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Phthalates are commonly used in plastic products in daily life. The endocrine-disrupting effects of phthalates have been widely reported. Accumulating evidence from human cohorts and lab animals indicate exposure to phthalates might impair neurodevelopment. However, the direct causal relationship and mechanism between phthalates with neurodevelopment and neurotoxicity have not been firmly established. We found that phthalates (i.e. DBP, DINP, BBP) disrupted the expression of estrogen receptors (esr1, esr2a, esr2b), and impaired neurogenesis in the brain of zebrafish during embryonic development. Moreover, the abnormal expression of estrogen receptors, especially esr2a, was partly rescued in zebrafish which exposed to phthalates, with the estrogen receptor antagonist tamoxifen. Hence, impaired neurogenesis of zebrafish exposed to phthalates was partly reversed by tamoxifen treatment. Moreover, our results show that induced pluripotent stem cells (iPSC)-derived human neurons exposed to phthalates triggered double-strand DNA breaks in vitro. Overall, this study demonstrates that exposure to phthalates affects neurodevelopment in zebrafish embryos and induces neurotoxicity in human neurons partly through disrupting the expression of estrogen receptors.

Lycium barbarum fruit (Goji or wolfberry) is widely used as a functional food as well as an herb in Chinese medicine. The polysaccharides of L. barbarum (LBPs) are well-known for immunomodulation and antioxidant activities. This study evaluated the yields, properties and bioactivities of LBPs extracted from L. barbarum fruit by different methods including hot water (HWE 100 °C), ultrasonic water (UWE 30–40 °C), subcritical water (SWE 110 °C), and ultrasound-enhanced subcritical water extraction (USWE 110 °C). The total LBP yield (80 min extraction followed by ethanol precipitation) was the highest from USWE (14% w/w) and the lowest from HWE (7.6%). The LBP from USWE also showed the highest protein (27.9%) and phenolic content (5.3%, w/w), and the highest antioxidant activities (scavenging radicals by DPPH and trolox equivalent antioxidant capacity assays; ferric reducing power). It also showed the highest immunoactivities in activating the phagocytosis and NO production of RAW264.7 macrophages. USWE was proven the most efficient method for extraction of LBPs. Temperature and ultrasound were two chief factors affecting the extraction yield, chemical properties and bioactivities of LBP.

Background The role of Kdr (VEGFR-2/Flk-1) in vascular formation has been well described, but the role of Flt1 (VEGFR-1) is not well studied and is generally considered as a decoy receptor for trapping VEGF. Methods The effects of VEGFR1/2 kinase inhibitor (VRI) and calycosin on Flt1 tyrosine kinase (TK) activity were evaluated by molecular docking, enzymatic inhibition assay, protein co-immunoprecipitation and siRNA gene knock-down analysis in HUVECs. Toxicities of the chemicals were examined using HUVECs viability. Their effects on angiogenesis and vessel formation were furthered studied in HUVECs in vitro and Tg ( fli-1 : EGFP ) zebrafish in vivo . The gene and protein expression of VEGF and VEGF receptors were investigated by quantitative RT-PCR and Western blot. Results VRI strongly inhibited physiological functions of both VEGF receptors and suppressed endothelial cell survival. This resulted in blood vessel loss in zebrafish embryos. Interestingly, calycosin co-treatment impeded VRI-induced blood vessel loss. Docking and kinase inhibition assay revealed that calycosin competed with VRI for the tyrosine kinase domain of Flt1 without affecting ATP binding. On the contrary, calycosin did not affect the interaction between VRI and Kdr-TK. Consistent with these results, calycosin counteracted the inhibition of Flt1-TK and PI3K phosphorylation induced by VRI in HUVECs. Further studies in vitro and in vivo showed that the minimizing effect of calycosin on VRI-mediated endothelial cytotoxicity was blocked by wortmannin (a PI3K inhibitor). The impeding effect of calycosin on VRI-induced blood vessel loss was absent in zebrafish embryos injected with Flt1 MO. Conclusions Flt1-tyrosine kinase (TK) activity contributed significantly in endothelial cells survival and vascular development during embryo angiogenesis in zebrafish by engaging PI3K/Akt pathway. General significance The roles of Flt1 activity in endothelial cell survival in physiological vascular formation may have been previously under-appreciated.

Crotamine is defensin-like cationic peptide from rattlesnake venom that possesses anticancer, antimicrobial, and antifungal properties. Despite these promising biological activities, toxicity is a major concern associated with the development of venom-derived peptides as therapeutic agents. In the present study, we used zebrafish as a system model to evaluate the toxicity of rhodamine B-conjugated (RhoB) crotamine derivative. The lethal toxic concentration of RhoB-crotamine was as low as 4 μM, which effectively kill zebrafish larvae in less than 10 min. With non-lethal concentrations (

In this study, seven strains of bacteria with polyhydroxyalkanoates (PHA)-producing ability (i.e. Bacillus cereus, Pseudomonas putida, Bacillus pumilus, Pseudomona huttiensis, Yersinia frederiksenii, Aeromonas ichthiosmia, and Sphingopyxis terrae) were isolated from various waste treatment plants in Hong Kong. Simultaneous wastewater treatment and PHA accumulation were successfully achieved in the bioreactors using isolated bacteria from different sludges. At the organic loading less than 13,000 ppm, more than 95% of chemical oxygen demand (COD) was removed by the isolated strains before the decrease of PHA accumulation. In addition, more than 95% of nitrogen removal was achieved by all isolated strains. In the bioreactors inoculated with single strains, the highest yields of poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxyvalerate) (PHV) were obtained in A. ichthiosmia (84 mg PHB/g) and B. cereus (69 mg/g), respectively. For the mixed culture, the highest yields of PHB and PHV were increased by 55% and 45% in the system inoculated with B. pumilus and A. ichthiosmia. The biologically synthesized PHA also showed the potential applications in drug delivery and tissue engineering. PHA-nanoparticles loaded with pyrene were successfully prepared by recombinant Escherichia coli. The results of in vitro drug release and biocompatibility tests revealed that nanoparticles could be used as safer dray carriers with high loading capacity and efficiency. After 20 days, the cells successfully grew on 90% of the PHA-aortic valve.

Ethnopharmacological relevance Fritillariae Bulbus (Beimu in Chinese) is derived from the bulbus of many Fritillaria species (family Liliaceae), which has been used as an antitussive herb in traditional Chinese medicine for more than 2000 years. Due to the complexity of plant origins and significant variations in chemical profiles, the characterization of the profile of the major bioactive constituents and its association with pharmacological activity are important for the quality control of Beimu herbs from different origins. Aim of the study This study aims to investigate the distribution of major bioactive isosteroidal alkaloids in Beimu herbs of different origins and its correlation with the tracheobronchial relaxant activity. Methods Quantification of 7 main bioactive 5α-cevanine isosteroidal alkaloids, including ebeiedine, ebeiedinone, hupehenine, isoverticine, verticine, verticinone and imperialine, in 23 Fritillaria species was performed using gas chromatography. The relaxant effect of different extracts of 4 commonly used Beimu herbs, namely Zhe-Beimu (F. thunbergii Miq.), Chuan-Beimu (F. cirrhosa D. Don), Hubei-Beimu (F. hupehensis Hsiao et K. C. Hsia) and Yi-Beimu (F. pallidiflora Schrenk), was evaluated using rat isolated tracheal and bronchial preparations pre-contracted with carbachol, the well established in vitro antitussive model. Results Amongst 23 Fritillaria species detected, significant variations of the types and quantities of 7 major isosteroidal alkaloids were determined, which served as an important indicator for the classification of different Beimu herbs with distinct geographic distributions. Based on the type and quantity of these alkaloids, different origins of Beimu could be clearly clustered into several subgroups by principal component analysis. Furthermore, both crude alkaloid and water extracts of all 4 Beimu herbs showed a dose-dependent tracheobronchial relaxation with different potencies. The total content of alkaloids (weight adjusted based on the activity of individual alkaloids) in Beimu extracts significantly correlated with their tracheobronchial relaxation effects (r2 > 0.9, p Conclusions The results demonstrated that the differences in chemical profile of major bioactive isosteroidal alkaloids and pharmacological activity of Beimu could be incorporated into a simple and unified method for quality control and potential prediction of activity of Beimu herbs from different origins.

Panax notoginseng saponins (PNS) are major active constituents of P. notoginseng. This study investigated the mechanisms of the hypocholesterolaemic and atheroscleroprotective effects of PNS. It was found that treatment of PNS (30 or 100 mg/kg/day, p.o.) could significantly reduce elevated serum total cholesterol, triacylglycerol, low-density lipoprotein cholesterol and atherogenic index induced by feeding rats with high cholesterol diet for 28 days. High-density lipoprotein cholesterol could also be increased by PNS treatment. Additionally, PNS significantly enhanced the hypercholesterolaemia-induced reduction of superoxide dismutase activity in the liver. In hypercholesterolaemic rats, PNS significantly improved the endothelium-dependent vasodilatation, acetylcholine-induced nitric oxide production and endothelial nitric oxide synthase mRNA expression in a dose-dependent manner. It also significantly reversed the hypercholesterolaemia-induced changes in mRNA expressions of cholesterol 7α-hydroxylase and peroxisome proliferator-activated receptor α but not HMG-CoA reductase. The findings provided evidence for using PNS to prevent the development of hypercholesterolaemia and atherosclerosis. The hypocholesterolaemic activity of PNS is likely to be resulted from inducing the biosynthesis of bile acids from cholesterol and promoting the β-oxidation of fatty acids in the liver. The vasoprotective function of PNS may be brought about by increasing the gene expression of eNOS in endothelial cells, its antioxidative activity, or both.

Consumption of functional foods for managing plasma cholesterol level has gained acceptance globally. The hypocholesterolaemic and vascular protective effects of the dried fruit of Crataegus pinnatifida, hawthorn (Shan Zha), were investigated in rats fed with normal diet, high cholesterol diet (HCD) or HCD plus Shan Zha 80% ethanolic extract treatment (30 or 100 mg/kg/day, p.o.) for 4 weeks. Shan Zha extract markedly reversed the increased plasma total cholesterol and high density lipoprotein cholesterol induced by HCD with a dose-dependent improvement on the atherogenic index. It also demonstrated good hepatoprotective function by reducing lipid content in the liver. The blunted endothelium-mediated aortic relaxation in HCD-fed rats was restored by high dosage of Shan Zha extract treatment. The current results showed that Shan Zha extract could provide its cholesterol lowering effect by up-regulating hepatic CYP7A1 mRNA expression which leads to enhanced bile acid biosynthesis. It is postulated that the hypocholesterolaemic effect is the primary beneficial effect given by Shan Zha extract; it then leads to other secondary beneficial effects such as vascular protective and hepatoprotective functions. Thus, Shan Zha extract could provide an overall improvement on the hepatic and vascular systems that may be important in relieving hypercholesterolaemia-related complications.

Gui-ling-gao (GLG), also known as turtle jelly, is a popular medicinal Chinese health/functional food prepared from several traditional Chinese medicinal herbs. The present study aimed at examining and explaining the anti-inflammatory properties of GLG by using lipopolysaccharide (LPS)-induced inflammation in splenocytes isolated from BALB/c mice. Additionally, the effects of GLG on mRNA and protein expressions of inducible nitric oxide synthase (iNOS) and pro-inflammatory cytokines in LPS-stimulated splenocyte proliferation were evaluated by real-time RT-PCR and Western blot assays. We demonstrated that GLG significantly inhibited LPS-induced splenocyte proliferation in a concentration-dependent manner. LPS-mediated up-regulations in the gene and protein expressions of iNOS, interleukin-1β (IL-1β) and tumour necrosis factor-alpha (TNF-α) were suppressed by GLG. These findings suggest for the first time that GLG exerts anti-inflammatory effects by inhibiting the expression of iNOS and pro-inflammatory cytokines such as IL-1β and TNF-α. The current study provides strong scientific evidence for the health-beneficial claim that consumption of GLG could help the body to modulate the immune system.

The dried ripe seed ofRaphanus sativusL., commonly known as radish seed (or Raphani Semen), is used as traditional Chinese medicine (TCM) to treat constipation, chronic tracheitis, and hypertension. The major active compounds in Raphani Semen are alkaloids, glucosinolates, brassinosteroids, and flavonoids. Fatty acids are its main nutritional contents. Raphani Semen has been demonstrated to have beneficial effects on hypertension, obesity, diabetes mellitus, constipation, and cough. So far, there is no report about the adverse/toxic effects of this herb on humans. However, Raphani Semen processed by roasting was reported to exhibit some adverse effects on mice. Additionally, erucic acid, the main fatty acid in Raphani Semen, was shown to enhance the toxicity of doxorubicin. Thus, Raphani Semen has a potential risk of causing toxicity and drug interaction. In summary, Raphani Semen is a valuable TCM herb with multiple pharmacological effects. More studies on Raphani Semen could help better understand its pharmacological mechanisms so as to provide clear scientific evidence to explain its traditional uses, to identify its therapeutic potential on other diseases, and to understand its possible harmful effects.

Background This study reported a comprehensive approach (comparing the extraction yields, chemical profiles, antioxidant properties and CYP450-inhibitory effects) to evaluated the effectiveness of various extraction methods [microwave-assisted extraction using water (MAE-W), heat reflux extraction using water (HRE-W), ultrasonic extraction using 70 % ethanol and ultrasonic extraction using ethanol (UE-E)] for Huang-Qin (HQ), the dried root of Scutellaria baicalensis Georgi. Results The HQ extraction efficiency by MAE-W was the best. The chemical profiles of extracts obtained using HRE-W and MAE-W were similar; whereas more flavones but less flavone glycosides were detected in the UE-E extract. There was no difference in the antioxidant properties among different extracts. In vitro human liver microsome assays illustrated that all extracts possessed herb–drug interaction potentials but the UE-E extract are shown with a potent interaction with CYP3A4-metabolized drugs. Conclusion MAE-W is a favorable method for the preparation of HQ extracts based on extraction yield, pharmacological properties and safety.

The prevalence of cognitive impairment and dementia caused by cerebrovascular disease is likely to increase with the global aging population. Vascular contributions to cognitive impairment and dementia (VCID) is a wide spectrum term used to include a diverse heterogeneous group of cognitive syndromes with vascular factors regardless of the cause of pathogenesis. VCID ranges from mild cognitive impairment to full-blown dementia with vascular dementia (VaD) as the most severe stage. It is further complexed by the coexistence of other forms of dementia such as Alzheimer’s disease (AD). Recent researches in the functions of the neurovascular unit (NVU) suggest that dysfunction of the NVU might be the cause of primary vascular events in the brain that leads to further neurodegeneration. In this review, we have briefly summarized various forms of VCID. There is currently no standard therapy for VCID or dementia. Given the fact that Traditional Chinese Medicine (TCM) has gained popularity worldwide, we also reviewed recent scientific and clinical findings on various antidementia TCM for the treatment of VCID, includingSalvia miltiorrhiza, Huperzia serrata, Ligusticum chuanxiong, Ginkgo biloba, Panax ginseng,and also TCM formula Sailuotong capsule (SLT) and Fufangdanshen tablets (FFDS).

Background Cycloartane triterpenoids exhibited anticancer effects. This study aims to identify any potential novel anticancer cycloartane triterpenoids from Cimicifuga foetida L. rhizome (Sheng ma) and the mode of actions. Methods Cycloartane triterpenoids were isolated from the C. foetida rhizome by a series of column chromatography and identified by IR, MS and NMR. Their anticancer effects on several human cancer cell lines, MCF-7, HepG2, HepG2/ADM, HeLa, and PC3, and normal human mammary epithelial cells MCF10A were investigated by colony formation and MTT assays. Morphological analysis of apoptosis induction was performed by acridine orange/ethidium bromide dual-staining and Hoechst 33258 nuclear staining. The cell-cycle profile and annexin V staining were evaluated by flow cytometry. Apoptosis were investigated by measuring changes in mitochondrial membrane potential and analyzing expression of cell cycle- and apoptosis-related proteins in MCF-7 cells by Western blotting. Results A novel cycloartane triterpenoid, 25-O-acetyl-7,8-didehydrocimigenol-3-O-β-d-(2-acetyl)xylopyranoside (ADHC-AXpn), together with the known 7,8-didehydrocimigenol-3-O-β-d-xylopyranoside (DHC-Xpn) were isolated. MCF-7 growth was significantly inhibited by ADHC-AXpn in a dose- and time-dependent manner (IC50: 27.81 µM at 48 h; P = 0.004 vs. control at 25 μM for 48 h treatment), and ADHC-AXpn was selectively cytotoxic for cancerous cells (MCF-7, HepG2/ADM, HepG2 and HELA cells) based on its higher IC50 values for normal cells MCF10A (IC50: 78.63 µM at 48 h) than for tumor cells. In MCF-7 cells, ADHC-AXpn induced G2/M cell cycle arrest by mediating cyclin-B1, and CDK1 and its phosphorylation; and induced apoptosis through the mitochondrial-mediated apoptotic pathway, with inhibition of Akt activation. As ADHC-AXpn suppressed phosphorylation of ERK1/2, Raf and Akt proteins in MCF-7 cells, its apoptotic effect might be associated with Raf/MEK/ERK signaling and Akt activation. Conclusions ADHC-AXpn significantly suppressed the growth of MCF-7 cells, induced mitochondrial apoptosis and cell-cycle arrest, and inhibited Raf/MEK/ERK signaling pathway and Akt phosphorylation.

An extract of Curcuma longa was tested in hypercholesterolemic rats to investigate its potential therapeutic effect on vascular conditions. Four experimental groups were used: normal diet (ND) control group, high cholesterol diet (HCD) group, and HCD subgroups supplemented with turmeric extract at 100 or 300 mg/kg of body weight (HCD100Tur and HCD300Tur groups, respectively). Turmeric extract was fed orally to animals, and dietary treatments lasted for 28 days. Hypercholesterolemia developed in the HCD, HCD100Tur, and HCD300Tur rats. Segments of the thoracic aorta were isolated, and an organ bath experiment was used to assess the vasorelaxation capability among all rats. Rats fed only HCD showed a marked decrease in acetylcholine-induced vasorelaxation compared with ND control rats. The HCD100Tur and HCD300Tur rats showed significant improvement in vasorelaxation compared with HCD rats. When vasorelaxation was induced by high concentrations of sodium nitroprusside, no differences in vasorelaxation were observed among the four groups of rats. A mechanistic study showed that HCD100Tur and HCD300Tur rats had significantly higher levels of the antioxidant enzymes superoxide dismutase and glutathione peroxidase than HCD rats. The transcript levels of heat shock protein 70 (hsp70), bcl2, bax-α, caspase (casp3), and glyceraldehyde 3-phosphate dehydrogenase in aortic tissues indicated that hypercholesterolemia significantly increased the expression of bax-α and casp3 but down-regulated bcl2 expression compared with the control group. Turmeric increased the expression of hsp70 and bcl2 but greatly reduced casp3 expression, indicating that turmeric improves vasorelaxation of the aorta in hypercholesterolemic rats by increasing antioxidant enzyme activities and likely suppressing apoptosis.

Ischemic heart disease is a major cause of death in the world. Common therapies, such as primary coronary angioplasty and thrombolysis, are applied to restore blood supply to the heart, limit infarct size and reduce mortality. However, the restoration of blood supply would generate reactive oxygen species in damaged sites of the myocardium, intensifying the damage to the cardiac tissues. Radix Scutellariae baicalensis (Huangqin) is a well-known herb in traditional Chinese medicine with high antioxidant power. In this study, extract of the dry root of Scutellaria baicalensis Georgi (Sb) was confirmed to have a high content of flavonoids and phenolic compounds. The cardioprotective effects of the Sb extracts (3, 30 and 300 mg/kg) were evaluated in myocardial ischemia-reperfusion injuried rats. The results showed that animals that had received five-day pretreatment of the Sb extract (30 mg/kg) had a significant reduction in myocardial infarct size and a marked increase in the activity of catalase in the liver. The Sb extract could additionally enhance acetylcholine-induced vasorelaxation. It was proposed that the Sb extract exerted its cardioprotection by stimulating the catalase activity and improving vascular elasticity.

Arctium lappa, commonly known as burdock, is being promoted/recommended as a healthy and nutritive food in Chinese societies. Burdock has been used therapeutically in Europe, North America and Asia for hundreds of years. The roots, seeds and leaves of burdock have been investigated in view of its popular uses in traditional Chinese medicine (TCM). In this review, the reported therapeutic effects of the active compounds present in the different botanical parts of burdock are summarized. In the root, the active ingredients have been found to "detoxify" blood in terms of TCM and promote blood circulation to the skin surface, improving the skin quality/texture and curing skin diseases like eczema. Antioxidants and antidiabetic compounds have also been found in the root. In the seeds, some active compounds possess anti-inflammatory effects and potent inhibitory effects on the growth of tumors such as pancreatic carcinoma. In the leaf extract, the active compounds isolated can inhibit the growth of micro-organisms in the oral cavity. The medicinal uses of burdock in treating chronic diseases such as cancers, diabetes and AIDS have been reported. However, it is also essential to be aware of the side effects of burdock including contact dermatitis and other allergic/inflammatory responses that might be evoked by burdock.

The hypocholesterolemic and atheroscleroprotective potentials of dietary consumption of hawthorn (dried fruit of Crataegus pinnatifida , Shan Zha) were investigated by monitoring plasma lipid profiles and aortic relaxation in Sprague–Dawley rats fed with either normal diet, high-cholesterol diet (HCD) or HCD supplemented with hawthorn powder (2%, w/w) (4 weeks). In HCD-fed rats, an increased plasma total cholesterol and LDL-cholesterol with a decreased HDL-cholesterol was observed, and consumption of hawthorn markedly suppressed the elevated total cholesterol and LDL-lipoprotein levels plus an increased HDL-cholesterol level. The blunted acetylcholine-induced, endothelium-dependent relaxation of isolated aortas of HCD-fed rats was improved by hawthorn. The development of fatty liver, an increased nitric oxide synthase (NOS) activity and an elevated oxidative stress (as estimated by the attenuated levels of anti-oxidant enzymes) associated with HCD were attenuated by hawthorn. Thus, the results demonstrated that hawthorn consumption provides overall beneficial effects on reversing HCD associated detrimental changes.

In Chinese communities, regular consumption of Chinese-medicated diets (CMD) (usually in the form of soup) is a traditional practice to promote health and prevent disease development. The overall improvement of health conditions is believed to be correlated with the anti-oxidant potentials of these herbs. Huangqin, roots of Scutellaria baicalensis Georgi (Lamiaceae), is one of the herbs commonly used in CMD. In this study, the anti-oxidant capacities of Huangqin extracts (water, ethanol and ether extracts) were evaluated and compared to commonly used CMD herbs, Heshouwu, roots of Polygonum multiflorum Thunb (Polygonaceae) and Renshen (or Ginseng), roots of Panax ginseng CA Meyer (Araliaceae). The anti-oxidant capacities were measured by using both cell-free assay [ferric reducing/anti-oxidant power (FRAP)] and biological methods [2,2'-azobis–(2-amidinopropane) (AAPH)-induced haemolysis assay and H2O2 -induced cell damage on H9C2 cells]. Additionally, the total phenolic content was measured using Folin-Ciocalteu methods. Water extract of Huangqin has the highest anti-oxidant activities compared to the ethanol and ether extracts. A positive relationship between the anti-oxidant effects and total phenolic contents of extracts was demonstrated. This shows that Huangqin could be an effective dietary anti-oxidant that can be consumed regularly as a functional food for the prevention of oxidant/free radical-related diseases.

Scutellaria baicalensis Georgi (Labiatae) (“huang qin”), Ligusticum chuanxiong Hort. (Umbelliferae) (“chuang xiong”), Panax notoginseng (Burk.) F.H. Chen (Araliaceae) (“san qi”), Uncaria rhynchophylla (Miq.) Jackson. (Rubiaceae) (“gou teng”), Rhokiola rosea L. (Crassulaceae) (“hong jing tian”) and Stephania tetrandra S. Moore (Menispermaceae) (“fang ji”) are commonly used traditional Chinese medicine (TCM) for hypertensive patients. The pharmacologically active compounds found in these TCMs are baicalin, ligustrazine, notoginsenoside R1, rhynchophylline, salidroside and tetrandrine, respectively, which possess antihypertensive properties with diverse cellular mechanisms. In this study, we attempted to evaluate a possible correlation of the antioxidative activities (using the DPPH assay) and the vaso-relaxation effects (using rat isolated thoracic aorta) of these compounds. In the antioxidative study, a relative order of free radical scavenging capacity (SR%) of baicalin ≥ tetradrine >> salidroside ≥ ligus...

We investigated the role(s) of monoamine oxidases (MAOs) on the altered 5-hydroxytryptamine (5-HT, serotonin)-induced tension development of the isolated umbilical artery of preeclamptic pregnancy of Chinese women. An enhanced 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy was observed when compared with that of normal pregnancy. The enhanced component of 5-HT-induced tension development was eradicated by clorgyline (a MAO-A inhibitor). Blockade of eNOS (endothelial isoform nitric oxide synthase) (N(omega)-nitro-L-arginine methyl ester), 5-HT transporter (citalopram), 5-HT receptor subtypes (5HT2B, SB 204741; 5-HT2C, RS 102221; 5-HT7, SB 269970), and endothelium denudation of the umbilical artery of normal pregnancy mimicked the enhanced 5-HT-induced tension development as observed in the preeclamptic tissues. In contrast, no apparent changes in 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy were observed with the same pharmacological manipulations. A decreased protein expression levels of MAO-A and eNOS (no iNOS and MAO-B expression was detected) and no change in caveolin-1 and 5-HT transporter expression were demonstrated in the umbilical artery (endothelium intact) lysate of preeclamptic pregnancy, compared to that of the umbilical artery of normal pregnancy. Thus, in the umbilical artery of preeclamptic pregnancy, a decrease of MAO-A and eNOS protein expression levels are probably associated with, or responsible for, the exaggerated 5-HT-induced tension development.

Sequential extraction with petroleum ether, chloroform–ethyl acetate, methanol, and water was carried out on 40 commonly used Chinese medicinal herbs. The antioxidant power of different fraction extracts was measured with both the DPPH assay and the FRAP assay. Ascorbic acid and butylated hydroxytoluene were used as two reference antioxidants. The total phenolic content of the extracts was measured by Folin-Ciocalteu method. The fraction extracts showed different antioxidant activity in the two assays; however, there was a significant positive correlation between the DPPH assay and the FRAP assay. Methanol fraction extracts generally demonstrated stronger antioxidant power and higher total phenolic content than did the others. The results suggest that Radix Sanguisorbae, Cortex cinnamomi, Herba taxilli, Semen arecae, and L., Cinnamomum cassia Presl ., Taxillus sutchuenensis (Lecomte) Danser, Areca catechu Linn. and Scutellaria baicalensis Georgi contain relatively high values in antioxidant power ...

A high performance liquid chromatographic method was developed and applied to the simultaneous determination of puerarin and danshensu in a traditional Chinese medicinal preparation containing Pueraria lobata and Salvia miltiorrhiza as its major herbal constituents. Elution was performed on a C18 column at 27°C with acetonitrile–water (0.5% acetic acid) at different proportions according to a time scheduled programme, and pumped at a flow rate of 1.0 mL/min. Indomethacin was used as an internal standard. High resolution was obtained between the analytes and the internal standard. The intra‐day and inter‐day precisions were better than 1.9% and 2.2%, respectively. The coefficient of determination for each analyte was above 0.997. It is a simple and sensitive analytical method with good accuracy and reproducibility.

Lobeliae chinensis. Herba (“ban bian lian”), Rheum officinale. Baill. (“da huang”), Sanguisorba officinalis. Linn. (“di yu”), Agrimonia pilosa. Ledeb. (“xian he cao”), and Paris polyphylla. Smith (“zhi hua tou”) are well-known traditional Chinese medicines. They are commonly used in traditional Chinese medicine formulae against cancer. In this study, the antioxidant and anticancer effects of water extracts of these herbs were investigated. In the antioxidant and anticancer studies, water extracts of di yu, xian he cao, and da huang were show to be the most antioxidative and had the highest growth inhibitory effect on human lung adenocarcinoma A549 cell and human breast cancer MCF-7 cell. By comparing their percentage free radical scavenging capacity (SR%) and percentage growth inhibition on A549 and MCF-7 cells, a positive linear relationship between antioxidant activity and anticancer effect of the five herbal water extracts was found. This suggested that the antioxidants of the herbal water extr...

Natural products are good sources of natural dietary antioxidants that are believed to protect the body against hepatotoxic effect induced by oxidative stress. Hedyotis diffusa Willd (Rubiaceae) (HDW) is a traditional Chinese medicinal herb that has been shown to possess a variety of antioxidant properties.The present study examines and explains the cell protective property of HDW water extract (WEHDW).2,2-Diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH) assay was used to measure the free radical scavenging property of WEHDW (0.001-10 mg/mL). The protective effect of WEHDW (0.3-10 mg/mL 2 h pretreatment) against hydrogen peroxide (H2O2, 200 μM for 6 h) induced cytotoxicity in human hepatic cells, LO2, was evaluated using cell viability assay and nuclear staining. The molecular pathway of WEHDW's effect was investigated by using Western blot assay.WEHDW had a 50% scavenging concentration (SC50) at 0.153 mg/mL in the DPPH assay. Exposure of LO2 cells to H2O2 resulted in apoptosis which could be markedly attenuated by pre-treating WEHDW in a concentration-dependent manner (0.5, 1, 3, 5, or 10 mg/mL) (all with p 0.001, versus control). Moreover, Hoechst (nuclear) staining showed that 1 mg/mL WEHDW could protect LO2 cells by attenuating apoptotic cell death mediated by H2O2. It was found that WEHDW reversed H2O2-induced activation of MEK/ERK pathway and H2O2-induced inhibition of P13-K/AKT/GSK3β pathway in LO2 cells.WEHDW may help to improve the antioxidant defense system, resulting in prevention of oxidative stress-related fatty liver diseases.

Ethnopharmacological relevance The fruit of Schisandra chinensis (Turcz.) Baill, has been traditionally used in management of liver diseases and ageing associated neurodegeneration. The bioactive compound from this medicinal plant would be valuable for its potential use in prevention and treatment of Parkinson׳s disease. Aim of the study The overall objective of the present study was to understand the neuroprotective effect of schisantherin A, a dibenzocyclooctadiene lignan from the fruit of S. chinensis (Turcz.) Baill, and to elucidate its underlying mechanism of action. Material and methods This study investigated the protective effect of schisantherin A against selective dopaminergic neurotoxin 6-hydroxydopamine (6-OHDA)-induced neural damage in human neuroblastoma SH-SY5Y cells and zebrafish models. Oxidative stress and related signaling pathways underlying the neuroprotective effect were determined by multiple biochemical assays and Western blot. Results Pretreatment with schisantherin A offered neuroprotection against 6-OHDA-induced SH-SY5Y cytotoxicity. Moreover, schisantherin A could prevent 6-OHDA-stimulated dopaminergic neuron loss in zebrafish. Our mechanistic study showed that schisantherin A can regulate intracellular ROS accumulation, and inhibit NO overproduction by down-regulating the over-expression of iNOS in 6-OHDA treated SH-SY5Y cells. Schisantherin A also protects against 6-OHDA-mediated activation of MAPKs, PI3K/Akt and GSK3β. Conclusion These findings demonstrate that schisantherin A may have potential therapeutic value for neurodegenerative diseases associated with abnormal oxidative stress such as Parkinson׳s disease.

In ischemic disorders such as chronic wounds and myocardial ischemia, there is inadequate tissue perfusion due to vascular insufficiency. Besides, it has been observed that prolonged use of anti-angiogenic agents in cancer therapy produces cardiovascular toxicity caused by impaired vessel integrity and regeneration. In the present study, we used VEGFR tyrosine kinase inhibitor II (VRI) to chemically induce vascular insufficiency in zebrafish in vivo and human umbilical vein endothelial cells (HUVEC) in vitro to further study the mechanisms of vascular morphogenesis in these pathological conditions. We also explored the possibility of treating vascular insufficiency by enhancing vascular regeneration and repair with pharmacological intervention. We observed that pretreatment of VRI induced blood vessel loss in developing zebrafish by inhibiting angiogenesis and increasing endothelial cell apoptosis, accompanied by down-regulation of kdr, kdrl and flt-1 genes expression. The VRI-induced blood vessel loss in zebrafish could be restored by post-treatment of calycosin, a cardiovascular protective isoflavone. Similarly, VRI induced cytotoxicity and apoptosis in HUVEC which could be rescued by calycosin post-treatment. Further investigation of the underlying mechanisms showed that the PI3K/AKT/Bad cell survival pathway was a main contributor of the vascular regenerative effect of calycosin. These findings indicated that the cardiovascular toxicity in anti-angiogenic therapy was mainly caused by insufficient endothelial cell survival, suggesting its essential role in vascular integrity, repair and regeneration. In addition, we showed that VRI-induced blood vessel loss in zebrafish represented a simple and effective in vivo model for studying vascular insufficiency and evaluating cancer drug vascular toxicities.

A new direct HPLC analytical method using evaporative light scattering detection coupled with a low-temperature adapter for the simultaneous determination of the major biologically active isosteroidal alkaloids in Bulbus Fritillariae, a commonly used antitussive traditional Chinese medicinal (TCM) herb, has been developed. The simultaneous separation of eight Fritillaria alkaloids was achieved on a reversed-phase C8 column with an isocratic mobile phase system consisting of acetonitrile-methanol-water (66.5:3.5:30, v/v) containing 0.006% triethylamine. This method provides good reproducibility and sensitivity for the quantification of six major isosteroidal alkaloids, namely peimissine, verticine, verticinone, imperialine, isoverticine and ebeiedine in different Fritillaria species with overall intra- and inter-day precision and accuracy of less than 11% and higher than 90%, respectively. The assay was successfully utilized to quantify the major biologically active alkaloids in five Fritillaria species. The results demonstrate that this method is simple, selective, and suitable for the quality control of this commonly used antitussive TCM herb, Bulbus Fritillariae. reserved.

The present paper describes the development of a most simple, sensitive, and specific gas chromatographic method to date, for the direct determination of seven major bioactive isosteroidal alkaloids, namely ebeiedine, ebeiedinone, ebeienine, hupehenine, isoverticine, verticine, verticinone and imperialine, in Fritillaria species, a commonly used antitussive traditional Chinese medicinal (TCM) herb. In the present study, a commercially available Supelco SAC-5 capillary column (30 m x 0.25 mm, 0.25 microm) specifically designed for the analysis of steroids was utilized for the direct determination of Fritillaria alkaloids. Calibration curves were obtained by spiking authentic compounds and the internal standard (solanidine) into herbal samples prior to extraction. Extraction was conducted simply by shaking the pre-alkalized diethyl ether solution (5.0 ml) containing dried herb (0.1 g) for 2 h. All calibration curves showed good linear regressions (r2>0.995) within test ranges. The assay was reproducible and accurate with the overall intra- and inter-day variation and accuracy of less than 10% and more than 90%, respectively. The developed GC method was successfully utilized to analyze seven major bioactive alkaloids in seven Fritillaria species, and the results demonstrate that this direct GC analytical method is suitable for the quality control of this commonly used antitussive TCM herb.

A method of precolumn derivatization GC with FID detection was developed for a simultaneous analysis of five major steroidal alkaloids of Fritillaria species, namely ebeiedine, ebeiedinone, verticine, verticinone and imperialine. Derivatization was carried out by trimethylsilylation of the hydroxyl-containing Fritillaria alkaloids to the corresponding trimethylsilylates with trimethylsilylimidazole. Reaction conditions were optimised and the alkaloids derivatives were characterised by on-line GC-MS. The validated GC method demonstrated a good linearity at the sampling ranges used. This analytical method is simple, convenient and reproducible. The developed assay was successfully applied to the determination of the major pharmacologically active alkaloids in three commonly used antitussive Fritillaria species: F. cirrhosa, F. thunbergii and F. pallidiflora.

Panax ginseng (Ginseng), Rhodiola rosea (Hong Jing Tian) and Schisandra chinensis (Wu Wei Zi) are well-known herbs in traditional Chinese medicine (TCM). Recently, there have been a number of studies on these three herbs. This review discusses their active components and major pharmacological effects. For P. ginseng, it has been shown to have an anti-inflammatory activity, affects pulmonary function and erectile dysfunction, improves cognition in patients with Alzheimer's disease and promotes sexual arousal in menopausal women as well as prevents cancer. For R. rosea, its effectiveness in alleviating depression and reducing fatigue is summarized in this review. Additionally, anti-cancer and other clinical effects of S. chinensis are also discussed. These three herbs are considered as adaptogens as they bear multiple functions and their effects were found to be very different in patients depending on the circumstances (age, gender, environment, diet, season, etc.). Thus, in most cases, the art of the TCM practitioner is to prescribe these herbs after a complete evaluation of overall heath status of the patients.

Background Panax notoginseng is commonly used for the treatment of cardiovascular diseases in China. The present study investigates the effects of three different saponin fractions (ie total saponins, PNS; protopanaxadiol-type saponin, PDS; and protopanaxatriol-type saponin, PTS) and two major individual ingredients (ie ginsenoside Rg1 and Rb1) from P. notoginseng on the endothelial inflammatory response in vitro and in vivo. Methods Recombinant human tumor necrosis factor-α (TNF-α) was added to the culture medium of human coronary artery endothelial cells (HCAECs) to induce an inflammatory response. A cell adhesion assay was used to determine the effect of the P. notoginseng saponin fractions on endothelial-monocyte interaction. The cell adhesion molecule (CAMs) expression, including ICAM-1 and VCAM-1, in the protein level on the surface of endothelial cells were measured by cellular ELISA. CAMs expression in mRNA level was also assayed by qRT-PCR in the HCAECs and the aorta of rat fed with high cholesterol diet (HCD). Western blotting was used to detect effect of the saponin fractions on CAMs protein expression in HCAECs. In addition, nuclear translocation of p65, a surrogate marker for NF-κB activation, was measured by immunostaining. Results Three saponin fractions and two individual ginsenosides exhibited the inhibitory effects on monocyte adhesion on TNF-α-activated HCAECs and expression of ICAM-1 and VCAM-1 at both mRNA and protein levels in vitro. The saponin fractions exhibited a similar trend of the inhibitory effects on the mRNA expression of CAMs in the aorta of HCD-fed rat in vivo. These inhibitory effect of saponin fractions maybe attribute partially to the suppression of the TNF-α-induced NF-κB activation. Conclusion Our data demonstrate that saponin fractions (ie PNS, PDS and PTS) and major individual ginsenosides (ie Rg1 and Rb1) have potential anti-atherogenic effects. Among the tested saponin fractions, PDS is the most potent saponin fraction against TNF-α-induced monocyte adhesion as well as the expression of adhesion molecules in vitro and in vivo.

Although zebrafish has become a popular animal model for drug discovery and screening, drug metabolism in zebrafish remains largely unknown. In this study, we probed the metabolic capability of zebrafish larvae with calycosin, one of the major isoflavone constituents of Radix Astragali that was previously demonstrated to be angiogenic in the zebrafish model. The metabolism of calycosin and accumulation of its metabolites in zebrafish larvae were determined using an LC-MS/MS method. Calycosin showed a slow but steady decrease from the culture medium as well as a steady accumulation in zebrafish larvae. Calycosin underwent major conjugation and minor oxidation in zebrafish larvae. A total of ten calycosin metabolites formed from glucuronidation, glucosylation, sulfation, oxidation or a combination of two of these metabolisms were identified, most of which were reported for the first time. Most metabolites increased steadily in the larvae over 24-h experimental period. The dominant phase II conjugation of calycosin in zebrafish larvae matched well with existing knowledge of isoflavone metabolism in mammalians. The findings shed a light in certain degree of similarity of phase II drug metabolism between zebrafish larvae and mammals and warrant further investigation on feasibility of adopting the zebrafish larvae as a whole-organism model for examining drug metabolism.

Grifola frondosa (Polyporaceae), maitake, is a widely consumed edible mushroom in some Asian countries. The fruit bodies and mycelia of maitake have shown different bioactive compounds with anticancer and other therapeutic properties.This study evaluated three chemically modified maitake polysaccharide-peptides' (MPSP) adjuvant effect (in vivo) and anticancer activity (in vitro growth inhibitory effect) compared with crude MPSP from G. frondosa.We investigated the possibility of enhancing the adjuvant effect and anticancer effect of crude MPSP by using simple chemical modification methods to convert crude MPSP to phosphorylated, acetylated or esterified MPSPs. The adjuvant effect and growth inhibitory effect were evaluated by C6 cell inoculated rat model with cyclophosphamide (CPA) treatment and in vitro cell viability assay, respectively.All four tested MPSPs showed significant adjuvant effect to CPA treatment on rats inoculated with C6 cancer cells. In addition, an obvious growth inhibitory effect was observed in C6 cancer cells but not in normal brain cells treated with various forms of MPSPs. Only phosphorylation could significantly (p 0.05) improve the adjuvant effect (in vivo) and growth inhibitory effect. A same rank order (phosphorylated MPSPesterified MPSP ≥ acetylated MPSP ≥ crude MPSP) of efficacy was observed in both the in vivo and in vitro assays.This study showed chemical phosphorylation could markedly enhance both adjuvant effects and growth inhibitory effects. This study demonstrated the feasibility of enhancing the efficacy of MPSP by using a simple chemical modification method, and this provides a foundation for future study in this area.

UNLABELLED The extract of Curcuma longa, better known as turmeric, was orally administered to experimental rats that were fed a high-cholesterol diet to investigate whether it could regulate plasma lipids and cholesterol levels and possibly improve hepatic conditions. With turmeric supplements, rats showed a significant decrease in total plasma cholesterol and low-density lipoprotein cholesterol but an increase in high-density lipoprotein cholesterol when compared with rats that were fed a high-cholesterol diet alone. Fatty liver developed in hypercholesterolemic rats with the high-cholesterol diet treatment, and this condition was markedly improved when rats were provided with turmeric supplements at 100 mg/kg or 300 mg/kg of body mass. The turmeric treatment resulted in a significant decrease in the total amount of hepatic lipid. Histological staining of liver tissues with Sudan III and hematoxylin showed that rats fed with a high-cholesterol diet alone had more and larger granular fat bodies than rats having turmeric extract supplementation in their high-cholesterol diet. Reverse-transcription polymerase chain reaction was used to assess the expression levels of enzymes involved in fat metabolism and cellular homeostasis in experimental rat livers. The results showed that rats fed a high-cholesterol diet supplemented with turmeric extract had a significant increase in the expression of cholesterol 7 α-hydroxylase, hemeoxygenase 1, and low-density lipoprotein receptors but a significant decrease in 3-hydroxy-3-methyl-glutaryl-CoA reductase level when compared with rats fed a normal or high-cholesterol diet, showing that turmeric prevents hypercholesterolemia and the formation of fatty liver by the modulation of expressions of enzymes that are important to cholesterol metabolism. PRACTICAL APPLICATION Turmeric may be considered a functional food for regulating plasma cholesterol levels and preventing the development of fatty liver in people who frequently consume a high-cholesterol diet.

Angiogenesis plays an important role in the development of neoplastic diseases such as cancer. Resveratrol and its derivatives exert antiangiogenic effects, but the mechanisms of their actions remain unclear. The aim of this study was to evaluate the antiangiogenic activity of resveratrol and its derivative trans-3,5,4'-trimethoxystilbene in vitro using human umbilical vein endothelial cells (HUVECs) and in vivo using transgenic zebrafish, and to clarify their mechanisms of action in zebrafish by gene expression analysis of the vascular endothelial growth factor (VEGF) receptor (VEGFR2/KDR) and cell-cycle analysis. trans-3,5,4'-Trimethoxystilbene showed significantly more potent antiangiogenic activity than that of resveratrol in both assays. In zebrafish, trans-3,5,4'-trimethoxystilbene caused intersegmental vessel regression and downregulated VEGFR2 mRNA expression. Trans-3,5,4'-trimethoxystilbene also induced G2/M cell-cycle arrest, most specifically in endothelial cells of zebrafish embryos. We propose that the antiangiogenic and vascular-targeting activities of trans-3,5,4'-trimethoxystilbene result from the downregulation of VEGFR2 expression and cell-cycle arrest at G2/M phase.

Folic acid supplementation provides beneficial effects on endothelial functions in patients with hyperhomocysteinemia. However, its effects on vascular functions under diabetic conditions are largely unknown. Therefore, the effect(s) of folic acid (5.7 and 71 microg/kg/day for 4 weeks) on aortic relaxation was investigated using obese/diabetic (+db/+db) mice and lean littermate (+db/+m) mice. Acetylcholine-induced relaxation in +db/+db mice was less than that observed in +db/+m mice. The reduced relaxation in +db/+db mice was restored by consumption of 71 microg/kg folic acid. Acetylcholine-induced relaxation (with and without folic acid treatment) was sensitive to N(G)-nitro-L-arginine methyl ester, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one, geldanamycin and triciribine. In addition, acetylcholine-induced relaxation was attenuated by resistin. The plasma level of resistin in +db/+db mice was sevenfold higher than that measured in +db/+m mice, and the elevated plasma level of resistin in +db/+db mice was reduced by 25% after treatment with 71 microg/kg folic acid. Folic acid slightly increased the ratio of reduced glutathione to oxidized glutathione in +db/+db mice. Moreover, folic acid caused a reduction in PTEN (phosphatase and tensin homolog deleted on chromosome 10) expression, an increase in the phosphorylation of endothelial nitric oxide synthase (eNOS(Ser1177)) and Akt(Ser473), and an enhanced interaction of heat shock protein 90 (HSP90) with eNOS in both strains, with greater magnitude observed in +db/+db mice. In conclusion, folic acid consumption improved blunted acetylcholine-induced relaxation in +db/+db mice. The mechanism may be, at least partly, attributed to enhancement of PI3K/HSP90/eNOS/Akt cascade, reduction in plasma resistin level, down-regulation of PTEN and slight modification of oxidative state.

Ethnopharmacological relevance Rheum officinale Baill. (Da Huang) is one of the herbs commonly used in traditional Chinese medicine formulae against cancer. The traditional decoction is similar to the water extract used in the present study. Aim of the study The water extract of Da Huang was investigated to see if it possesses anticancer effects through apoptotic pathways. Materials and methods Human lung adenocarcinoma A549 and human breast cancer MCF-7 cell lines were treated with different concentrations of Da Huang water extract at different time intervals. Growth inhibition was detected by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] and colony formation assays; apoptosis was detected by cell morphologic analysis, DNA fragmentation analysis and COMET assay. Results Da Huang water extract was found to have significant growth inhibitory effects on both A549 and MCF-7 cell lines with IC 50 values 620 ± 12.7 and 515 ± 10.1 μg/ml, respectively. Growth inhibitory effects were dose- and time-dependent. A significant decrease in cell number, DNA fragmentation and single DNA strand breakages were observed in the Da Huang water extract treated A549 and MCF-7 cells. Conclusions This suggests that the water extract of Da Huang exerts potential anticancer activity through growth inhibition and apoptosis on MCF-7 and A549 cells lines.

Puerarin, a major isoflavonoid compound from the Chinese herb, Ge-gen (Pueraria lobata), has effective treatment on myocardial and cerebral ischemia, glaucoma and sudden deafness in clinical setting in China. Our present work showed that puerarin (50, 150, 450 microM) concentration-dependently inhibited phenylephrine or KCl-induced contraction only in endothelium-intact rat aortic rings. In Ca(2+)-free solution, the antivasoconstriction of puerarin on phenylephrine was totally deprived. N (G)-nitro-L-arginine methyl ester, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, indomethacin and the three K(+) channel blockers, glibenclamide, tetraethylammonium and Ba(2+) displayed significant inhibitory effects on the antivasoconstriction of puerarin. 8-bromo-cGMP significantly strengthened the action of puerarin. Puerarin (10-160 microM) concentration-dependently induced the NO production in the rat aortic cells. These findings suggested that the antivasoconstriction elicited by puerarin is endothelium-dependent. NO/NO-cGMP pathway, PGI(2) and the opening of K(+) channels sensitive to glibenclamide, tetraethylammonium, and Ba(2+), which might be triggered by the extracellular Ca(2+) influx in the endothelium, appear to contribute to the antivasoconstriction of puerarin.

The cough and emetic reflexes involve a synchronized firing of motor neurones involved in respiratory control. Tachykinin NK1 receptor antagonists and 5-HT1A receptor agonists are examples of centrally acting drugs that reduce cough and emesis. In the present studies, therefore, we examined the possibility that other classes of drugs known to reducing cough have anti-emetic properties to prevent emesis induced by diverse challenges. We examined the potential of codeine (1-10 mg/kg), baclofen (1-10 mg/kg), scopolamine (0.3-10 mg/kg), diphenhydramine (1-10 mg/kg), imperialine (1-30 mg/kg) and verticine (0.3-3 mg/kg) to inhibit emesis induced by nicotine (5 mg/kg, s.c.), copper sulphate (120 mg/kg, intragastric), and provocative motion (4 cm horizontal displacement, delivered at 1 Hz) in Suncus murinus (house musk shrew). Only codeine had broad inhibitory properties (P0.05) to antagonize emesis induced by all challenges with ID50 values ranging from 1.2 to 2.3 mg/kg. Baclofen antagonized emesis induced by nicotine (maximum reduction was 44.9%, P0.05) and motion (maximum reduction was 97.3%, P0.01), but potentiated copper sulphate-induced emesis (maximum potentiation was 73.0%, P0.05). Scopolamine antagonized copper sulphate-induced emesis (maximum reduction was 61.2%, P0.05) and imperialine antagonized nicotine-induced emesis (maximum reduction was 30.2%, P0.01), but verticine potentiated motion-induced emesis (maximum potentiation was 60.0%, P0.05). Diphenhydramine did not significantly reduce emesis induced by any of the challenges (P0.05). In conclusion, codeine has broad inhibitory anti-emetic actions but a known ability to reduce coughing does not necessarily predict broad inhibitory anti-emetic properties.

Bulbus Fritillariae derived from plants of various Fritillaria species is the most commonly used antitussive traditional Chinese medicinal herb and is called Beimu. Herbs derived from similar and/or different species of Fritillaria are also used in Japan and Turkey as traditional or folk medicines. Isosteroidal alkaloids are the main bioactive ingredients in Fritillaria species. As the contents and structure types of these bioactive alkaloids vary in different Fritillaria species, quality control of these active principles in herbal Beimu is very important to ensure its safe and effective clinical use. This review describes the development of chromatographic analyses for the simultaneous qualitative and quantitative determination of the main bioactive Fritillaria isosteroidal alkaloids in herbal and biological samples. The recently developed direct HPLC-evaporative light scattering detection method is the most simple, selective and sensitive assay, and is readily used as a suitable quality control method for the analysis of the active principles of herbal Beimu.

Angiogenesis plays an important role in a wide range of physiological processes and many diseases are associated with the dysregulation of angiogenesis. The commonly used Chinese herbal medicine Radix Astragali (known as Huang qi in Chinese) is a potential candidate for treating this type of disease. Calycosin, a major isoflavonoid in Radix Astragali, was identified in our earlier study and shown to induce angiogenesis in human umbilical vein endothelial cells (HUVEC) in vitro and in zebrafish embryos in vivo. Using zebrafish as a testing model, we investigated the angiogenic effect of calycosin on the subintestinal vessels (SIVs) in zebrafish embryos. Our findings using transcriptional profiling by deep sequencing, and confirmed by quantitative real-time PCR (qPCR), demonstrate that calycosin modulated vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF) and ErbB signaling pathways. The inhibitory effects of calycosin-induced phenotypic responses by several pathway-specific inhibitors (VRI, SU5402, MEK1/2 Inhibitor, Wortmannin and LY294002) further identified the potential involvement of VEGF(R) and FGF(R) signaling pathways in the angiogenic activities of calycosin. We present a comprehensive framework of study using fluorescence microscopy, transcriptomics and qPCR to demonstrate the proangiogenic effects of calycosin in vivo. The data have elucidated the connection between morphological observations and genomic evidence, indicating the potential roles of several key signaling pathways in angiogenesis.