Integrated cell metabolomics and network pharmacology approach deciphers the anti-testosterone deficiency mechanisms of Bushen Zhuanggu Tang.

Testicular dysfunction is distinguished by a deficiency in testosterone levels, which can be attributed to the occurrence of oxidative stress injury in Leydig cells. The empirical prescription known as Bushen Zhuanggu Tang, developed by a highly experienced traditional Chinese medicine practitioner with six decades of clinical expertize, aligns with the traditional Chinese medicine principle of "kidney governing bone". Researchers have demonstrated that the administration of BSZGT can effectively enhance testosterone production. The objective of this study is to investigate the potential anti-testicular dysfunction effects of BSZGT and elucidate its underlying mechanism in an in vitro setting. Specifically, the impact of oxidative stress induced by H2O2 on the activity and testosterone levels of Leydig cells (TM3) was examined. Furthermore, the utilization of UPLC-QE-Qrbitrap-MS enabled the identification of the involvement of BSZGT in various metabolic pathways, including arginine biosynthesis, amino acyl-tRNA biosynthesis, Alanine, aspartate and glutamine metabolism, and Citrate Cycle, through the modulation of 25 distinct metabolites. Additionally, a network pharmacological analysis was conducted to investigate the pivotal protein targets associated with the therapeutic effects of BSZGT. The findings demonstrate the identification of six key proteins (CYP19A1, CYP1B1, ALOX5, ARG1, XDH, and MPO) that play a significant role in augmenting testicular function through their involvement in the ovarian steroid production pathway. In summary, our study presents a comprehensive research methodology that combines cell metabonomics and network pharmacology to enhance the discovery of new therapeutic agents for TD.

Study of the Mechanism of Astragali Radix in Treating Type 2 Diabetes Mellitus and Its Renal Protection Based on Enzyme Activity, Network Pharmacology, and Experimental Verification.

Astragali Radix (AR) is a common Chinese medicine and food. This article aims to reveal the active role of AR in treating Type 2 diabetes mellitus (T2DM) and its renal protective mechanism. The hypoglycemic active fraction was screened by α-glucosidase and identified by UPLC-QE-Orbitrap-MS spectrometry. The targets and KEGG pathway were determined through the application of network pharmacology methodology. Molecular docking and molecular dynamics simulation technology were used for virtual verification. Subsequently, a mouse model of T2DM was established, and the blood glucose and renal function indexes of the mice after administration were analyzed to further prove the pharmacodynamic effect and mechanism of AR in the treatment of T2DM. HA was determined as the best hypoglycemic active fraction by the α-glucosidase method, with a total of 23 compounds identified. The main active components, such as calycoside-7-O-ß-D-glucoside, methylnisoline, and formononetin, were revealed by network pharmacology. In addition, the core targets and the pathway have also been determined. Molecular docking and molecular dynamics simulation techniques have verified that components and targets can be well combined. In vivo studies have shown that AR can reduce blood sugar levels in model mice, enhance the anti-inflammatory and antioxidant activities of kidney tissue, and alleviate kidney damage in mice. And it also has regulatory effects on proteins such as RAGE, PI3K, and AKT. AR has a good therapeutic effect on T2DM and can repair disease-induced renal injury by regulating the RAGE/PI3K/Akt signaling pathway. This study provides ideas for the development of new drugs or dietary interventions for the treatment of T2DM.

Exploring Asthma Mechanism of Belamcanda Chinensis by "Dose-effect Weighted Coefficient" Network Pharmacology and Experiment Validation.

BACKGROUND: Asthma is a chronic inflammatory disease of the airways that seriously endangers human health. Belamcanda chinensis (BC), a traditional Chinese medicine, has been used to counteract asthma as it has been shown to possess anti-inflammatory and regulatory immunity properties. OBJECTIVE: The study aimed to investigate the mechanisms of action of BC in the treatment of asthma; a "dose-effect weighted coefficient" network pharmacology method was established to predict potential active compounds. METHODS: Information on the components and content of BC was obtained by UPLC-QEOrbitrap- MS spectrometry. Based on BC content, oral bioavailability, and molecular docking binding energy, dose-effect weighting coefficients were constructed. With the degree greater than average as the index, a protein-protein interaction (PPI) database was used to obtain the core key targets for asthma under dose-effect weighting. GO function and KEGG pathway analyses of the core targets were performed using DAVID software. Finally, MTT and ELISA assays were used to assess the effects of active components on 16HBE cell proliferation. RESULTS: The experimental results using the 16HBE model demonstrated BC to have a potential protective effect on asthma. Network pharmacology showed SYK, AKT1, and ALOX5 to be the main key targets, and Fc epsilon RI as the promising signaling pathway. Eight components, such as tectoridin, mangiferin, luteolin, and isovitexin were the main active compounds, Finally, we analyzed the LPS-induced 16HBE proliferation of each active ingredient. Based on the activity verification study, all five predicted components promoted the proliferation of 16HBE cells. These five compounds can be used as potential quality markers for asthma. CONCLUSION: This study provides a virtual and practical method for the simple and rapid screening of active ingredients in natural products.

Mechanism of action of Huangbaichen Sanwei formulation in treating T2DM based on network pharmacology and molecular docking.

Huangbaichen Sanwei formulation (HBCS) has been reported to have a good hypoglycemic effect, but its pharmacological mechanism of action remains unclear. We used network pharmacology and molecular docking to explore the potential mechanism of action of HBCS against type-2 diabetes mellitus (T2DM). Fifty-five active components from HBCS interfered with T2DM. Twenty-five core targets, such as AKT1, INS, INSR, MAPK1 were identified. Enrichment analyses showed that HBCS was involved mainly including insulin receptor signaling pathway, extracellular region, and insulin-like growth factor receptor binding and other biological processes; common targets had roles in treating T2DM by regulating diabetic cardiomyopathy and insulin resistance. Molecular docking verified that components combined with core targets. HBCS play a part in treating T2DM through multiple components and targets at the molecular level, which lays a theoretical foundation for research using HBCS to treat T2DM. The components, predicted targets, and T2DM targets of HBCS were searched through databases, and common targets were determined. Further screening of the core targets was conducted through the establishment of a protein -protein interaction network. The core targets were analyzed by Gene Ontology (GO) annotation utilizing the DAVID platform. And the enrichment of signaling pathways was explored by employing the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Cytoscape 3.9.1 was employed to construct a "TCM-components-core target-pathway" network. Autodock Vina was used to dock molecules to compare the binding activity of active molecules with targets.

Salvianolic Acid B Reduces the Inflammation of Fat Grafts by Inhibiting the NF-Kb Signalling Pathway in Macrophages.

BACKGROUND: Autologous fat grafting is a common method for soft tissue defect repair. However, the high absorption rate of transplanted fat is currently a bottleneck in the process. Excessive inflammation is one of the main reasons for poor fat transplantation. Salvianolic acid B (Sal-B) is a herbal medicine that shows promise for improving the effectiveness of fat transplantation. OBJECTIVE: The aim of this study was to improve fat graft survival by injecting Sal-B into fat grafts locally. METHODS: In vivo, 0.2 mL of Coleman fat was transplanted into nude mice along with Sal-B. The grafts were evaluated by histologic analysis at 2, 4, and 12 weeks posttransplantation and by microcomputed tomography at 4 weeks posttransplantation. In vitro ribonucleic acid sequencing, cell proliferation assays, anti-inflammatory activity assays, molecular docking studies, and kinase activity assays were performed in RAW264.7 cells to detect the potential mechanism. RESULTS: Sal-B significantly improved fat graft survival and attenuated adipose tissue fibrosis and inflammation. Sal-B also inhibited the polarization of M1 macrophages in fat grafts. In vitro, Sal-B inhibited the proliferation and activation of inflammatory pathways in RAW264.7 cells. In addition, Sal-B had an inhibitory effect on NF-κB (nuclear factor κ light polypeptide gene enhancer in B cells) signaling. This bioactivity of Sal-B may result from its selective binding to the kinase domain of the inhibitor of NF-κB kinase subunit ß. CONCLUSIONS: Sal-B could serve as a promising agent for improving the effect of fat transplantation by inhibiting the polarization of M1 macrophages through NF-κB signaling.

Screening of the Active Compounds against Neural Oxidative Damage from Ginseng Phloem Using UPLC-Q-Exactive-MS/MS Coupled with the Content-Effect Weighted Method.

The neuroprotective properties of ginsenosides have been found to reverse the neurological damage caused by oxidation in many neurodegenerative diseases. However, the distribution of ginsenosides in different tissues of the main root, which was regarded as the primary medicinal portion in clinical practice was different, the specific parts and specific components against neural oxidative damage were not clear. The present study aims to screen and determine the potential compounds in different parts of the main root in ginseng. Comparison of the protective effects in the main root, phloem and xylem of ginseng on hydrogen peroxide-induced cell death of SH-SY5Y neurons was investigated. UPLC-Q-Exactive-MS/MS was used to quickly and comprehensively characterize the chemical compositions of the active parts. Network pharmacology combined with a molecular docking approach was employed to virtually screen for disease-related targets and potential active compounds. By comparing the changes before and after Content-Effect weighting, the compounds with stronger anti-nerve oxidative damage activity were screened out more accurately. Finally, the activity of the selected monomer components was verified. The results suggested that the phloem of ginseng was the most effective part. There were 19 effective compounds and 14 core targets, and enriched signaling pathway and biological functions were predicted. After Content-Effect weighting, compounds Ginsenosides F1, Ginsenosides Rf, Ginsenosides Rg1 and Ginsenosides Rd were screened out as potential active compounds against neural oxidative damage. The activity verification study indicated that all four predicted ginsenosides were effective in protecting SH-SY5Y cells from oxidative injury. The four compounds can be further investigated as potential lead compounds for neurodegenerative diseases. This also provides a combined virtual and practical method for the simple and rapid screening of active ingredients in natural products.

Compound traditional Chinese medicine dermatitis ointment ameliorates inflammatory responses and dysregulation of itch-related molecules in atopic dermatitis.

BACKGROUND: Atopic dermatitis (AD) is a chronic inflammatory skin disease accompanied with itchy and scaly rash. Compound traditional Chinese medicine dermatitis ointment (CTCMDO) consists of a mixture of extracts from five plants, which had been used in AD treatment due to good anti-inflammatory and anti-allergic effects. MATERIALS AND METHODS: In this study, high-performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometer (LC/MS) were performed to analyze the active ingredients of CTCMDO in detail and to establish its HPLC fingerprint. Furthermore, the anti-inflammatory and antipruritic activities of CTCMDO were studied in the treatment of DNCB-induced AD in mice. RESULTS: A total of 44 compounds including phenylpropionic acid compounds, alkaloid compounds, curcumin compounds and lignans were identified via combined HPLC and LC/MS. A fingerprint with 17 common peaks was established. In AD-like mice, DNCB-induced scratching behavior had been suppressed in the treatment of CTCMDO in a dose-dependent manner. Furthermore, the detailed experimental results indicated that the AD can be effectively improved via inhibiting the production of Th1/2 cytokines in serum, reversing the upregulation of substance P levels of itch-related genes in the skin, and suppressing the phosphorylation of JNK, ERK, and p38 in the skin. CONCLUSION: This work indicated that CTCMDO can significantly improve AD via attenuating the pathological alterations of Th1/2 cytokines and itch-related mediators, as well as inhibiting the phosphorylation of mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB).

Dietary Intake of N-3 and N-6 Polyunsaturated Fatty Acids and Risk of Cancer: Meta-Analysis of Data from 32 Studies.

BACKGROUND: Large epidemiological studies have yielded conflicting results regarding the relationship between polyunsaturated fatty acids (PUFAs) and cancers. Here, we performed a meta-analysis to examine the link between dietary intake of n-3 and n-6 PUFAs and cancer risk. MATERIALS AND METHODS: We performed a search on PubMed, EMBASE, and the Cochrane Library. Studies that reported adjusted relative risk (RR) estimates with 95% confidence intervals (CI) for the associations of interest were included. RESULTS: Thirty-two studies involving 1,445,732 participants were included. Colorectal, breast and prostate cancer had been analyzed in our study. Specifically, for colorectal cancer, total n-3 PUFAs, marine n-3 PUFAs, α-linolenic acids (ALA) and n-6 PUFAs were not associated with the risk of it (RR 1.04, 95%CI 0.85-1.28; RR 0.99, 95%CI 0.89-1.09; RR 1.05, 95%CI 0.93-1.19; RR 1.02, 95%CI 0.94-1.11, respectively). For breast cancer, only marine n-3 PUFAs, but not total n-3 PUFAs, ALA, and n-6 PUFAs, was associated with a lower risk of it (RR 0.70, 95%CI 0.55-0.91). For prostate cancer, ALA and n-6 PUFAs also have no association with the risk of it. CONCLUSIONS: Most subtypes of PUFAs are probably not related to cancers. However, additional high-quality trials are warranted to corroborate the findings of this meta-analysis.

Screening of the Hepatotoxic Components in Fructus Gardeniae and Their Effects on Rat Liver BRL-3A Cells.

Fructus Gardeniae (FG) is a common Chinese medicine and food. However, the toxicity of FG has drawn increasing concern, especially its hepatotoxicity. The purpose of this study was to screen the hepatotoxic components of FG and evaluate their effects on rat liver BRL-3A cells. The chemical composition of FG was determined by HPLC-ESI-MS. CCK-8 assay was used to evaluate the cytotoxicity of ten chemical components from FG, and then the toxic components with significant inhibitory activity were selected for further study. The results showed that geniposide, genipin, genipin-1-gentiobioside, gardenoside, and shanzhiside all suppress cells viability. Apoptosis assays further indicated that geniposide and its metabolite genipin are the main hepatotoxic components of FG. Pretreatment of cells with geniposide or genipin increased the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). The activities of superoxide dismutase (SOD) and glutathione (GSH) were decreased, while the malondialdehyde (MDA) level was increased. The cell contents of tumor necrosis factor (TNF-α), interleukin-6 (IL-6), and nitric oxide (NO) were also increased. Molecular docking simulations were used to investigate the mechanism of FG-induced hepatotoxicity, revealing that geniposide and genipin bind strongly to the pro-inflammatory factor TNFR1 receptor of the NF-κB and MAPK signaling pathways. The obtained results strongly indicate that the hepatotoxicity of FG is caused by iridoids compounds. Genipin had the most significant hepatotoxic effect. These toxic substances destroy the cell antioxidant defense system, increasing inflammatory injury to the liver cells and leading to apoptosis and even necrosis. Thus, this study lays a foundation for toxicology research into FG and its rational application.

Screening and Isolation of Natural Antioxidants from Acer Ginnala Max by TLC-MS/MS Guided Bioautographic Method.

A rapid and simple method has been developed for the screening and identification of natural antioxidants from the leaves of Acer ginnala Maxim (AG). The process is that upon reaction with 1,1-diphenyl-2-picrylhydrazyl (DPPH), the white yellow spots of compounds with potential antioxidant effects will be significantly observed on the thin-layer chromatography (TLC), and possible structures will be presumed by the ESI/MS technique. Using the improved approach, 6 compounds in the AG extract were found to possess a potential antioxidant activity. They were speculated as quercetin-3-O-α-L-(3"-galloyl)-rhamnoside (1), quercetin-3-O-α-L-(2"-galloyl)-rhamnoside (2), quercetin-3-O-α-L-(2"-galloyl)-arabinopyranoside (3), acertannin (4), gallic acid (5), and methyl gallate (6). In addition, we were still found that compounds 2, 3, and 5 had favorable antioxidant activity from the scannogram of the DPPH reaction plate. As a result, the isolated 6 compounds structures were in accordance with the presumed structures. Furthermore, the free radical scavenging capacities of the available identified compounds were also investigated. Compounds 2, 3, and 5 showed significant DPPH.Scavenging capacities, with IC50 values of 2.83 µg/mL, 2.34 µg/mL, and 1.86µg/mL, respectively. The results indicated that this newly improved method could be widely applied for rapid screening and identification of natural antioxidants from Chinese herbal medicines.

Screening of the Active Component Promoting Leydig Cell Proliferation from Lepidium meyenii Using HPLC-ESI-MS/MS Coupled with Multivariate Statistical Analysis.

Lepidium meyenii is now widely consumed as a functional food and medicinal product, which is known as an enhancer of reproductive health. However, the specific chemical composition and mechanism of action for improving sexual function are unclear. The present study aims at screening and determining the potential compounds, which promote mouse leydig cells (TM3) proliferation. The partial least squares analysis (PLS) was employed to reveal the correlation between common peaks of high performance liquid chromatography (HPLC) fingerprint of L. meyenii and the proliferation activity of TM3. The results suggested that three compounds had good activities on the proliferation of TM3 and promoting testosterone secretion, there were N-benzyl-hexadecanamide, N-benzyl-(9z,12z)-octadecadienamide and N-benzyl-(9z,12z,15z)-octadecatrienamide which might be the potential bioactive markers related to the enhancing sexual ability functions of L. meyenii. The first step in testosterone synthesis is the transport of cholesterol into the mitochondria, and the homeostasis of mitochondrial function is related to cyclophilin D (CypD). In order to expound how bioactive ingredients lead to promoting testosterone secretion, a molecular docking simulation was used for further illustration in the active sites and binding degree of the ligands on CypD. The results indicated there was a positive correlation between the binding energy absolute value and testosterone secretion activity. In addition, in this study it also provided the reference for a simple, quick method to screen the promoting leydig cell proliferation active components in traditional Chinese medicine (TCM).

Screening and determination for potential acetylcholinesterase inhibitory constituents from ginseng stem-leaf saponins using ultrafiltration (UF)-LC-ESI-MS2.

INTRODUCTION: Previous studies have demonstrated that several ginsenosides have remarkable inhibitory effect on acetylcholinesterase (AChE). In the present study, ginseng stem-leaf saponins (GSLS) can improve learning and memory of Alzheimer's disease patients. However, much comprehensive information regarding AChE inhibition of GSLS and its metabolites is yet unknown. OBJECTIVE: The present study aims to screen and determine the potential of AChE inhibitors (AChEIs) from GSLS. METHODOLOGY: The active fraction of the GSLS detected in vitro AChE inhibition assays was selected as a starting material for the screening of the potential of AChEIs using ultrafiltration liquid chromatography coupled to electrospray ionisation tandem mass spectrometry (UF-LC-ESI-MS2 ). RESULTS: The results showed that 31 ginsenosides were identified with analysis using rapid resolution liquid chromatography with a diode array detector combined with electrospray ionisation tandem mass spectrometry (RRLC-DAD-ESI-MS2 ) from the active fraction, and there are 27 compounds with AChE binding activity. Among them, 11 ginsenosides were evaluated and confirmed using in vitro enzymatic assay, and ginsenosides F1 , Rd, Rk3 , 20(S)-Rg3 , F2 and Rb2 were found to possess strong AChE inhibitory activities. CONCLUSION: The proposed UF-LC-ESI-MS2 method was a powerful tool for the discovery of AChEIs from traditional Chinese medicine (TCM).

Rapid Screening and Characterization of Acetylcholinesterase Inhibitors from Yinhuang Oral Liquid Using Ultrafiltration-liquid Chromatography-electrospray Ionization Tandem Mass Spectrometry.

BACKGROUND: At present, approximately 17-25 million people in the world suffer from Alzheimer's disease (AD). The most efficacious and acceptable therapeutic drug clinically are the acetylcholinesterase inhibitors (AChEIs). Yinhuang oral liquid is a Chinese medicine preparation which contains AChEIs according to the literatures. However, no strategy has been presented for rapid screening and identification of AChEIs from Yinhuang oral liquid. OBJECTIVE: To develop a method for rapid screening and identification of AChEIs from Yinhuang oral liquid using ultrafiltration-liquid chromatography-electrospray ionization tandem mass spectrometry (UF-LC-ESI-MS/MS). MATERIALS AND METHODS: In this study, UF incubation conditions such as enzyme concentration, incubation time, and incubation temperature were optimized so as to get better screening results. The AChEIs from Yinhuang oral liquid were identified by high-performance liquid chromatography-ESI-MS and the improved Ellman method was used for the AChE inhibitory activity test in vitro. RESULTS: The results showed that Yinhuang oral liquid can inhibit the activity of AChE. We screened and identified seven compounds with potential AChE inhibitory activity from Yinhuang oral liquid, which provided experimental basis for the treatment and prevention of AD. CONCLUSION: The current technique was used to directly screen the active ingredients with acetylcholinesterase inhibition from complex traditional Chinese medicine, which was simple, rapid, accurate, and suitable for high-throughput screening of AChEI from complex systems. SUMMARY: A UF-LC-ESI-MS/MS method for rapid screening and identification of AChEIs from Yinhuang oral liquid was developedSeven compounds were screened and identified with potential AChE inhibitory activity from Yinhuang oral liquidIt provided experimental basis of Yinhuang oral liquid for the treating and preventing AD. Abbreviations used: (AD): Alzheimer's disease; (UF-LC-ESI-MS/MS): ultrafiltration-liquid chromatography-electrospray ionization tandem mass spectrometry; (AChEIs): acetylcholinesterase inhibitors.

[Application of intense pulsed light depilation in the auricular reconstruction with totally expanded skin].

OBJECTIVE: To investigate the application of intense pulsed light (IPL) depilation in the auricular reconstruction with totally expanded skin and its suitable parameter. METHODS: From February 2009 to September 2013, one tissue expander(100 or 150 milliliters) were implanted under the skin of mastoid in 126 patients who suffered from microtia. During the skin expanding, the hair on the expanded skin was depilated by JPL technique. The parameter, operation, interval, the depilation result and the complication were recorded. The autologous rib cartilage frameworks were implanted in the second phase of auricular reconstruction. RESULTS: The energy of IPL was 20 - 35 J/cm2 (mean 25. 3 J/cm2), the width of pulse was 26 - 30 ms ( mean 27. 9 ms), the interval between two operations was 4 weeks. Ninety-six patients were cured(76. 2%, 96/126). The total depilation efficient is 96. 8% (122/126). A fine banding blisters was found in one patient whose depilation areas was marked by black pen. Skin erythema was found in five patients and skin pigmentation was found in 7 patients. There were no skin necrosis and expander exposure in all patients. During 6 - 12 months of follow-up period, ninety-two patients were cured (73.0%, 92/126) and the total depilation efficient is 92. 9% (117/126). CONCLUSIONS: IPL depilation during the skin expanding is feasible and safe with careful procedure. The hairless expanded skin is enough to cover the framework, as well as the post-auricular area. The appearance of reconstructed ear is better without hair.

Simultaneous quantification and splenocyte-proliferating activities of nucleosides and bases in Cervi cornu Pantotrichum.

BACKGROUND: Cervi Cornu Pantotrichum has been a well known traditional Chinese medicine, which is young horn of Cervus Nippon Temminck (Hualurong: HLR). At present, the methods used for the quality control of Cervi Cornu Pantotrichum show low specificity. OBJECTIVE: To describe a holistic method based on chemical characteristics and splenocyte-proliferating activities to evaluate the quality of HLR. MATERIALS AND METHODS: The nucleosides and bases from HLR were identified by high performance liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI-MS), and six of them were chosen to be used for simultaneous HPLC quantification according to the results of proliferation of mouse splenocytes in vitro. RESULTS: In this study, eight nucleosides and bases have been identified. In addition, uracil, hypoxanthine, uridine, inosine, guanosine, and adenosine were chosen to be used for simultaneous HPLC quantification. Simultaneous quantification of these six substances was performed on ten groups of HLR under the condition of a TIANHE Kromasil C18 column (5 µm, 4.6 mm × 250 mm i.d.) and a gradient elution of water and acetonitrile. Of the ten groups, HLR displayed the highest total nucleoside contents (TNC, sum of adenosine and uracil, 0.412 mg/g) with the strongest splenocyte-proliferating activities. CONCLUSION: These results suggest that TNC (such as particularly highly contained adenosine and uracil) in HLR has a certain correlation with the activity of splenocyte-proliferating, and it may be used as a quality control for HLR. This comprehensive method could be applied to other traditional Chinese medicines to ameliorate their quality control.

[Alkaloids from cervi cornu pantotrichum and its effect on murine splenocytes proliferation].

OBJECTIVE: To study the alkaloids of Cervi Cornu Pantotrichum and its effect on murine splenocytes proliferation. METHODS: The constituents isolation and purification from Cervi Cornu Pantotrichum was carried out by reported column chromatography including Sephadex LH-20 and MCI (CHP20P) and their structures were elucidated on the basis of spectral compounds. The method of MTT was used to examine the effects of eight alkaloids and total alkaloids content (TAC) of Cervi Cornu Pantotrichum on murine splenocytes proliferation. RESULTS: Eleven compounds were isolated from Cervi Cornu Pantotrichum, and their structures were identified as follows: uracil (1), hypoxanthine (2), uridine (3) inosine (4), guanosine (5), 2'-deoxyguanosine (6), guanine (7), thymidine (8), thymine (9), cytidine (10) and adenosine (11). By the experiment of murine splenocytes proliferation activity in vitro, the results showed that the total alkaloids, uracil and adenosine had significantly promoted the proliferation of mouse spleen cells. CONCLUSION: Compounds 4 - 11 are isolated from Cervi Cornu Pantotrichum for the first time. The total alkaloids is one of the material basis of immunomodulatory effects of Cervi Cornu Pantotrichum, and uracil and adenosine are the most active.

Diosgenin inhibits the migration of human breast cancer MDA-MB-231 cells by suppressing Vav2 activity.

Diosgenin, a naturally occurring steroidal saponin, possess tumor therapeutic potential. However, the effect of diosgenin on cancer metastasis remains poorly understood. In this study, we performed in vitro experiments to investigate the inhibitory activity of diosgenin on human breast cancer MDA-MB-231 cell migration, and reveal the possible mechanism. Diosgenin caused a marked inhibition of cell migration in MDA-MB-231 cell by transwell assay. In addition, diosgenin significantly impacted MDA-MB-231 cell migratory behavior under real-time observation. We also found diosgenin significantly inhibited actin polymerization, Vav2 phosphorylation and Cdc42 activation, which might be, at least in part, attributed to the anti-metastatic potential of diosgenin. These findings reveal a new therapeutic potential of diosgenin for human breast cancer metastasis therapy.

Screening and determination for potential α-glucosidase inhibitory constituents from Dalbergia odorifera T. Chen using ultrafiltration-LC/ESI-MS(n).

In the present study, it was demonstrated that ethyl acetate soluble fraction partitioned from heartwood of Dalbergia odorifera T. Chen (HEF) had a remarkable inhibitory effect on α-glucosidase. Therefore HEF was selected as a starting material for screening the potential α-glucosidase inhibitors using ultrafiltration liquid chromatography/mass spectrometry (UF-LC/MS). Twenty-six compounds were identified with analysis of LC/MS. UF assay indicated that 18 compositions might be α-glucosidase inhibitors in HEF; eight of them were estimated for their α-glucosidase inhibitory activity, and the results showed that (2S)-liquiritigenin, (2S)-4',6-dihydroxy- 7-methoxyflavanone and isoliquiritigenin displayed obvious inhibition of yeast α-glucosidase. In addition, in order to control the quality of HEF, the content of five compounds in HEF was simultaneously determined for the first time. These results provide an important theoretical base for the further application of HEF to treat type 2 diabetes in the clinic and development of natural α-glucosidase inhibitors with low toxicity.

[Antioxidant activity constituents from root of Rubus crataegifolius].

OBJECTIVE: To study the antioxidant constituents from the root of Rubus crataegifolius. METHOD: The constituents isolation and purification from the root of R. crataegifolius was carried by reported column chromatography including silica gel, toyopearl, and their structures were elucidated on the basis of spectral compounds. DPPH method was used to evaluate the free radical scavenging activity of the isolated compounds. RESULT: Nine compounds were isolated from the root of R. crataegifolius, and their structures were identified as follow: euscaphic acid (1), kaempferol-3-O-beta-D-galactopyranoside (2), tormentic acid (3), 2alpha, 19alpha, 24-trihydroxyurs-12-ene-3-oxo-28-acid (4) , 2alpha-hydroxy-oleanolic acid (5), ursolic acid (6), daucosterol (7), beta-sitosterol (8) and polydatin (9). By experiment of antioxidant activity, the result showed compounds 2 and 9 revealed DPPH free radical scavenging rates were 95.60% and 75.23% at the concentration of 50 mg x L(-1). CONCLUSION: Compounds 1-8 were isolated from this plant for the first time, and compounds 2 and 9 showed the significant antioxidant activity.

[Study on the chemical constituents of aerial part of Ligusticum jeholense].

OBJECTIVE: To study the chemical constituents of the aerial part of Ligusticum jeholense. METHODS: The constituents were isolated by sillica gel column chromatography, Sephadex LH-20 column chromatography and their structures were elucidated by spectral analysis. RESULTS: Seven compounds were separated from the EtOH extracts. Their structures were identified as psoralen (1), beta-sitosterol (2), daucosterol (3), kaempferol-3-O-(2",4"-di-E-p-coumaroyl)-alpha-L-rhamnoside (4), kaempferol-3-O-beta-D-galactoside (5), quercetin-3-O-beta-D-galactoside (6), sucrose (7). CONCLUSION: Compounds 1, 4, 5 and 6 are isolated from the genus for the first time. Compounds 2, 3 and 7 are isolated from the aerial part of the plant for the first time.