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Background and Purpose Squalene epoxidase (SQLE) is a key enzyme involved in cholesterol biosynthesis, but increasing evidence reveals that SQLE is abnormally expressed in some types of malignant tumors, and the underlying mechanism remains poorly understood. Experimental Approach Bioinformatics analysis and RNA sequencing were applied to detect to differentially expressed genes in clinical HCC tumors. AnnexinV-FITC/PI, EdU assay, transwell, IHC staining, cytoskeleton F-actin filaments assay, RNA sequencing, dual-luciferase reporters and HE staining were evaluated to investigate the pharmacological effects and possible mechanisms of SQLE. Key Results We found that SQLE expression is specifically elevated in HCC tumors, correlating with poor clinical outcomes. SQLE promoted HCC growth, EMT, and metastasis both in vitro and in vivo. In contrast, silencing of SQLE expression prevented HCC development. Both RNA-seq and functional experiments revealed that the protumorigenic effect of SQLE on HCC is closely related to the activation of cellular TGF-β/SMAD signaling, but interestingly, the stimulatory effect of SQLE on TGF-β/SMAD signaling and HCC development is also critically dependent on STRAP, a serine and threonine kinase. SQLE expression is well correlated with STRAP in HCC, and further, to amplify TGF-β/SMAD signaling, SQLE even transcriptionally increased STRAP gene expression mediated by the trans-acting factor AP-2α. Finally, as a chemical inhibitor of SQLE, NB-598 markedly inhibited HCC cell growth and tumor development in mouse models. Conclusions and Implications Taken together, SQLE serves as an oncogene in HCC development by activating TGF-β/SMAD signaling, and targeting SQLE could be useful in drug development and therapy for HCC.

Fructus Psoralea is widely used to treat osteoporosis and skin inflammatory diseases. Because of the side effects on the liver, renal and cardiovascular systems, it is processed to salt-processed Fructus Psoraleae to meet the requirements of clinical use. However, the mechanisms involved in the transformation of the chemical components are unclear. In this study, ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry was used to analyze the chemical profiles of this herbal medicine and the chemical transformation mechanism involved during the salt processing was studied. A total of 83 compounds were identified. Principal component analysis and orthogonal partial least squares discriminate analysis were used to observe the distribution trend of all samples and visualize the difference. Raw and processed Fructus Psoraleae were clearly clustered into two groups. Furthermore, 17 marker compounds were identified as primary contributors to their differences based on t-test analysis (p 0.01) and orthogonal partial least squares discriminate analysis (variable importance for the projection 1). Finally, ultra-high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry was used to evaluate the quality of Fructus Psoraleae by simultaneous analysis of 13 components highly related to efficacy. There were variations in the contents of 13 chemicals of Fructus Psoraleae and salt-processed products. The results of untargeted and targeted metabolomics revealed that salt processing affected the chemical composition of Fructus Psoraleae.

Gardeniae Fructus, the dry fruit of Gardenia jasminoides Ellis, has been widely used for the treatment of different diseases. Although four types of processed Gardeniae Fructus products, characterized by differing effects, are available for clinical use, little is known regarding the respective processing mechanisms. In this study, ultra high performance liquid chromatography quadrupole time-of-flight mass spectrometry combined with multivariate statistical analysis was applied to characterize the chemical profiles of the differently processed Gardeniae Fructus products and to determine differences in their chemical compositions, thereby enabling us to identify those active compounds associated with the observed clinical effects. A total of 125 compounds were accordingly identified, among which, 56 were established as primary contributors to the significant differences (p < 0.01) between crude and processed Gardeniae Fructus, based on t-test analysis. Furthermore, the potential mechanisms underlying the chemical transformations that occur during processing were discussed. The findings of this study may not only contribute to the more effective quality control of Gardeniae Fructus but also provide basic information for elucidating the mechanisms underlying the changes in chemical constituents in response to processing, and provide a basis for further investigations of Gardeniae Fructus processing mechanisms. This article is protected by copyright. All rights reserved.

Rhein is an active component from Chinese herbal medicine. It can cause diarrhea by inhibiting Na+ , K+ -ATPase activity on intestinal epithelial cells, thus decreasing the re-absorption of Na+ from intestinal tract to blood. However, when this Na+ , K+ -ATPase inhibition was quantitated by colorimetric method that measures ATPase-catalyzed release of inorganic phosphorus, the data obtained are inconsistent and with great variation. To improve it, we developed a novel method by using inductively coupled plasma mass spectrometry (ICP-MS) to quantitate the amount of intracellular Rb+ . This method largely mimics the 86 RbCl tracer flux assay, but it uses non-radioactive RbCl as flux substrate. The results demonstrated that this method has better precision and accuracy than that of the conventional colorimetric method. More importantly, this method is free of radioactive substance, which is expected to be safer and more convenient than radioactive 86 RbCl tracer flux method. In conclusion, the ICP-MS method for Na+ , K+ -ATPase activity determination is novel and accurate. It can also provide a reference for studying the transport of other metal ions across membrane under biological conditions.

Rhein is an active component from Chinese herbal medicine. It can cause diarrhea by inhibiting Na

Bone defects remain a challenging problem for doctors and patients in clinical practice. Processed pyritum is a traditional Chinese medicine that is often used to clinically treat bone fractures. It contains mainly Fe, Zn, Cu, Mn, and other elements. In this study, we added the extract of processed pyritum to β-tricalcium phosphate and produced a porous composite TPP (TCP/processed pyritum) scaffold using digital light processing (DLP) 3D printing technology. Scanning electron microscopy (SEM) analysis revealed that TPP scaffolds contained interconnected pore structures. When compared with TCP scaffolds (1.35 ± 0.15 MPa), TPP scaffolds (5.50 ± 0.24 MPa) have stronger mechanical strength and can effectively induce osteoblast proliferation, differentiation, and mineralization in vitro. Meanwhile, the in vivo study showed that the TPP scaffold had better osteogenic capacity than the TCP scaffold. Furthermore, the TPP scaffold had good biosafety after implantation. In summary, the TPP scaffold is a promising biomaterial for the clinical treatment of bone defects.

Significance The two-dimensional (2D) magnetic semiconductor CrI3 hosts a variety of strong and tunable magneto-optical effects and allows for the development of novel magneto-optical devices. While the elastic magneto-optical effects in CrI3 are well understood, its recently discovered inelastic magneto-Raman effect remains to have case-specific interpretations varying upon the thickness of CrI3. We perform comprehensive Raman measurements on 2D CrI3 with polarization, temperature, layer number, and magnetic field dependence. We resolve a Davydov-split–induced N-fold multiplet in N-layer CrI3 and reveal the distinct magneto-Raman behaviors of individual phonons within the multiplet. Our results discover a layered-magnetism–coupled phonon scattering mechanism that explains the rich magneto-Raman effect in CrI3 of arbitrary thickness and elucidates the spin–phonon coupling physics in layered magnets., We used a combination of polarized Raman spectroscopy experiment and model magnetism–phonon coupling calculations to study the rich magneto-Raman effect in the two-dimensional (2D) magnet CrI3. We reveal a layered-magnetism–assisted phonon scattering mechanism below the magnetic onset temperature, whose Raman excitation breaks time-reversal symmetry, has an antisymmetric Raman tensor, and follows the magnetic phase transitions across critical magnetic fields, on top of the presence of the conventional phonon scattering with symmetric Raman tensors in N-layer CrI3. We resolve in data and by calculations that the first-order Ag phonon of the monolayer splits into an N-fold multiplet in N-layer CrI3 due to the interlayer coupling (N≥2) and that the phonons within the multiplet show distinct magnetic field dependence because of their different layered-magnetism–phonon coupling. We further find that such a layered-magnetism–phonon coupled Raman scattering mechanism extends beyond first-order to higher-order multiphonon scattering processes. Our results on the magneto-Raman effect of the first-order phonons in the multiplet and the higher-order multiphonons in N-layer CrI3 demonstrate the rich and strong behavior of emergent magneto-optical effects in 2D magnets and underline the unique opportunities of spin–phonon physics in van der Waals layered magnets.

Blood stasis syndrome (BSS) is characterized by blood retardation and is the major cause of some deadly diseases. Some factors that affect BSS have been identified. However, the small molecule that related to BSS is still largely unknown. Traditional Chinese Medicine (TCM), such as Sanleng and Ezhu, has been used for a long time in treating BSS and promising outcomes have been achieved. However, the mechanism of how they work is unclear. Thus, we constructed the Rat BSS model and treated them with Sanleng and Ezhu. Then, the liver dialysis of those rats was collected and the small molecule metabolites were analyzed by GC-MS based metabolomics approach. Our results showed after Sanleng and Ezhu treatment, several small molecule metabolites were significantly changed metabolites (VIP1 and P0.05). Pathway enrichment analysis also showed that Sanleng and Ezhu share the similar mechanism in treating BSS, such as regulating Glyoxylate and dicarboxylate metabolism pathway and energy metabolism. Besides, we also identified some key metabolites that were significantly correlated with BSS. In conclusion, those findings uncover the mechanism of Sanleng and Ezhu in treating BSS.

RATIONALE Crataegi Fructus (CF) is one of the most commonly used herbal medicines with a long history of clinical applications. CF is often processed to minimize gastric membrane irritation, although differently processed products can have different biological effects. The purpose of this study was to comprehensively identify the chemical composition of CF, determine the changes caused by processing, and elucidate the active constituents causing the clinical effects. This study aimed to define a theoretical basis for intensive mechanistic studies of CF processing and its reasonable clinical applications. METHODS An optimized ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC/QqTOFMS) method in positive and negative ion modes, coupled with multivariate statistical analyses, was developed for the identification and analysis of chemical components in raw and processed products of CF. RESULTS A total of 87 compounds were identified, including 61 marker compounds that were found to be primary contributors to the significant differences (p

Crataegi Fructus (CF) is widely used as a medicinal and edible material around the world. Currently, different types of processed CF products are commonly found in the market. Quality evaluation of them mainly relies on chemical content determination, which is time and money consuming. To rapidly and nondestructively discriminate different types of processed CF products, an electronic nose coupled with chemometrics was developed. The odour detection method of CF was first established by single-factor investigation. Then, the sensor array was optimised by a stepwise discriminant analysis (SDA) and analysis of variance (ANOVA). Based on the best-optimised sensor array, the digital and mode standard were established, realizing the odour quality control of samples. Meanwhile, mathematical prediction models including the discriminant formula and back-propagation neural network (BPNN) model exhibited good evaluation with a high accuracy rate. These results suggest that the developed electronic nose system could be an alternative way for evaluating the odour of different types of processed CF products.

Background: The manual identification of Fructus Crataegi processed products is inefficient and unreliable. Therefore, efficient identification of the Fructus Crataegis’ processed products is important. Objective: In order to efficiently identify Fructus Crataegis processed products with different odor characteristics, a new method based on an electronic nose and convolutional neural network is proposed. Methods: First, the original smell of Fructus Crataegis processed products is obtained by using the electronic nose and then preprocessed. Next, feature extraction is carried out on the preprocessed data through convolution pooling layer LCP1, convolution pooling layer LCP2 and a full connection layer LFC. Thus, the feature vector of the processed products can be obtained. Then, the recognition model for Fructus Grataegis processed products is constructed, and the model is trained to obtain the optimized parameters: filters F1 and F2, bias vectors B1, B2, B3, and B4, matrices M1 and M2. Finally, the features of the target processed products are extracted through the trained parameters to achieve accurate prediction. Results: The experimental results show that the proposed method has higher accuracy for the identification of Fructus Crataegis processed products, and is competitive with other machine learning based methods. Conclusion: The method proposed in this paper is effective for the identification of Fructus Crataegi processed products.

We use a combination of polarized Raman spectroscopy experiment and model magnetism-phonon coupling calculations to study the rich magneto-Raman effect in the two-dimensional (2D) magnet $\mathrm{CrI_3}$. We reveal a novel layered-magnetism-assisted phonon scattering mechanism below the magnetic onset temperature, whose Raman excitation breaks time-reversal symmetry, has an antisymmetric Raman tensor, and follows the magnetic phase transitions across critical magnetic fields, on top of the presence of the conventional phonon scattering with symmetric Raman tensors in $N$-layer $\mathrm{CrI_3}$. We resolve in data and by calculations that the 1st-order $A_g$ phonon of monolayer splits into a $N$-fold multiplet in $N$-layer $\mathrm{CrI_3}$ due to the interlayer coupling ($N$>=2) and that the phonons with the multiple show distinct magnetic field dependence because of their different layered-magnetism-phonon coupling. We further find that such a layered-magnetism-phonon coupled Raman scattering mechanism extends beyond 1st-order to higher-order multi-phonon scattering processes. Our results on magneto-Raman effect of the 1st-order phonons in the multiplet and the higher-order multi-phonons in $N$-layer $\mathrm{CrI_3}$ demonstrate the rich and strong behavior of emergent magneto-optical effects in 2D magnets and underlines the unique opportunities of new spin-phonon physics in van der Waals layered magnets., 16 pages, 4 figures + supplementary material

Crataegi Fructus and its processed products have been used as a traditional medicine for a long time, and numerous active components are responsible for their curative effects. However, a comprehensive and fast method for the quality control of its processed products is still lacking. In this study, two analytical methods based on color measurements and fingerprint analysis are established. In the color measurements, the color values of the peel and flesh of Crataegi Fructus were evaluated spectrophotometrically. Based on the results, a color reference range was established using percentiles, and the standard color difference value was established using the median color values. Then, the color values of Crataegi Fructus and its processed products were analyzed using Bayes linear discriminant analysis and mathematical functions were built in order to predict the degree of processing. Moreover, high-performance liquid chromatography fingerprint analysis was performed on a Hibar C18 column, and a high-performance liquid chromatography fingerprint pattern was obtained, from which nine peaks were identified. Chemometric methods were successfully applied to differentiate raw and processed Crataegi Fructus.

With the development of the Chinese transportation industry, the number of bridges has increased significantly, but this results in high pressure of structural maintenance and management. Bridge management system (BMS) is critical for efficient maintenance and ensured safety of bridge structures during long-term operation. Building information modeling (BIM) is an emerging technology with powerful visualization and informatization capability, making it an ideal tool for developing modern management systems. This paper introduces the development of a bridge management system based on the BIM technology. Industry Foundation Classes (IFC) and International Framework for Dictionaries (IFD) standards are studied and extended, and coding rules are proposed for the Chinese bridge industry. Also, a standard structural modeling method is proposed to fast build the bridge BIM model. Web-BIM oriented bridge management is proposed, and portable devices are introduced into the system. Collaborative management is realized for different users. The BIM-based maintenance management system is designed. Finally, a practical BIM-based BMS is established for a long-span cable-stayed bridge in China. This system integrates the BIM with the geographic information system (GIS) and contains information management, inspection management, technical condition evaluation, and enables users to cooperate with each other. Such a BMS could help to improve the management efficiency and ensure its normal operation, providing a useful platform for the maintenance of massive bridges in China.

Yi Huang decoction (YHD) has been used as one of famous traditional formula because of its unique effectiveness against gynecological diseases. YHD is composed of five herbs, including the rootstock of Dioscoma opposita Thunb. (Dioscoreaceae), the kernel of Etayale ferx Salisb. (Nymphaeaceae), the bark of Phellodendron chinense Schneid. (Rutaceae), the seed of Plantago asiatica L. (Plantaginaceae), and the seed of Ginkgo biloba L. (Ginkgoaceae). To effectively control the quality, the processing method for YHD was optimized by means of single factor test as well as orthogonal test in this study. A completely validated method based on HPLC coupled with diode array detector was performed on a Kromasil C(18) column at 30° with mobile phase of 0.1% aqueous phosphoric acid and acetonitrile. As a result, HPLC fingerprint on the basis of the chromatographic data from 32 batches of samples was obtained, which contained 44 common peaks. Among these common peaks, 6 peaks were identified as geniposidic acid, berberine hydrochloride, palmatine hydrochloride, phellodendrine chloride, magnoflorine, and verbascoside, respectively, based on their retention time relative to the standards. Meanwhile, the contents of these 6 compounds were also simultaneously examined. In sum, this study offered valuable information for the proper processing and quality control for YHD.

Estrogen replacement therapy (ERT) is utilized as a major regime for treatment of postmenopausal osteoporosis at present. However, long-term supplement of estrogen may cause uterine hyperplasia and hypertension leading to a high risk of endometrial cancer and breast cancer. Psoralea corylifolia L. has long been used as tonic and food additives in many countries. Previous studies had found two ingredients in P. corylifolia L.: bavachin and bakuchiol exhibited osteoblastic activity. The present study was designed to investigate the protective effect of bakuchiol and bavachin on ovariectomy-induced bone loss and explore the possible mechanism. In vivo, bakuchiol and bavachin could prevented estrogen deficiency-induced bone loss in ovariectomized rats without uterotrophic activity. In vitro studies suggested that bakuchiol and bavachin induced primary human osteoblast differentiation by up-regulating the Wnt signalling pathway. This study suggests that such a bone-protective role makes them a promising and safe estrogen supplement for the ERT.

Crataegi Fructus, an edible food, has been used as a traditional medicine to treat diseases for many years. There is substantial evidence that multiple constituents are responsible for the beneficial effects of Crataegi Fructus. To effectively control the quality of this herbal medicine, we developed an ultra high performance liquid chromatography with electrospray ionization tandem mass spectrometry protocol to simultaneously quantify ten compounds (chlorogenic acid, procyanidin B2, l-epicatechin, glucosylvitexin, vitexin-2-O-rhamnoside, vitexin, rutin, hyperoside, isoquercitrin, and quercetin) in Crataegi Fructus. Multiple-reaction monitoring was used for the quantification in the negative mode for 8 min. This proposed method is simple, reliable, sensitive, and specific. Further, the quantification parameters, including linearity, limit of detection, limit of quantification, precision, reproducibility, stability, and accuracy were optimized. The quality of the processed samples of Crataegi Fructus was evaluated using this method. Additionally, the method was successfully used to distinguish the medicinal components, including peel, kernel, and flesh. The data described in this study offer valuable information for the quality control and proper use of Crataegi Fructus.

In traditional Chinese medicine, raw and processed herbs are used to treat different diseases. Suitable quality assessment methods are crucial for the discrimination between raw and processed herbs. The dried fruit of Arctium lappa L. and their processed products are widely used in traditional Chinese medicine, yet their therapeutic effects are different. In this study, a novel strategy using high-performance liquid chromatography and diode array detection coupled with multivariate statistical analysis to rapidly explore raw and processed Arctium lappa L. was proposed and validated. Four main components in a total of 30 batches of raw and processed Fructus Arctii samples were analyzed, and ten characteristic peaks were identified in the fingerprint common pattern. Furthermore, similarity evaluation, principal component analysis, and hierachical cluster analysis were applied to demonstrate the distinction. The results suggested that the relative amounts of the chemical components of raw and processed Fructus Arctii samples are different. This new method has been successfully applied to detect the raw and processed Fructus Arctii in marketed herbal medicinal products.

In China, Semen Cassiae is used clinically to improve eyesight, relieve constipation, and to treat headache and dizziness. Prepared Semen Cassiae is obtained by stir-frying raw Semen Cassiae until it turned dark brown, micro dilatancy, and overflow aroma. After processing, the therapeutic effects change-the purgation effect is alleviated and the hepatoprotective effect is enhanced. To explore the changes in chemical compositions of Semen Cassiae after processing and clarify the material basis of the changed therapeutic effects, an ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry coupled with automated data analysis software and statistical strategy was developed. As a result, 53 compounds in raw Semen Cassiae and 43 compounds in prepared Semen Cassiae were found, a total of 55 chemical compounds were identified. Principle component analysis and t-test were processed by Markerview 1.2.1 software. Finally, 39 peaks were found to be the main contributors to the significant difference (p < 0.05) between raw and prepared Semen Cassiae. Compared with raw Semen Cassiae, 19 peaks showed a higher intensity in prepared Semen Cassiae, while the contents of 20 compounds in prepared Semen Cassiae were lower, most of which belonged to naphthopyrones glycosides and anthraquinone glycosides.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selectively triggers cancer cell death via its association with death receptors on the cell membrane, but exerts negligible side effects on normal cells. However, some non-small-cell lung carcinoma (NSCLC) patients exhibited resistance to TRAIL treatment in clinical trials, and the mechanism varies. In this study, we described for the first time that toosendanin (TSN), a triterpenoid derivative used in Chinese medicine for pain management, could significantly sensitize human primary NSCLC cells or NSCLC cell lines to TRAIL-mediated apoptosis both in vitro and in vivo, while showing low toxicity against human primary cells or tissues. The underlying apoptotic mechanisms involved upregulation of death receptor 5 (DR5) and CCAAT/enhancer binding protein homologous protein, which is related to the endoplasmic reticulum stress response, and is further associated with reactive oxygen species generation and Ca2+ accumulation. Surprisingly, TSN also induced autophagy in NSCLC cells, which recruited membrane DR5, and subsequently antagonized the apoptosis-sensitizing effect of TSN. Taken together, TSN can be used to sensitize tumors and the combination of TRAIL and TSN may represent a useful strategy for NSCLC therapy; moreover, autophagy serves as an important drug resistance mechanism for TSN.

Gardeniae Fructus (G.Fructus), the fruit of Gardenia jasminoides Ellis (Rubiaceae), is a commonly used traditional Chinese medicine (TCM) that has been used for the treatment of hepatitis, jaundice, hypersonic, diabetes and hematuria. Numerous researches have demonstrated that the major active constituents in G.Fructus were responsible for the majority of medical effects of this fruit and their quantification were important for the quality control of G.Fructus. However, in the current quality control standard, only geniposide was used as characteristic marker of G.Fructus, which could not reflect the overall quality of this fruit. In order to identify more chemical makers for improving the quality control standard and evaluate producing areas differentiation of G.Fructus, in the present study, a novel and sensitive high-performance liquid chromatography-diode array detector coupled to an electrospray tandem mass spectrometer (HPLC-DAD-ESI/MS) was developed for the simultaneous determination of 8 major constituents, including geniposidic acid (1), chlorogenic acid (2), genipin-1-β-gentiobioside (3), geniposide (4), genipin (5), rutin (6), crocin-1 (7), crocin-2 (8) in G.Fructus. Moreover, chemometric analysis techniques with principal component constituent analysis (PCA) and cluster analysis (CA) involved were introduced in statistical analysis of 8 investigated constituents in the 34 batches samples to discriminate the samples from different producing areas. The results indicated that the contents of the 8 major bioactive constituents in G.Fructus varied significantly among different producing areas. From results of the loading plot from PCA analysis, genipin-1-β-gentiobioside may have more influence in discriminating the sample from different producing areas, and which was found to be the most abundant bioactive component besides geniposide in all the 34 batches samples, suggesting that it should be added as chemical marker for further investigation on the pharmacological actions and the quality control of G.Fructus.

Recently, combination therapy with acupuncture and medicine as a practical strategy to treat diseases has gained increasing attention. The present study aimed to investigate whether acupuncture stimulation at ST.36 had a potential impact on the pharmacokinetics and tissue distribution of lignans. An HPLC-ESI/MS analytical method was established and successfully applied to a comparative study of drug concentration in plasma and tissues of three lignans. The parameters area under the plasma concentration-time curve from time zero to the final measurable point and from time zero to infinity, and peak concentration were significantly increased, with a prolonged mean residence time and a corresponding decrease in clearance in comparision with the Schisandra-alone group. Additionally, tissue concentrations of three lignans were improved in the group with acupuncture, especially in liver. The results indicated that acupuncture has a synergistic effect on the pharmacokinetics and tissue distribution of the three lignans, which could postpone their elimination, resulting in a longer blood circulating time in rat plasma and prolonged residence time in target tissues, leading to higher tissue concentration. The findings provide some scientific evidence for the mechanism of the combined use of acupuncture and herbal medicine. Furthermore, we suggest that acupuncture and its combination with herbal medicine should be investigated further as a possible adjuvant therapy in clinical treatment for liver injury.

Ethnopharmacological relevance Schisandra chinensis (SC) is a well-known traditional Chinese herbal medicine that has been used in clinical practices for thousands of years. However, the differences between the effects of unprocessed and vinegar-processed Schisandra chinensis (VSC) on cytochrome P450 (CYP450) activities are poorly understood. Aim of the study To evaluate the differences between processed and unprocessed SC on the metabolism of CYP1A2, CYP2E1 and CYP3A4 substrates in rats using a cocktail method based on a developed and validated HPLC method. We also investigate the influence of processing on the levels of CYP mRNA. Materials and methods Three probe substrates (theophylline, dapsone and chlorzoxazone) were delivered simultaneously into rats treated with single or multiple doses of processed or unprocessed SC extract. The plasma concentrations of the three probes were profiled by HPLC, and their corresponding pharmacokinetic parameters were calculated. Real-time RT-PCR was performed to determine the effects of processed and unprocessed SC on the mRNA expression of CYP1A2, CYP2E1 and CYP3A4 in the liver. Results Treatment with single or multiple doses of either extract of SC induced CYP3A4 enzyme activity and inhibited CYP1A2 enzyme activity in rats. Furthermore, the inhibitory effect of SC was more potent after vinegar processing than without vinegar processing. CYP2E1 enzyme activity was induced after treatment with a single dose but was inhibited after multiple doses. The mRNA expression results were in accordance with the pharmacokinetic results. Conclusions These results provide useful scientific data for the safe clinical application of either extract of SC in combination with other drugs, which should lack the side effects induced by other herb–drug interactions.

The ripened fruit of Schisandrae Chinensis Fructus has unique medical properties in Chinese medicine. It is commonly used after vinegar steaming. Vinegar steaming changes the color of Schisandrae Chinensis Fructus from red to black and enhances its acidic and astringent properties. Lignans are the well-investigated components in this herb. However, Schisandrae Chinensis Fructus is acidic in the theory of Chinese medicine, and whether vinegar processing changes its organic acid components remains largely unknown. In this study, the organic acids in this herb were derived by the method of methyl esterification, and further analyzed by gas chromatography with mass spectrometry. A total of 39 organic acid compounds were identified. Interestingly, Schisandrae Chinensis Fructus after vinegar processing showed a significant increase in the content of levulinic acid as compared to the unprocessed ones. Pharmacological experiments demonstrated that levulinic acid inhibited the contractility of isolated intestine and had an inhibitory effect on the excessive hyperfunction of small intestinal propulsion. Moreover, the extracts of vinegar-processed Schisandrae Chinensis Fructus had a stronger inhibitory on the excessive hyperfunction of small intestinal propulsion than that of unprocessed ones. Taken together, this study offers novel insight into the effect of Schisandrae Chinensis Fructus after vinegar processing.

Summary The ripe fruits of Schisandrae chinensis have a long history of use in traditional Chinese medicine to treat diseases and improve health. There is substantial evidence that lignan constituents are mainly responsible for the beneficial effects of this plant medicine. The amounts of the major bioactive lignans in this plant vary widely with species, habitat, and the collecting time, and as such, establishment of an HPLC fingerprint for quality control of this herbal medicine is of particular importance. To achieve this, ten batches of Fructus schisandrae chinensis were collected from Tieli, in China, and their chemical components were analyzed under optimized HPLC conditions. On the basis of the chromatographic data, a consistent HPLC fingerprint pattern containing 20 common peaks was obtained. Among these common peaks, six were identified as schizandrin, schizandrol B, schisantherin, deoxyschiandrin, γ-schizandrin B, and schizandrin C. On the basis of this HPLC fingerprint and principal-components ...

We have previously implicated reactive oxygen species oxygen (ROS) as a critical signal transducer in the upregulation of Na,K-ATPase by low K+ in MDCK cells, but how ROS mediate this process has not been well defined. We reported here that both of hydrogen peroxide (H2O2) and superoxide anion (O2*(-)) were rapidly produced at the early stage of low K+-treated MDCK cells. Further analysis revealed that NADP/NADPH oxidase-derived H2O2 was specifically involved in low K+-induced Na,K-ATPase alpha1 gene transcription as well as alpha1 and beta1 subunits expressions. Exogenous H2O2 even mimicked the stimulatory effect of low K+ on Na,K-ATPase alpha1 gene transcription. Low K+ triggered a H2O2-dependent ERK1/2 phosphorylation in MDCK cells, nonetheless, this ERK1/2 activation did not finally lead to the upregulation of Na,K-ATPase. Similar to previous findings that Na,K-ATPase beta1 gene transcription was mediated by Sp1, Na,K-ATPase alpha1 gene transcription in low K+-treated MDCK cells was also closely relevant to Sp1 participation, as confirmed by siRNA as well as PCR mutagenesis technologies. Furthermore, Sp1 activation was dependent on H2O2 generation triggered by low K+. Taken together, the data described in this study outlines an essential role of H2O2 and Sp1 in mediating the upregulation of Na,K-ATPase in MDCK cells by low external K+.

To study the cytotoxicity of four alkaloids: brucine, strychnine, brucine N-oxide and isostrychnine from nux vomica on SMMC 7721 cells and their possible mechanisms, MET assay was used to examine the growth inhibitory effects of these alkaloids. Brucine revealed the strongest growth inhibitory effect on SMMC-7721 cells. Furthermore, as directly observed under an inverted microscope, fluorescent microscope and transmission electronic microscope, brucine caused SMMC-7721 cell shrinkage, membrane blobbing, formation of apoptotic body as well as nucleus condensation, all of which are typical characteristics of apoptotic programmed cell death. In addition, brucine dose-dependently caused SMMC-7721 cells apoptosis via formation of subdipolid DNA and phosphatidylserine externalization, as evidenced by flow cytometry analysis. The brucine-induced apoptosis was partially attributed to the activation of caspase 3 as well as cyclooxygenase 2 inhibition, since neither caspase 3 specific inhibitor, z-DEVD-fmk nor was exogenous addition of prostaglandin E(2) able to completely abrogate the brucine-induced SMMC 7721 cell apoptosis. In sum, this paper indicate that the major alkaloids present in the seed of Strychnos nux-vomica are effective against SMMC-7721 cells proliferation, among which brucine proceeds SMMC-7721 cells death via apoptosis, probably through the participation of caspase 3 and cyclooxygenase 2.

Crataegi Fructus, an edible food, has been used as a traditional medicine to treat diseases for many years. There is substantial evidence that multiple constituents are responsible for the beneficial effects of Crataegi Fructus. To effectively control the quality of this herbal medicine, we developed an ultra high performance liquid chromatography with electrospray ionization tandem mass spectrometry protocol to simultaneously quantify ten compounds (chlorogenic acid, procyanidin B2, l-epicatechin, glucosylvitexin, vitexin-2-O-rhamnoside, vitexin, rutin, hyperoside, isoquercitrin, and quercetin) in Crataegi Fructus. Multiple-reaction monitoring was used for the quantification in the negative mode for 8 min. This proposed method is simple, reliable, sensitive, and specific. Further, the quantification parameters, including linearity, limit of detection, limit of quantification, precision, reproducibility, stability, and accuracy were optimized. The quality of the processed samples of Crataegi Fructus was evaluated using this method. Additionally, the method was successfully used to distinguish the medicinal components, including peel, kernel, and flesh. The data described in this study offer valuable information for the quality control and proper use of Crataegi Fructus.

In an attempt to dissect the mechanism of Strychnos nux-vomica, a commonly used Chinese folk medicine in the therapy of liver cancer, the cytotoxic effects of four alkaloids in Strychnos nux-vomica, brucine, brucine N-oxide, strychnine, and isostrychnine, on human hepatoma cells (HepG2) were screened by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrasolium bromide (MTT) assay. Brucine, among the four alkaloids, exhibited the strongest toxic effect, the mechanism of which was found to cause HepG2 cell apoptosis, since brucine caused HepG2 cell shrinkage, the formation of apoptotic bodies, DNA fragmentation, cell cycle arrest, as well as phosphatidylserine externalization, all of which are typical characteristics of apoptotic programmed cell death. Brucine-induced HepG2 cell apoptosis was caspase dependent, with caspase-3 activated by caspase-9. Brucine also caused the proteolytic processing of caspase-9. In addition, brucine caused depolarization of the mitochondrial membrane of HepG2 cells, the inhibition of which by cyclosporine A completely abrogated the activation of casapses and release of cytochrome c in brucine-treated HepG2 cells. These findings suggested a pivotal role of mitochondrial membrane depolarization in HepG2 cell apoptosis elicited by brucine. Furthermore, brucine induced a rapid and sustained elevation of intracellular [Ca2+], which compromised the mitochondrial membrane potential and triggered the process of HepG2 cell apoptosis. Finally, Bcl-2 was found to predominately control the whole event of cell apoptosis induced by brucine. The elevation of [Ca2+]i caused by brucine was also suppressed by overexpression of Bcl-2 protein in HepG2 cells. From the facts given above, Ca2+ and Bcl-2 mediated mitochondrial pathway were found to be involved in brucine-induced HepG2 cell apoptosis.

In traditional Chinese medicine, raw and processed herbs are used to treat different diseases. Suitable quality assessment methods are crucial for the discrimination between raw and processed herbs. The dried fruit of Arctium lappa L. and their processed products are widely used in traditional Chinese medicine, yet their therapeutic effects are different. In this study, a novel strategy using high-performance liquid chromatography and diode array detection coupled with multivariate statistical analysis to rapidly explore raw and processed Arctium lappa L. was proposed and validated. Four main components in a total of 30 batches of raw and processed Fructus Arctii samples were analyzed, and ten characteristic peaks were identified in the fingerprint common pattern. Furthermore, similarity evaluation, principal component analysis, and hierachical cluster analysis were applied to demonstrate the distinction. The results suggested that the relative amounts of the chemical components of raw and processed Fructus Arctii samples are different. This new method has been successfully applied to detect the raw and processed Fructus Arctii in marketed herbal medicinal products.

The ripe fruits of Gardenia jasminoides Ellis have been used as traditional Chinese medicine to treat diseases for a long history. Lines of evidence demonstrate that multiple active constituents are responsible for the therapeutic effects of this herbal medicine. However, effective methods for quality control of this herbal medicine are still lacking. In this study, a high-performance liquid chromatography (HPLC) fingerprint analysis was performed on a SinoChrom ODS-BP C18 column (4.6 mm × 250 mm, 5 μm) at 30°C with mobile phase of aqueous solution with 0.1% formic acid and acetonitrile. On the basis of the chromatographic data from 32 batches samples, the HPLC fingerprint pattern containing 27 common peaks was obtained. Among these common peaks, seven peaks were identified by the electrospray ionization-mass spectrometry as geniposidic acid, genipin-1-β-gentiobioside, chlorogenic acid, geniposide, rutin, crocin-1 and crocin-2 and the contents of these seven compounds were simultaneously determined. Finally, chemometric methods including hierarchical clustering analysis and principal component analysis were successfully applied to differentiate the samples from six producing regions. In sum, the data, as described in this study, offer valuable information for the quality control and proper use of Gardeniae Fructus.

Recently, combination therapy with acupuncture and medicine as a practical strategy to treat diseases has gained increasing attention. The present study aimed to investigate whether acupuncture stimulation at ST.36 had a potential impact on the pharmacokinetics and tissue distribution of lignans. An HPLC-ESI/MS analytical method was established and successfully applied to a comparative study of drug concentration in plasma and tissues of three lignans. The parameters area under the plasma concentration-time curve from time zero to the final measurable point and from time zero to infinity, and peak concentration were significantly increased, with a prolonged mean residence time and a corresponding decrease in clearance in comparision with the Schisandra-alone group. Additionally, tissue concentrations of three lignans were improved in the group with acupuncture, especially in liver. The results indicated that acupuncture has a synergistic effect on the pharmacokinetics and tissue distribution of the three lignans, which could postpone their elimination, resulting in a longer blood circulating time in rat plasma and prolonged residence time in target tissues, leading to higher tissue concentration. The findings provide some scientific evidence for the mechanism of the combined use of acupuncture and herbal medicine. Furthermore, we suggest that acupuncture and its combination with herbal medicine should be investigated further as a possible adjuvant therapy in clinical treatment for liver injury.

A sensitive, specific, convenient and endogenous interference-free microdialysis sampling method coupled with RRLC–MS was successfully developed and applied to the determination of protein-unbound curdione and curcumol in biological samples. Microdialysis probes were simultaneously inserted into the jugular vein toward heart and the median lobe near the center of liver of rats under anesthesia. The separation was accomplished on a Zorbax SB-C18 column (2.1 mm × 50 mm, 1.8 μm) with a gradient elution and chromatography was conducted with RRLC system. Analytes were detected by positive ion electrospray tandem mass spectrometry and quantified on the basis of extracted ion chromatography (EIC) peak area signal. The calibration curves were linear over the range of 3.3–213.2 ng/mL for curdione and 8.1–519.2 ng/mL for curcumol. All the validation data, such as accuracy, precision, stability and matrix effect were satisfactory and within the required limits. The validated method was successfully applied to the pharmacokinetic study of curdione and curcumol in rat blood and liver after oral administration of Rhizoma Curcumae extracts. The results could provide a meaningful basis for better understanding of the intracorporal process of Rhizoma Curcumae , which would be helpful for further study both in clinic and laboratory.

A new chemical component, cis-Z,Z'-3a.7a',7a.3a'-dihydroxyligustilide, was isolated from Angelica sinensis and its structure elucidated from its NMR and MS spectra and confirmed by X-ray single crystal diffraction analysis. We also explored the antioxidative properties of cis-Z,Z'-3a.7a',7a.3a'-dihydroxyligustilide on human umbilical vein endothelial cells (HUVECs) against injuries induced by hydrogen peroxide (H(2)O(2)) using an MTT assay and flow cytometry analysis. In addition, the activities of superoxide dismutase (SOD), malondialdehyde (MDA), lactate dehydrogenase (LDH), nitric oxide (NO) and reactive oxygen species (ROS) were determined. We found that cis-Z,Z'-3a.7a',7a.3a'-dihydroxyligustilide increased the viability of HUVECs injured by H(2)O(2) in a dose-dependent manner, reduced the apoptosis of HUVEC, and enhanced HUVEC proliferation. Our results demonstrated the remarkable in vitro antioxidative activities of this compound, indicating that it could be a potential antioxidant with protective effects against H(2)O(2)-induced HUVEC injuries.

To study the molecular mechanism of cyclooxygenase-2 (COX-2), one of effective ingredient of brucine, in inducing non-small cell lung cancer cell apoptosis.COX-2 promoter, transcription factor deletion mutants and COX-2 mRNA 3'-UTR-containing report plasmids were transfected with Renillia to non-small cell lung cancer A549 cell, in order to detect the activity of report gene luciferase and minimum cis-acting element of COX-2 promoter inhibited by brucine. The influence of brucine on IkappaB phosphorylation and the nuclear translocation of p65 were detected by immunoblotting assay.Brucine significantly suppressed LPS-induced COX-2 promoter activation, but revealed minor impact on COX-2 mRNA stability. NF-kappaB in the vicinity of COX-2 promoter-262 was an important cis-acting element of brucine for inhibiting the activity of COX-2 promoter. Brucine was found to inhibit the phosphorylation of IkappaBalpha as well as the nuclear translocation of p65.Brucine can improve A549 cells apoptosis by inhibiting the activity of NF-kappaB and the subsequent COX-2 gene expression.

OBJECTIVE To study the molecular mechanism of cyclooxygenase-2 (COX-2), one of effective ingredient of brucine, in inducing non-small cell lung cancer cell apoptosis. METHOD COX-2 promoter, transcription factor deletion mutants and COX-2 mRNA 3'-UTR-containing report plasmids were transfected with Renillia to non-small cell lung cancer A549 cell, in order to detect the activity of report gene luciferase and minimum cis-acting element of COX-2 promoter inhibited by brucine. The influence of brucine on IkappaB phosphorylation and the nuclear translocation of p65 were detected by immunoblotting assay. RESULT Brucine significantly suppressed LPS-induced COX-2 promoter activation, but revealed minor impact on COX-2 mRNA stability. NF-kappaB in the vicinity of COX-2 promoter-262 was an important cis-acting element of brucine for inhibiting the activity of COX-2 promoter. Brucine was found to inhibit the phosphorylation of IkappaBalpha as well as the nuclear translocation of p65. CONCLUSION Brucine can improve A549 cells apoptosis by inhibiting the activity of NF-kappaB and the subsequent COX-2 gene expression.

Psoralea Fructus, the dried and ripe fruit of Psoralea corylifolia L., have been used as traditional medicine. There is substantial evidence that multiple constituents are responsible for the beneficial effects of this medicine. To effectively control the quality of this herbal medicine, HPLC fingerprint analysis was performed on a SinoChrom ODS-BP column with mobile phase of a gradient prepared from H2O and CH3CN, which the conditions used for gradient elution were: 0-10 min, 5-45% CH3CN; 10-45 min, 45-70% CH3CN; 45-50 min, 70-100% CH3CN; 50-60 min, 100-100% CH3CN, and the flow rate was 1.0 ml/min. It was obtained on the basis of the chromatographic data from 28 batches of samples, which contained 26 common peaks and 13 peaks were identified by the electrospray ionization-mass spectrometry as psoralen, isopsoralen, isobavachin, neobavaisoflavone, bavachin, corylin, broussochalcone B, psoralidin, isobavachalcone, bavachinin, corylifol A, bavachalcone and backuchiol. The contents of these 13 compounds were also simultaneously examined. By using principal component analysis, 28 batches of samples collected from 6 producing locations with different collecting time were evaluated and differentiated. In summary, the data as described in this study offer valuable information for quality control and proper use of Psoralea Fructus.

To screen the anti-tumor effects of the four alkaloids: brucine, strychnine, brucine N-oxide and isostrychnine from the seed of Strychnos nux-vomica, MTT assay was used to examine the growth inhibitory effects of these alkaloids on human hepatoma cell line (HepG2). Brucine, strychnine and isostrychnine revealed significant inhibitory effects against HepG2 cell proliferation, whereas brucine N-oxide didn't have such an effect. In addition, brucine caused HepG2 cell shrinkage, membrane blebbing, apoptotic body formation, all of which are typical characteristics of apoptotic programmed cell death. The results of flow cytometric analysis demonstrated that brucine caused dose-dependent apoptosis of HepG2 cells through cell cycle arrest at G0/G1 phase, thus preventing cells entering S or G2/M phase. Immunoblot results revealed that brucine significantly decreased the protein expression level of cyclooxygenase-2, whereas increased the expression caspase-3 as well as the caspase-3-like protease activity in HepG2 cells, suggesting the involvement of cyclooxygenase-2 and caspase-3 in the pro-apoptotic effects exerted by brucine. Therefore, this paper indicate that the major alkaloids present in the seed of Strychnos nux-vomica are effective against HepG2 cells proliferation, among which brucine proceed HepG2 cells death via apoptosis, probably through the participation of caspase-3 and cyclooxygenase-2.

To further understand the purpose of the traditional processing method of the seeds of Strychnos nux-vomica L. (Loganiaceae) as well as analgesic and anti-inflammatory activities of brucine and brucine N-oxide extracted from this medicinal plant, various pain and inflammatory models were employed in the present study to investigate their pharmacological profiles. Both brucine and brucine N-oxide revealed significant protective effects against thermic and chemical stimuli in hot-plate test and writhing test. However, on different phases they exerted analgesic activities in formalin test. Brucine N-oxide showed stronger inhibitory effect than brucine in carrageenan-induced rat paw edema, both of them significantly inhibited the release of prostaglandin E2 in inflammatory tissue, reduced acetic acid-induced vascular permeability and the content of 6-keto-PGF1a in Freund's complete adjuvant (FCA) induced arthritis rat's blood plasma. In addition, brucine and brucine N-oxide were shown to reduce the content of 5-hydroxytryptamine (5-HT) in FCA-induced arthritis rat's blood plasma, while increase the content of 5-hydroxytryindole-3-acetic acid (5-HIAA) accordingly. These results suggest that central and peripheral mechanism are involved in the pain modulation and anti-inflammation effects of brucine and brucine N-oxide, biochemical mechanisms of brucine and brucine N-oxide are different even though they are similar in chemical structure.

To study the effect of chitosan clarification refining the decoction of Jiangtangtongmai by homogenous design method.The content of catalpol in the refined decoction by the alcohol-precipitation method and chitosan clarification method was determined by RP-HPLC.Catalpol can be well preserved by chitosan clarification method.Chitosan clarification method can replace alcohol-precipitation method in refining of the decoction of Jiangtangtongmai.