Assessment of the Impacts of Centipeda minima (L.) on Cell Viability, and Osteogenic Differentiation of Mesenchymal Stem Cell Spheroids.

Background and Objectives: Centipeda minima (L.) is a well-known and traditional pharmaceutical that has been utilized to treat different conditions controlling rhinitis, soothe pain, and decrease swelling. We assessed the impacts of Centipeda minima (L.) extricates (CMTs) on the osteogenic differentiation of cell spheroids made of human-bone-marrow-derived mesenchymal stem cells. Materials and Methods: Mesenchymal stem cells (MSCs) in spheroid 3D culture were generated and propagated in the presence of CMTs ranging from 0 to 1 µg/mL. Cell morphology was measured on Days 1, 3, 5, and 7. The quantitative cellular viability was evaluated on Days 1, 3, 5, and 7. Alkaline phosphatase activity assays were designed to measure the osteogenic differentiation of mesenchymal stem cell spheroids on Day 7. Alizarin Red S staining was performed to investigate the mineralization of cell spheroids on Days 7 and 14. Real-time polymerase chain reactions were used to measure the expression levels of RUNX2 and COL1A1 on Day 14. Western blot techniques were performed to identify the protein expression of Runt-related transcription factor 2 and type I collagen. Results: The control group's mesenchymal stem cells displayed a spheroid shape. There was no noticeable change in morphology with the addition of CMTs at final concentrations of 0.001, 0.01, 0.1, and 1 µg/mL compared with the untreated (control) group. The application of CMTs did not induce a significant change in cell viability. The relative alkaline phosphatase activity values in the 0.001, 0.01, 0.1, and 1 µg/mL CMT groups were 114.4% ± 8.2%, 130.6% ± 25.3%, 87.8% ± 3.4%, and 92.1% ± 6.8%, respectively, considering a control of 100% (100.0% ± 17.9%). On Day 14, calcium deposits were clearly observed in each group. The relative values of Alizarin Red S staining in the 0.001, 0.01, 0.1, and 1 µg/mL CMT groups were 100.1% ± 8.9%, 105.9% ± 0.0%, 109.7% ± 19.1%, and 87.0% ± 40.9%, respectively, considering a control of 100% (100.0% ± 28.7%). The addition of CMT significantly increased RUNX2 expression in the 0.01 µg/mL group and COL1A1 in the 0.001 and 0.01 µg/mL groups. Normalization of protein expression showed that the addition of CMTs significantly increased type I collagen expression in the 0.001, 0.01, and 1 µg/mL groups. Conclusions: In conclusion, CMTs influence the osteogenic differentiation of bone-marrow-derived mesenchymal stem cells and the use of CMTs may positively influence the osteogenic differentiation of cell spheroids.

Effects of noni on cellular viability and osteogenic differentiation of gingiva-derived stem cells demonstrated by RNA sequencing and quantitative PCR.

Noni fruit (Morinda citrifolia) has been widely used in traditional medicine across tropical and subtropical regions, and is now being paid more attention in Western medicine. The present study aimed to investigate the effects of noni extract on the change in the cellular morphology, maintenance of cellular viability and enhancement of osteogenic differentiation of stem cells. Stem cells obtained from gingiva were cultured where noni extracts existed at concentrations ranging from 10-200 ng/ml. Evaluations of cell morphology and cellular viability were performed. Alkaline phosphatase activity assays were performed to assess the osteogenic differentiation. Alizarin Red S staining was performed to evaluate the calcium deposits in the culture, with the addition of noni extract. Global gene expression was analyzed via next-generation mRNA sequencing. Gene ontology and pathway analyses were performed to determine the associated mechanisms. Validation procedures were performed via quantitative (q)PCR analysis. The addition of noni at concentrations ranging from 10-200 ng/ml did not produce significant morphological changes. There were significantly higher values of cellular viability, with the highest value at 100 ng/ml compared with the control (P<0.05). Furthermore, significantly higher values of alkaline phosphatase activity was noted in the 10 and 100 ng/ml groups compared with the 0 ng/ml group on day 7 (P<0.05). Alizarin Red S staining revealed calcium deposits in each group. In addition, the highest value for Alizarin Red S staining was observed at 100 ng/ml compared with the unloaded control (P<0.05). qPCR analysis demonstrated that the mRNA expression levels of RUNX2, BSP, OCN and COL1A1 increased following treatment with noni. Taken together, the results of the present study suggest that noni extract has enhancing effects on gingiva-derived mesenchymal stem cells, by enhancing cellular viability and osteogenic differentiation.

Evaluation of the Effects of Cuminum cyminum on Cellular Viability, Osteogenic Differentiation and Mineralization of Human Bone Marrow-Derived Stem Cells.

Background and Objectives: Cuminum cyminum L. has long been used in the treatment of various diseases in multiple geographical regions. This study was performed to determine the effects of C. cyminum methanolic extract (CCT) on the cellular viability, alkaline phosphatase activity and mineralization of human mesenchymal stem cells. Materials and Methods: Bone marrow-derived stem cells were cultured in the presence of CCT at concentrations of 0, 0.001, 0.01, 0.1 and 1 µg/mL. Evaluations of cell morphology were performed on days 1, 3, 7 and 14. Cellular viability was evaluated on days 1, 3, 5 and 7. On the 7th and 14th day, alkaline phosphatase activity measurements and Alizarin red S staining were conducted to assess the osteogenic differentiation of stem cells. A real-time polymerase chain reaction was used to determine the expression levels of RUNX2, BSP, OCN, COL2A1 and ß-catenin mRNAs. Results: Stem cells in the control group showed fibroblast-like morphology and the addition of CCT at 0.001, 0.01, 0.1 and 1 µg/mL did not generate noticeable changes in morphology compared with the untreated control group. The application of CCT did not produce significant changes in cellular viability or alkaline phosphatase activity compared with controls. Alizarin Red S staining was significantly increased with the application of CCT. Treatment with CCT increased the expressions of RUNX2, BSP and OCN. Conclusions: These results indicate that CCT enhanced the osteogenic differentiation of stem cells derived from bone marrow by regulating the expressions of RUNX2, BSP and OCN. Thus, the use of CCT may be applied to achieve beneficial effects on the mineralization of stem cells.

The Effects of Morinda citrifolia (Noni) on the Cellular Viability and Osteogenesis of Stem Cell Spheroids.

Background and objectives: Morinda citrifolia (Noni) has been widely used in herbal remedies to treat and prevent various kinds of diseases. We conducted this study to evaluate the effects of Noni extract on the maintenance of morphology, the improvement of cellular viability, and the enhancement of osteogenesis of stem cell spheroids. Materials and Methods: We cultured stem cell spheroids made with gingiva-derived stem cells in the presence of Noni extract at concentrations of 10, 100 and 200 ng/mL. We performed analysis of the cell morphology and changes in the cellular viability. We conducted alkaline phosphatase activity assays using a kit, and mineralization assays using an anthraquinone dye to evaluate the osteogenesis of stem cell spheroids with the addition of Noni extract. Results: The applied cells formed spheroids well, and the addition of Noni at 10, 100 and 200 ng/mL concentrations did not produce significant morphological changes. The quantitative values for cellular viability on Day 3 showed that the absorbance values at 450 nm were 0.314 ± 0.013, 0.318 ± 0.008, 0.304 ± 0.000 and 0.300 ± 0.011 for Noni at 0, 10, 100 and 200 ng/mL concentrations, respectively. The results of alkaline phosphatase activity with absorbance values at 405 nm were 0.189 ± 0.019, 0.174 ± 0.023, 0.192 ± 0.014 and 0.210 ± 0.062 for Noni at 0, 10, 100 and 200 ng/mL concentrations, respectively, on Day 4. There were significantly higher values of Alizarin Red S staining for Noni in the 10, 100 and 200 ng/mL groups, with the highest value at 100 ng/mL when compared with the unloaded control on Day 14. Conclusions: Based on these findings, we concluded that Noni extract might be applied for the enhanced osteogenic differentiation of stem cell spheroids.

The Electrical Isolation of the Left Atrial Posterior Wall in Catheter Ablation of Persistent Atrial Fibrillation.

OBJECTIVES: This study explored whether complete electrical isolation of the left atrial (LA) posterior wall improves the rhythm outcome of catheter ablation of persistent atrial fibrillation (AF). BACKGROUND: Although the STAR AF2 (Substrate and Trigger Ablation for Reduction of Atrial Fibrillation Trial Part II) proved no additional benefit of empirical extra-pulmonary vein (PV) LA ablation, the long-term recurrence rate after circumferential PV isolation (CPVI) alone remains high. METHODS: We randomly assigned 217 patients with persistent AF (83.1% men, age 58.7 ± 10.8 years, 73.3% long-standing persistent AF) to ablation with CPVI alone (CPVI group) or CPVI with a POsterior wall Box Isolation (POBI group). The endpoint of the POBI group was the elimination of the posterior atrial potentials by roof and posterior inferior lines and touch-up focal ablation. RESULTS: After a mean follow-up of 16.2 ± 8.8 months, the clinical recurrence rate did not significantly differ between the 2 groups (23.8% vs. 26.5%; p = 0.779) in the CPVI and POBI groups. The recurrence rate for atrial tachycardias (16.0% vs. 11.1%; p = 0.913) and cardioversion rates (6.7% vs. 13.7%; p = 0.093) to control clinical recurrences also did not significantly differ between the 2 groups. At the final follow-up, sinus rhythm was maintained without antiarrhythmic drug in 50.5% and 55.9% in the CPVI and POBI groups, respectively (p = 0.522). No significant difference was found in the major complication rates between the 2 groups, but the total ablation time was significantly longer in the POBI group (4,289 ± 1,837 s vs. 5,365 ± 2,358 s; p < 0.001). CONCLUSIONS: In patients with persistent AF, an empirical complete POBI did not improve the rhythm outcome of catheter ablation or influence the type of recurrent atrial arrhythmia. (Comparison of Circumferential Pulmonary Vein Isolation Alone Versus Linear Ablation in Addition to Circumferential Pulmonary Vein Isolation for Catheter Ablation in Persistent Atrial Fibrillation: Prospective Randomized Controlled Trial; NCT02721121).

Evaluation of fibroblast growth factor-2 on the proliferation of osteogenic potential and protein expression of stem cell spheroids composed of stem cells derived from bone marrow.

Fibroblast growth factor-2 (FGF-2) is reported to have various functions and is considered a key human mesenchymal stem cell mitogen, often supplemented to increase human mesenchymal stem cell growth rates. The purpose of this study was to evaluate the effects of FGF-2 on cellular viability and osteogenic differentiation using three-dimensional cell spheroids of stem cells. Three-dimensional cell spheroids were fabricated using concave silicon elastomer-based microwells in the presence of FGF-2 at concentrations of 0, 30, 60 and 90 ng/ml. Qualitative cellular viability was determined with a confocal microscope, and quantitative cellular viability was evaluated using a Cell Counting Kit-8 assay. Alkaline phosphatase activity and Alizarin Red S staining were used to assess osteogenic differentiation. Spheroids were well formed in silicon elastomer-based concave microwells on Day 1. The average spheroid diameters at Day 1 for FGF-2 at 0, 30, 60 and 90 ng/ml were 202.2±3.0, 206.6±22.6, 208.8±6.8 and 196.6±26.7 µm, respectively (P>0.05). The majority of the cells in the cell spheroids emitted green fluorescence. The relative Cell Counting Kit-8 assay values for FGF-2 at 0, 30, 60 and 90 ng/ml at Day 1 were 100.0±5.5, 101.8±8.8, 99.2±4.8 and 103.4±9.6% (P>0.05). The addition of FGF-2 at 60 ng/ml concentration produced the highest value for alkaline phosphatase activity. Mineralized extracellular deposits were evenly observed in each group, and the highest value was identified for FGF-2 groups at 60 ng/ml concentration for Alizarin Red S staining. Based on these findings, it was concluded that FGF-2 may increase alkaline phosphatase activity or Alizarin Red S staining, and further studies are needed to fully elucidate the mechanisms of FGF-2.

Label Adherence for Non-Vitamin K Antagonist Oral Anticoagulants in a Prospective Cohort of Asian Patients with Atrial Fibrillation.

PURPOSE: Label adherence for non-vitamin K antagonist oral anticoagulants (NOACs) has not been well evaluated in Asian patients with non-valvular atrial fibrillation (AF). The present study aimed to assess label adherence for NOACs in a Korean AF population and to determine risk factors of off-label prescriptions of NOACs. MATERIALS AND METHODS: In this COmparison study of Drugs for symptom control and complication prEvention of AF (CODE-AF) registry, patients with AF who were prescribed NOACs between June 2016 and May 2017 were included. Four NOAC doses were categorized as on- or off-label use according to Korea Food and Drug Regulations. RESULTS: We evaluated 3080 AF patients treated with NOACs (dabigatran 27.2%, rivaroxaban 23.9%, apixaban 36.9%, and edoxaban 12.0%). The mean age was 70.5±9.2 years; 56.0% were men; and the mean CHA2DS2-VASc score was 3.3±1.4. Only one-third of the patients (32.7%) was prescribed a standard dose of NOAC. More than one-third of the study population (n=1122, 36.4%) was prescribed an off-label reduced dose of NOAC. Compared to those with an on-label standard dosing, patients with an off-label reduced dose of NOAC were older (≥75 years), women, and had a lower body weight (≤60 kg), renal dysfunction (creatinine clearance ≤50 mL/min), previous stroke, previous bleeding, hypertension, concomitant dronedarone use, and anti-platelet use. CONCLUSION: In real-world practice, more than one-third of patients with NOAC prescriptions received an off-label reduced dose, which could result in an increased risk of stroke. Considering the high risk of stroke in these patients, on-label use of NOAC is recommended.

Improving the Microbial Safety of Fresh-Cut Endive with a Combined Treatment of Cinnamon Leaf Oil Emulsion Containing Cationic Surfactants and Ultrasound.

Endive is widely consumed in a fresh-cut form owing to its rich nutritional content. However, fresh-cut vegetables are susceptible to contamination by pathogenic bacteria. This study investigated the antibacterial activities of the combined treatment of cinnamon leaf oil emulsion containing cetylpyridinium chloride or benzalkonium chloride (CLC and CLB, respectively) as a cationic surfactant and ultrasound (US) against Listeria monocytogenes and Escherichia coli O157:H7 on endive. The combined treatment of CLC or CLB with US reduced the population of L. monocytogenes by 1.58 and 1.47 log colony forming units (CFU)/g, respectively, and that of E. coli O157:H7 by 1.60 and 1.46 log CFU/g, respectively, as compared with water washing treatment. The reduction levels of both pathogens were higher than those observed with 0.2 mg/ml sodium hypochlorite. In addition, the combined treatment showed no effect on the quality of the fresh-cut endive (FCE). In particular, the degree of browning in FCE was less for the treatment group than for the control and water washing treatment groups. Thus, cationic surfactant-based cinnamon leaf oil emulsions combined with US may be an effective washing treatment for the microbial safety of FCE.

Coffee Intake as a Risk Indicator for Tooth Loss in Korean Adults.

The aim of this study was to examine the association between coffee intake and tooth loss. This study hypothesized that the intake of coffee would increase the prevalence of tooth loss in Korean adults. Subject information was obtained from the Korea National Health and Nutrition Examination Survey conducted in 2010-2011. Sociodemographic and lifestyle variables, anthropometric and biochemical status, metabolic health and glucose tolerance status, as well as oral health behaviors were evaluated. The number of remaining teeth was negatively associated with the frequency of coffee intake (p-value < 0.05). Daily coffee consumers had significantly higher levels of body mass index (BMI), waist circumference (WC), total cholesterol, and low density lipoprotein cholesterol (LDL-C) (all p-value < 0.05). Individuals with less than 20 remaining teeth had higher BMI, WC, diastolic blood pressure, and LDL-C (all p-value < 0.05). Finally, participants who drank coffee on a daily basis were more likely to have fewer remaining teeth. The prevalence of having less than 20 remaining teeth was 69% higher in groups with daily coffee intake than those with coffee intake of less than once a month after adjustment for potential covariates (Odds Ratio [95% CI] = 1.69 [1.35, 2.13]). In conclusion, daily coffee consumption is closely associated with tooth loss in Korean adults.

Effects of Bambusa tulda on the proliferation of human stem cells.

To date, the effects of Bambusa tulda on stem cells have not been thoroughly assessed. The present study aimed to evaluate the effects of Bambusa tulda extract on the morphology and proliferative potential of human mesenchymal stem cells derived from the gingiva. The stem cells were cultured in a growth medium in the presence of Bambusa tulda methanolic extract (BBT) at concentrations ranging from 0.001 to 1%. Evaluation of cell morphology and cellular proliferation as well as immunofluorescent assays for collagen I were performed on days 1, 3, 5 and 7. Stem cells in the control group displayed a fibroblast-like morphology, and BBT treatment did not produce any noticeable morphological changes. However, application of 1% BBT produced a significant increase in cell proliferation. BBT, particularly at the concentration of 1%, also caused a noticeable increase of collagen I expression at day 1 and day 3. Based on these findings, it was concluded that BBT exerted beneficial effects on the proliferation of mesenchymal stem cells and enhanced collagen I expression at early time points.

Effects of Cimicifugae Rhizoma on the osteogenic and adipogenic differentiation of stem cells.

Cimicifugae Rhizoma, a herb with a long history of use in traditional Oriental medicine is reported to have anti-inflammatory, antioxidant, anti-complement and anticancer effects. The aim of the present study was to evaluate the effects of Cimicifugae Rhizoma extracts on the osteogenic and adipogenic differentiation of human stem cells derived from gingiva. Stem cells derived from gingiva were grown in the presence of Cimicifugae Rhizoma at final concentrations of 0.1, 1 and 10 µg/ml. Cell proliferation analyses were performed at day 15. For osteogenic differentiation experiments, the stem cells were cultured in osteogenic media containing ß-glycerophosphate, ascorbic acid-2-phosphate and dexamethasone, and osteogenic differentiation was evaluated by analysis of osteocalcin expression at 21 days. For adipogenic differentiation experiments, the stem cells were grown in adipogenic induction medium, and the adipogenic differentiation was evaluated by analysis of adipocyte fatty acid-binding protein at day 14. The cultures grown in the presence of 0.1 µg/ml Cimicifugae Rhizoma showed a significant increase in cellular proliferation at day 15 compared with the control group. The relative osteogenic differentiation in the presence of Cimicifugae Rhizoma for the 0.1, 1 and 10 µg/ml groups was 171.5±13.7, 125.6±28.7 and 150.5±9.0, respectively, when that of the untreated control group on day 21 was considered to be 100%. The relative adipogenic differentiation at day 14 of the 0.1, 1 and 10 µg/ml groups in the presence of Cimicifugae Rhizoma was 97.5±15.0, 102.9±12.8 and 87.0±6.8%, respectively when that of the untreated control group on day 14 was considered to be 100%. Within the limits of this study, Cimicifugae Rhizoma increased the proliferation of stem cells derived from the gingiva, and low concentrations of Cimicifugae Rhizoma may increase the osteogenic differentiation of stem cells.

Pulmonary vein reconnection predicts good clinical outcome after second catheter ablation for atrial fibrillation.

AIMS: Although electrically reconnected pulmonary veins (PV) are the main mechanism of atrial fibrillation (AF) recurrence, PV isolation (PVI) is well-preserved in certain patients who undergo a repeat procedure. We explored the association between PV reconnection and clinical outcomes after a second ablation. METHODS AND RESULTS: This observational cohort study included 143 patients (79.0% male, 56.1 ± 10.0 years old, 65.0% paroxysmal AF) who underwent a second procedure. Pulmonary vein isolation was well-maintained in 52 patients (PVP- group, 36.4%), although the remaining 91 patients showed PV reconnection (PVP+ group). After confirming PVI, we mapped non-PV triggers and conducted trigger ablation or additional linear ablation at redo-procedures. The proportion of females was higher (P = 0.030), and redo-ablation timing after the de novo procedure was later (P = 0.039) in the PVP- group than in the PVP+ group. Additional linear ablations were more likely to be performed in the PVP- group (90.4 vs. 61.5%, P < 0.001). During the 18.4 ± 10.2 month follow-up after the redo-ablation, the PVP+ group showed a lower clinical recurrence rate than the PVP- group (log-rank P = 0.011). The number of reconnected PVs was independently associated with a lower recurrence of AF after the redo-ablation in the total study population (HR 0.56, 95% CI 0.34-0.95, P = 0.032), particularly for patients with paroxysmal AF (HR 0.41, 95% CI 0.19-0.87, P = 0.021). CONCLUSION: Among patients who underwent redo-AF ablation, those with more PV reconnections showed better clinical outcomes than those with fewer PV reconnections. The mechanism of AF recurrence might be different in patients with lower numbers of PV reconnections during redo-procedures.

Challenging Achievement of Bidirectional Block After Linear Ablation Affects the Rhythm Outcome in Patients With Persistent Atrial Fibrillation.

BACKGROUND: It is not clear whether bidirectional block (BDB) of linear ablations reduces atrial fibrillation (AF) recurrence after radiofrequency catheter ablation. We hypothesized that BDB of linear ablation has prognostic significance after radiofrequency catheter ablation for persistent AF. METHODS AND RESULTS: Among 1793 consecutive patients in the Yonsei AF ablation cohort, this observational cohort study included 398 patients with persistent AF (75.6% male; age, 59.8±10.3 years) who underwent catheter ablation with a consistent ablation protocol of the Dallas lesion set: circumferential pulmonary vein isolation; cavotricuspid isthmus ablation (CTI); roof line (RL); posterior-inferior line (PIL); and anterior line (AL). BDB rates of de novo ablation lines were 100% in circumferential pulmonary vein isolation, 100% in CTI, 84.7% in RL, 44.7% in PIL, and 63.6% in AL. During 29.0±18.4 months of follow-up, 31.7% (126/398) of the patients showed clinical recurrence. Left atrial posterior wall (LAPW) isolation (BDBs of RL and PIL) was independently associated with lower clinical AF/atrial tachycardia recurrence (hazard ratio, 0.68; 95% CI, 0.47-0.98; P=0.041; log-rank, P=0.017), whereas BDBs of RL or AL were not (log-rank, P=0.178 for RL; P=0.764 for AL). Among 52 patients who underwent repeat procedures (23.0±16.1 months after de novo procedure), the BDB maintenance rates for CTI, RL, PIL, and AL were 94.2% (49 of 52), 63.5% (33 of 47), 62.1% (18 of 29), and 61.8% (21 of 34), respectively. CONCLUSIONS: Although PIL crosses the esophageal contact area, LAPW isolation is important for better clinical outcome in catheter ablation with a linear ablation strategy for patients with persistent AF.

Excessive Consumption of Green Tea as a Risk Factor for Periodontal Disease among Korean Adults.

This study was performed to assess the relationship between the amount of green tea that is consumed and periodontitis. It is based on data obtained from the Korea National Health and Nutrition Examination Survey, conducted between 2008 and 2010. A community periodontal index equal to code 3 was defined as moderate periodontitis, and code 4 was defined as severe periodontitis (n = 16,726). Consumption of green tea less than one cup per day was associated with a decreased prevalence of periodontal disease among Korean adults. The association between the consumption of green tea and periodontal disease was independent of various potential confounding factors, such as age, sex, body mass index, smoking, drinking, exercise, metabolic syndrome, frequency of tooth brushing per day, use of secondary oral products, the number of dental examination per year, diabetes, hypertension, and white blood cell count. Adjusted odds ratio and 95% confidence interval of no consumption was 1.360 (1.156, 1.601) when participants with consumption of two times per week ≤ x < 7 times per week was considered as a reference. However, consumption of one or more cups per day increased the prevalence of moderate and severe periodontitis. In conclusion, excessive consumption of green tea may be considered as a risk factor for periodontal disease among Korean adults.

Association between Consumption of Coffee and the Prevalence of Periodontitis: The 2008-2010 Korea National Health and Nutrition Examination Survey.

BACKGROUND: This study was performed to assess the relationship between the consumption of coffee and periodontitis using nationally representative data. METHODS: The data from the Korea National Health and Nutrition Examination Survey were used; the analysis in this study was confined to a total of 16,730 respondents over 19 years old who had no missing values for the consumption of coffee or outcome variables. A community periodontal index greater than or equal to code 3 was defined as periodontal disease. RESULTS: Consumption of coffee was significantly higher in the individuals with periodontitis in males. The odds ratios of the percentage of individuals with periodontitis tended to increase with the consumption of coffee. Adjusted odds ratios and their 95% confidence intervals of the male participants were 1, 1.131(0.792-1.617), 1.161(0.857-1.573), 1.053(0.805-1.379), 1.299(1.007-1.676), and 1.458(1.141-1.862) for once per month or less, once per month

Anti-inflammatory and cytotoxic effects of methanol, ethanol, and water extracts of Angelicae Dahuricae Radix.

Angelicae Dahuricae Radix has been used for the treatment of headaches, rhinitis, and colds in traditional medicine. Methanol, ethanol, and water extracts of Angelicae Dahuricae Radix were collected. A statistically significant reduction in the cellular viability of the mouse leukemic monocyte macrophage cell line was noted after treatment with water extracts of Angelicae Dahuricae Radix. Stimulation with lipopolysaccharides (LPS) for 24 h led to a robust increase in nitric oxide production, but Angelicae Dahuricae Radix at 400 µg/mL concentration significantly suppressed nitric oxide produced by the LPS-stimulated RAW 264.7 cells in 70% ethanol, absolute ethanol, 70% methanol, absolute methanol, and boiling water groups (P < 0.05). Pretreatment with absolute ethanol extract of Angelicae Dahuricae Radix suppressed the LPS-stimulated inducible nitric oxide synthase, interleukin-1ß, and cycloxygenase-2 expression. Angelicae Dahuricae Radix showed significant cytotoxic effects on the human adenocarcinoma cell line and keratin-forming cell line. (J Oral Sci 58, 125-131, 2016).

Low P-wave amplitude (<0.1 mV) in lead I is associated with displaced inter-atrial conduction and clinical recurrence of paroxysmal atrial fibrillation after radiofrequency catheter ablation.

AIMS: We hypothesized that P-wave amplitude in lead I is related to left atrial (LA) remodelling and inter-atrial conduction pattern, and has a predictive value for recurrence after radiofrequency catheter ablation (RFCA) among patients with paroxysmal atrial fibrillation (PAF). METHODS AND RESULTS: A total of 525 consecutive patients with PAF (76% male, 56 ± 12 years old) who underwent RFCA were included. We compared pre-procedural sinus rhythm electrocardiograms without antiarrhythmic drug effect with LA volume (CT), LA voltage (NavX), the earliest activation site (EAS) conduction pattern of LA, and clinical recurrence rate. P-wave amplitude in lead I was significantly lower in patients with recurrence than in those that remained in sinus rhythm (P < 0.001) during 21 ± 10-month follow-up. P-wave amplitude in lead I was linearly correlated with LA voltage (ß = 2.52, 95% CI 0.606-4.425, P = 0.010), LA conduction velocity (ß = 1.91, 95% CI 0.941-2.876, P < 0.001), and low septal displacement of EAS (ß = -1.67, 95% CI -2.352 to -0.996, P < 0.001). P-wave amplitudes <0.1 mV in lead I were independently associated with clinical recurrence of AF on multivariate Cox regression analysis (adjusted HR 2.163, 95% CI 1.307-3.581, P = 0.003). The integrated area under the curves was 0.705 (95% CI 0.655-0.755). CONCLUSION: Low P-wave amplitude (<0.1 mV) in lead I is related to LA remodelling and displaced inter-atrial conduction pattern to low septum, and independently predicts clinical recurrence after RFCA in patients with PAF.

Effect of Asiasari radix on osteoblastic differentiation of stem cells derived from gingiva.

OBJECTIVE: To examine the dose-dependent impact of Asiasari Radix (A. radix) on the cell viability, differentiation and mineralization of stem cells derived from gingiva. METHODS: Stem cells that were derived from gingiva were grown in the presence of A. radix at final concentrations that ranged from 0.001 to 10 µg/mL. The morphology of the cells was viewed under an inverted microscope and the analysis of cell proliferation was performed by using Cell Counting Kit-8 (CCK-8) on day 1. The alkaline phosphatase activity test was used to assess differentiation and Alizarin red S staining was used to assess mineralization of treated cells. RESULTS: The control group showed spindleshaped, fibroblast-like morphology and the shapes of the cells in 0.001, 0.01, 0.1, 1 and 10 µg/mL of A. radix were similar to that of the control group at day 1. The cultures growing in the presence of 0.001 µg/mL of A. radix at day 1 showed an increase in the CCK-8 value (P < 0.05). Cultures growing in the presence of 0.001 µg/mL of A. radix presented the highest value for alkaline phosphatase activity (P > 0.05). Mineralized extracellular deposits were observed after Alizarin Red S staining and the cultures grown in the presence of 0.001 µg/mL of A. radix showed the highest value for quantitative results for bound dye (P < 0.05). CONCLUSION: Within the limits of this study, A. radix influenced the proliferation of stem cells derived from the gingiva and low concentrations of A. radix might enhance osteogenic differentiation of the stem cells.

Evaluation of the effects of Cimicifugae Rhizoma on the morphology and viability of mesenchymal stem cells.

Cimicifugae Rhizoma is a traditional herbal medicine used to treat various diseases in Korea, China and Japan. Cimicifugae Rhizoma is primarily derived from Cimicifuga heracleifolia Komarov or Cimicifuga foetida Linnaeus. Cimicifugae Rhizoma has been used as an anti-inflammatory, analgesic and antipyretic remedy. The present study was performed to evaluate the extracts of Cimicifugae Rhizoma on the morphology and viability of human stem cells derived from gingiva. Stem cells derived from gingiva were grown in the presence of Cimicifugae Rhizoma at final concentrations that ranged from 0.001 to 1,000 µg/ml. The morphology of the cells was viewed under an inverted microscope and the analysis of cell proliferation was performed using a Cell Counting kit-8 (CCK-8) assay on days 1, 3, 5 and 7. Under an optical microscope, the control cells exhibited a spindle-shaped, fibroblast-like morphology. The shapes of the cells in the groups treated with 0.001, 0.01, 0.1, 1 and 10 µg/ml Cimicifugae Rhizoma were similar to the shapes in the control group. Significant alterations in morphology were noted in the 100 and 1,000 µg/ml groups when compared with the control group. The cells in the 100 and 1,000 µg/ml groups were rounder, and fewer cells were present. The cultures that were grown in the presence of Cimicifugae Rhizoma at a concentration of 0.001 µg/ml on day 1 had an increased CCK-8 value. The cultures grown in the presence of Cimicifugae Rhizoma at a concentration of 10 µg/ml on day 7 had a reduced CCK-8 value. Within the limits of this study, Cimicifugae Rhizoma influenced the viability of stem cells derived from the gingiva, and its direct application onto oral tissues may have adverse effects at high concentrations. The concentration and application time of Cimicifugae Rhizoma should be meticulously controlled to obtain optimal results.

Evaluation of the effects of Angelicae dahuricae radix on the morphology and viability of mesenchymal stem cells.

Angelicae dahuricae radix is a traditional herbal medicine used to treat various diseases in China and Korea, such as colds, headaches, rhinitis and psoriasis. Angelicae dahuricae radix has been used as an anti-inflammatory, analgesic, antipyretic and antioxidant remedy. This study was performed in order to evaluate the effects of the extracts of Angelicae dahuricae radix on the morphology and viability of mesenchymal stem cells derived from the gingiva. Mesenchymal stem cells derived from the gingiva were grown in the presence of Angelicae dahuricae radix at final concentrations that ranged from 0.001 to 100 µg/ml. The morphology of the cells was viewed under an inverted microscope, and the analysis of cell proliferation was performed with cell counting kit-8 (CCK-8) on days 1, 3 and 7. The cells in the control group had spindle-shaped, fibroblast-like morphology at days 1, 3 and 7 under optical microscopy. The shapes of the cells in 0.001, 0.01, 0.1, 1, 10 and 100 µg/ml Angelicae dahuricae radix were similar to the shapes of the cells in the control group. The relative values of the CCK-8 assays of 0.001, 0.01, 0.1, 1, 10, and 100 µg/ml Angelicae dahuricae radix were 102.5 ± 0.6, 133.3 ± 9.6, 148.4 ± 20.5, 147.7 ± 12.6, 132.3 ± 27.7 and 101.1 ± 4.6%, respectively, when the CCK-8 result of the control group on day 1 was considered to be 100%. There was a marginal increase in cell proliferation at 0.1 and 1 µg/ml groups at day 1; however, this did not achieve statistical significance (P=0.052). The relative values of the CCK-8 assays of 0.001, 0.01, 0.1, 1, 10 and 100 µg/ml Angelicae dahuricae radix were 96.5 ± 1.3, 89.3 ± 0.9, 90.3 ± 3.0, 84.8 ± 12.2, 92.3 ± 4.5 and 86.8 ± 11.7%, respectively, when the CCK-8 result of the control group on day 3 was considered to be 100% (P>0.05). The relative values of the CCK-8 assays of 0.001, 0.01, 0.1, 1, 10 and 100 µg/ml Angelicae dahuricae radix day 7 were 94.9 ± 22.3, 102.8 ± 22.1, 127.4 ± 7.4, 130.4 ± 1.3, 129.2 ± 10.8 and 124.8 ± 9.1%, respectively, when the CCK-8 result of the control group on day 7 was considered to be 100%, but there were no statistically significant differences among the groups (P>0.05). Within the limits of this study, Angelicae dahuricae radix at the tested concentrations did not produce statistically significant differences in the viability of stem cells derived from the gingiva.