[Development and research advances of iridoids from Valeriana jatamansi and their bioactivity].

Valeriana jatamansi (syn. V. wallichii), a traditional Chinese medicine recorded in Chinese Pharmacopeia (1977 and 2010 edition), has been used for treatment of a variety of conditions including sleep problems, obesity, nervous disorders, epilepsy, insanity, snake poisoning, eye trouble, and skin diseases. Also, it was used as an important substitute for the European V. officinalis, whose root preparation, popularly known as valerian, has been employed as a mild sedative for a long time. In recent years, much attention has been draw to the iridoids, one of the major bioactive constituents of V. jatamansi, leading to the discovery of a series of new iridoids with anti-tumor and neuroprotective activities. Their action machnism also has been discussed. This paper summerized the iridoids and their bioactivities from V. jatamansi in recent years, which could provide basic foundation for development and research of V. jatamansi.

[Metabolites of Aspergillus fumigatus].

Aspergillus fumigatus, a type of endophytic fungi from Erthrophleum fordii, was fermented with GPY culture medium. Fermented liquid and mycelium were extracted from fermented products after freezing and thawing treatment. After alcohol extraction, mycelium was extracted with ethyl acetate and n-butyl alcohol, respectively. According to the results of cytotoxity of tumor cells, ethyl acetate extracts were studied for their chemical constituents. Five diketopiperazine compounds were separated and purified with silica gel, MCI and Sephadex LH-20 column chromatography, reversed-phase chromatographic column and preparative HPLC, their structures were identified as cyclo- (R-Pro-R-Phe) (1), cyclo- (trans-4-OH-D-Pro-D-Phe) (2), cyclo- (R-Tyr-S-Ile) (3), cyclo-(R-Phe-S-Ile) (4), and cyclo-(R-Val-S-Tyr) (5) by using spectral methods.

Cytotoxic cassaine diterpenoid-diterpenoid amide dimers and diterpenoid amides from the leaves of Erythrophleum fordii.

Detailed phytochemical investigation from the leaves of Erythrophleum fordii resulted in the isolation of 13 compounds, including three cassaine diterpenoid-diterpenoid amide dimers (1, 3 and 5), and seven cassaine diterpenoid amides (6 and 8-13), together with three previously reported ones, erythrophlesins D (2), C (4) and 3beta-hydroxynorerythrosuamide (7). Compounds 1, 3 and 5 are further additions to the small group of cassaine diterpenoid dimers represented by erythrophlesins A-D. Their structures were determined by analysis of extensive one- and two-dimensional NMR experiments and ESIMS methods. Cytotoxic activities of the isolated compounds were tested against HCT-8, Bel-7402, BGC-823, A549 and A2780 human cancer cell lines in the MTT test. Results showed that compounds 1 and 3-5 exhibited significantly selective cytotoxic activities (IC(50)<10 microM) against these cells, respectively.

Thiodiketopiperazines produced by the endophytic fungus Epicoccum nigrum.

Thirteen new thiodiketopiperazines, epicoccin I (1), ent-epicoccin G (2), and epicoccins J-T (3-13), together with six known diketopiperazines (14-19), have been isolated from the endophytic fungus Epicoccum nigrum. The structures of 1, 2, and 10 were confirmed by X-ray crystallography, and the absolute configurations of 2, 4, 6, and 8 were assigned using Moshers' method. Compounds 2, 6, 12, and 17 showed potent activities in vitro against the release of beta-glucuronidase in rat polymorphonuclear leukocytes induced by platelet-activating factor, with IC(50) values of 3.07, 4.16, 4.95, and 1.98 microM, respectively. None of the 19 compounds exhibited detectable cytotoxic activities toward six tumor cell lines (A549, Be-l7402, BGC-823, HCT-8, HCT-116, and A2780) in the MTT assay.

Molecular Cloning of Human Interleukin-5 cDNA and Its Expression in E. coli.

hIL-5 cDNA was amplified through reverse transcription-polymerase chain reaction from peripheral blood lymphocytes induced with PMA and calcium ionophore A23187. The hIL-5 fragment was cloned into pUC18 and its sequence was identified to be hIL-5 cDNA sequence. The fragment which encodes hIL-5 mature polypeptide was amplified and cloned into pBV220 to express hIL-5 recombinant protein in E. coli, but there was no expected recombinant protein expressed. Only one clone expressed a 10kD peptide at high level. DNA sequencing showed that this clone had an adenosine deletion at 86th codon in hIL-5 cDNA and expressed a polypeptide of 93 amino acids at high level, and hIL-5 cDNA had not yet been expressed in E.coli successfully. These results suggested that two consecutive rare codons after 86th codon in hIL-5 cDNA could block polypeptide synthesis in E. coli. Through recombinant PCR technology the rare codons after 86th codon in hIL-5 cDNA were mutated into corresponding codons preferentially used in E. coli, and the mutated hIL-5 cDNA was highly expressed in pBVhIL5-H/DH5alpha to approximately 15% TCP after thermal induction. hIL-5 recombinant protein existed as inclusion bodies in E. coli. ELISA with a cross-reacting rat anti-mIL-5 monoclonal antibody confirmed the expressed product was hIL-5 recombinant protein.