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26.  Brazilian green propolis modulates inflammation, angiogenesis and fibrogenesis in intraperitoneal implant in mice 
Background
Chronic inflammatory processes in the peritoneal cavity develop as a result of ischemia, foreign body reaction, and trauma. Brazilian green propolis, a beeswax product, has been shown to exhibit multiple actions on inflammation and tissue repair. Our aim was to investigate the effects of this natural product on the inflammatory, angiogenic, and fibrogenic components of the peritoneal fibroproliferative tissue induced by a synthetic matrix.
Methods
Chronic inflammation was induced by placing polyether-polyurethane sponge discs in the abdominal cavity of anesthetized Swiss mice. Oral administration of propolis (500/mg/kg/day) by gavage started 24 hours after injury for four days. The effect of propolis on peritoneal permeability was evaluated through fluorescein diffusion rate 4 days post implantation. The effects of propolis on the inflammatory (myeloperoxidase and n-acetyl-β-D-glucosaminidase activities and TNF-α levels), angiogenic (hemoglobin content-Hb), and fibrogenic (TGF-β1 and collagen deposition) components of the fibrovascular tissue in the implants were determined 5 days after the injury.
Results
Propolis was able to decrease intraperitoneal permeability. The time taken for fluorescence to peak in the systemic circulation was 20 ± 1 min in the treated group in contrast with 15 ± 1 min in the control group. In addition, the treatment was shown to down-regulate angiogenesis (Hb content) and fibrosis by decreasing TGF-β1 levels and collagen deposition in fibroproliferative tissue induced by the synthetic implants. Conversely, the treatment up-regulated inflammatory enzyme activities, TNF-α levels and gene expression of NOS2 and IFN-γ (23 and 7 fold, respectively), and of FIZZ1 and YM1 (8 and 2 fold) when compared with the untreated group.
Conclusions
These observations show for the first time the effects of propolis modulating intraperitoneal inflammatory angiogenesis in mice and disclose important action mechanisms of the compound (downregulation of angiogenic components and activation of murine macrophage pathways).
doi:10.1186/1472-6882-14-177
PMCID: PMC4061536  PMID: 24886376
Water extract propolis; Cytokines; Macrophage activation; Fibrosis
27.  Perilla Extract improves gastrointestinal discomfort in a randomized placebo controlled double blind human pilot study 
Background
Gastrointestinal (GI) discomfort, e.g. bloating or rumbling, is a common symptom in otherwise healthy adults. Approximately 20% of the population, particularly women suffer from gastrointestinal discomfort and this affects quality of life. Recent studies discovered a link between the body and mind, called the gut-brain axis. Psychosocial factors, such as e.g. daily stress may cause altered gut physiology leading to ileum contractions and consequently gastrointestinal symptoms. In vitro and ex vivo studies clearly showed that a Perilla frutescens extract combines prokinetic, antispasmodic and anti-inflammatory effects. The aim of the intervention was to investigate the effects of the proprietary Perilla extract on GI discomfort in healthy subjects with gastrointestinal discomfort and reduced bowel movements in comparison to a placebo product.
Methods
The pilot study was performed according to a double-blind, randomized, placebo-controlled parallel design. Fifty healthy subjects with gastrointestinal discomfort and reduced bowel movements, 30-70 years, documented their GI symptoms, stool frequency and consistency daily during a 2-week run-in phase and a 4-week intervention phase with Perilla frutescens extract or placebo. GI symptoms were assessed on a 5-point scale daily and average scores over 14 days intervals were calculated.
Results
All GI symptoms were significantly improved over time by Perilla frutescens extract during the intervention phase (bloating: -0.44 ± 0.56, p = 0.0003; passage of gas: -0.30 ± 0.66, p = 0.0264; GI rumbling: -0.55 ± 0.87, p = 0.0014; feeling of fullness: -0.36 ± 0.72, p = 0.0152; abdominal discomfort: -0.54 ± 0.75, p = 0.004), whereas in the placebo group only abdominal discomfort was significantly improved (-0.31 ± 0.55, p = 0.0345). In the subgroup of women results were strengthened and a subscore out of bloating and abdominal discomfort was significantly improved against placebo (95%CI 0.003 to 0.77; p = 0.048).
Conclusion
The demonstrated effects of Perilla frutescens extract to improve GI complaints offer very promising results, taking into consideration the challenging set up of a nutritional human study with healthy subjects and in the area of digestive health, which is known for high placebo effects.
Trial registration number
NCT01931930 at ClinicalTrials.gov, Registration date 23rd August 2013.
doi:10.1186/1472-6882-14-173
PMCID: PMC4038823  PMID: 24885816
Gastrointestinal discomfort; Perilla frutescens; Bloating; Abdominal discomfort; Human study; Bowel movement; Passage of gas; Rumbling; Feeling of fullness
28.  Ginger extract diminishes chronic fructose consumption-induced kidney injury through suppression of renal overexpression of proinflammatory cytokines in rats 
Background
The metabolic syndrome is associated with an increased risk of development and progression of chronic kidney disease. Renal inflammation is well known to play an important role in the initiation and progression of tubulointerstitial injury of the kidneys. Ginger, one of the most commonly used spices and medicinal plants, has been demonstrated to improve diet-induced metabolic abnormalities. However, the efficacy of ginger on the metabolic syndrome-associated kidney injury remains unknown. This study aimed to investigate the impact of ginger on fructose consumption-induced adverse effects in the kidneys.
Methods
The fructose control rats were treated with 10% fructose in drinking water over 5 weeks. The fructose consumption in ginger-treated rats was adjusted to match that of fructose control group. The ethanolic extract of ginger was co-administered (once daily by oral gavage). The indexes of lipid and glucose homeostasis were determined enzymatically, by ELISA and/or histologically. Gene expression was analyzed by Real-Time PCR.
Results
In addition to improve hyperinsulinemia and hypertriglyceridemia, supplement with ginger extract (50 mg/kg) attenuated liquid fructose-induced kidney injury as characterized by focal cast formation, slough and dilation of tubular epithelial cells in the cortex of the kidneys in rats. Furthermore, ginger also diminished excessive renal interstitial collagen deposit. By Real-Time PCR, renal gene expression profiles revealed that ginger suppressed fructose-stimulated monocyte chemoattractant protein-1 and its receptor chemokine (C-C motif) receptor-2. In accord, overexpression of two important macrophage accumulation markers CD68 and F4/80 was downregulated. Moreover, overexpressed tumor necrosis factor-alpha, interleukin-6, transforming growth factor-beta1 and plasminogen activator inhibitor (PAI)-1 were downregulated. Ginger treatment also restored the downregulated ratio of urokinase-type plasminogen activator to PAI-1.
Conclusions
The present results suggest that ginger supplement diminishes fructose-induced kidney injury through suppression of renal overexpression of macrophage-associated proinflammatory cytokines in rats. Our findings provide evidence supporting the protective effect of ginger on the metabolic syndrome-associated kidney injury.
doi:10.1186/1472-6882-14-174
PMCID: PMC4047007  PMID: 24885946
Zingiber officinale Roscoe; Renoprotection; Anti-inflammation; Metabolic abnormalities
29.  Assessing the antibiotic potential of essential oils against Haemophilus ducreyi 
Background
Haemophilus ducreyi is the bacterium responsible for the genital ulcer disease chancroid, a cofactor for the transmission of HIV, and it is resistant to many antibiotics. With the goal of exploring possible alternative treatments, we tested essential oils (EOs) for their efficacy as antimicrobial agents against H. ducreyi.
Methods
We determine the minimum inhibitory concentration (MIC) of Cinnamomum verum (cinnamon), Eugenia caryophyllus (clove) and Thymus satureioides (thyme) oil against 9 strains of H. ducreyi using the agar dilution method. We also determined the minimum lethal concentration for each oil by subculturing from the MIC plates onto fresh agar without essential oil. For both tests, we used a 2-way ANOVA to evaluate whether antibiotic-resistant strains had a different sensitivity to the oils relative to non-resistant strains.
Results
All 3 oils demonstrated excellent activity against H. ducreyi, with MICs of 0.05 to 0.52 mg/mL and MLCs of 0.1-0.5 mg/mL. Antibiotic-resistant strains of H. ducreyi were equally susceptible to these 3 essential oils relative to non-resistant strains (p = 0.409).
Conclusion
E. caryophyllus, C. verum and T. satureioides oils are promising alternatives to antibiotic treatment for chancroid.
doi:10.1186/1472-6882-14-172
PMCID: PMC4048586  PMID: 24885682
Haemophilus ducreyi; Chancroid; Essential oil
30.  Antiviral activity and possible mode of action of ellagic acid identified in Lagerstroemia speciosa leaves toward human rhinoviruses 
Background
Human rhinoviruses (HRVs) are responsible for more than half of all cases of the common cold and cause billions of USD annually in medical visits and school and work absenteeism. An assessment was made of the cytotoxic and antiviral activities and possible mode of action of the tannin ellagic acid from the leaves of Lagerstroemia speciosa toward HeLa cells and three rhinoviruses, HRV-2, -3, and -4.
Methods
The antiviral property and mechanism of action of ellagic acid were evaluated using a sulforhodamine B assay and real-time reverse transcription-PCR (RT-PCR) with SYBR Green dye. Results were compared with those of the currently used broad-spectrum antiviral agent, ribavirin.
Results
As judged by 50% inhibitory concentration values, natural ellagic acid was 1.8, 2.3, and 2.2 times more toxic toward HRV-2 (38 μg/mL), HRV-3 (31 μg/mL), and HRV-4 (29 μg/mL) than ribavirin, respectively. The inhibition rate of preincubation with 50 μg/mL ellagic acid was 17%, whereas continuous presence of ellagic acid during infection led to a significant increase in the inhibition (70%). Treatment with 50 μg/mL ellagic acid considerably suppressed HRV-4 infection only when added just after the virus inoculation (0 h) (87% inhibition), but not before -1 h or after 1 h or later (<20% inhibition). These findings suggest that ellagic acid does not interact with the HRV-4 particles and may directly interact with the human cells in the early stage of HRV infections to protect the cells from the virus destruction. Furthermore, RT-PCR analysis revealed that 50 μg/mL ellagic acid strongly inhibited the RNA replication of HRV-4 in HeLa cells, suggesting that ellagic acid inhibits virus replication by targeting on cellular molecules, rather than virus molecules.
Conclusions
Global efforts to reduce the level of antibiotics justify further studies on L. speciosa leaf-derived materials containing ellagic acid as potential anti-HRV products or a lead molecule for the prevention or treatment of HRV infection.
doi:10.1186/1472-6882-14-171
PMCID: PMC4052798  PMID: 24885569
Human rhinovirus; Natural antiviral agent; Lagerstroemia speciosa; Ellagic acid; Tannin; Cytotoxicity; Selectivity; RNA replication
31.  A preliminary evaluation of antihyperglycemic and analgesic activity of Alternanthera sessilis aerial parts 
Background
Alternanthera sessilis is used by folk medicinal practitioners of Bangladesh for alleviation of severe pain. The objective of this study was to scientifically analyze the analgesic (non-narcotic) property of aerial parts of the plant along with antihyperglycemic activity.
Methods
Antihyperglycemic activity was measured by oral glucose tolerance tests. Analgesic (non-narcotic) activity was determined by observed decreases in abdominal writhings in intraperitoneally administered acetic acid-induced pain model in mice.
Results
Administration of methanol extract of aerial parts led to dose-dependent and significant reductions in blood glucose levels in glucose-loaded mice. At doses of 50, 100, 200 and 400 mg per kg body weight, the extract reduced blood sugar levels by 22.9, 30.7, 45.4 and 46.1%, respectively compared to control animals. By comparison, a standard antihyperglycemic drug, glibenclamide, when administered at a dose of 10 mg per kg body weight, reduced blood glucose level by 48.9%. In analgesic activity tests, the extract at the above four doses reduced the number of abdominal writhings by 27.6, 37.9, 41.4, and 44.8%, respectively. A standard analgesic drug, aspirin, reduced the number of writhings by 31.0 and 51.7%, respectively, when administered at doses of 200 and 400 mg per kg body weight.
Conclusion
The results validate the folk medicinal use of the plant to alleviate pain. At the same time, the antihyperglycemic activity result suggests that the plant may be a potential source for blood sugar lowering drug(s).
doi:10.1186/1472-6882-14-169
PMCID: PMC4035668  PMID: 24885344
Antihyperglycemic; Alternanthera sessilis; Glucose tolerance; Non-narcotic analgesic; Amaranthaceae
32.  The inhibitory effect of Mesembryanthemum edule (L.) bolus essential oil on some pathogenic fungal isolates 
Background
Mesembryanthemum edule is a medicinal plant which has been indicated by Xhosa traditional healers in the treatment HIV associated diseases such as tuberculosis, dysentery, diabetic mellitus, laryngitis, mouth infections, ringworm eczema and vaginal infections. The investigation of the essential oil of this plant could help to verify the rationale behind the use of the plant as a cure for these illnesses.
Methods
The essential oil from M. edule was analysed by GC/MS. Concentration ranging from 0.005 - 5 mg/ml of the hydro-distilled essential oil was tested against some fungal strains, using micro-dilution method. The plant minimum inhibitory activity on the fungal strains was determined.
Result
GC/MS analysis of the essential oil resulted in the identification of 28 compounds representing 99.99% of the total essential oil. A total amount of 10.6 and 36.61% constituents were obtained as monoterpenes and oxygenated monoterpenes. The amount of sesquiterpene hydrocarbons (3.58%) was low compared to the oxygenated sesquiterpenes with pick area of 9.28%. Total oil content of diterpenes and oxygenated diterpenes detected from the essential oil were 1.43% and 19.24%. The fatty acids and their methyl esters content present in the essential oil extract were found to be 19.25%. Antifungal activity of the essential oil extract tested against the pathogenic fungal, inhibited C. albican, C. krusei, C. rugosa, C. glabrata and C. neoformans with MICs range of 0.02-0.31 mg/ml. the activity of the essential oil was found competing with nystatin and amphotericin B used as control.
Conclusion
Having accounted the profile chemical constituent found in M. edule oil and its important antifungal properties, we consider that its essential oil might be useful in pharmaceutical and food industry as natural antibiotic and food preservative.
doi:10.1186/1472-6882-14-168
PMCID: PMC4057826  PMID: 24885234
Mesembryanthemum edule; Essential oil; GC/MS; Antifungal activity; Opportunistic fungi
33.  Enhancement of gefitinib-induced growth inhibition by Marsdenia tenacissima extract in non-small cell lung cancer cells expressing wild or mutant EGFR 
Background
Non-small cell lung cancer (NSCLC) expressed high levels of epidermal growth factor receptor (EGFR). Gefitinib (Iressa) has demonstrated clinical efficacy in NSCLC patients harboring EGFR mutations or refractory to chemotherapy. However, most of NSCLC patients are with wild type EGFR, and showed limited response to gefitinib. Therefore, to develop new effective therapeutic interventions for NSCLC is still required. Our previous study showed Marsdenia tenacissima extract (MTE) restored gefitinib efficacy in the resistant NSCLC cells, but whether MTE acts in the gefitinib-sensitive NSCLC cells is the same as it in the resistant one is unknown.
Methods
Dose response curves for gefitinib and MTE were generated for two sensitive NSCLC cell lines with mutant or wild type EGFR status. Three different sequential combinations of MTE and gefitinib on cell growth were evaluated using IC50 and Combination Index approaches. The flow cytometric method was used to detect cell apoptosis and cell cycle profile. The impact of MTE combined with gefitinib on cell molecular network response was studied by Western blotting.
Results
Unlike in the resistant NSCLC cells, our results revealed that low cytotoxic dose of MTE (8 mg/ml) combined gefitinib with three different schedules synergistically or additively enhanced the growth inhibition of gefitinib. Among which, MTE → MTE + gefitinib treatment was the most effective one. MTE markedly prompted cell cycle arrest and apoptosis caused by gefitinib both in EGFR mutant (HCC827) and wild type of NSCLC cells (H292). The Western blotting results showed that MTE → MTE + gefitinib treatment further enhanced the suppression of gefitinib on cell growth and apoptosis pathway such as ERK1/2 and PI3K/Akt/mTOR. This combination also blocked the activation of EGFR and c-Met which have cross-talk with each other. Unlike in gefitinib-resistant NSCLC cells, MTE alone also demonstrated certain unexpected modulation on EGFR related cell signal pathways in the sensitive cells.
Conclusion
Our results suggest that MTE is a promising herbal medicine to improve gefitinib efficacy in NSCLC regardless of EGFR status. However, why MTE acted differently between gefitinib-sensitive and -resistant NSCLC cells needs a further research.
doi:10.1186/1472-6882-14-165
PMCID: PMC4040364  PMID: 24884778
Marsdenia tenacissima extract (MTE); Gefitinib; Non-small cell lung cancer (NSCLC); Combination; EGFR related pathway
34.  Protective effects of pomegranate (Punica granatum) juice on testes against carbon tetrachloride intoxication in rats 
Background
Pomegranate fruit has been extensively used as a natural medicine in many cultures. The present study was aimed at evaluating the protective effects of pomegranate (Punica granatum) juice against carbon tetrachloride (CCl4)-induced oxidative stress and testes injury in adult Wistar rats.
Methods
Twenty eight Wistar albino male rats were divided equally into 4 groups for the assessment of protective potential of pomegranate juice. Rats of group I (control) received only vehicles and had free access to food and water. Rats of groups II and IV were treated with CCl4 (2 ml/kg bwt) via the intraperitoneal route once a week for ten weeks. The pomegranate juice was supplemented via drinking water 2 weeks before and concurrent with CCl4 treatment to group IV. Group III was supplemented with pomegranate juice for twelve weeks. The protective effects of pomegranate on serum sex hormones, oxidative markers, activities of antioxidant enzymes and histopathology of testes were determined in CCl4-induced reproductive toxicity in rats.
Results
Pomegranate juice showed significant elevation in testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) those depleted by the injection of CCl4. Activity levels of endogenous testesticular antioxidant enzymes; superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and glutathione reductase (GR) and glutathione (GSH) contents were increased while lipid peroxidation (LPO) and nitric oxide (NO) were decreased with pomegranate juice. Moreover, degeneration of germ and Leydig cells along with deformities in spermatogenesis induced after CCl4 injections were restored with the treatment of pomegranate juice.
Conclusion
The results clearly demonstrated that pomegranate juice augments the antioxidant defense mechanism against carbon tetrachloride-induced reproductive toxicity and provides evidence that it may have a therapeutic role in free radical mediated diseases.
doi:10.1186/1472-6882-14-164
PMCID: PMC4041339  PMID: 24884677
Punica granatum; Carbon tetrachloride; Oxidative stress; Testes; Rats
35.  Integrating complementary and alternative medicine into mainstream healthcare services: the perspectives of health service managers 
Background
Complementary and alternative medicine (CAM) is increasingly included within mainstream integrative healthcare (IHC) services. Health service managers are key stakeholders central to ensuring effective integrative health care services. Yet, little research has specifically investigated the role or perspective of health service managers with regards to integrative health care services under their management. In response, this paper reports findings from an exploratory study focusing exclusively on the perspectives of health service managers of integrative health care services in Australia regarding the role of CAM within their service and the health service managers rational for incorporating CAM into clinical care.
Methods
Health service managers from seven services were recruited using purposive and snowball sampling. Semi-structured interviews were conducted with the health service managers. The services addressed trauma and chronic conditions and comprised: five community-based programs including drug and alcohol rehabilitation, refugee mental health and women’s health; and two hospital-based specialist services. The CAM practices included in the services investigated included acupuncture, naturopathy, Western herbal medicine and massage.
Results
Findings reveal that the health service managers in this study understand CAM to enhance the holistic capacity of their service by: filling therapeutic gaps in existing healthcare practices; by treating the whole person; and by increasing healthcare choices. Health service managers also identified CAM as addressing therapeutic gaps through the provision of a mind-body approach in psychological trauma and in chronic disease management treatment. Health service managers describe the addition of CAM in their service as enabling patients who would otherwise not be able to afford CAM to gain access to these treatments thereby increasing healthcare choices. Some health service managers expressly align the notion of treating the whole person within a health promotion model and focus on the relevance of diet and lifestyle factors as central to a CAM approach.
Conclusions
From the perspectives of the health service managers, these findings contribute to our understanding around the rationale to include CAM within mainstream health services that deal with psychological trauma and chronic disease. The broader implications of this study can help assist in the development of health service policy on CAM integration in mainstream healthcare services.
doi:10.1186/1472-6882-14-167
PMCID: PMC4048459  PMID: 24885066
Integrative healthcare; Integrative medicine; Complementary and alternative medicine (CAM); Health services research; Chronic disease management; Psychological trauma management; Drug and alcohol rehabilitation
36.  An orally active immune adjuvant prepared from cones of Pinus sylvestris, enhances the proliferative phase of a primary T cell response 
Background
We have previously demonstrated that an alkaline extract of shredded pinecones yields a polyphenylpropanoid polysaccharide complex (PPC) that functions as an orally active immune adjuvant. Specifically, oral PPC can boost the number of antigen-specific memory CD8+ T cells generated in response to a variety of vaccine types (DNA, protein, and dendritic cell) and bias the response towards one that is predominately a T helper 1 type.
Methods
An immune response was initiated by intraperitoneal injection of mice with Staphylococcus enterotoxin B (SEB). A group of mice received PPC by gavage three times per day on Days 0 and 1. The draining lymph nodes were analyzed 48–96 h post-injection for the numbers of reactive T cells, cytokine production, the generation of reactive oxygen species, and apoptotsis.
Results
In this study we examined whether the ability of PPC to boost a T cell response is due to an effect on the proliferative or contraction phases, or both, of the primary response. We present data to demonstrate that oral PPC significantly enhances the primary T cell response by affecting the expansion of T cells (both CD4 and CD8) during the proliferative phase, while having no apparent effects on the activation-induced cell death associated with the contraction phase.
Conclusions
These findings suggest that PPC could potentially be utilized to enhance the T cell response generated by a variety of prophylactic and therapeutic vaccines designed to target a cellular response.
doi:10.1186/1472-6882-14-163
PMCID: PMC4051390  PMID: 24884568
Polyphenylpropanoid polysaccharide complex; Primary T cell response; Pine cone extract
38.  Grape seed extract prevents skeletal muscle wasting in interleukin 10 knockout mice 
Background
Muscle wasting is frequently a result of cancers, AIDS, chronic diseases and aging, which often links to muscle inflammation. Although grape seed extract (GSE) has been widely used as a human dietary supplement for health promotion and disease prevention primarily due to its anti-oxidative and anti-inflammative effects, it is unknown whether GSE affects muscle wasting. The objective is to test the effects of GSE supplementation on inflammation and muscle wasting in interleukin (IL)-10 knockout mice, a recently developed model for human frailty.
Methods
Male IL-10 knockout (IL10KO) C57BL/6 mice at 6 weeks of age were assigned to either 0% or 0.1% GSE (in drinking water) groups (n = 10) for 12 weeks, when skeletal muscle was sampled for analyses. Wild-type C57BL/6 male mice were used as controls.
Results
Tibialis anterior muscle weight and fiber size of IL10KO mice were much lower than wild-type mice. IL10KO enhanced nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling and inflammasome formation when compared to wild-type mice. Phosphorylation of anabolic signaling was inhibited, whereas muscle specific ubiquitin ligase, AMP-activated protein kinase (AMPK) and apoptotic signaling were up-regulated in IL10KO mice. GSE supplementation effectively rectified these adverse changes in IL10KO muscle, which provide an explanation for the enhanced muscle mass, reduced protein degradation and apoptosis in GSE supplemented mice compared to IL10KO mice without supplementation.
Conclusion
GSE supplementation effectively prevents muscle wasting in IL10KO mice, showing that GSE can be used as an auxiliary treatment for muscle loss associated with chronic inflammation and frailty.
doi:10.1186/1472-6882-14-162
PMCID: PMC4041050  PMID: 24884473
Apoptosis; Atrophy; Grape seed extract; IL-10; Inflammasome; Inflammation; Skeletal muscle; Wasting
39.  In vitro and ex-vivo cellular antioxidant protection and cognitive enhancing effects of an extract of Polygonum minus Huds (Lineminus™) demonstrated in a Barnes Maze animal model for memory and learning 
Background
Polygonum minus Huds.is a culinary flavouring that is common in South East Asian cuisine and as a remedy for diverse maladies ranging from indigestion to poor eyesight. The leaves of this herb have been reported to be high in antioxidants. Flavonoids which have been associated with memory, cognition and protection against neurodegeneration were found in P. minus.
Method
This study examined a P. minus aqueous extract (Lineminus™) for its antioxidant activity using the Oxygen Radical Absorbance Capacity (ORAC) assay, the ex vivo Cellular Antioxidant Protection of erythrocytes (CAP-e) assays and for potential anticholinesterase activity in vitro. Cognitive function and learning of Lineminus™ was evaluated using scopolamine induced cognition deficits in a Barnes maze, rodent model of cognition.
Results
The extract displayed in vitro antioxidant activity with a total ORAC value of 16,964 μmole TE/gram. Cellular antioxidant protection from free radical damage using the CAP-e assay, with an IC50 of 0.58 g/L for inhibition of cellular oxidative damage, was observed. The extract inhibited cholinesterase activity with an IC50 of 0.04 mg/ml with a maximum inhibition of 68%. In a rodent model of cognition using scopolamine induced cognition deficits in the Barnes maze, the extract attenuated scopolamine induced disruptions in learning at the higher dose of 100 mg/kg.
Conclusion
These data shows that P. minus possesses antioxidant and anticholinesterase activity and demonstrated enhanced cognition in vivo. The data suggest neuroprotective properties of the extract.
doi:10.1186/1472-6882-14-161
PMCID: PMC4036647  PMID: 24886679
Scopolamine; Antioxidant; Barnes maze; Polygonum minus; Cognition
40.  Yoga for managing knee osteoarthritis in older women: a pilot randomized controlled trial 
Background
Osteoarthritis (OA) is a common problem in older women that is associated with pain and disabilities. Although yoga is recommended as an exercise intervention to manage arthritis, there is limited evidence documenting its effectiveness, with little known about its long term benefits. This study’s aims were to assess the feasibility and potential efficacy of a Hatha yoga exercise program in managing OA-related symptoms in older women with knee OA.
Methods
Eligible participants (N = 36; mean age 72 years) were randomly assigned to 8-week yoga program involving group and home-based sessions or wait-list control. The yoga intervention program was developed by a group of yoga experts (N = 5). The primary outcome was the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) total score that measures knee OA pain, stiffness, and function at 8 weeks. The secondary outcomes, physical function of the lower extremities, body mass index (BMI), quality of sleep (QOS), and quality of life (QOL), were measured using weight, height, the short physical performance battery (SPPB), the Pittsburgh Sleep Quality Index (PSQI), the Cantril Self-Anchoring Ladder, and the SF12v2 Health Survey. Data were collected at baseline, 4 weeks and 8 weeks, and 20 weeks.
Results
The recruitment target was met, with study retention at 95%. Based on ANCOVAs, participants in the treatment group exhibited significantly greater improvement in WOMAC pain (adjusted means [SE]) (8.3 [.67], 5.8 [.67]; p = .01), stiffness (4.7 [.28], 3.4 [.28]; p = .002) and SPPB (repeated chair stands) (2.0 [.23], 2.8 [.23]; p = .03) at 8 weeks. Significant treatment and time effects were seen in WOMAC pain (7.0 [.46], 5.4 [.54]; p = .03), function (24.5 [1.8], 19.9 [1.6]; p = .01) and total scores (35.4 [2.3], 28.6 [2.1]; p = .01) from 4 to 20 weeks. Sleep disturbance was improved but the PSQI total score declined significantly at 20 weeks. Changes in BMI and QOL were not significant. No yoga related adverse events were observed.
Conclusions
A weekly yoga program with home practice is feasible, acceptable, and safe for older women with knee OA, and shows therapeutic benefits.
Trial registration
ClinicalTrials.gov: NCT01832155
doi:10.1186/1472-6882-14-160
PMCID: PMC4038088  PMID: 24886638
Yoga; Knee osteoarthritis; Symptom management; Older women
41.  Decrease of a specific biomarker of collagen degradation in osteoarthritis, Coll2-1, by treatment with highly bioavailable curcumin during an exploratory clinical trial 
Background
The management of osteoarthritis (OA) remains a challenge. There is a need not only for safe and efficient treatments but also for accurate and reliable biomarkers that would help diagnosis and monitoring both disease activity and treatment efficacy. Curcumin is basically a spice that is known for its anti-inflammatory properties. In vitro studies suggest that curcumin could be beneficial for cartilage in OA. The aim of this exploratory, non-controlled clinical trial was to evaluate the effects of bio-optimized curcumin in knee OA patients on the serum levels of specific biomarkers of OA and on the evaluation of pain.
Methods
Twenty two patients with knee OA were asked to take 2x3 caps/day of bio-optimized curcumin (Flexofytol®) for 3 months. They were monitored after 7, 14, 28 and 84 days of treatment. Pain over the last 24 hours and global assessment of disease activity by the patient were evaluated using a visual analog scale (100 mm). The serum levels of Coll-2-1, Coll-2-1NO2, Fib3-1, Fib3-2, CRP, CTX-II and MPO were determined before and after 14 and 84 days of treatment.
Results
The treatment with curcumin was globally well tolerated. It significantly reduced the serum level of Coll2-1 (p < 0.002) and tended to decrease CRP. No other significant difference was observed with the other biomarkers. In addition, curcumin significantly reduced the global assessment of disease activity by the patient.
Conclusion
This study highlighted the potential effect of curcumin in knee OA patient. This effect was reflected by the variation of a cartilage specific biomarker, Coll2-1 that was rapidly affected by the treatment. These results are encouraging for the qualification of Coll2-1 as a biomarker for the evaluation of curcumin in OA treatment.
Trial registration
NCT01909037 at clinicaltrials.gov
doi:10.1186/1472-6882-14-159
PMCID: PMC4032499  PMID: 24886572
Osteoarthritis; Cartilage; Collagen; Curcumin; Biomarker
42.  Positive patient experiences in an Australian integrative oncology centre 
Background
The purpose of this study was to explore the experiences of cancer patients’ utilising complementary and integrative therapies (CIT) within integrative oncology centres across Western Australia.
Methods
Across four locations 135 patients accessed CIT services whilst undergoing outpatient medical treatment for cancer. Of the 135 patients, 66 (61 ± 12 y; female n = 45; male n = 21) agreed to complete a personal accounts questionnaire consisting of open-ended questions designed to explore patients’ perceptions of CIT. All results were transcribed into nVivo (v9) and using thematic analysis, key themes were identified.
Results
Of the 66 participants, 100% indicated they would “recommend complementary therapies to other patients” and 92% stated “CIT would play a significant role in their future lifestyle”. A mean score of 8 ± 1 indicated an improvement in participants’ perception of wellbeing following a CIT session. Three central themes were identified: empowerment, support and relaxation. Fourteen sub-themes were identified, with all themes clustered into a framework of multifaceted views held by cancer patients in relation to wellbeing, role of significant others and control.
Conclusions
Exploration of patients’ experiences reveals uniformly positive results. One of the key merits of the environment created within the centres is patients are able to work through their cancer journey with an increased sense of empowerment, without placing them in opposition to conventional medical treatment. In order to effectively target integrative support services it is crucial to explore the experiences of patients in their own words and use those forms of expression to drive service delivery.
doi:10.1186/1472-6882-14-158
PMCID: PMC4032569  PMID: 24886476
Cancer; Patient experiences; Complementary therapy; Supportive services; Survivorship; Quality of life
43.  Wound healing and antibacterial properties of methanolic extract of Pupalia lappacea Juss in rats 
Background
Wound healing is a natural process that enables tissue repair after an injury. To shorten its duration and minimize associated complications, wounds are treated with medications. Currently there is a growing interest in the use of alternative wound dressing agents such as plant extracts. One plant used traditionally in wound treatment is Pupalia lappacea. In view of its use in wound care, we investigated the wound healing activities of 80% methanolic leave extract of Pupalia lappacea using excision, incision and dead space wound models. Also its effects on three common wound contaminants were investigated.
Methods
Excision wounds were created, contaminated with microbes and treated with ointments (10% and 20% w/w) prepared from Pupalia lappacea. Incision and dead space wounds were also created in rats which were subsequently dosed orally with the extract. The wound healing activities of Pupalia lappacea ointment on excision wound was assessed by rates of wound contraction and epithelialization as well as its antibacterial effects. The effects of Pupalia lappacea on incision and dead-space wounds were determined by the wound breaking strengths and weights of the granuloma tissues formed, respectively.
Results
Pupalia. lappacea ointments significantly (p < 0.05) accelerated wound healing with 20% ointment having the highest percentage wound contraction and rate of epithelialization. At 4, 7 and 14 days post treatment, mean total viable bacterial count of excision wounds of the extract treated groups were significantly (p < 0.05) lower compared against the control. Wound breaking strengths and weights of granuloma tissues formed in the extract treated groups were significantly (p < 0.05) higher than those of the control group. The minimum inhibitory concentration values obtained for the Pupalia lappacea extract against Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis were 9 mg/ml, 4 mg/ml and 3 mg/ml, respectively, while the corresponding minimum bactericidal concentrations were 10 mg/ml, 8 mg/ml and 7 mg/ml.
Conclusion
The results obtained showed that Pupalia. lappacea has good wound healing and antibacterial activities. These findings validate the use of this plant in traditional medicine for treatment of wounds.
doi:10.1186/1472-6882-14-157
PMCID: PMC4023168  PMID: 24886368
Pupalia lappacea; Wounds; Antibacterial; Ointments
44.  Polysaccharides from Liriopes Radix ameliorate streptozotocin-induced type I diabetic nephropathy via regulating NF-κB and p38 MAPK signaling pathways 
Background
The polysaccharides from Liriopes Radix (PSLR) has been indicated to ameliorate insulin signaling transduction and glucose metabolism. We aimed to investigate whether PSLR exerts an ameliorative effect on renal damage in diabetes induced by streptozotocin.
Methods
Diabetes was induced with STZ (60 mg/kg) by intraperitoneal injection in rats. Two weeks after STZ injection, rats in the treatment group were orally dosed with PSLR (200 and 300 mg/kg/day for 8 weeks. The normal rats were chosen as nondiabetic control group. Changes in renal function-related parameters in plasma and urine were analyzed at the end of the study. Kidneys were isolated for pathology histology, immunohistochemistry, and Western blot analyses.
Results
Diabetic rats exhibited renal dysfunction, as evidenced by reduced creatinine clearance, blood urea nitrogen and proteinuria, along with marked elevation in the ratio of kidney weight to body weight. All of these abnormalities were significantly reversed by PSLR. The histological examinations revealed amelioration of diabetes-induced glomerular pathological changes following treatment with PSLR. The less protein expressions of renal nephrin and podocin in diabetic rats were increased following treatment with PSLR. PSLR reduced the accumulation of ED-1-expressing macrophages in renal tissue of diabetic rats. PSLR almost completely abolished T cells infiltration and attenuated the expression of proinflammatory cytokines. PSLR treatments not only reduced the degradation of inhibitory kappa B kinase, but also downregulated the protein expression of nuclear factor kappa B (NF-κB) and p38 mitogen-activated protein kinase (MAPK) in diabetic kidney.
Conclusions
The results suggest that the renal protective effects of PSLR occur through improved glycemic control and renal structural changes, which are involved in the inhibition of NF-κB and p-38 MAPK mediated inflammation.
doi:10.1186/1472-6882-14-156
PMCID: PMC4041058  PMID: 24886259
45.  Target guided isolation, in-vitro antidiabetic, antioxidant activity and molecular docking studies of some flavonoids from Albizzia Lebbeck Benth. bark 
Background
Albizzia Lebbeck Benth. is traditionally important plant and is reported to possess a variety of pharmacological actions. The present research exertion was undertaken to isolate and characterized the flavonoids from the extract of stem bark of Albizzia Lebbeck Benth. and to evaluate the efficacy of the isolated flavonoids on in-vitro models of type-II diabetes. Furthermore, the results of in-vitro experimentation inveterate by the molecular docking studies of the isolated flavonoids on α-glucosidase and α-amylase enzymes.
Methods
Isolation of the flavonoids from the methanolic extract of stem bark of A. Lebbeck Benth was executed by the Silica gel (Si) column chromatography to yield different fractions. These fractions were then subjected to purification to obtain three important flavonoids. The isolated flavonoids were then structurally elucidated with the assist of 1H-NMR, 13C-NMR, and Mass spectroscopy. In-vitro experimentation was performed with evaluation of α-glucosidase, α-amylase and DPPH inhibition capacity. Molecular docking study was performed with GLIDE docking software.
Results
Three flavonoids, (1) 5-deoxyflavone (geraldone), (2) luteolin and (3) Isookanin were isolated from the EtOAc fraction of the methanolic extract of Albizzia lebbeck Benth bark. (ALD). All the compounds revealed to inhibit the α-glucosidase and α-amylase enzymes in in-vitro investigation correlating to reduce the plasma glucose level. Molecular docking study radically corroborates the binding affinity and inhibition of α-glucosidase and α-amylase enzymes.
Conclusion
The present research exertion demonstrates the anti-diabetic and antioxidant activity of the important isolated flavonoids with inhibition of α-glucosidase, α-amylase and DPPH which is further supported by molecular docking analysis.
doi:10.1186/1472-6882-14-155
PMCID: PMC4063241  PMID: 24886138
46.  Fei-Liu-Ping ointment inhibits lung cancer growth and invasion by suppressing tumor inflammatory microenvironment 
Background
Lung cancer is one of the leading causes of cancer-related mortality worldwide. Conventional chemotherapy and radiotherapy are the primary therapeutic methods for lung cancer with the use of combination therapies gaining popularity. The frequency and duration of treatment, as well as, managing lung cancer by targeting multiple aspects of cancer biology is often limited by toxicity to the patient. There are many naturally occurring anticancer agents that have a high degree of efficacy and low toxicity, offering a viable and safe approach for the treatment of lung cancer. The herbs traditionally used in Chinese medicine for anticancer treatment offer great potential to enhance the efficacy of conventional therapy. In this study, we evaluated the synergistic effects of Fei-Liu-Ping (FLP) ointment in treating lung cancer; a known anticancer Chinese herbal based formula.
Methods
In this study, A549 human lung carcinoma cell line and Lewis lung carcinoma xenograft mouse model were used. In addition, we utilized an in vitro co-culture system to simulate the tumor microenvironment in order to evaluate the molecular mechanisms of FLP treatment.
Results
FLP treatment significantly inhibited tumor growth in the Lewis lung xenograft by 40 percent, compared to that of cyclophosphamide (CTX) of 62.02 percent. Moreover, combining FLP and CTX inhibited tumor growth by 83.23 percent. Upon evaluation, we found that FLP treatment reduced the concentration of serum pro-inflammatory cytokines IL-6, TNF-α, and IL-1β. In addition, we also found an improvement in E-cadherin expression and inhibition of N-cadherin and MMP9. We found similar findings in vitro when we co-cultured A549 cells with macrophages. FLP treatment inhibited A549 cell growth, invasion and metastasis, in part, through the regulation of NF-κB and altering the expression of E-cadherin, N-cadherin, MMP2 and MMP9.
Conclusions
FLP exerts anti-inflammatory properties in the tumor microenvironment, which may contribute to its anticancer effects. FLP treatment may be a promising therapy for inflammation associated lung cancer treatment alone, or in combination with conventional therapies and may prevent lung cancer metastasis.
doi:10.1186/1472-6882-14-153
PMCID: PMC4036108  PMID: 24885825
Lung cancer; Fei-Liu-Ping ointment; Nuclear factor kappa light chain enhancer of activated B cells; Inflammation; Invasion; Epithelial mesenchymal transition; Cyclophosphamide
47.  Phytochemical constituents, nutritional values, phenolics, flavonols, flavonoids, antioxidant and cytotoxicity studies on Phaleria macrocarpa (Scheff.) Boerl fruits 
Background
The edible fruits of Phaleria macrocarpa (Scheff.) Boerl are widely used in traditional medicine in Indonesia. It is used to treat a variety of medical conditions such as - cancer, diabetes mellitus, allergies, liver and heart diseases, kidney failure, blood diseases, high blood pressure, stroke, various skin diseases, itching, aches, and flu. Therefore, it is of great interest to determine the biochemical and cytotoxic properties of the fruit extracts.
Methods
The methanol, hexane, chloroform, ethyl acetate, and water extracts of P. macrocarpa fruits were examined for phytochemicals, physicochemicals, flavonols, flavonoids and phenol content. Its nutritional value (A.O.A.C method), antioxidant properties (DPPH assay) and cytotoxicity (MTT cell proliferation assay) were also determined.
Results
A preliminary phyotochemical screening of the different crude extracts from the fruits of P. macrocarpa showed the presence secondary metabolites such as of flavonoids, phenols, saponin glycosides and tannins. The ethyl acetate and methanol extracts displayed high antioxidant acitivity (IC50 value of 8.15±0.02 ug/mL) in the DPPH assay comparable to that of the standard gallic acid (IC50 value of 10.8±0.02 ug/mL). Evaluation of cytotoxic activity showed that the crude methanol extract possessed excellent anti-proliferative activity against SKOV-3 (IC50 7.75±2.56 μg/mL) after 72 hours of treatment whilst the hexane and ethyl acetate extracts displayed good cytotoxic effect against both SKOV-3 and MDA-MB231 cell lines. The chloroform extract however, showed selective inhibitory activity in the breast cancer cell line MDA-MB231 (IC50 7.80±1.57 μg/mL) after 48 hours of treatment. There was no cytotoxic effect observed in the Ca Ski cell line and the two normal cell lines (MRC-5 and WRL-68).
Conclusion
The methanol extract and the ethyl acetate fraction of P. macrocarpa fruits exhibited good nutritional values, good antioxidant and cytotoxic activities, and merits further investigation to identify the specific compound(s) responsible for these activities.
doi:10.1186/1472-6882-14-152
PMCID: PMC4039986  PMID: 24885709
Cytoxicity; Phytochemicals; Nutritional values; Flavonoids; Flavonol; Phenol; Antioxidants; Phaleria macrocarpa (Scheff.) Boerl fruits or P. macrocarpa
48.  Polygonum viviparum L. induces vasorelaxation in the rat thoracic aorta via activation of nitric oxide synthase in endothelial cells 
Background
In the past several decades, Polygonum viviparum L. (PV) was reported to have antibacterial, antiulcer, antioxidant, antitumor, anti-inflammatory, and antiarthritic properties. The anti-inflammatory pathway was recently elucidated through cytosolic nuclear factor E2-related factor 2 (Nrf2) activation and heme oxygenase (HO)-1 protein expression. PV is a perennial herb and widely distributed in high-elevation mountain regions, such as the Tibetan Plateau. In Tibetan traditional medicine, PV is usually used to boost the blood circulation to dissipate blood stasis. Therefore, this study focused on how PV improves the vascular circulation and acts on vascular tissues.
Methods
In this study, we isolated aortas from Sprague-Dawley rats (male, weight about 250 ~ 350 g), and detected the effects of PV on phenylephrine (PE)-induced contraction and cyclic guanosine 3′,5′-monophosphate (cGMP) formation using aortic rings. In addition, human umbilical vein endothelial cells (HUVECs) were used to exam nitric oxygen (NO) synthase (NOS) activity by directly measuring NO production in the culture medium. Endothelial (e) NOS phosphorylation, and cytosolic Nrf2 and HO-1 expressions were measured using a Western blot analysis.
Results
PV dose-dependently relaxed PE-induced contractions in endothelial-intact but not -denuded aorta. The concentration to produce 50% relaxation was 22.04 ± 1.77 μg/ml. PV-induced vasorelaxation was markedly blocked by pretreatment with NG-nitro-L-arginine methyl ester (L-NAME), an NOS inhibitor, methylene blue (MB), a guanylyl cyclase inhibitor, and hemoglobin, an NO scavenger. PV increased cGMP formation; however, this effect was also suppressed by co-pretreatment with l-NAME, MB, hemoglobin, and Ca2+-free medium. In HUVECs, PV increased NO formation, which was greatly attenuated by NOS inhibitors (L-NAME and L-NMMA) and by removing extracellular Ca2+ and chelating intracellular Ca2+ with BAPTA-AM. In addition, PV promoted eNOS phosphorylation, Nrf2 degradation, and HO-1 protein expression according to a Western blot analysis.
Conclusions
The results suggest that PV possesses vasorelaxing action in an endothelium-dependent manner and works through activating Ca2+/calmodulin- dependent NO synthesis; when NO is released and then transferred to smooth muscle cells, NO activates guanylyl cyclase and increases cGMP formation, ultimately resulting in vasorelaxation. Thus, PV can be considered for application as a potential therapeutic approach for vascular-associated disorders.
doi:10.1186/1472-6882-14-150
PMCID: PMC4030279  PMID: 24885446
Polygonum viviparum L; Aorta; Vasorelaxation; cGMP; eNOS; HUVECs
49.  Quality of reporting on randomized controlled trials of acupuncture for stroke rehabilitation 
Background
Results from clinical studies on acupuncture for stroke rehabilitation are contradictory. The reason for the inconsistent findings especially lie in the transparency and accuracy of randomized controlled trials (RCTs) reports. This study aims to analyze the quality of reporting and its correlates in RCTs on acupuncture for stroke rehabilitation.
Methods
Quality of reporting for included papers was assessed against a subset of criteria adapted from the CONSORT 2010 statement and STRICTA. An overall quality score (OQS) and a combined key methodological index score (MIS) was calculated for each trial. Then, factors associated with OQS and MIS were identified.
Results
A total of 15 RCTs were included in full text. The median OQS based on the CONSORT statement and STRICTA was 8 and 12, respectively. The significant predictors for CONSORT OQS was funding source, for STRICTA was year of publication. With regard to the MIS, no variable was associated with improved methodological quality.
Conclusions
Our study found that the overall quality of reporting on RCTs of acupuncture for stroke rehabilitation was general or good. But some items’ reporting was found where information was insufficient or inadequate in most studies which needed substantial improvement.
doi:10.1186/1472-6882-14-151
PMCID: PMC4030573  PMID: 24885561
Acupuncture; Quality of reporting; Randomized controlled trials (RCTs); Stroke rehabilitation; The CONSORT statement; STRICTA
50.  A study of antioxidant activity, enzymatic inhibition and in vitro toxicity of selected traditional sudanese plants with anti-diabetic potential 
Background
Diabetes mellitus is a chronic metabolic disease with life-threatening complications. Despite the enormous progress in conventional medicine and pharmaceutical industry, herbal-based medicines are still a common practice for the treatment of diabetes. This study evaluated ethanolic and aqueous extracts of selected Sudanese plants that are traditionally used to treat diabetes.
Methods
Extraction was carried out according to method described by Sukhdev et. al. and the extracts were tested for their glycogen phosphorylase inhibition, Brine shrimp lethality and antioxidant activity using (DPPH) radical scavenging activity and iron chelating activity. Extracts prepared from the leaves of Ambrosia maritima, fruits of Foeniculum vulgare and Ammi visnaga, exudates of Acacia Senegal, and seeds of Sesamum indicum and Nigella sativa.
Results
Nigella sativa ethanolic extract showed no toxicity on Brine shrimp Lethality Test, while its aqueous extract was toxic. All other extracts were highly toxic and ethanolic extracts of Foeniculum vulgare exhibited the highest toxicity. All plant extracts with exception of Acacia senegal revealed significant antioxidant activity in DPPH free radical scavenging assay.
Conclusions
These results highly agree with the ethnobotanical uses of these plants as antidiabetic. This study endorses further studies on plants investigated, to determine their potential for type 2 diabetes management. Moreover isolation and identification of active compounds are highly recommended.
doi:10.1186/1472-6882-14-149
PMCID: PMC4017226  PMID: 24885334
Diabetes mellitus; Medicinal plants; Antioxidant activity; Glycogen phosphorylase; Brine shrimp

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