Antiperspirant effects and mechanism investigation of Mulisan decoction in rats based on plasma metabolomics.

CONTEXT: Mulisan decoction (MLS) is a classic formula of traditional Chinese medicine for treating hyperhidrosis. The mechanism remains unclear. OBJECTIVE: To investigate the antiperspirant effect and underlying mechanisms of MLS. MATERIALS AND METHODS: Fifty rats were divided into control, model, and three doses of MLS intervention groups (n = 10). Rats except for control group were induced diseases features of the applicable scope of MLS via i.p. reserpine (0.5 mg/kg/d) for 10 days. From day 11, MLS groups were administrated orally MLS at 0.6, 3, and 15 g/kg once a day for 14 days, respectively. After the last administration, sweating was induced in all rats via s.c. pilocarpine (25 mg/kg), the right hind foot of rats was stained, and sweat point numbers were observed. Rat serum was collected to detect IL-2, IL-6, IFN-γ, and TNF-α. Rat plasma was collected for endogenous metabolite analysis via UPLC-QE-Focus-MS. RESULTS: Rats treated with MLS presented a significant decrease in sweat point numbers (13.5%), increase in body weight (13.2%), and promotion in the balance of Th1/Th2 cytokine ratio via increasing IL-2 (38.3%), IFN-γ (20.1%), and TNF-α (22.0%) and decreasing IL-6 (24.7%) compared with the model group (p < 0.05). Plasma metabolomics disclosed 15 potential biomarkers related to model rats, of which two could be significantly reversed by MLS (p < 0.05). The involved pathways were pantothenate and CoA biosynthesis, and porphyrin metabolism. CONCLUSIONS: MLS demonstrated a good antiperspirant effect and metabolism improvement. These findings inspire more clinical study validation on immune improvement and antiperspirant effect.

Haimufang decoction, a Chinese medicine formula for lung cancer, arrests cell cycle, stimulates apoptosis in NCI-H1975 cells, and induces M1 polarization in RAW 264.7 macrophage cells.

BACKGROUND: Lung cancer has the highest morbidity and mortality in the world and novel treatment strategies are still needed. Haimufang decoction (HMF) is a patented clinical prescription of traditional Chinese medicine for lung cancer treatment. HMF is composed of four herbs and has been applied clinically in advanced cancer patients. However, its therapeutic mechanisms are still unclear. This study aims to elucidate the possible mechanisms of HMF for the treatment of lung cancer. METHODS: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay was applied for evaluating the proliferative effect of HMF in lung cancer cells and monocyte macrophage RAW264.7 cells. Flow cytometer was used to detect the effects of HMF on cell cycle and apoptosis, and western blotting was employed to explore the potential apoptotic mechanisms of HMF on lung cancer cells. For immunomodulatory effect, co-culture system was used to detect the activation of macrophage RAW264.7 cells when treated with HMF, and neutral red assay was used to measure the effect of HMF on the phagocytosis of the activated macrophages. Enzyme linked immunosorbent assay, flow cytometer, and immunofluorescence staining method were employed for the investigation on the underlying mechanisms of the immunomodulatory effect on RAW264.7 induced by HMF. RESULTS: HMF inhibited the proliferation, induced S phase cell cycle arrest, and stimulated apoptosis in lung cancer NCI-H1975 cells, while had negligible cytotoxicity on macrophage RAW264.7 cells. Moreover, HMF could activate macrophage RAW264.7 cells and promote the inhibition activity of RAW264.7 cells against lung cancer cells. And also, HMF activated macrophages and increased their phagocytic activity in a concentration-dependent manner. HMF increased the expression of macrophage activation marker CD40, the level of nitric oxide, the generation of intracellular reactive oxygen species, as well as M1 macrophages cytokines including tumor necrosis factor-α, interleukin-1ß, interleukin 12 p70, and interleukin 6. Further investigation showed that HMF induced M1 but not M2 phenotype polarization in RAW264.7 cells. CONCLUSIONS: HMF can mainly exert anticancer activity via (1) cytotoxicity to human lung cancer cells by proliferation inhibition, cell cycle arrest, and apoptosis induction; and also via (2) immunomodulation via macrophage cells activation and M1 phenotype polarization induction.

Antidiabetic potential of the ethyl acetate extract of Physalis alkekengi and chemical constituents identified by HPLC-ESI-QTOF-MS.

ETHNOPHARMACOLOGICAL RELEVANCE: The edible plant Physalis alkekengi (PA) is used in traditional medicine to treat diabetes. However, the anti-diabetic effects and constituents of the fruit and aerial parts of this plant have not been studied extensively. AIM OF THE STUDY: The purpose of this study was to investigate the antidiabetic potential of Physalis alkekengi and identify its chemical constituents. MATERIALS AND METHODS: In the present study, the in vitro glucose uptake capacity was tested using the 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG) assay in HepG2 cells. Secondly, the anti-diabetes effects of the ethyl acetate extracts of the aerial parts/fruit (EAP/EAF) of P. alkekengi were evaluated in high-fat diet-fed and streptozotocin-induced diabetic rats (seven groups, n = 7) daily at doses of 300 and 600 mg/kg for 28 days. Fasting blood glucose (FBG) was measured with a glucometer and the levels of total cholesterol (TC), triglyceride (TG), glycated serum protein (GSP), and fasting insulin (FINS) were measured by ELISA. Furthermore, insulin sensitivity index (ISI) and homeostasis model assessment-insulin resistance index (HOMA-IR) were calculated based on FBG and FINS. Changes in blood glucose concentration were assessed after an oral glucose challenge in diabetic rats treated with EAF and EAP extracts. In all assays, rosiglitazone, a current antidiabetic drug and insulin sensitizer, was also tested. Finally, the compounds in EAP were identified by HPLC-ESI-QTOF-MS analysis. RESULTS: EAP increased the uptake of 2-NBDG, a measure of direct glucose uptake, in HepG2 cells. Next, in diabetic rats treated with P. alkegenki extracts for 28 days, the levels of FBG, TC, TG and GSP and were lowered effectively, while FINS was increased significantly. EAP/EAF enhanced insulin sensitivity significantly as measured by ISI and HOMA-IR along with oral glucose tolerance test analysis. The EAP generally exerted the greatest effects. Lastly, a HPLC-ESI-Q-TOF-MS analysis identified 50 compounds, including 26 physalins, 10 flavonoids, and 9 phenolic acids, with 21 compounds found for the first time in P. alkekengi. CONCLUSIONS: The results support the merit of P. alkekengi as an antidiabetic herbal medicine or dietary supplement.

Development and Validation of Quantitative (1)H NMR Spectroscopy for the Determination of Total Phytosterols in the Marine Seaweed Sargassum.

Knowledge of phytosterol (PS) contents in marine algae is currently lacking compared to those in terrestrial plants. The present studies developed a quantitative (1)H NMR method for the determination of the total PSs in Sargassum. The characteristic proton signal H-3α in PSs was used for quantification, and 2,3,4,5-tetrachloro-nitrobenzene was used as an internal standard. Seaweed samples could be recorded directly after total lipid extraction and saponification. The results showed that the PS contents in Sargassum fusiforme (788.89-2878.67 mg/kg) were significantly higher than those in Sargassum pallidum (585.33-1596.00 mg/kg). The variable contents in both species suggested that fixed raw materials are very important for future research and development. Orthogonal projection to latent structures discriminant analysis was carried out in the spectral region of δ 3.00-6.50 in the (1)H NMR spectrum. S. fusiforme and S. pallidum could be separated well, and the key sterol marker was fucosterol.

Immunomodulatory N-acyl Dopamine Glycosides from the Icelandic Marine Sponge Myxilla incrustans Collected at a Hydrothermal Vent Site.

A chemical investigation of the sponge (Porifera) Myxilla incrustans collected from the unique submarine hydrothermal vent site Strytan, North of Iceland, revealed a novel family of closely related N-acyl dopamine glycosides. Three new compounds, myxillin A (1), B (2) and C (3), were isolated and structurally elucidated using several analytical techniques, such as HR-MS, 1D and 2D NMR spectroscopy. Myxillin A (1) and B (2)were shown to be structurally similar, composed of a dopamine moiety, but differ in the acyl chain length and saturation. The myxillin C (3) has a dehydrotyrosine moiety composing the same acyl chain and glycosylation as myxillin B (2). Myxillins A (1) and C (3) were tested for immunomodulating activity in an in vitro dendritic cell model. Dendritic cells matured and stimulated in the presence of myxillin A (1) secreted lower levels of IL-12p40, whilst dendritic cells matured and stimulated in the presence of myxillin C (3) secreted lower levels of IL-10 compared with dendritic cells matured and stimulated in the presence of the solvent alone. These opposing results indicate that the structural differences in the aromatic ring part of the molecules could have an impact on the immunological effects of dendritic cells. These molecules could, therefore, prove to be important in preventing inflammatory diseases on the one hand, and inducing a response to fight tumors and/or pathogens on the other hand. Further studies will be needed to confirm these potential uses.

Enterolacaciamine as a potential O-GlcNAcase activator from the leaves of Enterolobium cyclocarpum.

Enterolacaciamine (1), a new potential O-GlcNAcase activator, along with three known triterpenoid saponins, concinnoside B (2), concinnoside D (3), and julibroside A3 (4) was isolated from the leaves of Enterolobium cyclocarpum. Their structures were elucidated by chemical and spectroscopic methods (UV, MS, 1D and 2D NMR). Their effects on O-GlcNAcase activity were evaluated using O-GlcNAcase enzymatic assay. The results showed that compound 1 could obviously enhance the activity of O-GlcNAcase.

[Comparative analysis of trace elements in five marine-derived shell TCM using multivariate statistical analysis].

A comparable study were carried out by determination of trace elements on five marine-derived shell traditional Chinese medicine (TCM) (Ostreae Concha, Haliotidis Concha, Margaritifera Concha, Meretricis Concha, and Arcae Concha), which were recorded in the Chinese Pharmacopoeia (2010 version). Seven trace elements in 51 batches of this type of shell TCM were analyzed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), combined with principal component analysis (PCA) methods. The content of element Se, which exhibited significant differences among different drugs, could be used as a key element to distinguish this type of drugs. Meanwhile, the contents of elements Co, Cu, Mo, and Ba in Haliotidis Concha, Co and As in Margaritifera Concha, Mo and As in Meretricis Concha, Mo, As, and Ba in Arcae Concha, and Zn in Meretricis Concha were relatively stable. In the PCA plot, Arcae Concha and Meretricis Concha could be efficiently distinguished from Ostreae Concha together with Haliotidis Concha, and Margaritifera Concha. The results also showed a correlation with their medicinal function. In conclusion, trace elements in marine-derived shell TCM could not be neglected for their quality control.

[Identification of marine-derived shell TCM by near infrared spectroscopy].

The identification of five marine-derived shell traditional Chinese medicine (TCM) recorded in the Chinese Pharmacopoeia were studied. Using near infrared technology (NIR) combined with principal component analysis (PCA) methods, Ostreae Concha, Haliotidis Concha, and Margaritifera Concha could be efficiently distinguished from Meretricis Concha together with Arcae Concha. In the first principal components, Ostreae Concha exhibited obvious differences with high loadings in 4 236, 5 263, 7 142 cm(-1) concerning to the contents of CaCO3 and H2O in the samples. Arcae Concha and Meretricis Concha displayed significant differences with others in the second principal components, which can be illustrated by high loadings in 5 000 -4 430 cm(-1) areas. It is indicated that the second principal components might be related to organics which contained NH and CH groups, for example proteins. Meanwhile, our data showed a correlation between the function of these shell TCM and their distribution in the PCA plot. These results suggested that organic components in marine-derived shell TCM could not be neglected for their quality control.

Isolation of the molecular species of monogalactosyldiacylglycerols from brown edible seaweed Sargassum horneri and their inhibitory effects on triglyceride accumulation in 3T3-L1 adipocytes.

The chemical composition of monogalactosyldiacylglycerols (MGDGs) from brown alga Sargassum horneri and their inhibitory effects on lipid accumulation were investigated in this study. A total of 10 molecular species of MGDGs were identified using nuclear magnetic resonance, alkaline hydrolysis, gas chromatography-flame ionization detector, and high-performance liquid chromatography-tandem mass spectrometry methods. Individual molecular species of MGDGs, including (2S)-1-O-myristoyl-2-O-palmitoleoyl-3-O-ß-D-galactopyranosyl-sn-glycerol (1), (2S)-1-O-myristoyl-2-O-linoleyl-3-O-ß-D-galactopyranosyl-sn-glycerol (3), (2S)-1-O-palmitoyl-2-O-linolenoyl-3-O-ß-D-galactopyranosyl-sn-glycerol (5), (2S)-1-O-myristoyl-2-O-oleyl-3-O-ß-D-galactopyranosyl-sn-glycerol (7), (2S)-1-O-palmitoyl-2-O-palmitoleoyl-3-O-ß-D-galactopyranosyl-sn-glycerol (8), (2S)-1-O-palmitoyl-2-O-linoleyl-3-O-ß-D-galactopyranosyl-sn-glycerol (9), and (2S)-1-O-palmitoyl-2-O-oleyl-3-O-ß-D-galactopyranosyl-sn-glycerol (10), were then furnished using semi-preparative high-performance liquid chromatography, and their inhibitory effects on triglyceride (TG) accumulation and free fatty acid (FFA) levels in 3T3-L1 adipocytes were evaluated. Compounds 3 and 9 showed inhibitory effects on TG and FFA accumulation, with TG levels of 1.568 ± 0.2808 and 1.701 ± 0.1460 µmol/L and FFA levels of 0.149 ± 0.0258 and 0.198 ± 0.0229 mequiv/L, respectively, which were more effective than other compounds. The primary structure-activity relationship suggested that linoleyl [18:2(ω-6)] in the sn-2 position played an important role on triglyceride accumulation inhibition.

Laevinoids A and B: two diterpenoids with an unprecedented backbone from Croton laevigatus.

Chemical fractionation of the ethanolic extract of a Chinese herbal plant, Croton laevigatus, yielded laevinoids A (1) and B (2) with a new rearranged ent-clerodane scaffold named as laevinane. The structures of 1 and 2 were assigned on the basis of detailed spectroscopic analyses with their absolute configurations being established via single-crystal X-ray diffraction studies.

Chemical profiles and identification of key compound caffeine in marine-derived traditional Chinese medicine Ostreae concha.

To compare the chemical differences between the medicinal and cultured oyster shells, their chemical profiles were investigated. Using the ultra performance liquid chromatography-electron spraying ionization-mass spectrometry (UPLC-ESI-MS), combined with principal component analysis (PCA) and orthogonal projection to latent structures discriminant analysis (OPLS-DA), the discrimination of the chemical characteristics among the medicinal and cultured oyster shells was established. Moreover, the chemometric analysis revealed some potential key compounds. After a large-scale extraction and isolation, one target key compound was unambiguously identified as caffeine based on extensive spectroscopic data analysis (1D and 2D NMR, MS, and UV) and comparison with literature data.

Cytotoxic diterpenoids from Sapium insigne.

Chemical investigation into the twigs and leaves of Sapium insigne afforded seven new diterpenoids, sapinsignoids A-G (1-7), together with 10 known diterpenoids. The structures of 1-7 were assigned on the basis of detailed spectroscopic analysis and chemical degradation. Compounds 1-4 exhibited significant cytotoxicity against the A-549 tumor cell line (IC(50) 0.2-1.8 µM), while compounds 1-3 showed moderate cytotoxicity against the HL-60 cell line (IC(50) 2.7-6.5 µM).

Sesquiterpenes from Dysoxylum oliganthum and Dysoxylum excelsum.

Three new sesquiterpenes (1-3), (6R,7S,11R,10S)-15-hydroxy-sesquisabinene hydrate (1), (6R,7R,11S,10S)-15-hydroxy-sesquisabinene hydrate (2), and (6R,7R,10S)-15-hydroxy-zingiberenol (3), along with three known compounds, were isolated from the stems of Dysoxylum oliganthum; and three new isodaucane (salvionane) sesquiterpenes, namely isodauc-6-ene-10ß,14-diol (4), 4-epi-isodauc-6-ene-10ß,14-diol (5), and 4-epi-6α,10ß-dihydroxy-artabotrol (6) together with 15 known compounds were isolated from the twigs and leaves of D. excelsum. Their structures were established on the basis of extensive spectroscopic analysis and chemical shifts. The absolute configuration of C-10 in compounds 1-3 of a rare class was determined by using Snatzke's method.

Constituents of Trigonostemon heterophyllus.

Six new diterpenoids, 1-6, a new prenylated sesquiterpenoid, 7, and six known compounds were isolated from Trigonostemon heterophyllus. The structures of 1-7 were elucidated on the basis of NMR and MS analysis. The daphnane diterpenoids trigoheterins A (1) and B (2) possess a rare 4,6-oxetane moiety within this compound class, and trigoheteric acid methyl ester (6) is the first example of a 15,16-dinor-3,4-seco-cleistanthane. Trigoheterin E (5) exhibited cytotoxic activity against the HL-60 (IC50) 1.8 µM) and A-549 (IC50 10.0 µM) human cancer cell lines.

[Clinical observation on acupuncture combined with nerve block for treatment of lumbar disc herniation].

OBJECTIVE: To investigate the therapeutic effects of acupuncture combined with nerve block for treatment of lumbar disc herniation. METHODS: Ninety cases of lumbar disc herniation were randomly divided into three groups: acupuncture combined with nerve block group, acupuncture group and nerve block group, 30 cases in each group. The acupuncture combined with nerve block group was treated with acupuncture combined with nerve block therapy, L2-L5 Jiaji (EX-B 2), Zhibian (BL 54), Huantiao (GB 30), Yanglingquan (GB 34) etc. were selected for acupuncture, affected nerve root, sciatic nerve or common peroneal nerve were selected for nerve block with anti-inflammation-analgesic injection; the acupuncture group was treated with acupuncture only; the nerve block group was treated with nerve block only. After 4 weeks of treatment, the visual analogue scale (VAS), Oswestry lumbar dysfunction index (ODI) and modified MacNab standard were compared to evaluate the therapeutic effects of three groups. RESULTS: The VAS and ODI in all groups were significantly decreased after one week, two weeks and four weeks of treatment (all P<0.01); after one week of treatment, the scores of VAS and ODI in nerve block group and acupuncture combined with nerve block group were significantly lower than those of acupuncture group (P<0.05); after two weeks and four weeks of treatment, the scores of VAS and ODI in acupuncture combined with nerve block group were significantly lower than those of acupuncture group and nerve block group (P<0.05). The effective rate and excellent and good rate of the acupuncture combined with nerve block group were significantly higher than those of acupuncture group and nerve block group (both P<0.01). CONCLUSION: The nerve block therapy and acupuncture are effective methods for treatment of lumbar disc herniation, while it has a better effect when these two treatments are combined used.

Mesendanins A-J, limonoids from the leaves and twigs of Melia toosendan.

Ten new limonoids, namely, mesendanins A-J (1-10), together with 14 known compounds, have been isolated from the leaves and twigs of Melia toosendan. Their structures were established on the basis of spectroscopic data analysis.

Two novel sulfur compounds from the seeds of Raphanus sativus L.

The seeds of Raphanus sativus L., known as Lai-fu-zi in traditional Chinese Medicine, are always roasted before clinical use for avoiding nausea. During an investigation of the chemical difference between roasted and pre-roasted products, two novel sulfur-containing compounds, which mainly existed in the pre-roasted products, were isolated. Their structures and absolute configurations were established by spectroscopic and X-ray diffraction analysis.

A study on the antioxidant activity and tissues selective inhibition of lipid peroxidation by saponins from the roots of Platycodon grandiflorum.

Platycodi Radix is the root of Platycodon grandiflorum (Jacq.) A. DC and has been used as a traditional medicine in China. According to the theory of traditional Chinese medicine, Platycodi Radix (PR) possesses the character of lung meridian tropism (Guijing) and has selective effects on the lung and respiratory system. The aim of this study was to confirm the antioxidant effects of saponins from Platycodi Radix (PRS), with emphasis on its selective inhibition of lipid peroxidation in different tissues. The hydroxyl radical scavenging activity was determined by a H(2)O(2)/Fe(2+) system; a modified thiobarbituric acid reactive species assay was used to measure the lipid peroxide in rats' tissues. These antioxidant activities were compared to ascorbic acid (Vc). The results showed that PRS had antioxidant activities in various antioxidant systems. And the inhibition capability of lipid peroxidation of PRS and Vc were excellent, but differed greatly in different tissues. These results suggested that PRS had antioxidant effects and selective inhibition of lipid peroxidation. It indicated that the mechanism of Platycodi Radix to treat some diseases might be related to its antioxidant activity, especially its tissue selective effects. However, further study is needed.

Alkaloids from the leaves of Daphniphyllum subverticillatum.

Four new alkaloids, 17-hydroxydaphnigraciline (1), subdaphnidine A (2), daphnezomine L methyl ester (3), and 11-hydroxycodaphniphylline (4), along with 24 known analogues, were isolated from the leaves of Daphniphyllum subverticillatum. The structures of 1-4 were elucidated on the basis of spectroscopic methods and from chemical evidence. Daphnilongeridine (5) showed cytotoxicity against several tumor cell lines at IC(50) values in the range 2.4-9.7 microM and against the HMEC human microvascular endothelial cell line with an IC(50) of 2.7 microM.

[Determination of eight metal elements in Cichorium glandulosum Boiss et Huet by microwave digestion-FAAS].

The Cichorium glandulosum Boiss et Huet is a traditional Uighur natural herbal medicine, but has not been analyzed and studied in terms of its metal elements. In the experiment, the Cichorium glandulosum Boiss et Huet powder was digested with HNO3 by microwave digestion before determination. The eight metal elements, potassium, nickel, calcium, magnesium, iron, manganese, copper and zinc, in Cichorium glandulosum Boiss et Huet were determined by FAAS. The working conditions, accuracy and precision of the method were studied. The linear correlations of standard curves are good (r = 0.999 1-0.999 9). The recovery (n = 6) is 92.25%-110.5%, and the RSD (n = 6) is 0.7%-3.88%. The results showed that there were comparatively rich metal elements, among which are comparatively high calcium (65.84 mg x g(-1)), iron (24.38 mg x g(-1)), magnesium (278.17 mg x g(-1)) and potassium (18.50 mg x g(-1)), in Cichorium glandulosum Boiss et Huet, and the contents of other elements are nickel of 0.004 38 mg x g(-1), manganese of 0.52 mg x g(-1), copper of 0.016 5 mg x g(-1) and zinc of 0.18 mg x g(-1). This provided useful data for discussing the relationship between the content of the metal elements in Cichorium glandulosum Boiss et Huet and its clinical application in cardiovascular and osteoporosis disease.