miR-433 Inhibits Glioblastoma Progression by Suppressing the PI3K/Akt Signaling Pathway Through Direct Targeting of ERBB4.

Glioblastoma multiforme (GBM) is a highly malignant brain tumor where new biomarkers and drug targets are much needed in the oncology clinic. miR-433 was identified as a tumor-suppressing miRNA in several different types of human cancer. However, the integrative biology of miR-433 in GBM is still largely unknown. By analyzing the expression profiles of miR-433 in 198 patients with glioma at The Cancer Genome Atlas, we found that the miR-433 expression was decreased in glioma whereas the low expression of miR-433 was significantly associated with shorter overall survival. We then conducted in vitro studies and demonstrated that increased expression of miR-433 suppressed the proliferation, migration, and invasion of LN229 and T98G cells, two representative glioma cell lines. Further, using in vivo mouse model, we found that upregulation of miR-433 inhibited the tumor growth of glioma cells. To situate the integrative biology understanding of the action of miR-433 in glioma, we identified ERBB4 as a gene targeted directly by miR-433 in LN229 and T98G cells. Overexpressed ERBB4 rescued the phenotype caused by overexpression of miR-433. Finally, we showed that miR-433 suppressed the PI3K/Akt pathway in glioma cells. In conclusion, our study demonstrated that miR-433 could potentially act as a tumor suppressor for GBM and may serve as a potential therapeutic target for GBM. Further integrative biology and clinical translational research are warranted to evaluate miR-433 in GBM.

Integerrima A-E, phenylethanoid glycosides from the stem of Callicarpa integerrima.

Five new phenylethanoid glycosides integerrima A-E (1-5) were isolated from the stem of Callicarpa integerrima for the first time. Their structures were elucidated by extensive spectroscopic analyses. In addition, cytotoxicity, anti-adipogenic and antioxidant activities were evaluated. All the phenylethanoid glycosides would be nontoxic to the normal human hepatocytes LO-2 and pre-adipocytes 3T3-L1 cell lines, significantly promote the proliferation of normal hepatocytes, thus displaying the potential for hepatoprotective. Integerrima A (1), C (3) and D (4) exhibited selectively moderate cytotoxic activity against the hepatoma cell lines Bel-7402, with the IC50 value at 72.66, 80.43 and 84.88 µmol/L, respectively. Moreover, integerrima D (4) had significant activities on reducing lipid droplet formation, with the inhibition rate of 48.02% on the concentration of 200 µg/mL. Finally, the result of FRAP assays exhibited remarkable antioxidant activity in integerrima E (5), which was close to the positive control ascorbic acid with the concentration of 100 µg/mL.

Transcriptome and Metabolome Analysis Revealing the Improved ε-Poly-l-Lysine Production Induced by a Microbial Call from Botrytis cinerea.

ε-Poly-l-lysine (ε-PL) is a wide-spectrum antimicrobial agent, while its biosynthesis-inducing signals are rarely reported. This study found that Botrytis cinerea extracts could act as a microbial call to induce a physiological modification of Streptomyces albulus for ε-PL efficient biosynthesis and thereby resulted in ε-PL production (34.2 g/liter) 1.34-fold higher than control. The elicitors could be primary isolated by ethanol and butanol extraction, which resulted in more vibrant, aggregate and stronger mycelia. The elicitor-derived physiological changes focused on three aspects: ε-PL synthase, energy metabolism, and lysine biosynthesis. After elicitor addition, upregulated sigma factor hrdD and improved transcription and expression of pls directly contributed to the high ε-PL productivity; upregulated genes in tricarboxylic acid (TCA) cycle and energy metabolism promoted activities of citrate synthase and the electron transport system; in addition, pool enlargements of ATP, ADP, and NADH guaranteed the ATP provision for ε-PL assembly. Lysine biosynthesis was also increased based on enhancements of gene transcription, key enzyme activities, and intracellular metabolite pools related to carbon source utilization, the Embden-Meyerhof pathway (EMP), the diaminopimelic acid pathway (DAP), and the replenishment pathway. Interestingly, the elicitors stimulated the gene transcription for the quorum-sensing system and resulted in upregulation of genes for other antibiotic production. These results indicated that the Botrytis cinerea could produce inducing signals to change the Streptomyces mycelial physiology and accelerate the ε-PL biosynthesis. IMPORTANCE This work identified the role of microbial elicitors on ε-PL production and disclosed the underlying mechanism through analysis of gene transcription, key enzyme activities, and intracellular metabolite pools, including transcriptome and metabolome analysis. It was the first report for the inducing effects of the "microbial call" to Streptomyces albulus and ε-PL biosynthesis, and these elicitors could be potentially obtained from decayed fruits infected by Botrytis cinerea; hence, this may be a way of turning a biohazard into bioproduct wealth. This study provided a reference for application of microbial signals in secondary metabolite production, which is of theoretical and practical significance in industrial antibiotic production.

Effects of diluent gases on sooting transition process in ethylene counterflow diffusion flames.

The impacts of adding diluent gases (nitrogen, carbon dioxide, and helium) either to the fuel side or the oxidizer side on the sooting transition process in ethylene counterflow diffusion flames are investigated experimentally and numerically. A series of ethylene flames ranging from non-sooting to heavy-sooting are studied by gradually increasing the oxygen concentration on the oxidizer side. The optical method is used to analyze flame images, determining the sooting transition process. It is found that whether CO2 is added to the fuel side or the oxidizer side, the sooting transition process is delayed significantly. This process is slightly delayed when He is added to the fuel side, however, it is promoted when He is introduced to the oxidizer side. The numerical results show that in CO2-diluted flames, the mole fraction of the main soot precursors C2H2, C3H3, and C6H6 are reduced, which leads to the delay of soot formation. In addition, the H radical decreases while the OH radical increases, both of them are important for soot formation. In He-diluted flames, the concentration of C2H2, C3H3, and C6H6 decreased, as well as H and OH radicals. Moreover, adding He obviously changes the distribution area of products.

Comparison of the effects and prognosis of concurrent and staged resections for the treatment of resectable colorectal cancer liver metastasis.

OBJECTIVE: To compare the effects and prognosis of concurrent and staged resections for the treatment of resectable colorectal cancer liver metastasis (CRLM). METHODS: A prospective study was conducted on 118 patients with CRLM. The 59 cases in the observation group received concurrent resections, while the 59 cases in the control group received staged resections. The operation time, intraoperative blood loss, length of hospital stay, hospital cost, postoperative complications, 5-year survival rate and 3-year progression-free survival rate were recorded for all patients. Factors that affect the prognosis of CRLM patients were analyzed. RESULTS: The length of hospital stay, operation time, intraoperative blood loss, hospital cost were significantly lower in the observation group than in the control group (P<0.001). The two groups were equivalent with respect to postoperative complications, 5-year survival rate and 3-year progression-free survival rate (P>0.05). Independent risk factors affecting the prognosis of CRLM included the number of liver metastasis, whether resection is feasible after recurrence, and RAS genotype (P<0.05). CONCLUSION: Compared to staged resection for CRLM, concurrent resection has shorter operation time, less blood loss, and shorter length of hospital stay, while postoperative complications, long-term efficacy and survival benefits are comparable. Furthermore, the study has found that the number of liver metastasis, whether or not resection is feasible after recurrence, and RAS genotype are risk factors affecting the prognosis of CRLM.

YAP confers resistance to vandetanib in medullary thyroid cancer.

Medullary thyroid cancer (MTC) is the third most common thyroid cancer. RET (Rearranged in Transformation) gene mutations are considered as one of the major drivers of MTC. Vandetanib suppresses RET activity, and has shown promise in clinical trials. Unfortunately, acquired resistance to vandetanib has been observed in MTC, although the mechanism was largely unknown. We investigated the critical role of YAP (Yes-Associated Protein) on vandetanib resistance in MTC. For this, TT cells (medullary thyroid cancer cells) were treated with vandetanib for 3 months to generate a vandetanib-resistant cell line (TT-R). We investigated the role of YAP on vandetanib-resistance in TT-R cells by performing cell proliferation and colony formation assays, and examined the antitumor effects of YAP inhibitor and vandetanib in a mouse model of xenografted MTC. The TT-R cells displayed 6-fold higher IC50 to vandetanib than the TT cells. Overexpression of YAP resulted in resistance to vandetanib, whereas knockdown of YAP re-sensitized the TT-R cells to vandetanib. The YAP inhibitor synergized with vandetanib on tumor inhibition. Our results suggest that YAP plays an important role in acquired resistance to vandetanib in MTC, providing basis for combating MTC with YAP inhibitor and vandetanib.

Quassilactones A and B, structural characterization of a new class of norquassinoids from Brucea javanica.

Two novel norquassinoids possessing a unique ketal skeleton, designated quassilactones A (1) and B (2), were isolated from the fruits of Brucea javanica (Simaroubaceae). Their structures were established by extensive NMR and HR-ESI-MS spectroscopic analysis. The absolute configuration of 1 was determined through single-crystal X-ray crystallography, and that of 2 was assigned by comparing the calculated electronic and experimental circular dichroism with compound 1. In addition, their cytotoxic activities against three human cancer cell lines and their antimicrobial activities were evaluated.

Tolerogenic Dendritic Cells Generated with Tofacitinib Ameliorate Experimental Autoimmune Encephalomyelitis through Modulation of Th17/Treg Balance.

It is well known that dendritic cells (DCs) play a pivotal role in triggering self-specific responses. Conversely, tolerogenic DCs (tolDCs), a specialized subset, induce tolerance and negatively regulate autoreactive responses. Tofacitinib, a Janus kinase inhibitor developed by Pfizer for treatment of rheumatoid arthritis, is probable to be a promising candidate for inducing tolDCs. The aims of this study were to evaluate the effectiveness of tolDCs induced by tofacitinib in a myelin oligodendrocyte glycoprotein- (MOG-) specific experimental autoimmune encephalomyelitis (EAE) model and to investigate their effects on Th17/Treg balance in the animal model of multiple sclerosis (MS). Our results revealed that tofacitinib-treated DCs maintained a steady semimature phenotype with a low level of proinflammatory cytokines and costimulatory molecules. DCs treated by tofacitinib also induced antigen-specific T cells hyporesponsiveness in a concentration-dependent manner. Upon intravenous injection into EAE mice, MOG pulsed tolDCs significantly dampened disease activity, and adoptive cell therapy (ACT) disturbed Th17/Treg balance with a remarkable decrease of Th1/Th17 cells and an increase in regulatory T cells (Tregs). Overall, DCs modified by tofacitinib exhibited a typical tolerogenic phenotype, and the antigen-specific tolDCs may represent a new avenue of research for the development of future clinical treatments for MS.

A new quassinoid from fruits of Brucea javanica.

A new minor quassinoid named bruceine M (1), together with 12 known quassinoids (2-13), was isolated from the fruits of Brucea javanica (Simaroubaceae). Their structures were determined by interpretation of NMR and HR-ESI-MS data. The structures of the known compounds were confirmed by comparison of their spectral data with those reported in the literatures. In vitro cytotoxicity of the isolated compounds against cancer cell lines Bel-7404, MCF-7 and A549 was evaluated. Compounds 4, 6 and 9 exhibited significant growth inhibitory activity against three cancer cell lines.

A new flavanocoumarin from the root of Flemingia philippinensis.

One new flavanocoumarin, flemicoumarin A (1) was isolated from the EtOAc-soluble partition of the root of Flemingia philippinensis, along with three known compounds, namely 4,2'-epoxy-4',5-dihydroxy-7,5'-dimethoxy-3-phenylcoumarin (2), kaempferol 6-C-glucoside (3) and dracocephaloside (4). The structure of compound 1 was elucidated on the basis of its 1D, 2D NMR, CD and MS data. The structures of the known compounds were identified by comparison of their spectroscopic data with those reported in the literature. Compounds 1-4 exhibited inactivity against MCF-7, A549 and Hep-G2 human cancer cell lines in vitro by MTT colorimetric assay.

Two new lignans from the stems of Schisandra bicolor.

Two new lignans, named zuihonins E (1) and F (2), were isolated from the stems of Schisandra bicolor Cheng var. tuberculata Law. The structures of the new lignans were elucidated on the basis of extensive spectroscopic analysis, including 1D, 2D NMR, and MS experiments, and their absolute stereochemistry was determined by circular dichroism spectrum. Compounds 1 and 2 did not inhibit the growth of hepatoma carcinoma cell (HepG2), lung carcinoma cell (A549), and human breast carcinoma (MCF-7) cell lines.

Interaction of pseudolaric acid B with the colchicine site of tubulin.

We purified pseudolaric acid B (PAB) from the root and stem bark of Pseudolarix kaempferi (Lindl.) Gorden. Confirming previous findings, we found that the compound had high nanomolar IC50 antiproliferative effects in several cultured cell lines, causing mitotic arrest and the disappearance of intracellular microtubules. PAB strongly inhibited tubulin assembly (IC50, 1.1 µM) but weakly inhibited the binding of colchicine to tubulin, as demonstrated by fluorescence and with [³H]colchicine. Kinetic analysis demonstrated that the mechanism of inhibition was competitive, with an apparent K(i) of 12-15 µM. Indirect studies demonstrated that PAB bound rapidly to tubulin and dissociated more rapidly from tubulin than the colchicine analog 2-methoxy-5-(2',3',4'-trimethoxyphenyl)tropone, whose complex with tubulin is known to have a half-life of 17s at 37 °C. We modeled PAB into the colchicine site of tubulin, using the crystal structure 1SA0 that contains two αß-tubulin heterodimers, both bound to a colchicinoid and to a stathmin fragment. The binding model of PAB revealed common pharmacophoric features between PAB and colchicinoids, not readily apparent from their chemical structures.