Effects of flavonoids extracted from the whole plant of Patrinia Villosa (Thunb) Juss in a rat model of chronic pelvic inflammation

Purpose: To investigate the effects of total flavonoids (PLV) extracted from the  whole plant of Patrinia Villosa (Thunb.) Juss (PTJ) in a rat model of chronic pelvic inflammation.Methods: An orthogonal test design was employed to optimize the extraction  conditions of PLV via reflux extraction by ethanol. Rats were randomly divided into control group, model group and PLV groups. An absorbable gelatin sponge with  pathogens was inserted into the cervix of the rat to establish a pelvic inflammatory model. The PLV groups were orally administered PLV at doses of 25, 50 and 100 mg/kg for eight days. Enzyme-linked immunosorbent assay (ELISA) was used for the determination of inflammatory cytokines in rat serum and the culture  supernatant of RAW264.7 cells. Real-time reverse transcription polymerase chain reaction (real-time PCR) was employed to determine mRNA levels.Results: The optimum extraction conditions for PLV by orthogonal test were  obtained: extraction time (120 min), ratio of liquid to raw material (20 mL/g) and ethanol concentration (50 %). By treating with PLV, the levels of TNF-α, IL-6 and IL-1β significantly decreased (p < 0.01), while IL-10 level significantly increased (p < 0.01) in the serum of chronic pelvic inflammatory rats and LPS-stimulated  macrophages. In addition, a similar trend was observed in the mRNA levels of LPS-stimulated macrophages treated with PLV.Conclusion: PLV showes significant anti-inflammatory effects on chronic pelvic inflammation. The potential mechanism is related to regulating the expression of inflammatory factorsKeywords: Patrinia Villosa (Thunb.) Juss, Total flavonoids, Chronic pelvic  inflammation, Inflammatory cytokine

Regulation of Bestrophin Cl Channels by Calcium: Role of the C Terminus

Human bestrophin-1 (hBest1), which is genetically linked to several kinds of retinopathy and macular degeneration in both humans and dogs, is the founding member of a family of Cl− ion channels that are activated by intracellular Ca2+. At present, the structures and mechanisms responsible for Ca2+ sensing remain unknown. Here, we have used a combination of molecular modeling, density functional–binding energy calculations, mutagenesis, and patch clamp to identify the regions of hBest1 involved in Ca2+ sensing. We identified a cluster of a five contiguous acidic amino acids in the C terminus immediately after the last transmembrane domain, followed by an EF hand and another regulatory domain that are essential for Ca2+ sensing by hBest1. The cluster of five amino acids (293–308) is crucial for normal channel gating by Ca2+ because all but two of the 35 mutations we made in this region rendered the channel incapable of being activated by Ca2+. Using homology models built on the crystal structure of calmodulin (CaM), an EF hand (EF1) was identified in hBest1. EF1 was predicted to bind Ca2+ with a slightly higher affinity than the third EF hand of CaM and lower affinity than the second EF hand of troponin C. As predicted by the model, the D312G mutation in the putative Ca2+-binding loop (312–323) reduced the apparent Ca2+ affinity by 20-fold. In addition, the D312G and D323N mutations abolished Ca2+-dependent rundown of the current. Furthermore, analysis of truncation mutants of hBest1 identified a domain adjacent to EF1 that is rich in acidic amino acids (350–390) that is required for Ca2+ activation and plays a role in current rundown. These experiments identify a region of hBest1 (312–323) that is involved in the gating of hBest1 by Ca2+ and suggest a model in which Ca2+ binding to EF1 activates the channel in a process that requires the acidic domain (293–308) and another regulatory domain (350–390). Many of the ∼100 disease-causing mutations in hBest1 are located in this region that we have implicated in Ca2+ sensing, suggesting that these mutations disrupt hBest1 channel gating by Ca2+

LC/MS Guided Isolation of Alkaloids from Lotus Leaves by pH-Zone-Refining Counter-Current Chromatography

The traditional methods used in natural product separation primarily target the major components and the minor components may thus be lost during the separation procedure. Consequently, it’s necessary to develop efficient methods for the preparative separation and purification of relatively minor bioactive components. In this paper, a LC/MS method was applied to guide the separation of crude extract of lotus (Nelumbo nucifera Gaertn.) leaves whereby a minor component was identified in the LC/MS analysis. Afterwards, an optimized pH-zone-refining CCC method was performed to isolate this product, identified as N-demethylarmepavine. The separation procedure was carried out with a biphasic solvent system composed of hexane-ethyl acetate-methyl alcohol-water (1:6:1:6, v/v) with triethylamine (10 mM) added to the upper organic phase as a retainer and hydrochloric acid (5 mM) to the aqueous mobile phase eluent. Two structurally similar compounds – nuciferine and roemerine – were also obtained from the crude lotus leaves extract. In total 500 mg of crude extract furnished 7.4 mg of N-demethylarmepavine, 45.3 mg of nuciferine and 26.6 mg of roemerine with purities of 90%, 92% and 96%, respectively. Their structures were further identified by HPLC/ESI-MSn, FTICR/MS and the comparison with reference compounds

3D experimental study on a cylindrical floating breakwater system

The objective of the present study is to investigate the performance of a cylindrical floating breakwater system based on 3D experimental tests. The experiments were carried out in the wave basin (36 m*60 m*1.5 m)of the Ocean University of China. The cylindrical floating breakwater system consists of 10cylindrical floating breakwater units and 10mesh cages with balls in them, connected by 18 connectors and moored by a taut mooring system. The wave transmission coefficients, reflection coefficients, dissipation coefficients and motion responses of the floating breakwater are measured in both oblique and beam sea conditions. It is found that with the increase of the wavelength, both of the wave transmission coefficients and motion response amplitude of the FB system suffers an increase before it reaches its peak value, followed by a decrease trend. It can be concluded from the experiments that the proposed FB system has a satisfactory performance and it can be used to a wide range of sea conditions

Organocatalyzed Asymmetric Reaction Using α-Isothiocyanato Compounds

Organocatalyzed asymmetric reaction using α-isothiocyanato compounds has received much attention in the past 5 years, and significant progress has been made for three types of isothiocyanato compounds, including α-isothiocyanato amides, esters, and phosphonates. This chapter covers the recent advances of α-isothiocyanato compounds in the organocatalytic asymmetric reaction