Auricular Acupressure on Breast Pain Among Breastfeeding Mothers Receiving Gentle Hand Techniques: A Randomized, Single-Blind, Sham-Controlled Trial.

BACKGROUND: Auricular acupressure influences pain reduction in patients with diseases. However, study results on its influence on breast pain, common among breastfeeding mothers, are insufficient. RESEARCH AIM: This study aimed to explore the effectiveness of auricular acupressure on breast pain among breastfeeding mothers who also received a standard gentle hand technique. METHODS: This was a placebo-controlled study wherein auricular acupressure was provided for 4 weeks to 52 breastfeeding mothers consulting a local breastfeeding clinic. In the intervention group, auricular acupressure was applied to specific acupoints, including Shenmen, central rim, breast, and endocrine, which are related to breast pain and postpartum lactation. For the placebo control group, auricular acupressure was applied to acupoints not related to breast pain. The degree of breast pain was assessed using a numeric rating scale and a pressure algometer. RESULTS: After 4 weeks of auricular acupressure, numeric rating scale pain scores did not show a significant change. However, the pressure pain threshold for the upper left (Z = -2.202, p = .028) and upper right (t = 2.613, p = .012) areas of the right breast increased significantly in the intervention group. CONCLUSION: This study employed subjective and objective measurements to evaluate the efficacy of auricular acupressure in alleviating breast pain. The intervention shows potential as a nursing measure. Further research is required to determine the optimal intervention duration and frequency, particularly for breastfeeding parents with severe pain, and to evaluate long-term outcomes.

The Effects of Auricular Acupressure on Menopausal Symptoms, Stress, and Sleep in Postmenopausal Middle-Aged Women: A Randomized Single-Blind Sham-Controlled Trial.

INTRODUCTION: Middle-aged women in the postmenopausal period experience menopause symptoms, stress, and poor sleep quality due to hormonal changes. Nonetheless, most of them recognize such symptoms as an aging process without receiving proper treatment, and there are few nonpharmacologic interventions available. METHODS: This was a randomized single-masked, sham-controlled trial. For the intervention group, a vaccaria seed was applied to the auricular points of Shenmen, anterior lobe, adrenal glands, central rim, and endocrine that are related to the menopause symptoms, stress, and sleep while applying a seed to the auricular points not related to the forementioned symptoms to the control group. The Menopause Rating Scale (MRS), Perceived Stress Scale (PSS), heart rate variability, and electroencephalogram (EEG) were measured before the intervention, 4 weeks after the intervention, and 8 weeks after the intervention. Actigraphy was measured with a Fitbit, and the Pittsburgh Sleep Quality Index (PSQI) was measured before and after the intervention. The study was registered with the World Health Organization International Clinical Trials Registry Platform (KCT0007364). RESULTS: The MRS showed significant differences over time in the intervention group (F, 22.057; P < .001). There was a significant difference over time in the PSS (F, 22.576; P < .001), stress index measured by heart rate variability (F, 14.027; P = .001), and antistress quotient of the right brain measured by EEG (F, 4.865; P = .033). Sleep quality, measured by the PSQI (t = -4.050, P < .001), and sleep efficiency measured by actigraphy (t = 5.996, P < .001) were found to be significantly different over time in the intervention group. DISCUSSION: This study demonstrated that auricular acupressure is effective in improving menopause symptoms, stress, and sleep in postmenopausal middle-aged women. Therefore, auricular acupressure may be a useful nonpharmacologic intervention for alleviating these symptoms in this population.

Xanol Promotes Apoptosis and Autophagy and Inhibits Necroptosis and Metastasis via the Inhibition of AKT Signaling in Human Oral Squamous Cell Carcinoma.

Angelica keiskei Koidzumi (A. keiskei) is used as a traditional medicine, anti-aging agent, and health food, as well as to restore vitality. Xanthoangelol (xanol), a prenylated chalcone, is the predominant constituent of A. keiskei. Oral squamous cell carcinoma (OSCC), the most common malignancy, has a high proliferation rate and frequent metastasis. However, it is unknown whether xanol has anti-OSCC effects on apoptosis, autophagy, and necroptosis. In the present study, we purified xanol from A. keiskei and demonstrated that it suppressed cell proliferation and induced cytotoxicity in human OSCC. Xanol triggered apoptotic cell death by regulating apoptotic machinery molecules but inhibited necroptotic cell death by dephosphorylating the necroptotic machinery molecules RIP1, RIP3, and MLKL in human OSCC. We also found that xanol inhibited the PI3K/AKT/mTOR/p70S6K pathway and induced autophagosome formation by enhancing beclin-1 and LC3 expression levels and reducing p62 expression levels. Furthermore, we showed that xanol prevented the metastatic phenotypes of human OSCC by inhibiting migration and invasion via the reduction of MMP13 and VEGF. Finally, we demonstrated that xanol exerted anticancer effects on tumorigenicity associated with its transformed properties. Taken together, these findings demonstrate the anticancer effects and biological mechanism of action of xanol as an effective phytomedicine for human OSCC.

The effects of auricular acupressure on blood pressure, stress, and sleep in elders with essential hypertension: a randomized single-blind sham-controlled trial.

AIMS: Reduction of blood pressure and improvement of stress and sleep disorders in patients with essential hypertension were important factors. This study aimed to examine the effects of auricular acupressure on blood pressure, stress, and sleep quality among elderly with essential hypertension in Korea. METHODS AND RESULTS: This study was a single-blind, randomized, placebo-controlled study. The experimental group (n = 23) received 8 weeks of auricular acupressure intervention on specific acupoints (superior triangular fossa, Shenmen, kidney, heart, and occiput) related to blood pressure, pulse rate, stress, and sleep, whereas the control group (n = 23) received auricular acupressure on non-specific acupoints. The primary outcomes were the systolic and diastolic blood pressure measured weekly. The secondary outcomes were the pulse rate, heart rate variability, actigraphy measured through Fitbit, and the Pittsburgh sleep quality index (PSQI) were measured before and after the experiment. There were statistically significant differences between two groups in systolic blood pressure (F = 5.67, P = 0.022), diastolic blood pressure (17.53, P < 0.001), and pulse rate (F = 6.78, P = 0.013) over time. Stress index (χ2 = 2.12, P = 0.040) and sleep efficiency (χ2 = 3.57, P = 0.001) were also significantly different before and after the experiment. However, there was no significant difference in the PSQI. CONCLUSION: The findings showed that auricular acupressure leads to improvements in blood pressure, stress, and sleep in elderly people with essential hypertension. Therefore, auricular acupressure can be used as an alternative nursing intervention for hypertension prevention, stress, and sleep management. REGISTRATION: WHO ICTRP KCT0007364.

Suffruticosol A elevates osteoblast differentiation targeting BMP2-Smad/1/5/8-RUNX2 in pre-osteoblasts.

The Paeonia suffruticosa ANDR. (P. suffruticosa) is commonly used in traditional medicine for various purposes. Suffruticosol A (Suf-A), isolated from P. suffruticosa, is a beneficial compound with antibiofilm, antivirulence, and anti-inflammatory properties. The aim of the present study was to investigate the biological effects of Suf-A on osteogenic processes in pre-osteoblasts. It was determined here in that Suf-A (>98.02%), isolated from P. suffruticosa, showed no cytotoxicity at 0.1-30 µM; however, it induced cytotoxicity at 50-100 µM in pre-osteoblasts. Suf-A increased osteogenic alkaline phosphatase activity and expression levels of noncollagenous proteins. Adhesion and trans-migration on the extracellular matrix were potentiated by Suf-A, but not by wound-healing migration. Suf-A did not affect autophagy or necroptosis during osteoblast differentiation. We found that Suf-A increased runt-related transcription factor 2 (RUNX2) levels and mineralized matrix formation. RUNX2 expression was mediated by Suf-A-induced BMP2-Smad1/5/8 and mitogen-activated protein kinase signaling, as demonstrated by Noggin, a BMP2 inhibitor. These results suggest that Suf-A is a potential natural osteogenic compound.

Trifloroside Induces Bioactive Effects on Differentiation, Adhesion, Migration, and Mineralization in Pre-Osteoblast MC3T3E-1 Cells.

Gentianae Scabrae Radix is used in traditional medicine and is known to possess bioactive compounds, including secoiridoid glycosides, flavonoids, lignans, and triterpenes. Trifloroside (TriFs) is a secoiridoid glycoside known for its antioxidant activity; however, its other effects have not been studied. In the present study, we investigated the biological effects of TriFs isolated from the roots of Gentianae Scabrae Radix using pre-osteoblast MC3T3E-1 cells. No cellular toxicity was observed with 1 µM TriFs, whereas 5-100 µM TriFs showed a gradual increase in cell viability. Alkaline phosphatase staining and microscopic observations revealed that 1-10 µM TriFs stimulated osteogenic activity during early osteoblast differentiation. Trifloroside also increased mineral apposition during osteoblast maturation. Biochemical analyses revealed that TriFs promoted nuclear RUNX2 expression and localization by stimulating the major osteogenic BMP2-Smad1/5/8-RUNX2 pathway. Trifloroside also increased p-GSK3ß, ß-catenin, p-JNK, and p-p38, but not Wnt3a, p-AKT, and p-ERK. Moreover, TriFs increased the MMP13 levels and promoted cell migration and adhesion. In contrast, TriFs-induced osteoblast differentiation and maturation had negligible effects on autophagy and necrosis. Our findings suggest that TriFs induces osteogenic effects through differentiation, adhesion, migration, and mineral apposition. Therefore, TriFs is suggested as a potential drug target in osteoblast-mediated bone diseases.

Suffruticosol B Is an Osteogenic Inducer through Osteoblast Differentiation, Autophagy, Adhesion, and Migration.

Suffruticosol B (Suf-B) is a stilbene found in Paeonia suffruticosa ANDR., which has been traditionally used in medicine. Stilbenes and their derivatives possess various pharmacological effects, such as anticancer, anti-inflammatory, and anti-osteoporotic activities. This study aimed to explore the bone-forming activities and mechanisms of Suf-B in pre-osteoblasts. Herein, >99.9% pure Suf-B was isolated from P. suffruticosa methanolic extracts. High concentrations of Suf-B were cytotoxic, whereas low concentrations did not affect cytotoxicity in pre-osteoblasts. Under zero levels of cytotoxicity, Suf-B exhibited bone-forming abilities by enhancing alkaline phosphatase enzyme activities, bone matrix calcification, and expression levels with non-collagenous proteins. Suf-B induces intracellular signal transduction, leading to nuclear RUNX2 expression. Suf-B-stimulated differentiation showed increases in autophagy proteins and autophagosomes, as well as enhancement of osteoblast adhesion and transmigration on the ECM. These results indicate that Suf-B has osteogenic qualities related to differentiation, autophagy, adhesion, and migration. This also suggests that Suf-B could have a therapeutic effect as a phytomedicine in skeletal disorders.

Effects of Scoparone on differentiation, adhesion, migration, autophagy and mineralization through the osteogenic signalling pathways.

Scoparone (SCOP), an active and efficient coumarin compound derived from Artemisia capillaris Thunb, has been used as a traditional Chinese herbal medicine. Herein, we investigated the effects of SCOP on the osteogenic processes using MC3T3-E1 pre-osteoblasts in in vitro cell systems. SCOP (C11 H10 O4 , > 99.17%) was purified and identified from A. capillaries. SCOP (0.1 to 100 µM concentrations) did not have cytotoxic effects in pre-osteoblasts; however, it promoted alkaline phosphatase (ALP) staining and activity, and mineralized nodule formation under early and late osteogenic induction. SCOP elevated osteogenic signals through the bone morphogenetic protein 2 (BMP2)-Smad1/5/8 pathway, leading to the increased expression of runt-related transcription factor 2 (RUNX2) with its target protein, matrix metallopeptidase 13 (MMP13). SCOP also induced the non-canonical BMP2-MAPKs pathway, but not the Wnt3a-ß-catenin pathway. Moreover, SCOP promoted autophagy, migration and adhesion under the osteogenic induction. Overall, the findings of this study demonstrated that SCOP has osteogenic effects associated with cell differentiation, adhesion, migration, autophagy and mineralization.

[Effect of Auriculotherapy on Musculoskeletal Pain: A Systematic Review and Meta-Analysis].

PURPOSE: The aim of this study was to evaluate the effect of auriculotherapy on musculoskeletal pain in adults. METHODS: A total of 885 studies were retrieved from nine databases (PubMed, Scopus, CINAHL, Web of Science, Ovid Medline, Cochrane Library, RISS, KMbase, and KISS). Sixteen studies were selected for meta-analysis, which satisfied the inclusion criteria and the evaluation of risk of bias. Demographic data, auriculotherapy types, intervention characteristics, auricular points, and outcomes related to pain (subjective pain scale, and amount of analgesic) were extracted from all included studies. The effect size of auriculotherapy was analyzed through comprehensive meta analysis 3.0, and the presence of publication bias was analyzed through a funnel plot and Egger's regression. RESULTS: The results of the meta-analysis (n = 16) revealed that the auriculotherapy was significantly superior to the control group on present pain in adults (Hedges' g = -0.35, 95% Confidence Interval [CI] = -0.55~-0.15). According to the results of subgroup analysis, the effect size of auricular acupuncture therapy (Hedges' g = 0.45, 95% CI = -0.75~-0.15) was higher than the auricular acupuncture (Hedges' g = 0.27, 95% CI = -0.53~0.00): the longer the intervention period, the greater the effect size. CONCLUSION: In this study, auriculotherapy demonstrates a significant reduction in musculoskeletal pain in adults. Therefore, it is necessary to refine the curriculum to include auriculotherapy as a nursing intervention to relieve musculoskeletal pain in adults and encourage its use in clinical settings.

Calycosin-7-O-ß-Glucoside Isolated from Astragalus membranaceus Promotes Osteogenesis and Mineralization in Human Mesenchymal Stem Cells.

Stem cells have received attention in various diseases, such as inflammatory, cancer, and bone diseases. Mesenchymal stem cells (MSCs) are multipotent stem cells that are critical for forming and repairing bone tissues. Herein, we isolated calycosin-7-O-ß-glucoside (Caly) from the roots of Astragalus membranaceus, which is one of the most famous medicinal herbs, and investigated the osteogenic activities of Caly in MSCs. Caly did not affect cytotoxicity against MSCs, whereas Caly enhanced cell migration during the osteogenesis of MSCs. Caly increased the expression and enzymatic activities of ALP and the formation of mineralized nodules during the osteogenesis of MSCs. The osteogenesis and bone-forming activities of Caly are mediated by bone morphogenetic protein 2 (BMP2), phospho-Smad1/5/8, Wnt3a, phospho-GSK3ß, and phospho-AKT, inducing the expression of runt-related transcription factor 2 (RUNX2). In addition, Caly-mediated osteogenesis and RUNX2 expression were attenuated by noggin and wortmannin. Moreover, the effects were validated in pre-osteoblasts committed to the osteoblast lineages from MSCs. Overall, our results provide novel evidence that Caly stimulates osteoblast lineage commitment of MSCs by triggering RUNX2 expression, suggesting Caly as a potential anabolic drug to prevent bone diseases.

The effects of auricular acupressure on the sleep of the elderly using polysomnography, actigraphy and blood test: Randomized, single-blind, sham control.

AIM: This study was conducted to examine the effects of auricular acupressure on sleep in elderly people with sleep disorders. METHODS: This was a randomized, double-blind, sham-controlled study. The participants aged over 65 years old were randomly assigned to the experimental group (n = 21) and the sham control group (n = 21). The participants in the experimental group and the sham control group received auricular acupressure on sleep-disorder-related points or to sleep-disorder-unrelated points, respectively. The intervention was implemented for a total of eight weeks. To validate the effects of the treatment, polysomnography with the Alice portable sleep diagnostic system; actigraphy with Fitbit Alta; and melatonin, serotonin, and cortisol blood tests were conducted. RESULTS: Non-Rapid Eye Movement sleep stage 3 duration change (Z = -2.187, p = .029) and Non-Rapid Eye Movement sleep stage 3 ratio change (Z = -2.423, p = .014), measured by polysomnography, of the experimental group showed a significant increase over time compared to the sham control group. CONCLUSIONS: Auricular acupressure applied for eight weeks was found to be effective in increasing Non-Rapid Eye Movement sleep stage 3 duration and Non-Rapid Eye Movement sleep stage 3 ratio among sleep stages of the elderly. Consequently, it showed that auricular acupressure can be used as a proven nursing intervention method for sleep disorder in elders to increase deep sleep duration and ratio.

The effects of auricular acupressure on stress, anxiety, and depression of outpatient nurses in South Korea.

BACKGROUND AND PURPOSE: Outpatient nursing is an occupation with high emotional labor and job stress. The purpose of this study was to examine the effects of auricular acupressure on stress, anxiety, and depression of outpatient nurses. MATERIALS AND METHODS: The study design was a randomized controlled trial. Participants were 54 outpatient nurses, divided into an experimental (n = 28) and a control group (n = 26). Auricular acupressure using vaccaria seeds was administered to the experimental group, while placebo auricular acupressure using vaccaria seeds was administered to the placebo group. Sessions continued for 5 weeks. Outcome measurements included blood tests such as cortisol and serotonin. RESULTS: There was a statistically significant difference in depression scores (t = 3.111, p = 0.002) and serotonin levels (t = 6.304, p < 0.001) between the experimental and placebo groups. Auricular acupressure decreased depression scores, and the depression-related physiological index, serotonin, also changed significantly. CONCLUSION: Auricular acupressure using vaccaria seeds was effective in decreasing depression in outpatient nurses.

Renal Protective Effect of Beluga Lentil Pretreatment for Ischemia-Reperfusion Injury.

MATERIALS AND METHODS: Mice were divided into four groups: normal, untreated, low- (2 mg), and high-dose (8 mg) beluga lentil treatment groups. Beluga lentil was orally administered for 2 weeks, followed by bilateral renal ischemia for 20 min and reperfusion for 30 min. Blood samples and kidney tissues were collected and analyzed to investigate renal function, histopathology, epithelial and endothelial cell damage, apoptosis, oxidative stress, and inflammatory responses. RESULTS: The pretreated groups maintained renal function, with significantly lower blood urea nitrogen (BUN) and creatinine levels, compared with the other groups. The histopathological analysis showed reduced proximal tubule injury and decreased injury-related molecule (kidney injury molecule 1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL)) secretion in the pretreated groups compared with the other groups. Terminal deoxynucleotidyl transferase dUTP nick-end labeling- (TUNEL-) positive cells and the secretion of apoptosis-related molecules (Fas and caspase 3) were significantly reduced in the pretreated groups compared with the other groups. The pretreated groups showed positive microvessel-associated gene (cluster of differentiation (CD31)) expression and negative adhesion molecule (intracellular adhesion molecule 1 (ICAM-1)) expression. An antioxidant effect was observed in the pretreatment groups, with reduced malonaldehyde (MDA) expression and increased antioxidant enzyme (superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and glutathione peroxidase (GPx)) secretion. In the pretreated groups, F4/80+ macrophages and CD4+ T cell infiltration were inhibited and proinflammatory cytokine (interleukin- (IL-) 1ß, IL-6, and tumor necrosis factor- (TNF-) α) levels decreased; however, the levels of anti-inflammatory cytokines (transforming growth factor- (TGF-) ß, IL-10, and IL-22) increased. CONCLUSIONS: Beluga lentil pretreatment demonstrated protective effects against I/R-induced renal damage, via antiapoptotic, anti-inflammatory, and antioxidant activities.

TMARg, a Novel Anthraquinone Isolated from Rubia cordifolia Nakai, Increases Osteogenesis and Mineralization through BMP2 and ß-Catenin Signaling.

BACKGROUND: Plant extracts have long been regarded as useful medicines in the treatment of human diseases. Rubia cordifolia Nakai has been used as a traditional medicine, as it has pharmacological properties such as antioxidant and anti-inflammatory activity. However, the biological functions of TMARg, isolated from the roots of R. cordifolia, in osteoblast differentiation remain unknown. This study was performed to investigate the pharmacological effects and intracellular signaling of TMARg in the osteoblast differentiation of pre-osteoblast MC3T3-E1 cells and mesenchymal precursor C2C12 cells. METHODS: Cell viability was evaluated using an MTT assay. Early and late osteoblast differentiation was examined by analyzing the activity of alkaline phosphatase (ALP), and by staining it with Alizarin red S (ARS). Cell migration was determined by using migration assays. Western blot analysis and immunocytochemical analysis were used to examine the intracellular signaling pathways and differentiation proteins. RESULTS: In the present study, TMARg showed no cytotoxicity and increased the osteoblast differentiation in pre-osteoblasts, as assessed from the alkaline phosphate (ALP) staining and activity and ARS staining. TMARg also induced BMP2 expression and increased the p-smad1/5/8-RUNX2 and ß-catenin pathways in both MC3T3-E1 and C2C12 cells. Furthermore, TMARg activated mitogen-activated protein kinases (MAPKs) and increased the cell migration rate. In addition, the TMARg-mediated osteoblast differentiation was suppressed by BMP and Wnt inhibitors with the downregulation of BMP2 expression. CONCLUSION: These findings demonstrate that TMARg exerts pharmacological and biological effects on osteoblast differentiation through the activation of BMP2 and ß-catenin signaling pathways, and suggest that TMARg might be a potential phytomedicine for the treatment of bone diseases.

Effect of Differential Thermal Drying Conditions on the Immunomodulatory Function of Ginger.

Thermal drying is a common process used in the food industry for the modification of agricultural products. However, while various studies have investigated the alteration in physiochemical properties and chemical composition after drying, research focusing on the relationship between different dehydration conditions and bioactivity is scarce. In the current study, we prepared dried ginger under nine different conditions by varying the processing time and temperature and compared their immunomodulatory effects. Interestingly, depending on the drying condition, there were significant differences in the immunestimulating activity of the dried ginger samples. Gingers processed at 50°C 1h displayed the strongest activation of macrophages measured by TNF-α and IL-6 levels, whereas, freezedried or 70°C- and 90°C-dried ginger showed little effect. Similar results were recapitulated in primary bone marrow-derived macrophages, further confirming that different dehydration conditions can cause significant differences in the immune-stimulating activity of ginger. Induction of ERK, p38, and JNK signaling was found to be the major underlying molecular mechanism responsible for the immunomodulatory effect of ginger. These results highlight the potential to improve the bioactivity of functional foods by selectively controlling processing conditions.

Regulation of Th17 Cytokine-Induced Osteoclastogenesis via SKI306X in Rheumatoid Arthritis.

This study aimed to investigate the regulatory effect of SKI305X, a mixed extract of three herbs, in T helper (Th)17 cytokine-induced inflammation and joint destruction in rheumatoid arthritis (RA). Synovial fibroblasts were isolated from RA patients and cultured with Th17 cytokines including interleukin (IL)-17, IL-21, and IL-22 and SKI306X, and tumor necrosis factor (TNF)-, IL-1, and receptor activator of nuclear factor kappa-Β ligand (RANKL) expression and production were investigated using real-time PCR and ELISA of culture media. After peripheral blood (PB) cluster of differentiation (CD)14+ monocytes were cultured in media supplemented with Th17 cytokines and SKI306X, tartrate-resistant acid phosphatase positive (TRAP+) multinucleated giant cells (mature osteoclasts) were enumerated and gene expression associated with osteoclast maturation was assessed via real-time PCR analysis. After PB monocytes were co-cultured with IL-17-stimulated RA synovial fibroblasts in the presence of SKI306, osteoclast differentiation was assessed. When RA synovial fibroblasts were cultured with IL-17, IL-21, and IL-22, TNF-, IL-1, and RANKL expression and production were increased; however, SKI306X reduced cytokine expression and production. When PB monocytes were cultured in media supplemented with Th17 cytokines, osteoclast differentiation was stimulated; however, SKI306X decreased osteoclast differentiation and osteoclast maker expression. When PB monocytes were co-cultured with IL-17-stimulated RA synovial fibroblasts, osteoclast differentiation was increased; however, SKI306X decreased osteoclast differentiation and osteoclast maker expression. SKI306X reduced Th17 cytokine-induced TNF-, IL-1, and RANKL expression and osteoclast differentiation, providing novel insights into adjuvant therapy for regulating inflammation and joint destruction in RA.

Effects of resveratrol on Th17 cell-related immune responses under tacrolimus-based immunosuppression.

BACKGROUND: We previously reported that tacrolimus (Tac) does not decrease T helper 17 cells (Th17) response in kidney transplantation. In this study, we evaluated whether Resveratrol (Resv) has immunosuppressive effects by decreasing Th17 responses in Tac-based immunosuppression. METHODS: We investigated the effects of Resv under Tac-treatment conditions, on CD4+ T cell differentiation to Th17 cells in peripheral blood mononuclear cells (PBMCs), and proliferation of CD4+ T cells co-cultured with human renal proximal tubular epithelial cells (HRPTEpiCs). The effects of Resv on Th17 cells were tested in the murine skin transplant model. RESULTS: In PBMCs, Tac did not but combination of Tac and Resv further suppressed Th17 immune response. In the co-culture study, combination of Resv to Tac significantly decreased HRPTEpiC-induced T cell proliferation compared to Tac alone. Resv treatment in the Jurkat cell induced the expression of AMP-activated protein kinase and suppressed the expression of mammalian target of rapamycin (mTOR), suggesting blocking Th17 pathway by Resv. In the murine skin transplant model, combination of Resv to Tac significantly prolonged skin graft survival accompanied by the suppression of Th17 cells, compared to either the Tac-alone or control groups. CONCLUSION: The results of our study suggest that Resv provides additional immunosuppressive effects to Tac by suppressing effector CD4+ T cells, especially Th17 cells, in the transplantation setting.

Quercetin, a Plant Polyphenol, Has Potential for the Prevention of Bone Destruction in Rheumatoid Arthritis.

We investigated the immune-regulatory function of quercetin, in interleukin (IL)-17-produced osteoclastogenesis in rheumatoid arthritis (RA). RA fibroblasts-like synoviocytes (RA-FLS) were stimulated with IL-17, and the mRNA expression and secretion of receptor activator of nuclear factor kappa-B ligand (RANKL) were detected by real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. CD14+ monocytes (osteoclast precursors) were stimulated with IL-17, RANKL, with/without quercetin, and tartrate-resistant acid phosphatase activity was evaluated to assess osteoclast differentiation. Osteoclast differentiation was investigated after coculturing IL-17-stimulated RA-FLS and Th17 cells with monocytes. CD4+ T cells were cocultured with quercetin under Th17-inducing conditions, and their differentiation to Th17 cells and Treg cells was determined by flow cytometry analysis. We found that IL-17 stimulated RA-FLS to produce RANKL and quercetin decreased the IL-17-induced RANKL protein levels. Quercetin decreased the IL-17-produced activation of mammalian target of rapamycin, extracellular signal-regulated kinase and inhibitor of kappa B-alpha. When monocytes were stimulated with IL-17, macrophage colony-stimulating factor or RANKL, mature osteoclasts were formed, and quercetin decreased this osteoclastogenesis. When monocytes were cultured with IL-17-prestimulated RA-FLS or Th17 cells, osteoclasts were produced, and quercetin decreased this osteoclast differentiation. In Th17-differentiation conditions, quercetin suppressed Th17 cell and the production of IL-17, but quercetin did not affect Treg cells. Quercetin inhibits IL-17-stimulated RANKL production in RA-FLS and IL-17-stimulated osteoclast formation. Quercetin reduces Th17 differentiation. Quercetin could be an additional therapeutic option for bone destructive processes in RA.

N-acetyl-l-cysteine controls osteoclastogenesis through regulating Th17 differentiation and RANKL production in rheumatoid arthritis.

BACKGROUND/AIMS: This study aimed to determine the regulatory role of N-acetyl-l-cysteine (NAC), an antioxidant, in interleukin 17 (IL-17)-induced osteoclast differentiation in rheumatoid arthritis (RA). METHODS: After RA synovial fibroblasts were stimulated by IL-17, the expression and production of receptor activator of nuclear factor κ-B ligand (RANKL) was determined by real-time polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA). Osteoclastogenesis was also determined after co-cultures of IL-17-stimulated RA synovial fibroblasts, Th17 cells and various concentrations of NAC with monocytes. After human peripheral CD4+ T cells were cultured with NAC under Th17 condition, IL-17, interferon γ, IL-4, Foxp3, RANKL, and IL-2 expression and production was determined by flow cytometry or ELISA. RESULTS: When RA synovial fibroblasts were stimulated by IL-17, IL-17 stimulated the production of RANKL, and NAC reduced the IL-17-induced RANKL production in a dose-dependent manner. NAC decreased IL-17-activated phosphorylation of mammalian target of rapamycin, c-Jun N-terminal kinase, and inhibitor of κB. When human peripheral blood CD14+ monocytes were cultured with macrophage colony-stimulating factor and IL-17 or RANKL, osteoclasts were differentiated, and NAC reduced the osteoclastogenesis. After human peripheral CD4+ T cells were co-cultured with IL-17-pretreated RA synovial fibroblasts or Th17 cells, NAC reduced their osteoclastogenesis. Under Th17 polarizing condition, NAC decreased Th17 cell differentiation and IL-17 and RANKL production. CONCLUSION: NAC inhibits the IL-17-induced RANKL production in RA synovial fibroblasts and IL-17-induced osteoclast differentiation. NAC also reduced Th17 polarization. NAC could be a supplementary therapeutic option for inflammatory and bony destructive processes in RA.

Muscat Bailey A grape stalk extract ameliorates high-fat diet­induced obesity by downregulating PPARγ and C/EPBα in mice.

Muscat Bailey A grape stalk is an organic waste produced in marked amounts during the vinification of grapes. Previous studies have indicated that grape stalk is rich in bioactive phenolic compounds, and exhibits antioxidant and UV­protective activities. However, its effects on obesity and obesity­associated disorders have not yet been investigated. The effects of grape stalk extract on improving metabolic features were examined using a high­fat diet (HFD)­induced obesity mouse model. Oral administration of 200 mg/kg/day grape stalk extract over 16 weeks markedly prevented HFD­induced obesity, hepatic steatosis, diabetic symptoms and the risk of developing cardiovascular disease. Furthermore, grape stalk extract prevented oxidative stress and inflammation caused by HFD in mice. The beneficial effect may be associated with CCAAT/enhancer­binding protein α and peroxisome­proliferator­activated receptor Î³ down-regulation in liver tissue. Collectively, the results of the present study indicated that grape stalk extract may be a potent functional food ingredient for preventing obesity, hepatic steatosis and type 2 diabetes.