Novel strategy for quality consistency evaluation of Chinese medicine “YIQING” tablet that combines the simultaneous quantification and screening of ten bioactive constituents

The UV characteristics for different categories compounds in complex traditional Chinese medicines and herbal preparations usually vary. Thus, to achieve the integral analysis of multiwavelength fingerprint characteristics, we introduced a novel strategy of multiwavelength total fusion profiling. The simultaneous separation and quantification of multiple components by reversed‐phase high‐performance liquid chromatography coupled with diode array detection was developed in an effective, accurate, and reliable way. Furthermore, a 2,2‐diphenyl‐1‐picrylhydrazyl radical (DPPH) scavenging assay was set up to detect and screen the bioactivity of similar‐structure constituents (aloe‐emodin, rhein, emodin, chrysophanol, baicalein, wogonin, baicalin, wogonoside, berberine hydrochloride, and jatrorrhizine hydrochloride). Moreover, the high‐performance liquid chromatography DPPH assay was developed to monitor the relationship between the biological activity and the spatial structure, the number of hydroxyl groups, the concentration of the analytes in samples. The result of qualitative classification for 15 batches of “YIQING” tablets using principle component analysis was consistent with the quantitative fingerprint assessment using the average linear quantitative fingerprint method. Therefore, chemometrics, multiwavelength total fusion profiling in conjunction with average linear quantitative fingerprint method and antioxidant activity can control the quality of traditional Chinese medicines/herbal preparations comprehensively and practically.     

Simultaneous quantification of multiple active components from Xiexin decoction in rat plasma by LC‐ESI‐MS/MS: application in pharmacokinetics

A sensitive and specific liquid chromatography–electrospray ionization–tandem mass spectrometric (LC‐ESI‐MS/MS) method was developed and validated to simultaneously quantify 11 active compounds (coptisine, jatrorrhizine, berberine, palmatine, baicalin, baicalein, wogonoside, wogonin, rhein, emodin and aloeemodin) from Xiexin decoction (XXD) in rat plasma. Plasma samples extracted by a single‐step protein precipitation procedure were separated using the gradient mode on a Dikma ODS‐C₁₈ column. Selected reaction monitoring scanning was employed for quantification with switching electrospray ion source polarity between positive and negative modes in a single run. Calibration curves offered satisfactory linearity (r > 0.995) at linear range of 0.47–60 ng/mL for coptisine, jatrorrhizine, berberine and palmatine, 15–1930 ng/mL for baicalin, 20–2560 ng/mL for baicalein, 14–1790 ng/mL for wogonoside, 0.57–72.8 ng/mL for wogonin, 10–1280 ng/mL for rhein, 0.6–76.8 ng/mL for emodin and 3.0–384 ng/mL for aloeemodin. The intra‐ and interday precisions were less than 10.2% in terms of relative standard deviation (RSD), and the accuracies were within ±10.84% in terms of relative error (RE). It was successfully applied to the evaluation of pharmacokinetics after single oral doses of XXD were administered to rats. Copyright © 2010 John Wiley & Sons, Ltd.     

Simultaneous Determination of Ten Active Components in Chinese Medicine “Huang-Lian-Shang-Qing” Tablets by High-Performance Liquid Chromatography Coupled with Photodiode Array Detection

A simple and sensitive high-performance liquid chromatography coupled with a photodiode array detection (HPLC-PAD) method was investigated for the simultaneous determination of ten components (rutin, berberine, palmatine, baicalin, wogonoside, baicalein, wogonin, emodin, chrysophanol, and physcion) in “Huang-Lian-Shang-Qing” (HLSQ) tablets. The method was optimized and the mobile phase composed of methanol (A)-3% phosphoric acid (B) was used to elute the targets in a gradient elution mode. All the calibration curves, precisions, and recoveries were good. Then, this method was successfully used to determine the ten compounds in 33 batches of HLSQ tablets for quality control of this medicinal product.     

Simultaneous determination of six herbal components in intestinal perfusate by high‐performance liquid chromatography

An effective, accurate and reliable HPLC with UV detection method was developed and validated for quantitation of six components: baicalin, berberine hydrochloride, quercetin, kaempferol, isorhamnetin and baicalein in intestinal perfusate using rotundin as an internal standard. The chromatographic separation was performed on a Welchrom‐C₁₈ column (250 × 4.6 mm i.d. with 5.0 µm particle size) with a mobile phase consisting of acetonitrile, water, phosphoric acid and triethylamine (30:70:0.2:0.1,v/v) at a flow rate of 1.0 mL/min and a UV detection at 270 nm. The method had a chromatographic run time of 30 min and excellent linear behavior over the investigated concentration ranges observed with the values of r higher than 0.99 for all the analytes. The lower limit of quantification of the analytical method was 0.09 µg/mL for berberine hydrochloride, quercetin, kaempferol and baicalein and 0.18 µg/mL for baicalin and isorhamnetin. The intra‐ and inter‐day precisions measured at three concentration levels were all less than 10% for all analytes. The bias ranged from ?6.91 to 4.33%. The validated method has been successfully applied to investigate the rat intestine absorption profiles of baicalin, berberine hydrochloride, quercetin, kaempferol, isorhamnetin and baicalein. Copyright © 2009 John Wiley & Sons, Ltd.     

Calcium cycling proteins in heart failure, cardiomyopathy and arrhythmias

A growing body of evidence, including studies using genetically engineered mouse models, has shown that Ca2+ cycling and Ca2+-dependent signaling pathways play a pivotal role in cardiac hypertrophy and heart failure. In addition, recent studies identified that mutations of the genes encoding sarcoplasmic reticulum (SR) proteins cause human cardiomyopathies and lethal ventricular arrhythmias. The regulation of Ca2+ homeostasis via the SR proteins may have potential therapeutic value for heart diseases such as cardiomyopathy, heart failure and arrhythmias.     

Pressurized hot water extraction of berberine, baicalein and glycyrrhizin in medicinal plants

Pressurized hot water extraction (PHWE) using a laboratory made system was applied for the extraction of thermally labile and reasonably polar components such as berberine in coptidis rhizoma, glycyrrhizin in radix glycyrrhizae/liquorice and baicalein in scutellariae radix. PHWE was carried out dynamically at a flow of 1 ml/min, temperature between 95 and 140 °C, an applied pressure of 10-20 bar and extraction time of 40 min. Extraction by PHWE was found to give efficiencies comparable to Soxhlet extraction for baicalein in scutellariae radix and sonication for berberine in coptidis rhizoma, and glycyrrhizin in radix glycyrrhizae. Effects of ethanol added into the water used in PHWE were explored. Pressurized liquid extraction (PLE) with methanol as solvent was used for extraction of baicalein in scutellariae radix. The marker compounds present in the various medicinal plant extracts were determined by gradient elution HPLC.     

Simultaneous quantification of twelve active components in Yiqing granule by ultra-performance liquid chromatography: application to quality control study

An ultra‐performance liquid chromatography (UPLC) method coupled with photodiode array (PDA) detection has been developed and validated for simultaneous quantification of 12 active components (berberine, palmatine, jatrorrhizine, aloe‐emodin, rhein, emodin, chrysophanol, physcion, baicalin, baicalein, wogonoside and wogonin) in Yiqing granule. Optimum separation were achieved on a C₁₈ column (50 × 2.1 mm i.d., 1.7 µm particle) through a 7.5 min gradient delivery of a mixture of A (acetonitrile) and B (0.1% aqueous phosphoric acid containing 1.8 mmol/L sodium dodecyl sulfonate and 10% acetonitrile, v/v) at a flow rate of 0.3 mL/min at 30°C. Because of the different UV characteristics of these components, three detection wavelengths were used for quantitative analysis. All of the analytes showed good linearity (r of >0.999). The method was validated for repeatability, precision, stability, accuracy and selectivity. The validated method was applied to quality control of Yiqing granule from different production batches. Copyright © 2010 John Wiley & Sons, Ltd.     

Novel Therapeutic Effects of Pterosin B on Ang II-Induced Cardiomyocyte Hypertrophy

Pathological cardiac hypertrophy is characterized by an abnormal increase in cardiac muscle mass in the left ventricle, resulting in cardiac dysfunction. Although various therapeutic approaches are being continuously developed for heart failure, several studies have suggested natural compounds as novel potential strategies. Considering relevant compounds, we investigated a new role for Pterosin B for which the potential life-affecting biological and therapeutic effects on cardiomyocyte hypertrophy are not fully known. Thus, we investigated whether Pterosin B can regulate cardiomyocyte hypertrophy induced by angiotensin II (Ang II) using H9c2 cells. The antihypertrophic effect of Pterosin B was evaluated, and the results showed that it reduced hypertrophy-related gene expression, cell size, and protein synthesis. In addition, upon Ang II stimulation, Pterosin B attenuated the activation and expression of major receptors, Ang II type 1 receptor and a receptor for advanced glycation end products, by inhibiting the phosphorylation of PKC-ERK-NF-κB pathway signaling molecules. In addition, Pterosin B showed the ability to reduce excessive intracellular reactive oxygen species, critical mediators for cardiac hypertrophy upon Ang II exposure, by regulating the expression levels of NAD(P)H oxidase 2/4. Our results demonstrate the protective role of Pterosin B in cardiomyocyte hypertrophy, suggesting it is a potential therapeutic candidate.     

Cross talk between cell-cell and cell-matrix adhesion signaling pathways during heart organogenesis: implications for cardiac birth defects.

The anterior-posterior and dorsal-ventral progression of heart organogenesis is well illustrated by the patterning and activity of two members of different families of cell adhesion molecules: the calcium-dependent cadherins, specifically N-cadherin, and the extracellular matrix glycoproteins, fibronectin. N-cadherin by its binding to the intracellular molecule beta-catenin and fibronectin by its binding to integrins at focal adhesion sites, are involved in regulation of gene expression by their association with the cytoskeleton and through signal transduction pathways. The ventral precardiac mesoderm cells epithelialize and become stably committed by the activation of these cell-matrix and intracellular signaling transduction pathways. Cross talk between the adhesion signaling pathways initiates the characteristic phenotypic changes associated with cardiomyocyte differentiation: electrical activity and organization of myofibrils. The development of both organ form and function occurs within a short interval thereafter. Mutations in any of the interacting molecules, or environmental insults affecting either of these signaling pathways, can result in embryonic lethality or fetuses born with severe heart defects. As an example, we have defined that exposure of the embryo temporally to lithium during an early sensitive developmental period affects a canonical Wnt pathway leading to beta-catenin stabilization. Lithium exposure results in an anterior-posterior progression of severe cardiac defects.     

Chemopreventive effects of natural dietary compounds on cancer development

Chemoprevention, a relatively new and promising strategy to prevent cancer, is defined as the use of natural dietary compounds and/or synthetic substances to block, inhibit, reverse, or retard the process of carcinogenesis. The chemopreventive effects elicited by these natural dietary compounds are believed to include antioxidative, anti-inflammatory activity, induction of phase II enzymes, apoptosis, and cell cycle arrest. Many mechanisms have been shown to account for the anticarcinogenic actions of natural dietary compounds; attention has recently been focused on intracellular-signaling cascades as common molecular targets for various chemopreventive natural dietary compounds. In this critical review, we will summarize current knowledge on natural dietary compounds that act through the signaling pathways and modulate gene expression to induce detoxifying enzymes, programmed cell death, anti-inflammatory, and anti-proliferative effects, thus providing evidence for these substances in cancer chemopreventive action (128 references).     

Phenolic compounds in Brassica vegetables

Phenolic compounds are a large group of phytochemicals widespread in the plant kingdom. Depending on their structure they can be classified into simple phenols, phenolic acids, hydroxycinnamic acid derivatives and flavonoids. Phenolic compounds have received considerable attention for being potentially protective factors against cancer and heart diseases, in part because of their potent antioxidative properties and their ubiquity in a wide range of commonly consumed foods of plant origin. The Brassicaceae family includes a wide range of horticultural crops, some of them with economic significance and extensively used in the diet throughout the world. The phenolic composition of Brassica vegetables has been recently investigated and, nowadays, the profile of different Brassica species is well established. Here, we review the significance of phenolic compounds as a source of beneficial compounds for human health and the influence of environmental conditions and processing mechanisms on the phenolic composition of Brassica vegetables.     

Expression of MMP2 and MMP9 (Gelatinases A and B) in Human Colon Cancer Cells

Matrix metalloproteinases (MMPs) are a family of zinc-dependent neutral endopeptidases, collectively capable of degrading essentially all matrix components. Elevated levels of distinct MMPs are detected in tumor tissue or serum of patients with advanced cancer, and they are the major prognostic indicators in cancer. Inhibition of MMPs has been explored as a therapeutic goal for almost two decades. Nitric oxide (NO), a free radical plays an important role in signaling pathways in regulation of MMP expression. In the present study, we demonstrated the role of exogenous NO levels in the regulation of MMP2 and MMP9 (gelatinases A and B) in colon cancer cell line WiDr and its inhibition with emodin (a naturally occurring anthraquinone).     

UPLC‐ESI/MS determination of 17 active constituents in two categorized formulas of traditional Chinese medicine,Sanhuang Xiexin Tang and Fuzi Xiexin Tang: application in comparing the differences in decoctions and macerations

A rapid and sensitive UPLC‐ESI/MS method was established and validated to determine 17 active constituents (aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, benzoylmesaconine, berberine, palmatine, jatrorrhizine, coptisine, baicalein, wogonin, baicalin, wogonoside, emodin, aloe‐emodin and rhein) in Sanhuang Xiexin Tang (SXT) and Fuzi Xiexin Tang (FXT), which are two classic compound recipes from Xiexin Tang categorized formulas in traditional Chinese medicien. The separation was performed on a UPLC BEH C₁₈ column gradient eluted using acetonitrile and 0.1% formic acid as mobile phase. ESI/MS was operated in positive ([M + H]⁺) in selected ion recording mode for analysis of alkaloids and flavones, while in negative ([M ? H]^?) selected ion recording mode for anthraquinones. All of the 17 constituents exhibited good linearity in a relatively wide concentration ranges with the lowest limits of detection of 0.38 ng/mL. All of the relative standard deviation values of intra‐ and inter‐precisions and stabilities of 17 constituents were within 5%. The method was successfully applied to determine 17 active constituents in decoctions and macerations of SXT and FXT. The results indicated that different preparative methods resulted in significant diversity in concentrations of the 17 analytes. Herb–herb interaction appeared between aconitum alkaloids in Aconiti Lateralis Radix Preparata and another three herbs. Copyright © 2013 John Wiley & Sons, Ltd.     

固相萃取/液相色谱-质谱/质谱法测定山银花中5种主要黄酮苷元的含量

该文通过含有盐酸的乙醇溶液回流水解并提取,HLB固相萃取柱净化,液相色谱-质谱/质谱法检测,建立了山银花中槲皮素、木犀草素、山萘酚、芹菜素和黄芩素5种黄酮苷元含量的测定方法。实验以芦丁、木犀草苷、紫云英苷、野漆树苷和黄芩苷5种黄酮苷为代表开展研究,山银花样品经50%的乙醇溶液(含10%浓盐酸)回流2 h水解黄酮苷,同时对黄酮苷元进行提取,HLB固相萃取柱净化,采用Mightysil RP-18色谱柱分离,液相色谱-质谱/质谱法检测(电喷雾离子源、多反应监测模式、负离子扫描),外标法定量测定水解后的5种黄酮苷元含量。方法的定量下限(S/N=10)为0.005 g/kg(槲皮素),0.01 g/kg(木犀草素和芹菜素)和0.05 g/kg(山萘酚和黄芩素)。在0~1.0 g/kg范围内,5种黄酮苷元的线性相关系数均大于0.995;在山银花样品中对待测物进行3种加标水平的回收实验(加标水平相当于水解后槲皮素和木犀草素含量为:0.10、0.20、0.40 g/kg,山萘酚、芹菜素和黄芩素含量为:0.05、0.10、0.20 g/kg),方法的平均回收率70.4%~104%;相对标准偏差为4.0%~12%。该方法实现了山银花中多种主要黄酮苷元含量的同时测定,且对研究山银花药效及与黄酮类化合物的关系具有重要意义。     

Advanced development in analysis of phytochemicals from medicine and food dual purposes plants used in China

The concept of "Let food be thy medicine and medicine be thy food" was widely accepted for thousand years. It is now well known that some foods and food components have beneficial physiological and psychological effects. In China, the Ministry of Health of the People's Republic of China announced 87 items of materials, including 81 plants, could be used as both medicine and food. Increasing consumer demand for safety and health benefits, it is of critical importance to have high-quality and comprehensive data on bioactive compounds (phytochemicals) in these materials. In this review, we summarized the advanced development (2006-2010) in analysis of phytochemicals from medicine and food dual purposes plants (MFDPP) used in China.     

Cell integrity signaling and response to stress in fission yeast

Cellular responses to external signals are regulated by conserved mitogen-activated protein (MAP) kinase signaling cascades. These pathways are triggered by a vast range of stimuli. They phosphorylate numerous proteins, produce significant changes in the gene expression, and regulate diverse processes ranging from proliferation and differentiation to apoptosis in all eukaryotic cells. Three conserved MAP kinase signaling pathways have been identified in the fission yeast Schizosaccharomyces pombe. In this article, we present an overview of two of those pathways that regulate the response of fission yeast to stress and maintain cell integrity. The structure of these signaling modules and the function of the pathways, including the regulation by endogenous inhibitors, are discussed.     

Engineering modular protein interaction switches by sequence overlap

Many cellular signaling pathways contain proteins whose interactions change in response to upstream inputs, allowing for conditional activation or repression of the interaction based on the presence of the input molecule. The ability to engineer similar regulation into protein interaction elements would provide us with powerful tools for controlling cell signaling. Here we describe an approach for engineering diverse synthetic protein interaction switches. Specifically, by overlapping the sequences of pairs of protein interaction domains and peptides, we have been able to generate mutually exclusive regulation over their interactions. Thus, the hybrid protein (which is composed of the two overlapped interaction modules) can bind to either of the two respective ligands for those modules, but not to both simultaneously. We show that these synthetic switch proteins can be used to regulate specific protein-protein interactions in vivo. These switches allow us to disrupt an interaction with the addition or activation of a protein input that has no natural connection to the interaction in question. Therefore, they give us the ability to make novel connections between normally unrelated signaling pathways and to rewire the input/output relationships of cellular behaviors. Our experiments also suggest a possible mechanism by which complex regulatory proteins might have evolved from simpler components.