Two undescribed benzopyran derivatives from Ranunculus sceleratus L. with their in vitro anti-inflammatory activity in lipopolysaccharide-activated RAW 264.7 cells.

Previous studies on Ranunculus sceleratus L. have shown the existence of coumarins and their anti-inflammatory effect. Phytochemical work was conducted to investigate the bioactive compounds, leading to the isolation of two undescribed benzopyran derivatives, namely ranunsceleroside A (1) and B (3), together with two known coumarins (2, 4) from the whole plant of R. sceleratus L. All compounds were structurally identified by extensive spectroscopic analysis and then investigated for their inhibitory effect on nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6) production induced by lipopolysaccharide (LPS) in RAW 264.7 murine macrophages, repectively. As a result, compound 1-4 presented inhibitory effects on the production of NO, TNF-α, IL-1ß, and IL-6 in a concentration-dependent manner, which provides a potential chemical basis for the traditional use of R. sceleratus L. as an anti-inflammatory plant.

[Simultaneous determination of eight active components in Chrysanthemum indicum by HPLC].

This study is aimed to establish a high-performance liquid chromatography (HPLC) method for simultaneous determination of chlorogenic acid, caffeic acid, 1,3-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, luteolin-7-O-beta-D-glucoside, 3,4-dicaffeoylquinic acid, linarin and luteolin in Chrysanthemum indicum. The separation was carried out on a Shim pack VP-ODS (4.6 mm x 250 mm, 5 microm) column eluting with mobile phases of methanol (A) and water containing 0.3% phosphoric acid (B) in gradient mode (0-9 min, 85% -80% B; 9-12 min, 80% -70% B; 12-15 min, 70% -65% B; 15-20 min, 65% -60% B; 20-23 min, 60% -55% B; 23-29 min, 55% -54.4% B; 29-32 min, 54.4% -45% B; 32-37 min, 45% -5% B; 37-45 min, 5% -85% B) at the flow rate of 1.0 mL x min(-1). The column temperature was 35 degrees C and the detection wavelength was set at 326 nm. The good separation of chlorogenic acid, caffeic acid, 1,3-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, luteolin-7-O-beta-D-glucoside, 3,4-dicaffeoylquinic acid, linarin and luteolin was achieved within 40 min. Calibration curves of the eight effective components showed good linear relationship (r > 0.999 5, n = 7). The average recoveries were within 97.03%-102.3% (RSD < 2.0%, n = 6). The method is simple, accurate and repeatable and can be used for the quality control of Ch. indicum.

Review on Research Progress and Prospects of Cicada Flower, Isaria cicadae (Ascomycetes).

Cicada flower, Isaria cicadae Miq., has been a traditional Chinese medicine for approximately 1600 years. Many works on its identification, bioactivities, and clinical use against some disorders have been published, but some inaccuracies and inconsistencies need to be further clarified. In combination with our > 20 years of research and application of cicada flower and examination of the literature and patents published in recent years, this article summarizes and reviews the life cycle and taxonomy, genome size and mating type, molecular systematic classification and cultivation, active ingredients, and pharmacological functions of I. cicadae.

Clinical Randomized Controlled Study of Acupuncture Treatment on Children with Autism Spectrum Disorder (ASD): A Systematic Review and Meta-Analysis.

This study aimed to summarize the effectiveness and safety of acupuncture in the treatment of autism spectrum disorder (ASD) through literature analysis and evaluation. All studies were retrieved from various databases as follows: English databases, such as PubMed, Cochrane Library, Ovid, and Web of Science, and Chinese databases, such as China National Knowledge Infrastructure (CNKI), WanFang Data (WF), and Technology Periodical Database (VIP). The Cochrane Collaboration's Bias Risk Assessment Scale was used to assess the studies' risk of bias. The effects of acupuncture treatment for ASD were determined using the following indicators: childhood autism rating scale (CARS), autism behavior check list (ABC), Reynell developmental language scale (RDLS), and functional independence measure of children (WeeFIM). The risk map of bias of these studies' quality and the meta-analysis results of the indicators was prepared with RevMan 5.2 software. Finally, 16 studies were included, five of which were in English and 11 were in Chinese. The 16 studies included 1332 patients. The CARS results for subgroup analysis were as follows: acupuncture subgroup (MD = -2.65, 95% CI (-3.22, -2.07)) and acupuncture plus massage subgroup (MD = -10.35, 95% CI (-11.34, -9.36)). The ABC results were as follows: (MD = -6.70, 95% CI (-9.10, -4.29)). The analysis results of sensory, relating, language, body and object use, and social/self-help in the subitems of ABC were as follows: sensory (MD = -2.67, 95% CI (-2.90, -2.44)), relating (MD = -3.28, 95% CI (-3.55, -3.02)), language (MD = -2.45, 95% CI (-2.73, -2.16)), body and object use (MD = -1.19, 95% CI (-1.38, -1.00)), and social/self-help (MD = -2.09, 95% CI (-2.30, -1.89)). For the analysis results of comprehension and expression ages in the subitems of RDLS, the comprehension age results were as follows: (MD = 0.08, 95% CI (-0.06, 0.22), P = 0.27). Those of expression age were as follows: (MD = 0.15, 95% CI (0.04, 0.26), P=0.009). The WeeFIM results were as follows: (MD = 3.70, 95% CI (2.38, 5.02)). This study suggested that acupuncture could effectively treat ASD. However, acupuncture methods and prescriptions at this stage remain heterogeneous, and acupuncture treatment operations require standardization. Studies using rigorous and standard research designs are needed to draw stronger conclusions about the advantages of using acupuncture to treat children and adolescents with ASD.

Preventive effects of total flavonoids of Litsea coreana leve on hepatic steatosis in rats fed with high fat diet.

AIM OF THE STUDY: To evaluate the protective effects of total flavonoids of Litsea Coreana leve (TFLC) on rat high fat diet-induced hepatic steatosis model. MATERIALS AND METHODS: Rats were given either a high fat diet alone or the same diet plus TFLC for 4 weeks. RESULTS: TFLC improved liver histology with reduced serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as well as decreased the over accumulation lipids in serum and liver. TFLC increased serum levels of leptin and insulin, while decreased serum TNFalpha level in high fat diet fed rat. Furthermore, TFLC was found increased the expression of peroxisome proliferator-activated receptor alpha (PPARalpha) in high fat diet fed rat liver. These benefits were associated with increased superoxide dismutase (SOD) and decreased malondialdehyde (MDA) in high fat diet fed rat liver. CONCLUSIONS: TFLC exerts protective effects against hepatic steatosis in rats fed with high fat diet possibly through its antioxidant actions, improving the adipocytokines release and increasing the expression of PPARalpha.

Effect of total flavonoids of Chrysanthemum indicum on the apoptosis of synoviocytes in joint of adjuvant arthritis rats.

Chrysanthemum is a traditional Chinese medicine used in China to treat inflammatory diseases. The total flavonoids Chrysanthemum indicum (TFC) were extracted from the dried bud of Chrysanthemum indicum. Our previous study had demonstrated that TFC was a new class of effective anti-inflammation, analgesia and immunoloregulation agents. In this study, we established an adjuvant arthritis (AA) model by injection of Freund's Complete Adjuvant (FCA) to investigate the effect of TFC on the apoptosis of synoviocytes in AA Rats. Synoviocytes isolated from knee joint of rats were treated with different doses of TFC in vitro. Synoviocytes proliferation was measured by MTT assay, and DNA fragmentations were evaluated on agarose gel electrophoresis. The levels of caspase-3 cleaved fragments were analyzed by Western blot. The annexin V stain assay was used to explore the inhibition of caspase-3 on the amelioration of synoviocytes apoptosis. The results showed that TFC inhibited the proliferation of synoviocytes. Electrophoresis showed higher ladders of DNA bands in the TFC group. Cleaved fragments of caspase-3 were increased significantly. Furthermore, the apoptotic synoviocytes were markedly decreased by the caspase-3 specific inhibitor. These results suggest that TFC could induce synoviocytes apoptosis and suppress proliferation of synoviocytes in adjuvant-induced arthritis rats.

Preliminary study of total flavonoids from Litsea coreana Levl. on experimental adjuvant-induced arthritis in rats.

Litsea coreana Levl., a traditional Chinese medicine, has long been used for its diverse benefits such as detoxification and detumescence. Total flavonoids from Litsea coreana Levl. (TFLC) are the effective fraction of L. coreana. This study was designed to investigate the anti-inflammatory effects and mechanisms of TFLC against Feund's complete adjuvant (FCA)-induced arthritis in rats. Arthritis was evaluated by secondary paw swelling, polyarthritis index, body weight and histopathologic analysis. Con A- or LPS-stimulated splenocyte proliferation and cytokine (IL-1 and IL-2) production were assessed by MTT assay and activated mouse cell proliferation assay, respectively. The results indicate that therapeutic administration of TFLC (50, 100, 200 mg/kg, ig x 12 days) could significantly suppress secondary arthritis in rats with adjuvant-induced arthritis (AA). In vivo, TFLC (50, 100, 200 mg/kg, ig x 12 days) augmented splenocyte proliferation and increased IL-2 production in splenocytes, while reduced IL-1 activity in peritoneal macrophages (PM(Phi)) of AA rats. In vitro, TFLC at concentrations from 0.005 to 50 microg/ml exerted the same immunoregulatory effects on AA rats as those in vivo. In addition, an attractive feature of TFLC lies in its apparent lack of toxicity. These results suggest that TFLC without toxicity has a significant anti-arthritic effect on AA rats which could be associated with its anti-inflammatory and immunomodulatory properties.

Protective effect of total flavonoids from Bidens bipinnata L. against carbon tetrachloride-induced liver injury in mice.

Bidens bipinnata L. is well known in China as a traditional Chinese medicine. This study was designed to evaluate the hepatoprotective activity of the total flavonoids of B. bipinnata L. (TFB) against carbon tetrachloride (CCl4)-induced acute liver injury in mice and to determine its mechanism of action. Oral administration of TFB at doses of 50, 100 and 200 mg kg(-1) for 7 days significantly reduced the elevated relative values of liver weight, serum transaminases (alanine aminotransferase and aspartate aminotransferase) and the hepatic morphologic changes induced by CCl4 in mice. In addition, TFB markedly inhibited CCl4-induced lipid peroxidation and enhanced the activity of the antioxidant enzymes superoxide dismutase and glutathione peroxidase. Moreover, pretreatment with TFB suppressed nitric oxide production and nuclear factor-kappaB activation in CCl4-treated mice. The results suggest that TFB has significant hepatoprotective activity and its mechanism is related, at least in part, to its antioxidant properties. Further research is required to investigate the detailed mechanism of the protective effect of TFB on acute liver injury.

Analysis of radical scavenging active components in the fermented mycelia of Ophiocordyceps formosana.

The rates of 1,1-diphenyl-2-picryhydrazyl (DPPH)-free radical scavenging and hydroxyl radical eliminating were employed as indexes to qualitatively and quantitatively analyse the radical scavenging activity of the extracts with different solvents from fermented mycelia of Hirsutella huangshanensis RCEF0868, the anamorph of Ophiocordyceps formosana (FMOF). The results showed that both the aqueous extract and the methanol extract had significant radical scavenging activity. The half maximal inhibitory concentrations (IC50) of the aqueous extract on DPPH-free radical and hydroxyl radical are 0.85 and 1.37 mg/mL, respectively, while the IC50 of methanol extract is 1.23 and 2.91 mg/mL, respectively. The combined liquid chromatography-diode array detector-high resolution mass spectrometer(HRMS) analysis and activity determination revealed that the molecular formulas of three radical scavengers in aqueous extract were C6H14O6(1), C14H17N5O8(2) and C20H33N5O9(3). In addition to the compound 1, the methanol extract had another three radical scavengers as C30H22O11(4), C30H22O10(5) and C30H18O10(6). All the six kinds of components exhibited the DPPH radical scavenging activity, and compounds 1, 4 and 5 also showed the hydroxyl radical eliminating activity at the same time. By querying the database of natural products and reference to the relevant reports, compound 3 may be a new entity. The others were respectively cordycepic acid, succinoadenosine, 4α-oxyrugulosin, rugulosin and skyrin and, among them, succinoadenosine and 4α-oxyrugulosin were found in entomogenous fungi for the first time.

Identification and determination of myriocin in Isaria cicadae and its allies by LTQ-Orbitrap-HRMS.

A hybrid linear ion trap-quadrupole-Orbitrap high-resolution mass spectrometry (LTQ-Orbitrap-HRMS) was used to qualitatively and quantitatively analyse the myriocin in Isaria cicadae and its allies. The samples were prepared with 95% methanol for 30 min by ultrasonic-assisted extraction. The target compound was purified by ODS solid-phase extraction (SPE) column. The enriched samples were identified by mass spectrometry. The results showed that the contents of myriocin in both wild and artificial Isaria cicadae were below the detection limit, while a strain of Ophiocordyceps longissima and Cordyceps cicadae Shing (Dujiaolong), both closely related to the Isaria cicadae, and its asexual mycelia are rich in myriocin. It suggests that it may be wrong to consider C. cicadae as I. cicadae's teleomorph in Genbank or Mycobank in many published reports based on chemical classification, and the species rich in myriocin is probably not Isaria cicadae.

Immunomodulating effects of fractioned polysaccharides isolated from Yu-Ping-Feng-Powder in cyclophosphamide-treated mice.

Yu-Ping-Feng-Powder (YP), a traditional Chinese medicine prescription, is widely applied in China for the cure and prevention of diseases related to immunodeficiency. To test whether the fractioned polysaccharides (YPF-P) isolated from YP have immunomodulating activities, the effects of YPF-P on cyclophosphamide (Cy)-treated mice were studied in relation to phagocytosis of macrophage, splenocyte proliferation, and humoral, and cellular immunity parameters. It was found that YPF-P enhances phagocytic activity, augments ConA- or LPS-stimulated T cell proliferation, increases the quantitative haemolysis of SRBC (QHS) and delayed-type hypersensitivity reaction (DTH) to dinitrofluorobenzene. Hence, YPF-P restored the immuno-competence suppressed by Cy. YPF-P also augmented IL-2 and IFN-gamma production, but failed to increase IL-4 production, which indicates that there is high probability that it enhance Th1 function. These results suggested that YPF-P has immunomodulating effects and that the polysaccharides constitute one of the active components of YP.

Nutritional Composition and Bioactive Constituents of Artificial Culture of Ophiocordyceps longissima (Ascomycetes).

In this study, the compositions of Ophiocordyceps longissima mycelia, synnemata, and fruiting bodies were first analyzed in order to clarify its chemical basis for development as a health food or medicine. We found that the contents of crude protein, polysaccharides, and macroelements were highest in mycelia, whereas effective components, including mannitol, ergosterol, adenosine, inosine, Zn, and Se, were lowest in mycelia. Polysaccharide, mannitol, and ergosterol levels in synnemata (2.33, 4.54, and 0.66 g/100 g, respectively) were similar to those in fruiting bodies, but was significantly different from those of mycelia (4.79, 1.77, and 0.43 g/100 g). Trehalose content in fruiting bodies (2.15 g/100 g) was >4 times higher than that in synnemata (0.5 g/100 g). Adenosine content in fruiting bodies (0.024 g/100 g) was 3-4 times higher than that of synnemata, whereas inosine, cytosine, guanosine, and uridine (0.093, 0.145, 0.053, and 0.073 g/100 g) were highest in synnemata. Cu and Se were lower in mycelia (3.30 × 10⁻4 and 3.1 × 10⁻5 g/100 g) than in synnemata (1.85 × 10⁻³ and 7.2 × 10⁻5 g/100 g) and fruiting bodies (1.67 × 10⁻³ and 4.3 × 10⁻5 g/100 g). As, Hg, Cd, and Pb in mycelia were under the limit of edible fungus health standard; Pb was not found, but Cd, Hg, and As were detected in fruiting bodies. These findings suggest that O. longissima could be utilized in different culture methods according to market demand and might be a possible health food or medicinal resource.

Orcinol glucoside produces antidepressant effects by blocking the behavioural and neuronal deficits caused by chronic stress.

This study focused on the antidepressant potential of orcinol glucoside (OG) and its possible mechanisms of action. We established a depressed rat model using 3 consecutive weeks of chronic unpredictable mild stress (CUMS). The antidepressant-like effect of OG was revealed using the sucrose preference test, the open field test, the forced swimming test (FST), and the tail suspension test (TST). The activity of the hypothalamic-pituitary-adrenal (HPA) axis was evaluated by detecting the serum corticosterone (CORT) concentrations and mRNA expression of corticotrophin-releasing hormone (CRH) in the hypothalamus. The protein expression levels of brain-derived neurotrophic factor (BDNF) and total phosphorylated-ERK1/2 were detected by western blot. The results showed that OG treatment (1.5, 3, or 6mg/kg) alleviated the depression-like behaviour of rats under CUMS, as indicated by the increased sucrose preference and the decreased immobility in both the FST and TST, although the rearing frequency in the open field test increased only in the group that received the lowest dose (1.5mg/kg OG). Rats that received OG treatment exhibited reduced serum CORT levels and CRH mRNA expression in the hypothalamus, suggesting that the hyperactivity of the HPA axis in CUMS rats was reversed by OG treatment. Moreover, OG treatment upregulated the protein levels of BDNF and phosphorylated-ERK1/2 in the hippocampus, even above control levels. Our findings suggest that OG improved depressive behaviour in CUMS rats by downregulating HPA axis hyperactivity and increasing BDNF expression and ERK1/2 phosphorylation in the hippocampus.

Protective effects of total flavonoids from Litsea coreana on alcoholic fatty liver in rats associated with down-regulation adipose differentiation-related protein expression.

Alcoholic fatty liver (AFL) is a reversible condition, but it can potentiate the development of alcoholic hepatitis and even cirrhosis by increasing oxidant generation, which is one of the key pathogenic factors and could result in alcoholic liver disease (ALD). Total flavonoids from Litsea coreana (TFLC), an active component extracted from Litsea coreana leve, have been shown to have therapeutic effects on hyperlipidemia. The present study was to evaluate the protective effects of TFLC on alcoholic fatty liver (AFL) in rats, and investigate the potential mechanism. An AFL model in rats was established by intaking different doses of alcohol (concentration from 5% to 40%) over 12 weeks. Serum levels of TG, TC, LDL-C, HDL-C, TNF-α, insulin, and glucose were measured, histopathologic changes were determined, and expression of adipose differentiation-related protein (ADRP) in the liver were evaluated by Western blotting and immunohistochemistry, respectively. The results showed that treatment with TFLC resulted in decreased serum levels of TG, TC, LDL-C, TNF-α, glucose and insulin, as well as improved liver index. Morphological evaluation revealed rats in model group developed a severe steatosis, but the severities of liver steatosis were effectively ameliorated in TFLC (200 and 400 mg/kg) treated groups. Expression of hepatic ADRP were increased in model group, and suppressed in TFLC treated groups. These results suggest that TFLC had a protective effect on AFL rats; the mechanism may be involved in regulation serum lipid profiles via down-regulation of hepatic expression of ADRP in AFL rats.

Antidiabetic effects of total flavonoids from Litsea Coreana leve on fat-fed, streptozotocin-induced type 2 diabetic rats.

This study was initiated to determine the possible antidiabetic effects of total flavonoids of Litsea Coreana leve (TFLC), an alcohol extract from the dried leaves of Litsea Coreana leve, on type 2 diabetic rats. Male Sprague-Dawley rats (n = 40, 160-180 g) were divided into two groups and fed with normal chow diet (Normal Control group) or high-fat diet (HFD) for a period of 4 weeks. After 4 weeks of dietary manipulation, the HFD-fed rats were injected with 30 mg/kg streptozocin (STZ) to induce diabetes 72 hours after STZ injection. These diabetic rats were randomly divided into 3 groups (n = 10): Diabetic Control group, Diabetic + TFLC group and Diabetic + PIO group. Diabetic + TFLC group and Diabetic + PIO group were orally administered with 400 mg/kg TFLC or 10 mg/kg pioglitazone (all suspended in 0.5% CMC-Na) respectively for 6 weeks. All rats were examined for body weight, serum and hepatic biochemical indices, content of malondialdehyde (MDA), activities of superoxide dismutase (SOD) and pathological changes in liver and pancreas, as well as protein tyrosine phosphatase 1B (PTP1B) expression in liver. The diabetic rats became obese, insulin resistant, hyperglycemic and hyperlipidemic. Treatment with TFLC showed a significant increase in insulin sensitivity, serum HDL-C level and SOD activities, meanwhile marked decrease in body weight, serum FFA, TC, TG, LDL-C, CRP, MDA content. TFLC also attenuated pathologic alterations in liver and pancreatic islet. Furthermore, TFLC was found to decrease the expression of PTP1B in diabetic rat liver. These results suggested that TFLC could ameliorate hyperglycemia, hyperlipoidemia, inflammation and oxidation stress, as well as insulin resistance of type 2 diabetic rats.

Preventive effects of a fractioned polysaccharide from a traditional Chinese herbal medical formula (Yu Ping Feng San) on carbon tetrachloride-induced hepatic fibrosis.

OBJECTIVES: The study was to investigate the prevention effects and possible mechanism of Yu Ping Feng San fractioned polysaccharide (YPF-P) on CCl(4)-induced liver fibrosis in rats. METHODS: YPF-P was prepared from root of Astragalus membranaceus, rhizome of Atractylodes macrocephaia and root of Raidix saposhnikoviae, and compared with polysaccharide from root of Astragalus membranaceus (AP). Hepatic fibrosis was induced by subcutaneous injection with carbon tetrachloride twice weekly for 12 weeks in Sprague-Dawley rats. YPF-P, AP and colchicine were administered intragastrically daily to carbon tetrachloride-treated rats. Histopathological changes of the liver and hepatic stellate cells were evaluated by Masson staining and transmission electron microscopy, respectively. Markers of fibrosis were determined by radioimmunoassay, biochemistry assay and ELISA. The mRNA expressions of tissue inhibitor of metalloproteinase-1 (TIMP-1), matrix metalloproteinase-13 (MMP-13), procollagen I and collagen III were detected by RT-PCR. KEY FINDINGS: YPF-P dose-dependently alleviated the degree of liver fibrosis and inhibited hepatic stellate cell transformation into myofibroblast-like cells, markedly reduced the elevated levels of hyaluronic acid, laminin, type IV collagen, type III procollagen, hydroxyproline and transforming growth factor beta-1, suppressed procollagen I, collagen III and TIMP-1 expression, and improved the TIMP-1/MMP-13 ratio. MMP-13 expression was only promoted moderately by YPF-P. Compared with AP, YPF-P showed more potency on most markers except laminin, type IV collagen and MMP-13 mRNA. CONCLUSIONS: YPF-P prevented the progress of rat liver fibrosis induced by carbon tetrachloride and had a more potent preventative effect. The preventative effect may be associated with the ability of YPF-P to inhibit the synthesis of matrix collagen and balance the TIMP/MMP system.