo-sporioides which belonged to Glomerellaceae family. This fungus showed highest pro-duction of E2.2 when incubated in potato dextrose broth with initial pH value of the medium at 5, and was incubated for 15 days. Phloroglucinol was found to better enhance E2.2 production.
Conclusions: Colletotrichum gloeosporioides found in P. macrocarpa plant is prom-ising as a potential alternative source of E2.2.

Objective:To identify and isolate the chemical compounds of Phaleria macrocarpa (P.macrocarpa) fruit ethanolic extract.Methods:Dried fruit of P.macrocarpa was extracted with 90％ ethanol and partitioned between n-hexane/H2O and ethyl acetate/H2O.The organic layer was fractionated by various stationary phase and identified by using combined data of ID [(proton nuclear magnetic resonance (NMR),carbon-13 NMR)],2D-NMR (heteronuclear multiplequantum correlation and heteronuclear multiple-bond correlation),and mass spectrum.Results:Purification of n-hexane and ethyl acetate fractions from ethanolic extract of P.macrocarpa fruit resulted in isolation of nine compounds.Conclusions:A new compound was isolated and identified as glyceryl pentacosanoate.Also,two xanthones,which are 1,7-dihydroxy-3,6-dimethoxyxanthone and 1,6,7-trihydroxy-3-methoxyxanthone,are firstly reported to be isolated from P.macrocarpa.

Objective To isolate new endophytic fungus from Phaleria macrocarpa (P. macrocarpa) that is able to produce E2.2 compound. Methods Endophytic fungi were isolated from P. macrocarpa. Morphological and molecular identification was done to determine the species of the endophytic fungus. High performance liquid chromatography was used to determine the ability of this fungus to produce E2.2 compound and to quantify the total yield of E2.2 from fungal fermentation. Fermentation process was optimized by observing suitable medium, pH and length of fermentation process. Phloroglucinol and gallic acid addition were examined to determine the effect of each compound on E2.2 production. Results One endophytic fungus was successfully isolated from P. macrocarpa plant. Morphological and molecular identification showed that it was a Colletotrichum gloeosporioides which belonged to Glomerellaceae family. This fungus showed highest production of E2.2 when incubated in potato dextrose broth with initial pH value of the medium at 5, and was incubated for 15 days. Phloroglucinol was found to better enhance E2.2 production. Conclusions Colletotrichum gloeosporioides found in P. macrocarpa plant is promising as a potential alternative source of E2.2.

Objective To study the regulation of trombinol on thrombopoietin, an essential regulator of thrombocyte production. Methods Effect of trombinol on thrombopoietin regulation was evaluated at the mRNA and protein levels in human hepatoma HepG2 cells. The mRNA expressions were revealed by PCR and real-time PCR, while the protein expressions were analyzed using western blotting and human ELISA kit. Statistical differences between the test were determined by student's t-test with P < 0.05 was considered statistically significant. Results Trombinol significantly increased the expression of thrombopoietin at the level of mRNA and protein secretion in HepG2 cell lines. Trombinol with the concentration of 15 μg/mL, positively induces 2.5-fold of thrombopoietin expression. Up-regulation of GABP, a transcription factor of thrombopoietin, is suggested to be involved in cellular regulatory mechanisms of trombinol. Here, our result shows convincing evidence that trombinol affects the thrombopoietin productions in vitro. This molecular explanation of thrombopoietin's stimulating function is in line with the traditional use of Psidium guajava for treatment of diseases involving thrombocytopenia. Conclusions Thrombopoietin stimulating function of trombinol could be potentially considered as one of alternative treatment for thrombocytopenia-related cases, including post chemotherapy shock, dengue fever and liver failure.

Objective To prove the molecular mechanisms of Mahkota Dewa (Phaleria macrocarpa) in suppressing proliferation of human retinoblastoma cells through suppression of cell cycle's gene-regulators expression. Methods In this study, the molecular mechanism of anti-tumor effect of fractioned extract of Phaleria macrocarpa (DLBS1425) in human retinoblastoma cells Y-79 was investigated by measuring the tumor cells viability, the assessment of population profiles of tumor cells in the cell cycle, and the mRNA concentration of p16, p21, p53, cyclin D, cyclin E, and E2F. Results DLBS1425 showed an inhibition effects towards proliferation of Y-79 cell line. Inhibition of proliferation was shown by suppression of cell cycle progression. DLBS1425 downregulated cyclin E, a G1 phase regulator gene of cell cycle, in dose-dependent manner without affecting p53–p21 pathway. In the other word, DLBS1425 inhibits cell proliferation through suppression of cyclin E independently towards conventional proliferation pathway. Conclusions Our results suggest that DLBS1425 is a potential anticancer agent which targets genes involved in cell proliferation in human retinoblastoma cells which make it pharmacologically ideal for the prevention and/or treatment of retinoblastoma cancer.

Objective To identify and isolate the chemical compounds of Phaleria macrocarpa (P. macrocarpa) fruit ethanolic extract. Methods Dried fruit of P. macrocarpa was extracted with 90% ethanol and partitioned between n-hexane/H

Objective To verify that Proliverenol has a potential ability in protecting cells from ethanol-induced hepatotoxicity. Methods Activity of Proliverenol against ethanol-induced apoptosis was evaluated at mRNA and protein levels in HepG2 cell exposed to Proliverenol for 1 and 3 h. Results Proliverenol conferred hepatoprotective activity through increasing cell survival up to 53%–69% via up-regulation of APEX1 DNA repair enzyme for 3.0–4.7 fold and down-regulating of nuclear factor-κB, tumor necrosis factorα and caspase-8 expression, allowing them to prevent 4.5–6.9 fold of alanine aminotransferase (ALT) leakage in HepG2 cells. Our finding revealed that Proliverenol repressed expression of ALT, which is significantly important as possible alternative mechanism for increased blood transaminase activities. In addition, the result also showed that caspase-8 pathway seemed to be involved in the molecular pathway rather than directly inducing mitochondrial damage. Conclusions The data support our hypothesis that Proliverenol has a potential ability in protecting cells from ethanol-induced hepatotoxicity. We propose that Proliverenol provides hepatoprotective activity through up-regulating expression of APEX1 that repress DNA fragmentation, and down-regulating expression of nuclear factor-κB, tumor necrosis factorα and caspase-8, which therefore repress ALT leakage and its expression.

Objective To characterize proteins and other nutrients in striatin (DLBS0333), a bioactive protein fraction isolated from snakehead fish (Channa striata) and to investigate its wound healing activity. Methods Proteins and other constituents in striatin were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, two dimension electrophoresis, immunoblotting assay, spectroscopy and high performance liquid chromatography. The wound healing activity of striatin was studied in vitro using 3T3 fibroblast cells and in vivo using wound-induced animal model. Various parameters related to wound healing process were evaluated. Results Striatin contained four major bioactive proteins with approximate molecular weight of 8.3, 10.9, 15.4 and 16.7 kDa. In addition to proteins, striatin also contained amino acids (10 essential and 7 non essential amino acids), fatty acids (palmitic acid, oleic acid, stearic acid, linoleic acid, arachidonic acid), vitamins (vitamin A, vitamin B