Determination and Distribution Study of Pogostone in Rat Tissues by Ultra-Fast Liquid Chromatography

Purpose: To develop and validate a rapid, sensitive and reliable ultra-fast liquid chromatography (UFLC) method with photodiode array (PDA) detection for the determination of pogostone (PO) in rat tissues using honokiol as internal standard (IS).Methods: Rats were randomly divided into two groups (intravenous administration group and oral administration group) and given of a single dose of 10 mg/kg PO by intravenous administration and oral administration, respectively. After intravenous injection, the rats were sacrificed at 15, 60 and 360 min, while rats, after oral administration, were euthanasized at 30, 90 and 360 min, respectively. For the analysis of the preparation, optimal chromatographic conditions were determined using Acquity UPLC BEH C18 column with acetonitrile-water containing 0.1 % formic acid (55:45, v/v) as the mobile phase, at a flow rate of 400 μL/min. UV detection wavelength was set at 310 nm with temperature maintained at 30 °C.Results: Good linear relationship of calibration curve (r > 0.9984) was achieved over the range of 0.1 - 40 μg/mL for all the tissue samples. The limit of quantification (LOQ) and limit of detection (LOD) were 0.1 and 0.05 μg/mL, respectively. This method proved to have good precision, accuracy, stability, extraction recovery and matrix effect for tissue distribution studies of PO in rats.Conclusion: The developed method is suitable for tissue distribution studies in rats following intravenous and oral administration of PO at a dose of 10 mg/kg.Keywords: Ultra-fast liquid chromatography, Tissue distribution, Pogostone, Honokiol, Rat

Small inhibitor of Bcl-2, HA14-1, selectively enhanced the apoptotic effect of cisplatin by modulating Bcl-2 family members in MDA-MB-231 breast cancer cells

Inhibition or downregulation of Bcl-2 represents a new therapeutic approach to by-pass chemoresistance in cancer cells. Therefore, we explored the potential of this approach in breast cancer cells. Cisplatin and paclitaxel induced apoptosis in a dose-dependent manner in MCF-7 (drug-sensitive) and MDA-MB-231 (drug-insensitive) cells. Furthermore, when we transiently silenced Bcl-2, both cisplatin and paclitaxel induced apoptosis more than parental cells. Dose dependent induction of apoptosis by drugs was enhanced by the pre-treatment of these cells with HA14-1, a Bcl-2 inhibitor. Although the effect of cisplatin was significant on both cell lines, the effect of paclitaxel was much less potent only in MDA-MB-231 cells. To further understand the distinct role of drugs in MDA-MB-231 cells pretreated with HA14-1, caspases and Bcl-2 family proteins were studied. The apoptotic effect of cisplatin with or without HA14-1 pre-treatment is shown to be caspase-dependent. Among pro-apoptotic Bcl-2 proteins, Bax and Puma were found to be up-regulated whereas Bcl-2 and Bcl-x(L) were down-regulated when cells were pretreated with HA14-1 followed by paclitaxel or cisplatin. Enforced Bcl-2 expression in MDA-MB-231 cells abrogated the sensitizing effect of HA14-1 in cisplatin induced apoptosis. These results suggest that the potentiating effect of HA14-1 is drug and cell type specific and may not only depend on the inhibition of Bcl-2. Importantly, alteration of other pro-apoptotic or anti-apoptotic Bcl-2 family members may dictate the apoptotic response when HA14-1 is combined with chemotherapeutic drugs

Hormonal regulation of ovarian bursa fluid in mice and involvement of aquaporins.

In rodent species, the ovary and the end of oviduct are encapsulated by a thin membrane called ovarian bursa. The biological functions of ovarian bursa remain unexplored despite its structural arrangement in facilitating oocytes transport into oviduct. In the present study, we observed a rapid fluid accumulation and reabsorption within the ovarian bursa after ovarian stimulation (PMSG-primed hCG injection), suggesting that the ovarian bursa might play an active role in regulating local fluid homeostasis around the timing of ovulation. We hypothesized that the aquaporin proteins, which are specialized channels for water transport, might be involved in this process. By screening the expression of aquaporin family members (Aqp1-9) in the ovarian tissue and isolated ovarian bursa (0, 1, 2 and 5 h after hCG injection), we found that AQP2 and AQP5 mRNA showed dynamic changes after hCG treatment, showing upregulation at 1-2 h followed by gradually decrease at 5 h, which is closely related with the intra-bursa fluid dynamics. Further immunofluorescence examinations of AQP2 and AQP5 in the ovarian bursa revealed that AQP2 is specifically localized in the outer layer (peritoneal side) while AQP5 localized in the inner layer (ovarian side) of the bursa, such cell type specific and spatial-temporal expressions of AQP2 and 5 support our hypothesis that they might be involved in efficient water transport through ovarian bursa under ovulation related hormonal regulation. The physiological significance of aquaporin-mediated water transport in the context of ovarian bursa still awaits further clarification

The alexithymia traits and personality characteristics in patients with irritable bowel syndrome

2012-2013 > Academic research: refereed > Publication in refereed journalVersion of RecordPublishe

Synthesis and Characterization of Monodispersed Copper Colloids in Polar Solvents

A chemical reduction method for preparing monodispersed pure-phase copper colloids in water and ethylene glycol has been reported. Owing to the reduction property of ethylene glycol, the reaction rate in ethylene glycol is higher than that in water. In addition, the amount of reducing agent can be reduced largely. Ascorbic acid plays roles as reducing agent and antioxidant of colloidal copper, due to its ability to scavenge free radicals and reactive oxygen molecules. Thermogravimetric results reveal that the as-prepared copper nanoparticles have good stability, and they begin to be oxidized at above 210 °C. Polyvinyl pyrrolidone works both as size controller and polymeric capping agents, because it hinders the nuclei from aggregation through the polar groups, which strongly absorb the copper particles on the surface with coordination bonds

Water imbibition of shale and its potential influence on shale gas recovery-a comparative study of marine and continental shale formations

A large volume of fracturing fluid is pumped into a well to stimulate shale formation. The water is imbibed into the reservoir during this procedure. The effect of the imbibed water on gas recovery is still in debate. In this work, we study the spontaneous imbibition of water into marine shale samples from the Sichuan Basin and continental shale samples from Erdos Basin to explore the fluid imbibition characteristics and permeability change during water imbibition. Comparison of imbibition experiments shows that shale has stronger water imbibition and diffusion capacity than relatively higher permeability sandstone. Once the imbibition stops, water in shale has stronger ability to diffuse into deeper matrix, the water content in the main flow path decreases. Experiments in this study show that marine shale has stronger water imbibition capacity than continental shale. The permeability of continental shale decreases significantly with increasing imbibition water volume; however, the permeability of marine shale decreases at first and increases after a certain imbibition time. The induced fracture is obvious in the marine shale. SEM analysis shows that the relationship between the clay mineral and organic matter of continental shale is much more complex than that of marine shale, which may be the key factor restricting the water imbibition because the flow path is trapped by swelled clay minerals. Through this study, we concluded that whether gas recovery benefits from water imbibition depends on three aspects: 1) the diffusion ability of liquid into matrix; 2) the new cracks introduced by imbibed water; and 3) the formation sensibility. This study is useful for optimizing fracture fluids and determining the best flow-back method. (C) 2016 Elsevier B.V. All rights reserved

Inhibition of B16 melanoma growth and metastasis in C57BL mice by vaccination with a syngeneic endothelial cell line

<p>Abstract</p> <p>Background</p> <p>Key role of angiogenesis in tumor growth and metastasis based on accumulating evidence and recent progress of immunotherapy have led us to investigate vaccine therapy targeting tumor angiogenesis.</p> <p>Methods</p> <p>C57BL/6J mice were vaccinated with a syngeneic endothelial cell line Tpit/E by subcutaneous injection once a week. Prior to ninth vaccination, the mice were challenged with B16/F10 melanoma cells by subcutaneous inoculation on the back for the tumor growth model or by tail venous injection for the lung metastasis model. Development of subcutaneous tumor and lung metastasis was monitored by computed tomography scanning, which enabled accurate evaluation with the minimized sacrifice of mice.</p> <p>Results</p> <p>Vaccination with Tpit/E cells inhibited subcutaneous tumor growth and appearance of lung metastasis compared to control. Survival period was elongated in the Tpit/E vaccination in both of the two models. We also obtained hybridomas secreting specific antibodies to Tpit/E cells from a mouse vaccinated with the cells, indicating that specific immune response to the syngeneic endothelial cells was elicited.</p> <p>Conclusion</p> <p>These results suggest that vaccination with an autologous endothelial cell line may be effective against melanoma.</p

Erroneous attribution of relevant transcription factor binding sites despite successful prediction of cis-regulatory modules

<p>Abstract</p> <p>Background</p> <p><it>Cis</it>-regulatory modules are bound by transcription factors to regulate gene expression. Characterizing these DNA sequences is central to understanding gene regulatory networks and gaining insight into mechanisms of transcriptional regulation, but genome-scale regulatory module discovery remains a challenge. One popular approach is to scan the genome for clusters of transcription factor binding sites, especially those conserved in related species. When such approaches are successful, it is typically assumed that the activity of the modules is mediated by the identified binding sites and their cognate transcription factors. However, the validity of this assumption is often not assessed.</p> <p>Results</p> <p>We successfully predicted five new <it>cis</it>-regulatory modules by combining binding site identification with sequence conservation and compared these to unsuccessful predictions from a related approach not utilizing sequence conservation. Despite greatly improved predictive success, the positive set had similar degrees of sequence and binding site conservation as the negative set. We explored the reasons for this by mutagenizing putative binding sites in three <it>cis</it>-regulatory modules. A large proportion of the tested sites had little or no demonstrable role in mediating regulatory element activity. Examination of loss-of-function mutants also showed that some transcription factors supposedly binding to the modules are not required for their function.</p> <p>Conclusions</p> <p>Our results raise important questions about interpreting regulatory module predictions obtained by finding clusters of conserved binding sites. Attribution of function to these sites and their cognate transcription factors may be incorrect even when modules are successfully identified. Our study underscores the importance of empirical validation of computational results even when these results are in line with expectation.</p