Electroacupuncture reduces myocardial infarct size and improves post-ischemic recovery by invoking release of humoral, dialyzable, cardioprotective factors.

Previous studies have shown that electroacupuncture (EA) can induce cardioprotection against ischemia-reperfusion (IR) injury, but its mechanisms are incompletely understood. We have previously shown that several other forms of remote preconditioning of the heart work, at least in part, via the release of circulating cardioprotective factors into the bloodstream, that can be dialyzed and subsequently shown to reduce IR injury in isolated hearts. We used the same methods to assess whether EA leads to similar humoral cardioprotection. EA rabbits were subjected to 60 min of bilateral stimulation at the Neiguan point, following which their blood was drawn, dialyzed, and used to perfuse hearts in Langendorff preparation and subsequently subjected to 60 min of global ischemia and 120 min of reperfusion. Compared to controls, dialysate from EA animals led to significant reduction in infarct size and improved functional recovery. The degree of cardioprotection was no different to that seen in animals randomized to receive remote preconditioning using transient limb ischemia (4 cycles of 5 min ischemia/5 min reperfusion). These results suggest that EA recapitulates the cardioprotection achieved by remote preconditioning, by similarly leading to release of circulating cardioprotective factors.

Comprehensive separation and analysis of alkaloids from Stephania yunnanensis by counter-current chromatography coupled with liquid chromatography tandem mass spectrometry analysis.

The polar compounds such as alkaloid compounds are important bioactive components in traditional Chinese medicines. In present study, a comprehensive method for separation and analysis of polar compounds from the polar fraction of traditional Chinese medicine Stephania yunnanensis was established. Both the major components and minor components were analyzed by counter-current chromatography combined with liquid chromatography tandem mass spectrometry (LC-MS(n)). From 50 mg polar fraction of crude extract, 15.2mg corydine and 4.8 mg stepharine with purities over 90% were successfully separated via a polar solvent system n-butanol: methanol: water (4:1:5, v/v) with 10 mM NaOH as an additive in the lower phase, in one step operation. Their structures were further identified by 1H NMR and FTICR-MS. Besides, three minor components were identified by HPLC-MS(n) based on the fragmentation behavior of the purified compounds.

Two-dimensional counter-current chromatography: 1st traditional counter-current chromatography, 2nd acid-base elution counter-current chromatography.

An on-line column-switching counter-current chromatography (CCC) with solid-phase trapping interphase is presented in this paper. The large volume injection is avoided using solid-phase trapping interphase. Thereby, totally different chemical composition biphasic solvent system can be utilized to enhance system orthogonality. In the present work, a 140 mL-capacity CCC instrument was used in the first dimension, and a parallel three-coil CCC centrifuge (40 mL each coil) was used in the second dimensional separation allowing three injections at the same time. With biphasic solvent system composed of n-hexane: ethyl acetate: methanol: water (1:1:1:1, v/v), five well-separated fractions were obtained in the first dimension. Two fractions were online concentrated and further separated in the second dimension with solvent system composed of methyl tert-butyl ether: acetonitrile: water (2:2:3, v/v), where trifluoroacetic acid (10 mM) was added to the upper organic phase as a retainer and triethylamine (10mM) to the aqueous mobile phase as an eluter. Four hydroxyanthraquinones were successfully separated and purified from Chinese medicinal plant Rheum officinale in only one step.

Preparative separation of isoquinoline alkaloids from Stephania yunnanensis by pH-zone-refining counter-current chromatography.

In this paper, five isoquinoline alkaloids were successfully separated from a crude extract of Stephania yunnanensis using pH-zone-refining counter-current chromatography in single-step. With a two-phase solvent system composed of methyl-tert-butyl ether (MtBE)-acetonitrile-water (2:2:3, v/v) where triethylamine (10 mM) was added to the upper organic phase as a retainer and hydrochloric acid (5 mM) to the aqueous mobile phase as an eluter. From 1.4 g crude extract, 68.7 mg isocorydine, 78.2 mg corydine, 583.4 mg tetrahydropalmatine, 36.3 mg N-methylasimilobine, and 47.3 mg anonaine were separated with purities over 90%. Their structures were identified by (1)H NMR, (13)C NMR, ESI-MS data.

Screening of antioxidant phenolic compounds in Chinese rhubarb combining fast counter-current chromatography fractionation and liquid chromatography/mass spectrometry analysis.

In this paper, an effective method combing fast elution-extrusion counter-current chromatography (CCC) and LC/MS for rapid screening of antioxidative phenolic compounds in Chinese Rhubarb is presented. An integrated three-coil CCC column (40 mL each coil) was used to accomplish the optimization of biphasic liquid system. In a single run (approximately 40 min), the solvent system composed of n-hexane/ethyl acetate/methanol/water (1:1:1:1, v/v) was selected as optimum CCC liquid system for fast fractionation of the crude ethanol extract. With a 140 mL-capacity CCC instrument, 100 mg Chinese Rhubarb extract was separated under the optimized conditions, producing six fractions in only 100 min. The quantities of each fraction were approximately 15 mg. In addition, each fraction was subjected to antioxidant activity assay and characterized by LC/MS analysis. Fifty compounds, including phenolic acids, phenolic glucosides and hydroxyanthraquinones, were detected by LC/MS/MS analysis. As a result, gallic acid together with Fr I showed excellent antioxidant activity, which was well consistent with previous studies and exhibited great potential for natural drug discovery program of the present method.

Ionic liquid-based microwave-assisted extraction of phenolic alkaloids from the medicinal plant Nelumbo nucifera Gaertn.

An ionic liquid-based microwave-assisted extraction (ILMAE) approach has been successfully applied to the effective extraction of the phenolic alkaloids present in samples of the medicinal plant Nelumbo nucifera Gaertn. The ionic liquids investigated comprised a range of four anions, four 1-alkyl-3-methylimidazolium derivatives differing in hydrophobic chain length. The results indicate that varying the anion has apparent effects on the overall extraction efficiency. In addition, the influence of some microwave parameters, such as irradiation power, extraction time and solid-liquid ratio, are also investigated. Under the optimized conditions, the proposed approach has been evaluated in comparison with the conventional heat-reflux extraction (HRE) and regular MAE. The reduction of the extraction times (from 2h to 90s) and remarkable higher efficiency (20-50% improved) supports the suitability of the proposed approach. In addition, the proposed method is validated by the recovery, correlation coefficient (R(2)), and reproducibility (RSD, n=5), which are in the range of 98-105%, 0.9994-0.9998, and 1.2-5.4%, respectively.

Overexpressing GSH1 and AsPCS1 simultaneously increases the tolerance and accumulation of cadmium and arsenic in Arabidopsis thaliana.

The goal of this study was to develop transgenic plants with increased tolerance for and accumulation of heavy metals and metalloids from soil by simultaneous overexpression of AsPCS1 and GSH1 (derived from garlic and baker's yeast) in Arabidopsis thaliana. Phytochelatins (PCs) and glutathione (GSH) are the main binding peptides involved in chelating heavy metal ions in plants and other living organisms. Single-gene transgenic lines had higher tolerance to and accumulated more Cd and As than wild-type. Compared to single-gene transgenic lines, dual-gene transformants exhibited significantly higher tolerance to and accumulated more Cd and As. One of the dual-gene transgenic lines, PG1, accumulated twice the amount of Cd as single-gene transgenic lines. Simultaneous overexpression of AsPCS1 and GSH1 led to elevated total PC production in transgenic Arabidopsis. These results indicate that such a stacking of modified genes is capable of increasing Cd and As tolerance and accumulation in transgenic lines, and represents a highly promising new tool for use in phytoremediation efforts.