[Two new steroidal alkaloids from ripe berries of Solanum nigrum].

Two previously undescribed steroidal alkaloids, compounds 1-2, along with two known ones(3-4), were isolated from the 80% ethanol extract of ripe berries of Solanum nigrum by chromatographic methods, including silica gel, ODS, and HPLC. Based on spectroscopic and chemical evidence, including IR, NMR, and HR-ESI-MS data, the structures of the isolated compounds were identified as 12ß,27-dihydroxy solasodine-3-O-ß-D-glucopyranoside(1), 27-hydroxy solasodine-3-O-ß-D-glucopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-ß-D-glucopyranoside(2), solalyraine A(3), and 12ß,27-dihydroxy solasodine(4). Compounds 1-2 were tested for their potential effects against the proliferation of A549 cells, which revealed that compounds 1-2 had weak cytotoxic activity.

Steroidal alkaloids from Solanum nigrum and their cytotoxic activities.

Eight undescribed, along with five known steroidal alkaloids were isolated from Solanum nigrum L., a plant used in traditional Chinese medicine. Their structures were elucidated by NMR, HR-ESI-MS, and IR spectroscopy. Two compounds displayed an unusual structure in steroidal alkaloids with an open E-ring and without an F-ring present. To evaluate their bioactivities, nine compounds were selected to intervene five human cancer cell lines including H1299, HepG2, HeLa, HCT116, and MCF7 respectively. All compounds exhibited inhibitory effects for the five cell lines, revealing potential anti-tumor activities from Solanum nigrum.

Steroidal glycoalkaloids from Solanum lyratum inhibit the pro-angiogenic activity of A549-derived exosomes.

In this study, seven previously undescribed steroidal glycoalkaloids, compounds 1-7, were isolated from Solanum lyratum, along with two known ones (8 and 9). Comprehensive spectroscopy techniques were used to determine their structures. Although 1-8 only showed a weak inhibitory effect on the proliferation of the tumor-derived vascular endothelial cells, however, in a former study we found both total steroidal glycoalkaloids from Solanum lyratum (TSGS) and 9 significantly inhibited tumor angiogenesis and its mechanism was linked to its ability to interfere with cell membrane lipid rafts. Lipid rafts are closely related to the functions of tumor-derived exosomes, a vital factor in cancer progression. Thus, we investigated the impacts of TSGS and 9 on the functions of A549-derived exosomes. Our results indicated that A549-derived exosomes can significantly enhance the angiogenesis abilities of human umbilical vein endothelial cells, whereas the intervention of TSGS or 9 significantly inhibited this activity of A549-derived exosomes. These findings suggest that TSGS and 9 exert anti-tumor angiogenesis by inhibiting the pro-angiogenic activity of A549-derived exosomes.

An-te-xiao capsule inhibits tumor growth in non-small cell lung cancer by targeting angiogenesis.

An-te-xiao capsule consists of total alkaloids from the dried whole plantof Solanum lyratum, and showed antitumor effects in our previous study. However, its inhibitory effect on multiple non-small cell lung cancer (NSCLC) cell lines and the underlying mechanisms have not been elucidated clearly. This study sought to investigate the inhibitory effects of An-te-xiao capsule on three main types of NSCLC cell lines (A549, NCI-H460, and NCI-H520) in vitro and in vivo and the underlying mechanisms of action including its potential anti-angiogenesis effects. An-te-xiao capsule showed no acute oral toxicity in mice, and significantly prolonged survival time in a mouse model of Lewis tumor xenograft. The inhibition of A549, NCI-H460, and NCI-H520 cells by An-te-xiao capsule was reflected in its effects on tumor growth, histopathological changes, tumor microvessel density (MVD), cell cycle regulatory proteins, and cell apoptosis. In vitro, An-te-xiao capsule repressed migration, invasion, and tube formation of tumor-derived vascular endothelial cells (Td-ECs), which were obtained using a co-culture system, in the presence or absence of vascular endothelial growth factor (VEGF) at safe concentrations selected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Moreover, An-te-xiao capsule inhibited the secretion of VEGF by A549 cells in the co-culture system and suppressed the phosphorylation of VEGF receptor 2 (VEGFR2). Taken together, An-te-xiao capsule has potential for treating NSCLC.

[Research and application of quality standard for standard decoction of Chrysanthemi Flos].

Standard decoction of traditional Chinese medicine (TCM) is prepared by standardized process, and can be used as references to evaluate the quality of dosage forms such as decoction and dispensing granules. In order to determine the quality evaluation method for standard decoction of Chrysanthemi Flos and investigate its application, 10 batches of white chrysanthemum of Hangzhou were collected to prepare the standard decoction of white chrysanthemum of Hangzhou with standardized process parameters. Parameters such as traits, relative density, pH value, extraction ratio, transfer rate and fingerprint were selected as the indexes for quality evaluation. The established quality evaluation method for standard decoction of Chrysanthemi Flos was applied in the detection of two types of commercial Chrysanthemum dispensing granules. The results showed that the standard decoction of Chrysanthemi Flos was a clear yellow-brown aqueous solution; the relative density was 1.007-1.011; the pH value was between 5.37-5.56; the average extraction ratio was 23.6%, ranging from 19.93% to 29.69%; the average transfer ratewas 56.2% in terms of chlorogenic acid, 57.4% in terms of luteoloside and 30.6% in terms of 3,5-O-dicaffeoylquinic acid. Fingerprint similarity was between 0.864-0.989.The method showed good precision, stability and repeatability in fingerprint analysis, indicating reliable and representative results for standard decoction of Chrysanthemi Flos, and it can be used to evaluate and standardize other dosage forms.

[Calcium mobilizing effect of hawthorn leaf procyanidins in vascular endothelial cells].

The hawthorn leaves have the effect of activating blood, removing blood stasis, regulating qi through the veins, dissolving turbidity and lowering lipid. Procyanidinis is one of its main active components and plays an important role in regulating vasoactivity. Previous studies showed that the regulating effect of procyanidins was related to its regulation on nitric oxide secretion from vascular endothelial cells, and this effect was dependent on the extracellular calcium concentration, suggesting that the changes in intracellular calcium ion concentration in endothelial cells may play a key role in this process. However, the research on this issue is still insufficient so far. This study is aimed to observe the effect of hawthorn leaf oligomeric procyanidins (HLP) on calcium mobilization of vascular endothelial cells, and investigate the underlying mechanism. Human umbilical vein endothelial cells (HUVEC) were cultured in vitro and labeled with Fura-2. HUVEC were treated with HLP at concentrations of 6.25, 12.5, 25 and 50 mg·L⁻¹, and the intracellular calcium concentrations were measured with a living cell microscope for 30 min. HLP increased the intracellular calcium concentration of HUVEC in a concentration dependent manner; and the intracellular calcium concentrations in 25 and 50 mg·L⁻¹ HLP groups were significantly higher than that in the normal group. With the use of calcium-free incubation buffer, addition of calcium chelating agent EGTA in incubation buffer, or use of inhibitors for sodium calcium exchanger, the effect of HLP was significantly inhibited. On the other hand, the effect of HLP could also be weakened by inhibiting the calcium release from the intracellular storage. In conclusion, these results suggest that HLP can elicit calcium mobilization in vascular endothelial cells, which may be one of the mechanisms for its vascular modulatory activity; and this calcium mobilizing effect may be achieved through promoting both extracellular calcium influx and intracellular calcium release, additionally the former may be related to activating the reverse transport of Na⁺-Ca²âº exchangers on the cell membrane.

Study design and implementation for population pharmacokinetics of Chinese medicine: An expert consensus.

Although many population pharmacokinetics (PPK) researches have been conducted on chemical drugs, few have been in the field of Chinese medicine (CM). Each ingredient in CMs possesses different pharmacokinetic characteristics, therefore, it is important to develop methods of PPK studies on them to identify the differences in CM drug safety and efficacy among the population subgroups and to conduct quantitative studies on the determinants of CM drug concentrations. To develop an expert consensus on study design and implementation for PPK of CM, in August 2013, 6 experts in the field of PPK, CMs pharmacology, and statistics discussed problems on the PPK research protocol of CMs, and a consensus was reached. The medicines with toxicity and narrow therapeutic windows and with wide range of target population or with frequent adverse reactions were selected. The compositions with definite therapeutic effects were selected as indices, and specific time points and sample sizes were designed according to standard PPK design methods. Target components were tested through various chromatography methods. Total quantity statistical moment analysis was used to estimate PPK parameters of each component and PPK models reflecting the trend of CMs (which assists in reasonable adjustments on clinical dosage). This consensus specifies the study design and implementation process of PPK. It provides guidance for the following: post-marketing clinical studies, in vivo investigations related to the metabolism in different populations, and development and clinical adjustment of dosages of CMs.

[Advantages of population pharmacokinetics and its application in the field of traditional Chinese medicine].

The accurate medical treatment is based on the information of the genome, which is the best treatment for the patients. Population pharmacokinetic study can be formulated according to the individual differences of patients to the dose, in the accurate medical model which has a unique advantage. At present, there are many problems such as adverse drug reaction in Chinese traditional medicine, and it is necessary to introduce a group of medicine on the basis of precise medical treatment. However, due to the different characteristics of traditional Chinese medicine and chemical medicine, it is necessary to combine the population pharmacokinetics, genetics and statistical methods to establish a research method which is in line with the characteristics of Chinese medicine. The key scientific problem is to make clear the active components of Chinese medicine metabolism of the drug metabolic enzyme gene, and pay attention to the analysis of the polymorphism of the overall role of drug metabolism enzymes in the human body. Clear key scientific issues and break through the bottleneck, so as to achieve the precise medical treatment, to international.

[Expert consensus on population pharmacokinetics of Chinese medicine ( draft version for comments)].

In population pharmacokinetic (PPK) research of Chinese medicines with narrow therapeutic windows of toxicity, or when the target population is not homogeneous, or when there are frequent adverse reactions to parenterally administered Chinese medicine, select those that have a definite therapeutic effect, and in which the compositions of the toxic substances compositions are known, for study, and use complete PPK sampling design to take samples at specific time points. Use gas chromatography, HPLC, and LC-MS methods for the detection of target components. Finally, use total quantity statistical moment analysis, to account for each component of the PPK parameters. Thus, PPK model can reflect the overall trend of Chinese medicine, to provide the basis for reasonable clinical dosage adjustments.

[Study on the chemical constituents of the PTP 1B inhibitory effective parts of Paeoniae Rubra Radix].

OBJECTIVE: To study the chemical constituents of the PTP 1B inhibitory effective parts of Paeoniae Rubra Radix. METHODS: The effective part of Paeoniae Rubra Radix was enriched by Sephadex LH-20. Compounds were isolated by various column chromatography and their structures were elucidated through spectroscopic analysis. RESULTS: Thirteen compounds were identified as: benzoylpaeoniflorin(1), albiflorin(2), paeoniflorigenone(3), methyl gallate(4), adenosine(5),2-amino adenosine(6), 3,3'-Di-O-methylellagic acid(7), 3,3',4-trimethyl ellagic acid(8), dihydrokaempferol(9), glycerol(10), dibutyl phthalate(11). CONCLUSION: Compounds 5-11 are obtained from this plant for the first time.

Population pharmacokinetics of naringin in total flavonoids of Drynaria fortunei (Kunze) J. Sm. in Chinese women with primary osteoporosis.

OBJECTIVE: To evaluate the effect of covariates on the pharmacokinetic profiles of naringin in the total flavonoids of Drynaria fortunei (Kunze) J. Sm. in the Qianggu Capsule () by evaluating Chinese women with primary osteoporosis. METHODS: A total of 98 female patients from the communities of Jingshan, Beixinqiao, Jiaodaokou, Chaoyangmen, and Donghuamen in Beijing, China, aged 40 to 80 years, were included in this study. Blood samples were collected before and 0.5, 1, 2, 3, 4, 6, 8, 10, 12, and 24 h after a single oral dose of Qianggu Capsule. The concentration in blood samples from 32 patients before and 0.5, 1, 2, 3, and 4 h after drug administration were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, and full set of pharmacokinetic data was analyzed with nonlinear mixed-effect modeling (NONMEM) software. The mean of population parameters clearance (C1), central distribution volume (V), absorption rate constant (Ka1), inter-compartmental clearance (C2), peripheral distribution volume (V2) were set as parameters and estimated through base model, covariate model, and final model. Age, height, weight, blood urea nitrogen (BUN), serum creatinine (Scr), alanine transaminase (ALT), aspartate transaminase (AST), hyperlipidemia, Liver (Gan) Kidney (Shen) yin insufficiency (GSYI), Kidney (Shen) yang insufficiency (SYI) were set as covariates. RESULTS: The relationships between these parameters and covariates were analyzed. The results showed that C1 was the main parameter influenced by the selected covariates among the population parameters, and the relationships between the covariates and C1 were analyzed, among the selected covariates hyperlipidemia was identified as significant covariate of C1. CONCLUSION: The pharmacokinetic behaviors of naringin are altered with hyperlipidemia in Chinese women with primary osteoporosis.

[Normative research on bacteriostasis and relieving itching external therapeutic function of kochiae fructus].

OBJECTIVE: To normalize bacteriostasis and relieving itching external therapeutic function of Kochiae Fructus. METHODS: Itching guinea pig model caused by histamine, itching mice model, eczema guinea pig model caused by OVA, and inhibitory effect on pathogens in vitro were used to observe the itching threshold, symptoms and other related physiological index, as well as the inhibitory effect on the normal skin fungi by water extraction of Kochiae Fructus to evaluate the external therapeutic function of Kochiae Fructus. RESULTS: The itching threshold of guinea pig itching model treated by water extraction of Kochiae Fructus at high, medium and low three dosage level, could be significantly increased when compared with negative control group (P < 0.05 or P < 0.01); Red speckle of OVA guinea pig model treated by water extraction of Kochiae Fructus at high, medium and low three dosage level, could be significantly decreased when compared with negative control group (P < 0.05 or P < 0.01); The number of itching and total time of itching within 30 minutes of mice model caused by R-glycose anhydride treated by water extraction of Kochiae Fructus at high, medium and low three dosage level, could be significantly decreased when compared with negative control group (P < 0.05 or P < 0.01); Several common skin fungi could be significantly inhibited by the water extraction of Kochiae Fructus. MIC of the water extraction of Kochiae Fructus on Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, Trichophyton violaceum, and Trichophyton schoenleini were 3.12%, 0.78%, 0.78%, 0.78%, 0.78%, respectively. CONCLUSION: Kochiae fructus has the effect of bacteriostasis and relieving itching.

[Experimental study of the total flavonoid in Hypericum perforatum on depression].

OBJECTIVE: To investigate pharmacological effects of the total flavonoid in Hypericum perforatum on depression. METHOD: Experimental depression was induced by subcutaneous injection of reserpine in mice. The concentration of monoamine transmitters including 5-HT and NE, the activity of monoamine oxidase (MAO) in brain and reserpine-induced symptoms of depression, such as ptosis, attenuation of autonomous activity, behavioral despair, acquired helplessness and sleep, were measured respectively to evaluate the effects of the total flavonoid in H. perforatum on the depression. RESULT: The total flavonoid in H. perforatum significantly decreased the activity of MAO, inhibited the ptosis and the attenuation of autonomous behavior induced by reserpine respectively. The levels of 5-HT and NE were also attenuated by the total flavonoid in H. perforatum remarkably. In addition, the total flavonoid in H. perforatum was shown to inhibit behavioral despair and acquired helplessness and to prolong the sleep time in the mice. Following the treatment with the total flavonoid in H. perforatum, 5-THP, at the dosage without any side-effects, caused the tremble in the mice. CONCLUSION: The results indicate that total flavonoid in H. perforatum can significantly inhibit the depression.

Effect of combination of extracts of ginseng and ginkgo biloba on acetylcholine in amyloid beta-protein-treated rats determined by an improved HPLC.

AIM: To determine the concentration of acetylcholine (ACh) in amyloid beta-protein (Abeta) treated rats and offer a method determining ACh as well. METHODS: A 1-month combination of extrats of ginseng and ginkgo biloba (Naoweikang) ig administration to rats was performed daily after bilateral injection of Abeta(1-40) (4 g/L, 1 microL for each side) into hippocampus. After decollation, homogenizing, and centrifuging and extracting, a high pressure liquid chromatographic (HPLC) method using electrochemical detection (ECD) combined with two immobilized enzyme reactors was used to determine ACh in rat whole brain. RESULTS: With a mobile phase consisting of disodium hydrogen orthophosphate, tetramethylammonium chloride (TMACl), octanesulfonic acid sodium salt (OSA) and "Reagent MB" at a final pH of 8.0, ACh was determined while removing the interfering choline in less than 10 min at a flow rate of 0.35 mL/min on a platinum (Pt) working electrode at a potential of +300 mV vs a solid-state palladium (Pd) reference electrode. Linear regression analysis of peak area vs concentration demonstrated linearity in the 28.01 to 1400.06 microg/L injection range. The r-value was 0.9978. The limit of detection (LOD) is 0.28 ng on column. ACh in whole brain decreased by 20.34 % (from 162.1+/-32.7 to 134.7+/-14.0 microg/L, P<0.05) after bilateral injection of Abeta into rat hippocampus. After Naoweikang administration (31 and 15.5 mg/kg, respectively), ACh increased by 19.97 % (from 134.7+/-14.0 to 161.6+/-26.2 microg/L, P<0.05) and 18.56 % (from 134.7+/-14.0 to 159.7+/-22.9 microg/L, P<0.05), respectively. CONCLUSION: Naoweikang significantly increased the level of ACh in whole brain of Abeta treated rats. And a sensitive, selective and reliable method for routinely determining ACh in rat whole brain was established in this study.