[A new secoiridoid from Cornus officinalis].

The chemical constituents from Cornus officinalis were isolated and purified by various techniques such as macroporous adsorption resin, silica gel, octadecylsilyl(ODS), Sephadex LH-20 column chromatography and preparative high-performance liquid chromatography(HPLC). The structures of the isolates were determined by a combination of spectroscopic techniques such as high-resolution electrospray ionization mass spectrometry(HR-ESI-MS), one-dimensional(1D) and two-dimensional(2D) nuclear magnetic resonance(NMR) spectroscopy. Ten compounds were isolated from the aqueous extract of C. officinalis and identified as(±)-cornuscone(1),(-)-(Z)-4-hydroxy-3-methoxyphenylpropene 4-O-ß-L-xylopyranosyl-(1→6)-ß-D-glucopyranoside(2), kaempferol 3-O-ß-D-glucopyranoside(3), kampferol(4), myricetin(5), trifolin(6), quercetin 3-O-ß-D-glucopyranoside(7), quercetin 3-O-ß-D-glucuronide-6″-methyl ester(8), quercetin 3-O-ß-D-glucuronide-6″-ethyl ester(9) and pyrogallol(10). Compound 1 is a new secoiridoid, named(±)-cornuscone with a rare methyl substitution at the C-1 position. The anti-inflammatory activity of 1 was evaluated in lipopolysaccharide(LPS)-induced RAW264.7 cells in mice. The results showed the median inhibition concentration(IC_(50)) of 1 was(31.15±1.29)µmol·L~(-1), which demonstrated that the anti-inflammatory activity of 1 was significantly superior to that of indomethacin [IC_(50) value of(48.32±1.66)µmol·L~(-1)].

Existing knowledge on Euphorbia fischeriana Steud. (Euphorbiaceae): Traditional uses, clinical applications, phytochemistry, pharmacology and toxicology.

ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia fischeriana Steud. (Euphorbiaceae) is a perennial herb distributed in grassland, hill slopes or gravel hillside, with average altitude of 100-600 m. The whole grass of E. fischeriana is toxic with roots used as folk medicine to treat Zhushui, dyspepsia, abdominal distension, abdominal pain, cough, as well as external applications such as cure of scabies and tuberculosis of lymph nodes. AIM OF THE REVIEW: This systematic review aims to provide a detailed and in-depth summary about the reported advances in traditional uses, clinical applications, phytochemistry, pharmacology and toxicity of E. fischeriana, so as to offer fresh ideas and broader vision and insights for subsequent studies. MATERIALS AND METHODS: Various scientific data bases such as CNKI, Elsevier, Google Scholar, Pubmed, Science Direct, SciFinder Scholar and Web of Science were searched to collect information about E. fischeriana. Other relevant literatures were searched in 'Flora of China Editorial Committee', ancient books, Ph.D and Masters' Dissertation to get more data of E. fischeriana. RESULTS: A total of 241 chemical constituents have been identified from the roots of E. fischeriana, including diterpenoids, triterpenoids, meroterpenoids, acetophenones, flavonoids, coumarins, steroids, phenolic acids, tannins, etc. Various pharmacological activities have been demonstrated, especially anti-tumor, antibacterial, anti-inflammatory, antiviral and anti-leukemia activities. Moreover, different investigations about clinical uses and toxicology of E. fischeriana indicated that attention should be paid to its usage and dosage. CONCLUSION: The researches of E. fischeriana are excellent, but gap still remains. As a poisonous traditional Chinese medicine, there are not enough studies on the toxicity of E. fischeriana. In addition, scholars' research on the pharmacological mechanism of E. fischeriana focuses more on the anti-tumor activity, which can be broadened in the future. Presumably, chemical constituents and biological activities of diterpenoids and trace meroterpenoids in E. fischeriana deserve further research in-depth in the future, in order to provide low toxicity and high efficiency lead compounds. Meanwhile, further studies on other medicinal aspects may lay a foundation for the comprehensive development and utilization of E. fischeriana.

Cloning and analysis of the photoreceptor genes Oxwc-1 and Oxwc-2 from ethnopharmacological fungus Ophiocordyceps xuefengensis.

Ophiocordyceps xuefengensis is an ethnopharmacological fungus with broad pharmacological properties. Light is a critical environmental factor for the stromata formation and development of many fungi. In this study, photomorphogenesis and blue light receptor genes were studied using a strain of O. xuefengensis. Light represses vegetative growth, but conidia linked to stromata can be observed under both light and dark conditions. Light and dark conditions had little effect on the accumulation of polysaccharides and adenosine. The genes Oxwc-1 and Oxwc-2 encoding photoreceptors of O. xuefengensis were cloned and predicted to possess polypeptides of 937 and 525 amino acids, respectively. A phylogenetic analysis based on fungal WC-1/2 supported OxWC-1 and OxWC-2 were photoreceptor. The expression of both the Oxwc-1 and Oxwc-2 genes reached a maximum after receiving light stimulation for 15 min, which might relate to the inhibition of stromata growth.

Indole-based alkaloids from Ophiocordyceps xuefengensis.

Seven undescribed indole-based alkaloids, xuefengins A-D and xuefenglasins A-C, were isolated from natural Ophiocordyceps xuefengensis, along with six known alkaloids. Their structures were elucidated by comprehensive spectroscopy, with absolute configurations confirmed by comparison with calculated electronic circular dichroism spectra. Eleven of the isolates were tested for cytotoxicity against the U937, NB4, MCF-7, Hep G2, and A549 cancer cell lines. Two compounds exhibited moderate activities, with IC50 values of 2.83-25.68 µM and 1.54-12.16 µM. Further pharmacological studies showed that these two compounds inhibit cell proliferation by inducing apoptosis, and decreasing p38 and caspase-3 levels in A549 cells.

Behaviors of Glioblastoma Cells in in Vitro Microenvironments.

Glioblastoma (GBM) is the most malignant and highly aggressive brain tumor. In this study, four types of typical GBM cell lines (LN229, SNB19, U87, U251) were cultured in a microfabricated 3-D model to study their in vitro behaviors. The 3-D in vitro model provides hollow micro-chamber arrays containing a natural collagen interface and thus allows the GBM cells to grow in the 3-D chambers. The GBM cells in this model showed specific properties on the aspects of cell morphology, proliferation, migration, and invasion, some of which were rarely observed before. Furthermore, how the cells invaded into the surrounding ECM and the corresponding specific invasion patterns were observed in details, implying that the four types of cells have different features during their development in cancer. This complex in vitro model, if applied to patient derived cells, possesses the potential of becoming a clinically relevant predictive model.

The pharmacological properties of Ophiocordyceps xuefengensis revealed by transcriptome analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: The Yao ethnic group in Xuefeng Mountains area have used Xuefeng cordyceps, the caterpillar-fungus complex of Ophiocordyceps xuefengensis, for treating a variety of diseases for long. Just like some other cordyceps, O. xuefengensis, which is identified as the sister taxon of O. sinensis in 2013, also seems to have broad pharmacological properties, not only enhancing human immunity, anti-bacteria, anti-virus, but also anti-tumor. However, investigation of the medicinal fugal species O. xuefengensis can be found only in few literature records since its pharmacological and therapeutic use is mainly in traditional Yao communities by local healers. AIM OF THE STUDY: The aim of this study is to collect samples of Xuefeng cordyceps and isolate the strain of O. xuefengensis, to determine bioactive components and evaluate the anti-tumor activity, to obtain the gene expression profile of O. xuefengensis and reveal its pharmacological properties by de novo transcriptome analysis. Accordingly, we attempt to provide information and give a comprehensive understanding of this mysterious medicinal fugal species from traditional Yao communities of China. MATERIAL AND METHODS: Bioactive components were determined with HPLC-DAD-Q-TOF-MS technology; in vitro anti-tumor activity against 6 cell lines was evaluated using standard MTT assay; transcriptome analysis was done by de novo sequencing; unique genes were functionally profiled basing on Gene Ontology Database and the targeted genes were examined by blast. RESULTS: Trace cordycepin, an anti-tumor agent, was detected in O. xuefengensis water extract. To some extent, the raw water extract of O. xuefengensis showed in vitro anti-tumor activity, against A549, HepG2, MCF-7, PC-3 and Raji cell lines. A total of 94,858 transcripts and 49,001 unique genes were obtained, amongst, 43.4% unique genes were matched with those of O. sinensis. Not all supposed genes related to cordycepin biosynthetic pathways were found by transcriptome analysis. CONCLUSION: According to the gene expression profile, O. xuefengensis is very close to medicinal fungus O. sinensis. Raw water extract of O. xuefengensis, to a certain degree, could inhibit the growth of tumor cells, indicating that this fungus could be a new resource for the exploration of anti-tumor drug.

A new cordycepin-producing caterpillar fungus Ophiocordyceps xuefengensis with artificial infection to the host, cultivation of mycelia and stromata.

Caterpillar fungi have numerous pharmacological and therapeutic applications in traditional medicine, due to a variety of active chemical constituents, such as cordycepin and adenosine. It is imperative to discover new resource for artificial cultivation and biometabolite production since the traditional natural species are endangered. In this study, a new strain HACM 001 was isolated and identified as Ophiocordyceps xuefengensis by rDNA-ITS sequencing. This strain showed the potential of artificial infection to caterpillar larvae leading to mummification, as well as fermentation mycelia in liquid culture and cultivation stromata in solid medium. Eight nucleosides and nucleobases, especially cordycepin and adenosine, were determined and analyzed with HPLC-DAD-Q-TOF-MS/MS technology. Cordycepin was detected in all forms of present O. xuefengensis strain at different contents, among which the highest content (37.1 µg/g) appeared in the stromata cultivated on solid medium. The content of adenosine in mycelia and stromata, respectively, reached 1155 µg/g and 1470 µg/g. Therefore, O. xuefengensis might be an alternative source for obtaining artificial fungus-caterpillar-larvae complex and producing cordycepin and adenosine.

Ribosomal S6 Kinase 2 (RSK2) maintains genomic stability by activating the Atm/p53-dependent DNA damage pathway.

Ribosomal S6 Kinase 2 (RSK2) is a member of the p90(RSK) family of serine/threonine kinases, which are widely expressed and respond to many growth factors, peptide hormones, and neurotransmitters. Loss-of function mutations in the RPS6KA3 gene, which encodes the RSK2 protein, have been implicated in Coffin-Lowry Syndrome (CLS), an X-linked mental retardation disorder associated with cognitive deficits and behavioral impairments. However, the cellular and molecular mechanisms underlying this neurological disorder are not known. Recent evidence suggests that defective DNA damage signaling might be associated with neurological disorders, but the role of RSK2 in the DNA damage pathway remains to be elucidated. Here, we show that Adriamycin-induced DNA damage leads to the phosphorylation of RSK2 at Ser227 and Thr577 in the chromatin fraction, promotes RSK2 nuclear translocation, and enhances RSK2 and Atm interactions in the nuclear fraction. Furthermore, using RSK2 knockout mouse fibroblasts and RSK2-deficient cells from CLS patients, we demonstrate that ablation of RSK2 impairs the phosphorylation of Atm at Ser1981 and the phosphorylation of p53 at Ser18 (mouse) or Ser15 (human) in response to genotoxic stress. We also show that RSK2 affects p53-mediated downstream cellular events in response to DNA damage, that RSK2 knockout relieves cell cycle arrest at the G2/M phase, and that an increased number of γH2AX foci, which are associated with defects in DNA repair, are present in RSK2-deficient cells. Taken together, our findings demonstrated that RSK2 plays an important role in the DNA damage pathway that maintains genomic stability by mediating cell cycle progression and DNA repair.

Significantly upregulated TACSTD2 and Cyclin D1 correlate with poor prognosis of invasive ductal breast cancer.

The tumor-associated calcium signal transducer 2 (TACSTD2) gene has been reported to be highly expressed in many types of human epithelial cancers, and is associated with tumor metastasis and poor prognosis. The aims of the present investigation were to analyze the TACSTD2 and Cyclin D1 expression at the mRNA and protein levels and to assess its prognostic significance in invasive ductal breast cancer (IDC). The expressions of TACSTD2 and Cyclin D1 in IDC tissues were consistently higher than those in the tumor-adjacent non-malignant tissues by a one-step real-time polymerase chain reaction and immunohistochemistry (P<0.001 and P=0.023, respectively). The statistical analysis of clinicopathologic characteristics and immunohistochemistry by the χ(2) test showed that the high expression of TACSTD2 in IDC was correlated to histological grade (P=0.023), P53 status (P=0.042), Cyclin D1 status (P<0.001), lymph node metastasis (P<0.001), distant metastasis (P=0.004) and TNM staging (P<0.001). Kaplan-Meier survival and Cox regression analyses were performed to evaluate the prognosis of IDC. These analyses also showed that a high TACSTD2 expression (P=0.003), a high Cyclin D1 expression (P=0.041), and lymph node metastasis (P=0.006) were independent prognosis factors. Collectively, our studies demonstrated that the high expression of TACSTD2 correlates with a poor prognosis in IDC.