Five new diketopiperazines, prenylcyclotryprostatin B (1), 20-hydroxycyclotryprostatin B (2), 9-hydroxyfumitremorgin C (3), 6-hydroxytryprostatin B (4), and spirogliotoxin (5), were isolated from the marine-derived fungus Aspergillus fumigatus YK-7, along with nine known compounds, 6-14. Their structures were elucidated by spectroscopic methods, and their antiproliferative effects on human leukemic monocyte lymphoma U937 and human prostate cancer PC-3 cell lines were assessed in vitro. Compounds 10, 12, and 13 exhibited significant cell growth-inhibitory activities against U937 cell line, with the IC(50) values of 1.8, 0.2, and 0.5 μM, respectively.
Fusion expression is a promising strategy for the production of bioactive peptides in Escherichia coli. In this study, we constructed a new recombinant expression plasmid containing the coding sequence of 56-residue B1 domain of streptococcal protein G (GB1). For easy purification and cleavage of the recombinant proteins, except GB1, an engineered hexahistidine and tobacco etch virus (TEV) protease recognition sites were included in the fusion sequence. Next, we cloned the coding sequence of human epidermal growth factor (hEGF) into this new plasmid and produced the recombinant hEGF in E. coli. The bioactive hEGF is a 53-amino acid peptide and is stabilized by three intramolecular disulphide bonds. Compared with glutathione Stransferase, thioredoxin and small ubiquitin-related modifier, GB1 greatly improved the expression and solubility of hEGF. Moreover, the recombinant hEGF bound to the nickel nitrilotriacetic acid resin column, was easily cleaved by TEV protease and the free hEGF was released. The results showed that this new plasmid was appropriate for recombinant production of small bioactive peptides, such as hEGF, which contains a high proportion of hydrophobic residues and intramolecular disulphide linkages.
A new cytotoxic dimeric naphthopyrone, aurasperone H (1), together with eight related known polyketides (2-9) was isolated from a marine-derived fungus Aspergillus niger 2HL-M-8. The structure of new compound 1 was elucidated on the basis of its spectroscopic data (1D, 2D NMR and CD). Compound 1 exhibited moderate inhibitory activity against the human lung adenocarcinoma A549 and the human leukaemia HL-60 cell lines. Compound 5 displayed significant in vitro antiproliferative activity against HL-60 cell line with an IC50 value of 0.8 μM.
In vivo engineering of hepatic tissue based on primary hepatocytes offers new perspectives for the treatment of liver diseases. However, generation of thick, three-dimensional liver tissue has been limited by the lack of vasculature in the tissue-engineered constructs. Here, we used collagen hydrogel as a matrix to generate engineered hepatic units to reconstitute three-dimensional, vascularized hepatic tissue in vivo. Hepatocytes harvested from Sprague-Dawley rats were mixed with liquid type I collagen, concentrated Dulbecco's modified Eagle's medium (2 x), and hepatocyte maintenance medium to create hepatocyte/collagen hydrogel constructs. The constructs were then dissociated into cylindrical hepatic units (diameter/height: 2000-4000 microm/500-1000 microm). Stacking of hepatic units under the subcutaneous space resulted in significant cell engraftment, with the formation of large fused hepatic system (more than 0.5 cm thickness) containing blood vessels. In contrast, only less cell engraftment could be achieved when hepatocytes were transplanted in a manner of whole constructs. Functional maintenance of the engineered hepatic tissue was confirmed by the expression of liver-specific mRNA and proteins. The engineered hepatic tissue has the ability to respond to the regenerative stimulus. In conclusion, large hepatic tissue containing blood vessels could be engineered in vivo by merging small hepatic units. This approach for tissue engineering is simple and represents an efficient way to engineer hepatic tissue in vivo.
Background and Aim. Adipose derived mesenchymal stem cells (ADMSCs) may be an attractive source for acute and chronic liver injury because they are abundant and easy to obtain. We aim to investigate the efficacy of ADMSCs transplantation in the acute liver failure (ALF) caused by carbon tetrachloride (CCl4) in mice. Methods. ADMSCs were isolated from inguinal fat pads of enhanced green fluorescent protein (EGFP) transgenic mice and their surface markers and differentiation potential were analyzed. ALF models were established by infusion of CCl4 and divided into two groups: control group; EGFP-ADMSCs transplantation group. The restoration of biological functions of the livers receiving transplantation was assessed via a variety of approaches such as survival rates, live function parameters, histological localization of EGFP-ADMSCs, and Immunofluorescence analysis. Results. ADMSCs were positive for CD105, CD44 but negative for CD45, CD34 and had adipogenic, osteogenic differentiation potential. The survival rate of transplantation group significantly increased compared to PBS group. Furthermore, the transplanted cells were well integrated into injured livers and produced albumin, cytokeratin-18. Conclusion. Direct transplantation of ADMSCs is an effective treatment for ALF. The transplanted ADMSCs exhibit the potential to differentiate into hepatocyte-like cells in the injured livers.
Two new anthraquinones, emodin-3-O-sulphate (1) and citreorosein-3-O-sulphate (2), as well as five known anthraquinones, were isolated from a marine-derived fungus Penicillium oxalicum. The structures of these compounds were determined by spectroscopic methods (1D and 2D NMR, HR-ESI-MS).
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