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A method was developed for the preparative separation of two alkaloids from the crude extract of the radix of Rauvolfia verticillata (Lour.) Baill. in a single run. The two-phase solvent system composed of petroleum ether–ethyl acetate–methanol–water (5:5:2:8, v/v), where triethylamine (40 mmol/L) was added to the upper organic phase as the stationary phase and hydrochloric acid (10 mmol/L) was added to the lower aqueous phase as the mobile phase, was selected for this separation by pH-zone-refining counter-current chromatography (PZRCCC). For the preparative separation, the apparatus was rotated at a speed 850 rpm, while the mobile phase was pumped into the column at 2 mL/min. As a result, 112 mg of reserpine and 21 mg of yohimbine were obtained from 3 g of crude extract in a single run. The analysis of the isolated compounds was determined by high-performance liquid chromatography (HPLC) at 230 nm with purities of over 91.0%, and the chemical identification was carried out by the data of electrospray ionization–mass spectrometry (ESI–MS) and nuclear magnetic resonance (NMR) spectroscopy. The technique introduced in this paper is an efficient method for preparative separation of reserpine and yohimbine from devil pepper radix. It will be beneficial to utilize medicinal materials and also useful for the separation, purification, and pharmacological study of Chinese herbal ingredients.

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Abstract  

The complex of [Tb2(p-ClBA)6(PHEN)2] [(p-ClBA: p-chlorobenzoate and PHEN: 1,10-phenanthroline) was prepared and characterized by elemental analysis and IR spectroscopy. The thermal behavior of [Tb2(p-ClBA)6(PHEN)2] in dynamic nitrogen atmosphere was investigated by TG-DTG, SEM and IR techniques. By the kinetic method of processing thermal analysis data put forward by Malek et al., it is defined that the kinetic model for the first-step thermal decomposition is SB(m,n). The activation energy E and the pre-exponential factor lnA for this step reaction are 164 kJ mol-1 and 32.80, respectively. The lifetime equation at mass loss of 10% was deduced as lnτ=(-33.0569+20512.36/T by isothermal thermogravimetric analysis.

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Journal of Thermal Analysis and Calorimetry
Authors: W. Liu, S. Kim, J. Lopez, B. Hsiao, M. Keating, I.-H. Lee, B. Landes, and R. Stein

Abstract  

In this study, the stepwise isothermal crystallization or thermal fractionation of Ziegler—Natta and metallocene based polyethylenes (ZN-PE and m-PE) with two kinds of branch lengths (ethyl and hexyl) and branch compositions were studied using simultaneous synchrotron small-angle X-ray scattering (SAXS)/wide-angle X-ray diffraction (WAXD) and differential scanning calorimetry (DSC). The crystal long period and the invariant were determined by SAXS, and the variations of crystal unit cell parameters and the degree of crystallinity were determined by WAXD. The arithmetic mean length (Ln), the weightedmean length (Lw) and the broadness index (Lw/Ln) of the studied polyethylenes were previously determined by DSC. Results from these studies were interpreted using the model of branch exclusion, which affects the ability of the chain-reentry into the crystal phase. Multiple SAXS peaks and step-change in crystallinity change (WAXD) were seen during heating, which corresponded well with the crystal thickness distribution induced by stepwise crystallization. The effects of the heterogeneity of the 1-olefin branch length and the distribution on the crystal long period and the invariant as well as the degree of crystallinity were discussed.

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The aphid Sitobion avenae F. is one of the most harmful pests of wheat growth in the world. A primary field screening test was carried out to evaluate the S. avenae resistance of 527 wheat landraces from Shaanxi. The results indicated that 25 accessions (4.74%) were resistant to S. avenae in the three consecutive seasons, of which accession S849 was highly resistant, and seven accessions were moderately resistant. The majority of S. avenae resistant accessions come from Qinling Mountains. Then, the genetic variability of a set of 33 accessions (25 S. avenae resistant and 8 S. avenae susceptible) originating from Qinling Mountains have been assessed by 20 morphological traits and 99 simple sequence repeat markers (SSRs). Morphological traits and SSRs displayed a high level of genetic diversity within 33 accessions. The clustering of the accessions based on morphological traits and SSR markers showed significant discrepancy according to the geographical distribution, resistance to S. avenae and species of accessions. The highly and moderately resistant landrace accessions were collected from the middle and the east part of Qinling Mountains with similar morphology characters, for example slender leaves with wax, lower leaf area, and high ear density. These S. avenae resistant landraces can be used in wheat aphid resistance breeding as valuable resources.

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The forward and reverse cDNA subtractive libraries before and after the toxic effect of α-amanitin were constructed by suppression subtractive hybridization and randomly selected clones from each subtractive library were screened by PCR and dot blot hybridization. A total of 85 genes with altered expression were finally identified, with 41 genes from the forward library and 44 from the reverse library. Subsequently, the antagonistic effects of candidate traditional Chinese medicines were evaluated based on the genetic transcription levels of the genes with significant altered expression, including Catnβ, Flt3-L, IL-7r and Rpo2-4. The results indicated that Silybum marianum (L.) Gaert and Ganoderma lucidum had significant down-regulated effects on the transcription level of Catnβ that was up-regulated by α-amanitin, and the two herbs also up-regulated the transcription levels of Flt3-L and Rpo2-4. Silybum marianum (L.) had significant up-regulated effects on the IL-7r that was down-regulated by α-amanitin. These preliminary studies suggested that Silybum marianum (L.) and Ganoderma lucidum were effective antagonists against the toxicity of α-amanitin.

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Shewanella putrefaciens supernatant was found to increase the virulence factors of Vibrio parahaemolyticus by efficiently degrading its acylhomoserine lactone (AHL). To further reveal the regulation mechanism and its key degrading enzyme, a potential AHL-degrading enzyme acylase (Aac) from S. putrefaciens was cloned, and the influences of temperature, pH, protein modifiers, and metals on Aac were tested. Aac was significantly influenced by temperature and pH, and exhibited the highest AHL-degrading activity at temperatures of 37 °C and pH of 8. Mg2+ and Fe2+ can further increase the AHL-degrading activity. 10 mM EDTA inhibited its activity possibly by chelating the co-factors (metals) required for Aac activity. Tryptophan and arginine were identified as key components for Aac activity that are critical to its AHL-degrading activity. This study provides useful information on Aac and for V. parahaemolyticus control.

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Compound danshen preparations (CDPs) are used clinically for the treatment of cardiovascular and cerebrovascular diseases. By using the quantitative analysis of multi-components by single-marker (QAMS) method, sixteen compounds (danshensu, protocatechuic acid, protocatechuicaldehyde, caffeic acid, rosmarinic acid, lithospermic acid, notoginsenoside R1, salvianolic acid B, ginsenoside Rg1, ginsenoside Re, salvianolic acid A, salvianolic acid C, ginsenoside Rb1, ginsenoside Rd, cryptotanshinone, and tanshinone IIA were quantified on an ACQUITY ultraperformance liquid chromatography (UPLC) HSS T3 column (2.1 × 100 mm, 1.8 μm) with the mobile phase consisting of 0.1% formic acid aqueous solution (A) and acetonitrile (B) using a gradient elution at the flow rate of 0.30 mL/min in 30 min at 30°C, which was also validated by UPLC-diode array detection (DAD) and UPLC-electrospray ionization multistage/mass spectrometry (ESI-MS/MS) for assuring the feasibility and accuracy. Tested by robustness experiment under slightly changeable conditions, the stability of relative correction factor (RCF) proved to be stable, with RSDs below 5.69%, except for notoginsenoside R1 with relative standard deviation (RSD) 7.83%. This reliable and convenient QAMS method resolved the problem of standard substance insufficiency and improved the quality assessment of preparations consisting of complex compounds with different chemical structures, such as CDPs.

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Cereal Research Communications
Authors: Y.P. Jing, D.T. Liu, X.R. Yu, F. Xiong, D.L. Li, Y.K. Zheng, Y.F. Hao, Y.J. Gu, and Z. Wang

The objective of the present study was to understand the developmental regularity of wheat endosperm cells at different Days After Pollination (DAP) using microscopic and histochemical methods. Resin semi-thin sections of the endosperm and the enzymatically dissociated Starchy Endosperm Cells (SECs) were observed under a light microscope. The results showed that: (1) SECs were irregular-shaped and had two types of starch granules: large oval-shaped A-type starch granules and small spherical B-type starch granules. (2) The growth shape of SECs was referred to as S-curve and the fastest cell growth period was at 16–24 DAP. (3) The largest increase and growth of A-type starch granules were mainly at 4–16 DAP. B-type starch granules increased rapidly after 16 DAP and made up over 90% of the total starch granules in SEC during the late stage of endosperm development. (4) The nuclei of SEC deformed and degenerated during the middle and late stages of endosperm development and eventually disappeared. However, starch granules still increased and grew after the cell nuclei had degenerated. The investigations showed the development regularity of starch endosperm cells and starch granules, thereby improving the understanding of wheat endosperm development.

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Cereal Research Communications
Authors: N. Zhang, R.Q. Pan, J.J. Liu, X.L. Zhang, Q.N. Su, F. Cui, C.H. Zhao, L.Q. Song, J. Ji, and J.M. Li

Plants with deficiency in Gibberellins (GAs) biosynthesis pathway are sensitive to exogenous GA3, while those with deficiency in GAs signaling pathway are insensitive to exogenous GA3. Thus, exogenous GA3 test is often used to verify whether the reduced height (Rht) gene is involved in GAs biosynthesis or signaling pathway. In the present study, we identified the genetic factors responsive to exogenous GA3 at the seedling stage of common wheat and analyzed the response of the plant height related quantitative trait loci (QTL) to GA3 to understand the GAs pathways the Rht participated in. Recombinant inbred lines derived from a cross between KN9204 and J411 with different response to exogenous GA3 were used to screen QTL for the sensitivity of coleoptile length (SCL) and the sensitivity of seedling plant height (SSPH) to exogenous GA3. Two additive QTL and two pairs of epistatic QTL for SCL were identified, meanwhile, two additive QTL and three pairs of epistatic QTL for SSPH were detected. For the adult plant height (PH) investigated in two environments, six additive QTL were identified. Three QTL qScl-4B, qSsph-4B and qPh-4B were mapped in one cluster near the functional marker Rht-B1b. When PH were conditional on SSPH, the absolute additive effect value of qPh-4B and qPh-6B were reduced, suggesting that the Rhts in both two QTL were insensitive to exogenous GA3, while the additive effect values of qPh-2B, qPh-3A, qPh-3D and qPh-5A were not significantly changed, indicating that the Rhts in these QTL were sensitive to exogenous GA3, or they were not expressed at the seedling stage.

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