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To investigate the possible molecular mechanism of total glycosides of Chishao (Radix Paeoniae Rubra) (TG-RPR) on proliferation and apoptosis of hepatocellular carcinoma cells.The proliferation of TG-RPR on HepG2 cells was detected using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The apoptosis of HepG2 cells was measured by annexin V-FITC/double staining. The phosphatase and tensin homolog deleted on chromosome ten (PTEN) / phosphatidylinositol 3-kinase (PI3K) / protein kinase B (Akt) signaling pathway was evaluated by Western Blot and reverse transcription-polymerase chain reaction (RT-PCR).TG-RPR can up-regulation the expression of pro-apoptotic factors such as PTEN and BCL2-Associated X (Bax), down-regulation the expression of anti-apoptotic factors including B-cell lymphoma-2 (Bcl-2), PI3K, and Akt.TG-RPR significantly inhibits the proliferation of HepG2 cells in a dose-dependent manner and promotes apoptosis. These results demonstrated TG-RPR has significant inhibitory effect on HepG2 cells. These results identify a critical role of TG-RPR in proliferation and apoptosis of HepG2 cells via modulating PTEN/PI3K/Akt signaling pathway. TG-RPR may offer a promise as a potential pharmaceutical therapy for hepatocellular carcinoma.

Cervical cancer is considered the fourth most common malignant disease in women. Recently, tannin from Spatholobi Caulis (TTS) has been shown to have potent anticancer and antiproliferative characteristics in a few preliminary studies. This experiment used 3D microfluidic, flow cytometry, and gene chip technology to study the efficacy and mechanism of action of TTS, as well as molecular docking technology to study the effect of drugs on related proteins. The cell survival rates of the five groups measured by the 3D microfluidic chip were 94%, 85%, 64%, 55%, and 42%, respectively. With the increase in drug concentration, the cell survival rate gradually decreased. Apoptosis rates detected in the five groups were 2.12%, 15.87%, 33.40%, 41.13%, and 55.10%, respectively. These data suggest that TTS can promote cell apoptosis. The percentages of cells in the G0/G1 phase were 43.39%, 55.07%, 59.57%, 64.56%, and 67.39% in the five groups, respectively. TTS was demonstrated to inhibit the conversion of cells from G0/G1 to S phase and G2/M phase and inhibit gene and protein synthesis to block cell proliferation. TTS can effectively modulate pathogenic proteins. The results confirmed the efficacy of TTS against HeLa cells and that TTS can be used as an adjunct in cervical cancer prevention and treatment.

Ethnopharmacological relevance Glycyrrhiza is the dry root and rhizome of the leguminous plant, Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat. or Glycyrrhiza glabra L., which was firstly cited in Shennong's Herbal Classic in Han dynasty and was officially listed in the Chinese Pharmacopoeia, has been widely used in China during the past millennia. Licoflavone is the major component of Glycyrrhiza with anti-ulcer activity. The present study is based on clarifying the anti-ulcer effect of licoflavone, aiming at elucidating the possible molecule mechanisms of its action for treating gastric ulcer rats induced by acetic acid. Materials and methods Rats were divided into 7 groups, and drugs were administered from on the day after the onset of gastric ulcer (day 3) until day 11 of the experiment once daily continuously. The plasma were analyzed by high-performance liquid chromatography combined with time-of-flight mass spectrometry (HPLC/ESI-TOF-MS), significant different metabolites were investigated to explain its therapeutic mechanism. Furthermore, quantitative real time polymerase chain reaction (RT-PCR) analysis was performed to detect the expression of RNA in stomach tissue for verifying the above results. Results Licoflavone can effectively cure the gastric ulcer, particularly the middle dose group. According to the statistical analysis of the plasma different metabolites from each groups and the expression of genes in tissues, sixteen significant different metabolites, including histamine, tryptophan, arachidonic acid, phingosine-1-phosphate etc., contributing to the treatment of gastric ulcer were discovered and identified. In RT-PCR analysis, the results of the expression of RNA were corresponded with what we discovered. Conclusions Our study indicated licoflavone plays the role of treating gastric ulcer by regulating inflammation mediators and amino acid metabolism. We demonstrated that metabolomics technology combined with gene technology is a useful tool to search different metabolites and to dissect the potential mechanisms of traditional Chinese medicine (TCM) in treating gastric ulcer.

Cirsium setosum (Willd.) MB. has been reported to exert significant anti-hemorrhagic, anti-inflammation, antimicrobial, sedative and detoxicating efficacy. It has been widely used to treat gastrointestinal bleeding, uterine bleeding, infectious hepatitis and cardiovascular disease in China. Recent studies have shown that flavonoids are the main active components in C. setosum. Nevertheless, to the best of our knowledge, there is no report concerning the simultaneous determinations and pharmacokinetics of constituents in C. setosum flavonoids in rat plasma. In this study, a rapid, sensitive and selective triple quadrupole liquid chromatography-mass spectrometry method was developed to determine eight analytes from the flavonoids of C. setosum in rat plasma. In addition, the pharmacokinetic study of the eight analytes in rats after oral administration of C. setosum flavonoids was successfully completed through this method. According to the pharmacokinetic parameters of the eight analytes, rutin, naringin, quercetin, acacetin, wogonin were the long-acting components of the C. setosum flavonoids, with long elimination time and high bioavailability. Of note, the method developed in this study fills a blank in pharmacokinetic studies of C. setosum flavonoids. Our findings provide valuable views on the understanding of the absorption mechanism of C. setosum flavonoids and their clinical efficacy.

As a traditional Chinese medicine (TCM) with a usage history of over 2,000 years in China, Spica Schizonepetae possesses definite clinical activity in the treatment of non-small cell lung cancer (NSCLC). However, its active ingredients and mechanism of action remain unclear at present. The further exploration of its active components and underlying mechanism will provide a basis for the development of candidate anti-tumor drugs. Our previous study explored the chemical constituents of Spica Schizonepetae extract (SSE). On this basis, molecular networking technology was applied in analyzing the QTOF-MS/MS data of rat plasma after intragastric administration of SSE using the GNPS database platform. A total of 26 components were found, including 9 proterotype components and 17 metabolites, which revealed the potential active ingredients of SSE. Later, the Lewis lung cancer mouse model was established, and the inhibition rate and histopathological sections were used as the indicators to investigate the anti-tumor effect of SSE, whereas the body weight, survival rate, thymus index and spleen index served as the indicators to explore the pharmacological effects of SSE on improving mouse immunity. The results showed that SSE had comparable anti-tumor efficacy to cisplatin, which enhanced the immunity, improved the quality of life, and extended the survival time of lung cancer mice. Furthermore, human A549 lung tumor cells were selected to explore the mechanism of SSE in treating NSCLC based on cell metabonomics. After data mining by the MPP software, 23 differential endogenous metabolites were identified between SSE and tumor groups. Moreover, results of pathway enrichment analysis using the MetaboAnalyst 4.0 software indicated that these metabolites were mainly enriched in four metabolic pathways (p < 0.1). By adopting the network pharmacology method, the metabolic pathways discovered by cell metabolomics were verified against the ChEMBL, STITCH, UniProt and TCGA databases, and differences in the underlying mechanism between cells and humans were found. It was proved that SSE affected the metabolism of purine, arachidonic acid and histidine to exert the anti-tumor efficacy. Furthermore, the multi-target, multi-pathway, and immunoenhancement mechanism of SSE in anti-tumor treatment was revealed, which provided a scientific basis for new drug development and the rational application of Spica Schizonepetae in clinic.

This study was designed to elucidate the chemical composition and anti-cancer effects of Schizonepeta tenuifolia’s ethanol extracts. Microfluidic technology was used in the study of Schizonepeta tenuifolia from 9 different geographic regions. The ethanol extracts were examined with HPLC to establish their Fingerprints in order to analyze the relationship between the spectrum and efficacy index through Grey Correlation software, and a rapid HPLC-Q-TOF/MS method was established. The result shows that chromatographic peaks of the 19, 6, 11, 16, 18th are the representative diosmetin, luteoloside, hesperidin, luteolin, and apigenin. The 10, 12, 20th peaks may be naringenin-7-O-glucuronide or quercitrin, rosmarinate or acetylcorynoline, and 5,7-dihydroxy-6,4-dimethoxy flavone. The major chemical composition of Schizonepeta tenuifolia was found to have the anti-lung-tumor effects. A new method was established for the quality control of traditional Chinese medicine.

In order to further clarify the rational use of different medicinal parts of Schizonepeta, microfluidic technology was used in this study to investigate the differences in drug efficacy against lung cancer in vitro. The ethanol extracts were examined with HPLC to establish their fingerprints in order to analyze the relationship between the spectrum and efficacy index through Grey Correlation software, and a rapid HPLC-Q-TOF/MS method was established. The result in vitro shows that the effect and components of different medicinal parts had a certain differences, and apoptosis and necrosis rate from big to small in turn is leaf, flower, root, stem. The chromatographic peaks of the 26, 12, 2, 6 and 15th are the luteolin, icynaroside, rosmarinic, caffeic acid, and hesperidin, while the 20 and 10th may be dan phenolic acid L and benzoic acid. On the one hand, preliminary study reflects that the root of Schizonepeta tenuifolia may be developed into the medicinal parts in future. On the other hand, the major chemical composition of S. tenuifolia was found to have the anti-lung-tumor effects. This new method was established for the quality control and the rational use of different parts of traditional Chinese medicine.

Ethnopharmacological relevance Metabolomics is the comprehensive assessment of endogenous metabolites of a biological system in a holistic context, and its property consists with the global view of Traditional Chinese Medicine (TCM). Shuihonghuazi Formula (SHHZF) has been used for liver cancer early treatment in clinical for more than thirty years, but its mechanism remains unclear completely. This paper was designed to explore the therapeutic effects of SHHZF on liver cancer and its metabolomic characters. Materials and methods All the rats were given diethylnitrosamine (DEN) at the dosage of 70 mg/kg for 14 weeks. From the 7th weeks, SHHZF was given to the rats which lasted for 10 weeks. Therapeutic effects of SHHZF was compared with that of cyclophosphamide (CTX). High performance liquid-chromatography/electrospray-ionization time of flight mass spectrometer (HPLC/ESI-TOF-MS) combined with pattern recognition approaches including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), was integrated to approximate the comprehensive metabolic signature and discover differentiating metabolites by Agilent MPP 12.1. The changes in metabolic profiling in plasma were restored to their baseline values after SHHZF treatment according to the PLS-DA score plots. Results The results indicated that 23 ions as “differentiating metabolites”. The alterations in those metabolites were associated with perturbations in fatty acid and bile acid metabolism, in response to liver cancer through immune and nervous system. And SHHZF could increase the uptake and utilization of linoleic acid and oleic acid, increase arachidonic acid-like substance content and enhance organism immunity of liver cancer rats. And it also could increase the translation from phosphatidylethanolamine (PE) to phosphatidylcholine (PC), linoleic acid metabolism and inhibits abnormal metabolism of bile acid. Conclusions The mechanism of therapeutic effects of SHHZF on liver cancer by adjusting the activities of PE N-methyl transferase (PEMT), Lysophospholipase D, methylenetetrahydrofolate reductase (MTHFR) and lysophospholipase was elucidated by the method of metabonomics for the first time.

Qizhiweitong particles (QZWT) which is derived from the Sinisan decoction in Shang Han Za Bing Lun, composed of Bupleurum chinenis, Paeonia obovata, Citrus aurantium L., Glycyrrhiza uralensis Fisch., Cyperus rotundus and Rhizoma Corydalis is a traditional Chinese medicine (TCM) treating gastrointestinal diseases. It have been used in clinical for years. It have been used in clinical for years. According to previous research, Bupleurum chinenis, Citrus aurantium, Cyperus rotundus in QZWT play the role of promoting gastric peristalsis, which consist of complex chemical constituents. The aim of this study is to probe the multiple effective components with gastrointestinal prokinetic efficacy in QZWT and investigate the multitarget integrated adjustment mechanism of QZWT curing atropine-induced gastrointestinal motility dysfunction mice.One hundred and thirty two male mice were randomly divided into 11 groups, including control group, model group, Domperidone group, Mosapride group, QZWT group and six components groups. With gastric retention rate, rate of small intestine propulsion, serum content of GAS and MTL as indexes to evaluate the curing effect on gastrointestinal movement disorders caused by atropine in mice. A serum metabonomics method based on the ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) had been established to investigate the mechanism of QZWT and these components, and PCA and PLS-DA have been used to distinguish different groups and found potential biomarkers.Four components from six present good prokinetic effects, including Bupleurum Polysaccharide, Citrus aurantium flavonoid, Citrus aurantium essential oil and Cyperus rotundus flavonoids. These components and QZWT regulate 5 potential biomarkers in the body, and primarily involved in 5 metabolic pathways. These potential biomarkers possess direct or indirect connections, each biomarker regulated by multiple components, each component adjusting multiple targets, and QZWT is nearly the sum of its components.This experiment deepened our understanding of insufficient gastrointestinal dynamics, confirmed that QZWT treating gastrointestinal disorders was through multicomponent, multitarget ways. These results fully reflect the multiple targets synergy characteristics of TCM.

To optimize conditions for extracting total flavonoids from Schizonepeta tenuifolia by the method of analytical hierarchy process with the correlation method of dose-effect.Take the amount and concentration of ethanol,the extraction time and times as examine factors, with the inhibition rate of Caco-2 human cloning colonic cancer cell as index to optimize the optimum conditions of total flavonoids from Schizonepeta tenuifolia by orthogonal experiment design, and analyze the Pearson correlation coefficient between the total flavonoids content, hesperidin content, extraction rate and pharmacodynamics indexes through SPSS 19.0 software to determine the dose-effect comparative indicators, and then analytic hierarchy process is adopted to define the weight coefficients range of the content of total flavonoids and hesperidin.The best extraction process conditions were as follows:15 times the amount of 75% ethanol, water circumfluence extracting for 3 times,each time for 2 hours; The weight coefficient ranges of the content of total flavonoids and hesperidin were 0.7500-0.9000 and 0.1000-0.2500 respectively.Using the comprehensive scores of the content of total flavonoids and hesperidin, which is calculated by weight coefficient ranges that based on the analytical hierarchy process,to replace the inhibition rate as the indicator to optimize the optimum extraction process is scientific and reasonable. This optimization process is simple and practical,which provides a new method for selecting the best extraction indicators to get effective ingredients from traditional Chinese medicine.

To establish the HPLC fingerprint of flavonoids of Phyllanthi Fructus from different habitats.HPLC method was adopted. The flavonoids composition of Phyllanthi Fructus from 10 different habitats was determined on an Agilent C, chromatographic column with 0. 5% formic acid water (A)-acetonitrile (B) as the mobile phase in gradient elution under the wavelength of 254 nm. The HPLC fingerprints of flavonoids composition of Phyllanthi Fructus were established to evaluate the qualitiy of them.The HPLC fingerprints of flavonoids composition of Phyllanthi Fructus from 10 different habitats were established. 18 common peaks were found and the similarities of them were more than 0. 90 except the ones from Guangxi and Guangdong.The method is simple, accurate and repeatable. It can be used for research and quality control of the effective components in Phyllanthi Fructus.

Two new C21 steroidal glycosides were isolated from Cynanchum wallichii Wight. Their structures were elucidated as caudatin-3-O-β-d-glucopyranosyl-(1 → 4)-β-d-oleandropyranosyl-(1 → 4)-β-d-cymaropyranosyl-(1 → 4)-β-d-digitoxopyranoside (1) and caudatin-3-O-β-d-glucopyranosyl-(1 → 4)-β-d-cymaropyranosyl-(1 → 4)-β-d-oleandropyranosyl-(1 → 4)-β-d-cymaropyranosyl-(1 → 4)-β-d-digitoxopyranoside (2) by spectroscopic methods including 1D and 2D NMR experiments.

To establish the chromatography-efficacy relation method for analyzing the anti-inflammatory activity of Qizhi Weitong particles, in order to lay a foundation for quality control and pharmacodynamic evaluation of traditional Chinese medicine compounds.On the basis of a full-time multi-wavelength fusion fingerprint of Qizhi Weitong particles, the latin hypercube sampling was used to divide six herbs in Qizhi Weitong particles into groups of different proportions to determine their inhibition ratios of TNF-alpha, IL-6 and NO released by LPS-induced RAW264. 7 cells. Pharmaeodynamic data and chemical information of HPLC fingerprints of each group were analyzed with the gray correlation method to get the anti-inflammatory effect of each chromatographic peak, and then fitted with BP neural network to establish the chromatography-efficacy relation.There were 25 peaks closely related to the anti-inflammatory activity. With the 25 peaks as input items, the 3-BP network was adopted to establish the neural network model for anti-inflammatory effect of Qizhi Weitong particles.With an error of less than 7%, the model could better fit with the complicated non-linear relation of the compound, and applied in studying the chromatography-efficacy relation. In this study on the chromatography-efficacy relation, a new method is established to evaluate the anti-inflammatory activity of Qizhi Weitong particles. It is of practical significance as an effective approach for controlling quality and exploring the material basis for efficacy of traditional Chinese medicine compounds.

To control the quality of Qizhiweitong granules with the all-time multi-wavelength fusion fingerprint quantification as the major technique.Agilent TC-C18 (4.6 mm x 250 mm, 5 microm) chromatographic column was adopted, with 0.02% formic acid water-acetonitrile as the mobile phase for linear gradient elution. The flow rate was 1 mL x min(-1), column temperature was 30 degrees C, and detector wavelength was 230, 254, 283 nm. Matlab was adopted for all-time multiple-wavelength fusion for data in dif format.A good relationship was shown for albiflorin in 56.5-452 mg x L(-1) (r = 0.999 8), paeoniflorin in 107-856 mg x L(-1) (r = 0.999 8), licorice glycoside in 73.4-687 mg x L(-1) (r = 0.999 8), naringin in 109-872 mg x L(-1) (r = 0.999 8), neohesperidin in 48.0-384 mg L(-1) (r = 0.999 8), and glycyrrhizic acid in 38.6-308 mg x L(-1) (r = 0.999 8), with recoveries of 0.999 8.The method is simple, accurate and highly reproducible, and can provide basis for quality control of Qizhiweitong granules.